No full-text available
To read the full-text of this research,
you can request a copy directly from the authors.
Acetobacter methanolicus sp. nov., an Acidophilic Facultatively Methylotrophic Bacterium
Abstract
A new species, Acetobacter methanolicus, is described. The strains investigated were isolated from sludge and from a yeast fermentation process in which methanol was the sole source of carbon and energy. A total of about 140 phenotypic features were tested. The strains proved to be acidophilic and facultatively methylotrophic, and they differed from other Acetobacter species by growing well on methanol, glucose, gluconate, 2,3-butanediol, and caproic acid as sole sources of carbon and energy. Ethanol was “overoxidized” only at initial concentrations of <0.5%. Lactate was oxidized very weakly, but it was not utilized as a sole carbon source for growth. Yeast extract or pantothenic acid was essential for growth. The specific epithet of the proposed new species refers to its isolation from media in which methanol was the sole source of carbon. The deoxyribonucleic acid base composition of type strain MB58 (= IMET 10945) is 62.3 mol% guanine plus cytosine.
... The genus Acidomonas was introduced for the facultatively methylotrophic bacterium, Acetobacter methanolicus Uhlig et al. 1986(Urakami et al. 1989). However, the generic name was not accepted for a long time (Swings 1992, Sievers et al. 1994. ...
... 3.1. Acidomonas methanolica (Uhlig et al. 1986) Urakami, Tamaoka, Suzuki and Komagata 1989. emend. ...
... emend. Yamashita, Uchimura and Komagata 2004 Basonym: Acetobacter methanolicus Uhlig, Karbaum and Steudel 1986. For the characteristics of the species, refer to Uhlig et al. (1986), Urakami et al. (1989), Yamashita et al. (2004). ...
... The genus Acidomonas was introduced for the facultatively methylotrophic bacterium, Acetobacter methanolicus Uhlig et al. 1986(Urakami et al. 1989). However, the generic name was not accepted for a long time (Swings 1992, Sievers et al. 1994. ...
... 3.1. Acidomonas methanolica (Uhlig et al. 1986) Urakami, Tamaoka, Suzuki and Komagata 1989. emend. ...
... emend. Yamashita, Uchimura and Komagata 2004 Basonym: Acetobacter methanolicus Uhlig, Karbaum and Steudel 1986. For the characteristics of the species, refer to Uhlig et al. (1986), Urakami et al. (1989), Yamashita et al. (2004). ...
This book, written by leading international authorities in the field, covers all the basic and applied aspects of acetic acid bacteria. It describes the importance of acetic acid bacteria in food industry by giving information on the microbiological properties of fermented foods as well as production procedures. Special attention is given to vinegar and cocoa, which are the most familiar and extensively used industrial applications of acetic acid bacteria. This book is an essential reference to all scientists, technologists, engineers, students and all those working in the field of food science and technology.
... a Data from Lisdiyanti et al. (2002), and Yamashita et al. (2004). b Contradictory reports were published for the growth on methanol by A. pasteurianus, A. lovaniensis, and A. pomorum (Gosselé et al., 1983b;Uhlig et al., 1986;Urakami et al., 1989;Sokollek et al., 1998a;Cleenwerck et al., 2002) (see text in section Acetobacter). ...
... Methylotrophic acetic acid bacteria were isolated from a septic methanol yeast process (Steudel et al., 1980) and from sludge and described by Uhlig et al. (1986) as a new species, Acetobacter methanolicus, at present classified as Acidomonas methanolica (Urakami et al., 1989) (Table 1). ...
... Contradictory data were reported concerning the growth on methanol or utilization of methanol by A. pasteurianus, A. lovaniensis, A. pomorum, and A. malorum strains (Gosselé et al., 1983b;Uhlig et al., 1986;Urakami et al., 1989;Sokollek et al., 1998a;Cleenwerck et al., 2002). Ambiguity about growth on methanol and utilization of methanol can be ascribed to unclear definitions of assimilation and utilization of methanol, and growth on methanol. ...
Introduction and General CharacteristicsAcetic acid bacteria comprise a widespread group of Gram-negative, obligately aerobic rods. They occur mainly in sugary, acidic and alcoholic habitats and have been studied extensively, since they can play a positive, neutral or detrimental role in foodstuffs and beverages. Some species of the Acetobacteraceae play a key role in the industrial manufacture of vinegar. The following genera belong to this family: Acetobacter (type genus), Acidomonas, Asaia, Gluconacetobacter, Gluconobacter and Kozakia. The names Acetobacter and Gluconobacter are known in literature since 1898 and 1935, respectively, whereas the other genus names were published after 1989.All members of the Acetobacteraceae are obligately aerobic and their metabolism is strictly respiratory with oxygen as the terminal electron acceptor. A common feature of the acetic acid bacteria (with the exception of Asaia) is the aerobic oxidation of ethano ...
... The genus name Acidomonas was validly published for acidophilic, methanol-utilizing bacteria (Urakami et al., 1989), incorporating Acetobacter methanolicus Uhlig et al. 1986. Acidomonas methanolica is the type and only species reported so far in this genus. ...
... The revival of Acetobacter methanolicus was once proposed (Sievers et al., 1994), but another study justified the description of the genus Acidomonas . However, since the work of Uhlig et al. (1986) and Urakami et al. (1989), few Acidomonas strains have been isolated until now. In addition, this taxon is problematic, as some characteristics differ in the descriptions by Uhlig et al. (1986) and Urakami et al. (1989) and only four Acidomonas methanolica strains are available from culture collections worldwide. ...
... However, since the work of Uhlig et al. (1986) and Urakami et al. (1989), few Acidomonas strains have been isolated until now. In addition, this taxon is problematic, as some characteristics differ in the descriptions by Uhlig et al. (1986) and Urakami et al. (1989) and only four Acidomonas methanolica strains are available from culture collections worldwide. They are the type strain NRIC 0498 T (=DSM 5432 T =JCM 6891 T = LMG 1668 T =MB58 T ) (Steudel et al., 1980;Uhlig et al., 1986), LMG 1667, LMG 1669 and LMG 1735. ...
The genus Acidomonas and the species Acidomonas methanolica were recharacterized by using the type strain (NRIC 0498 T ), three reference strains and 10 methanol-utilizing bacteria that were isolated from activated sludge from three different sewage-treatment plants in Tokyo. Based on 16S rDNA sequences, all strains formed a single cluster within the Acetobacteraceae that was clearly different from the genera Acetobacter, Gluconobacter, Gluconacetobacter, Asaia and Kozakia. The 14 strains were identified as a single species, Acidomonas methanolica, by DNA-DNA similarities, showed DNA G+C contents that ranged from 62 to 63 mol% and had Q-10 as the major quinone, accounting for > 87 % of total ubiquinones. Cells of Acidomonas methanolica had a single polar flagellum (or occasionally polar tuft flagella); this differs from a previous study that described peritrichous flagella. Oxidation of acetate was positive for all strains, but oxidation of lactate was weakly positive and varied with strains. Dihydroxyacetone was not produced from glycerol. Pantothenic acid was an essential requirement for growth. The strains tested grew at mostly the same extent at pH 3.0-8.0. Therefore, Acidomonas methanolica should be regarded as acidotolerant, not acidophilic. The descriptions of the genus Acidomonas and the species Acidomonas methanolica Urakami, Tamaoka, Suzuki and Komagata 1989 are emended with newly obtained data.
... Acidomonas methanolica MB58 (former name: Acetobacter methanolicus MB58) was originally isolated as a facultatively methylotrophic bacterium from a septic methanolprocessing mixture in Germany (Uhlig et al., 1986). Acidomonas methanolica is now recognized as a unique acetic acid bacterium capable of assimilating both methanol and ethanol. ...
... A k-mer size of 39 among different minimum-overlapping lengths gave the best results in terms of N 50 (the median contig length), resulting in 546 contigs with total 3 690 031 bp (coverage: 90.2 x). The A. methanolica genome had 65.4% G + C content, which was relatively higher than that previously reported (Uhlig et al., 1986). The draft genome sequence contained 3270 protein-coding genes, which were extracted using METAGENEANNOTATOR (MGA) (Noguchi et al., 2008), and functions of the gene products were annotated by comparison with similar proteins detected by Basic Local Alignment Search Tool (BLAST) (Altschul et al., 1997). ...
Acidomonas methanolica (former name Acetobacter methanolicus) is a unique acetic acid bacterium capable to grow on methanol as a sole carbon source. Here we report the draft genome sequencing of A. methanolica type strain MB58, showing that it contains 3,270 protein-coding genes, including the genes involved in oxidation of methanol such as mxaFJGIRSACKL and hxlAB, and oxidation of ethanol such as adhAB and adhS. This article is protected by copyright. All rights reserved.
... Recently, Kishimoto et al. (13) have reported Acidobacterium capsulatum, a new acidophilic heterotrophic bacterium isolated from acidic mineral environments. A facultatively methylotrophic bacterium designated as Acidomonas methanolica is also a mesophilic acidophlle (28)(29)(30). ...
... The isolates and Acidiphilium species (6,9,11,18,36) are definitely distinguished from Acidobacterium capsulatum in cellular fatty acid composition, quinone system, G+C content of DNA, and genetic relatedness (13). Acidomonas methanolica (28)(29)(30) is similar to our isolates and Acidiphilium species in ubiquinone type and some other respects. However, the former can be differentiated from the latter in oxidase positive, hydroxy fatty acids with 3-OH C16:0 and 2-OH C16:0 acids, and low G + C content of DNA and DNA homology (13,30), and further in peculiar utilization pattern of the sole carbon and energy sources and requirement of yeast extract or pantothenic acid as a growth factor. ...
Six strains of acidophilic chemoorganotrophic bacteria from acid mine drainage were studied in their taxonomic aspects. They were gram negative, aerobic, mesophilic, oxidase negative, catalase positive, urease positive, nonsporeforming, and rod-shaped. Carotenoid and bacteriochlorophyll a were formed. Two strains had a polar flagellum and other two strains fimbriae. They used a wide variety of organic compounds for growth, but did not use ferrous iron, elemental sulfur, and thiosulfate as the sole energy source. Acetate was inhibitory to growth. Growth was enhanced by adding high concentrations of glucose or complex organic compounds such as trypticase soy (BBL) and yeast extract. Methanol was utilized as the sole source of carbon and energy. The major ubiquinone was Q-10. The major cellular fatty acid was straight-chain unsaturated C18:1 acid. The hydroxy acid was 3-OH C14:0 acid. The DNA base composition was 66.2 to 68.1mol% guanine plus cytosine. The isolates showed relatively low levels of genetic similarity to Acidiphilium cryptum and Acidiphilium organovorum. On the basis of the phenotypic, chemotaxonomic, and genotypic characters, we conclude the isolates as a new species, for which we propose Acidiphilium multivorum sp. nov. The type strain is AIU 301 (JCM 8867).
... Acidomonas methanolica (formerly Acetobacter methanolicus) strain MB58 is a facultative methylotroph for which methanol is the sole source of carbon and energy (Uhlig et al. 1986). A. methanolica contains both methanol-oxidation and ethanol-oxidation systems, and regulates the expression of two respiratory chains with the carbon sources for the growth (Matsushita et al. 1992). ...
Pyrroloquinoline quinone (PQQ)-dependent dehydrogenases (quinoproteins) of acetic acid bacteria (AAB), such as the membrane-bound alcohol dehydrogenase (ADH) and the membrane-bound glucose dehydrogenase, contain PQQ as the prosthetic group. Most of them are located on the periplasmic surface of the cytoplasmic membrane, and function as primary dehydrogenases in cognate substance-oxidizing respiratory chains. Here, we have provided an overview on the function and molecular architecture of AAB quinoproteins, which can be categorized into six groups according to the primary amino acid sequences. Based on the genomic data, we discuss the types of quinoproteins found in AAB genome and how they are distributed. Our analyses indicate that a significant number of uncharacterized orphan quinoproteins are present in AAB. By reviewing recent experimental developments, we discuss how to characterize the as-yet-unknown enzymes. Moreover, our bioinformatics studies also provide insights on how quinoproteins have developed into intricate enzymes. ADH comprises at least two subunits: the quinoprotein dehydrogenase subunit encoded by adhA and the cytochrome subunit encoded by adhB, and the genes are located in a polycistronic transcriptional unit. Findings on stand-alone derivatives of adhA encourage us to speculate on a possible route for ADH development in the evolutional history of AAB. A combination of bioinformatics studies on big genome sequencing data and wet studies assisted with genetic engineering would unravel biochemical functions and physiological role of uncharacterized quinoproteins in AAB, or even in unculturable metagenome.
... It could not be isolated from flowers and fruits. Acidomonas strains isolated mostly from sludge (Uhlig, et al., 1986;Urakami et al., 1989). Yamada et al., (1999) were isolated sixty-four of AAB from Indonesian sources such as fruits, flowers and fermented foods and identified them as Acetobacter strains (fortyfive isolates), Gluconacetobacter strains (eight isolates) and ...
... A. methanolica is an acidophilic facultatively methylotrophic bacterium growing on mineral media of a pH of 4.0-4.5, containing methanol, glycerol or glucose as the only carbon source [92]. Unlike other AAB, A. methanolica metabolizes methanol through the ribulose phosphate pathway. ...
Agro-industrial by-products and wastes pose serious, widespread problems with considerable economic and environmental consequences in developed countries. However, many of the by-products contain large amounts of sugars that make them potentially excellent raw materials for the biotechnological production of added value products; in particular, by-products from perishables such as fruits can be highly useful for this aim. The growing significance and demand for gluconic acid have promoted an interest in integrating both issues as a strategy for the revalorization of these resources.
The pertinence of this strategy can be better understood by examining the properties of gluconic acid and its derivatives and their uses and production methods, especially biotechnological methods, to update the existing reviews on the topic.
Future advances in this direction may be promoted by the development of genetically modified organisms for the generation of new technological processes and the optimization of existing ones. Particular attention is paid to acetic acid bacteria.
... Increases in abundance of methanotrophic bacteria may be an early indication of increases in methane contamination. It should also be noted that there were increases in the abundance of WPS-2 taxa in MSA+ samples, and this group has been shown to co-reside with methanotrophs, suggesting WPS-2 populations might be utilizing the derivatives of methane oxidation (Uhlig et al., 1986;Nogales and Moore, 2001;Fuss and Smock, 2003;Sharp et al., 2012;Grasby et al., 2013). While high-throughput 16S rRNA gene sequencing provided insight into the potential impacts of fracking on microbial communities in the headwater stream ecosystems, future studies should address the functional capacity and metabolic response of these microbial communities to environmental perturbations associated with fracking. ...
Hydraulic fracturing and horizontal drilling have increased dramatically in Pennsylvania Marcellus shale formations, however the potential for major environmental impacts are still incompletely understood. High-throughput sequencing of the 16S rRNA gene was performed to characterize the microbial community structure of water, sediment, bryophyte, and biofilm samples from 26 headwater stream sites in northwestern Pennsylvania with different histories of fracking activity within Marcellus shale formations. Further, we describe the relationship between microbial community structure and environmental parameters measured. Approximately 3.2 million 16S rRNA gene sequences were retrieved from a total of 58 samples. Microbial community analyses showed significant reductions in species richness as well as evenness in sites with Marcellus shale activity. Beta diversity analyses revealed distinct microbial community structure between sites with and without Marcellus shale activity. For example, operational taxonomic units (OTUs) within the Acetobacteracea, Methylocystaceae, Acidobacteriaceae, and Phenylobacterium were greater than three log-fold more abundant in MSA+ sites as compared to MSA- sites. Further, several of these OTUs were strongly negatively correlated with pH and positively correlated with the number of wellpads in a watershed. It should be noted that many of the OTUs enriched in MSA+ sites are putative acidophilic and/or methanotrophic populations. This study revealed apparent shifts in the autochthonous microbial communities and highlighted potential members that could be responding to changing stream conditions as a result of nascent industrial activity in these aquatic ecosystems.
... x Tanticharoenia x x as Acetobacter) was first isolated in sludges of a yeast fermentation (Uhlig et al., 1986). Among the bacteria corresponding to other families, the genus Bacillus was highly represented with 19% of the reads followed by Terriglobus with lower percentage. ...
a b s t r a c t Acetic acid bacteria (AAB) usually develop biofilm on the aireliquid interface of the vinegar elaborated by traditional method. This is the first study in which the AAB microbiota present in a biofilm of vinegar obtained by traditional method was detected by pyrosequencing. Direct genomic DNA extraction from biofilm was set up to obtain suitable quality of DNA to apply in culture-independent molecular tech-niques. The set of primers and TaqMan e MGB probe designed in this study to enumerate the total AAB population by Real Time e PCR detected between 8 Â 10 5 and 1.2 Â 10 6 cells/g in the biofilm. Pyrose-quencing approach reached up to 10 AAB genera identification. The combination of culture-dependent and culture-independent molecular techniques provided a broader view of AAB microbiota from the strawberry biofilm, which was dominated by Ameyamaea, Gluconacetobacter, and Komagataeibacter genera. Culture-dependent techniques allowed isolating only one genotype, which was assigned into the Ameyamaea genus and which required more analysis for a correct species identification. Furthermore, biofilm visualization by laser confocal microscope and scanning electronic microscope showed different dispositions and cell morphologies in the strawberry vinegar biofilm compared with a grape vinegar biofilm.
... Stock cultures were maintained on methanol-containing minimal agar slopes at 4 "C or in 30% (v/v) glycerol at -20 "C. The defined growth medium was that described by Uhlig et al. (1986), containing methanol (1 %, v/v). ...
Membranes of the acidophilic methylotroph Acetobucter methanolicus contained only b-and c-type cytochrome and a CO-binding b-type cytochrome. An azide-sensitive oxidase that oxidizes cytochrome c and ascorbatel TMPD was solubilized from the membrane with a mixture of CHAPS and Zwittergent3-12 (1.7 fold increase in specific activity with 32% yield). The solubilized oxidase is unusually stable with respect to high ionic strength (200 mM-NaC1) and stable between pH 4.0 and 6.8. Of the two soluble c-type cytochromes from A. methanoficus only the typical class I cytochrome (cytochrome cH) was a good substrate, as was equine cytochrome c. The oxidase was partially purified by anion-exchange chromatography but further purification proved impossible. The yield with respect to equine cytochrome c oxidation was l8%, with a 22-fold purification, but during purification most of the activity with respect to cytochrome cH and TMPD was lost. Neutral phospholipids had little effect on activity of the oxidase but the charged phospholipids phosphatidylglycerol and phosphatidylserine stimulated activity up to about fourfold. It proved impossible to incorporate the oxidase into active lipoprotein vesicles. During the purification process the pH optimum for oxidation of cytochrome cH was unchanged (pH 5.6) whereas that for oxidation of equine cytochrome changed from pH 9.5 to 7.5 and the sensitivity of the oxidase to azide changed from non-competitive to competitive during the purification process. The partially-purified oxidase contained only b-type cytochrome, some of which was CO-reactive. It is proposed that the oxidase is a cytochrome co type of oxidase that loses its cytochrome c component during the purification process and is only able to oxidize c-type cytochromes if these can be formed into a 'reconstituted' cytochrome co with the partially-purified oxidase.
... It could not be isolated from flowers and fruits. Acidomonas strains isolated mostly from sludge (Uhlig, Karbaum, & Steudel, 1986;Urakami, Tamaoka, Suzuki, & Komagata, 1989). Yamada et al. (1999) were isolated sixty-four of AAB from Indonesian sources such as fruits, flowers and fermented foods and identified them as Acetobacter strains (fortyfive isolates), Gluconacetobacter strains (eight isolates) and Gluconobacter strains (eleven isolates). ...
Acetic acid bacteria (AAB) comprise a group of gram-negative or gram-variable, ellipsoidal to rod-shaped cells that have an obligate aerobic metabolism with oxygen as the terminal electron acceptor. In the first classification of AAB, two main genera were determined as Acetobacter and Gluconobacter, but nowadays twelve genera are recognized and accommodated to the family Acetobacteraceae, the Alphaproteobacteria: Acetobacter, Gluconobacter, Acidomonas, Gluconacetobacter, Asaia, Kozakia, Swaminathania, Saccharibacter, Neoasaia, Granulibacter, Tanticharoenia and Ameyamaea. Isolation, purification, identification and preservation of AAB are very difficult. Phenotypic methods based on physiological abilitiesies have been used for identification of AAB by using various media. These phenotypic properties have now been complemented or replaced by molecular techniques, which are DNA and RNA based techniques.AAB are widespread in nature on various plants (fruits, cereals, herbs, etc.). They are important microorganisms in food industry because of their ability to oxidize many types of sugars and alcohols to organic acids as end products during fermentation process. The best known industrial application of AAB is vinegar production. This group of bacteria is also used in cellulose and sorbose production. On the other hand, the oxidizing ability of AAB could have spoilage effect in some products such as in wine. The aim of the present review is to introduce the importance of AAB in food industry by showing their current taxonomy, enumeration, isolation and identification methods, isolation sources and beneficial effects in food production systems.
... Group 1 (Methylobacillus strains), group 2 (Protomonas and Methylo,bacterium strains), group 3 (Ancylobacter strains), group 4 (Hyphomicrobium strains), group 5 (Xanthobacter strains), group 6, group 8 (Paracoccus strains), and group 9 (Thiobacillus strain) methanol-utilizing bacteria (16,18,19,23,24) were cultivated in medium B broth (21). Group 7 (Acetobacter strains) methanol-utilizing bacteria (18,19,69) were cultivated in medium C broth. Medium C contained 3.0 g (NH4)2SO4, 4.O g KH2PO4, O.2 g MgSO4. ...
The isoprenoid compounds in gram-negative methanol-, methane-, and methylamine-utilizing bacteria were investigated. All strains tested contained ubiquinone, but none contained menaquinone. The ubiquinone types were Q-8, Q-9, or Q-10. The so-called obligate methylotrophs and methanotrophs (genus Methylobacillus, Methylophaga, Methylomonas, Methylococcus, and Methylovibrio) contained ubiquinone Q-8. The Hyphomicrobium strains contained Q-9. The other facultative methylotrophs and methylamine-utilizing bacteria contained Q-10. A large amount of squalene occurred in the Methylobacillus, Methylophaga, Methylomonas, and Methylococcus strains which utilize one-carbon compounds via the ribulose monophosphate pathway. The Protomonas extorquens and Methylobacterium organophilum strains contained a large amount of sterols (Hop-22(29):ene and Hopan-22-ol), carotenoid pigments, and a small amount of squalene. The Hyphomicrobium strains contained a small amount of squalene and Hop-22(29)-ene. The Xanthobacter strains contained a large amount of carotenoid pigments (zeaxanthin, zeaxanthin monorhamnoside, and zeaxanthin dirhamnoside). The Protomonas and Methylobacterium strains were unique in the existence of sterols and large amounts of total isoprenoid compounds, 4.68 to 7.97 mg/g of dry cell. The distribution of squalene, sterols, quinones, and carotenoid pigments conforms with the morphological, physiological, and other chemo-taxonomic characteristics in gram-negative methanol-, methane-, and methylamine-utilizing bacteria.
... Acidomonas, acidophilic monad). The type species is Acidomonas methanolica (Uhlig et al. 1986) Urakami , Tamaoka, Suzuki, and Komagata comb. nov. ...
A new genus of acidophilic, facultatively methylotrophic bacteria is described. These organisms are gram-negative, nonsporeforming, nonmotile, and rod shaped and grow at pH 2.0 to 5.5. These characteristics are unique among the methanol-utilizing bacteria. The deoxyribonucleic acid base composition is 63 to 65 mol% guanine plus cytosine. Acetobacter methanolicus TK 0705(T) (T = type strain) is a typical strain in this group. These bacteria are distinguished from type and representative strains of Acetobacter, Gluconobacter, Acidiphilium, and Thiobacillus on the basis of deoxyribonucleic acid-deoxyribonucleic acid homology. A new genus, Acidomonas, is proposed to include this group of methylotrophic bacteria. The type species of the genus Acidomonas is Acidomonas methanolica comb. nov., with type strain TK 0705 (= IMET 10945).
... ACIDOMONAS Urakami et al., 1989emend. Yamashita et al., 2004 Acidomonas methanolica Yeast fermentation (Uhlig et al., 1986) process Urakami et al., 1989 High acid vinegar (Sievers et al., 1992) fermentation Yamada et al., 1997 Gluconacetobacter hansenii Vinegar (Gosselé et al., 1983) Yamada et al., 1997emend. Lisdiyanti et al., 2006 (Boesch et al., 1998) Buchanan et al. (1966). ...
The exploitation of acetic acid bacteria (AAB) has a long history in fermentation processes and now represents an emerging
field in biotechnological applications, especially with regard to the biosynthesis of useful chemicals with a potentially
high economic value and, in food science, through the standardization of microbiological processes for the manufacture of
both vinegar and other fermented beverages.
... For instance, members of the genera, Asaia have infected patients with cystic fibrosis, Granulibacter has infected patients with CGD, and Gluconobacter infected a patient with a history of intravenous drug use, and another with cystic fibrosis [8]. A. methanolica is an acidotolerant, facultatively methylotrophic, aerobic, gram-negative, catalase-positive, urease-positive, and oxidase-positive, non-spore-forming, nonmotile, rodshaped bacteria that is 0.8 to 1.0 by 1.5 to 3.0 micrometers, occurring singly, or rarely in pairs [9]. Initially categorized in 1986 as Acetobacter methanolicus, the organism, was reclassified to the genus Acidomonas in 1989 by Urakami et al. based upon its methyltrophic capabilities, which were absent in bacteria of the genus Acetobacter [10,11]. ...
Purpose:
Adenitis for which no causative organism can be isolated is a common occurrence in patients with chronic granulomatous disease (CGD). Here we identify Acidomonas methanolica as a pathogen associated with adenitis in a patient with CGD.
Methods:
The causative pathogen was obtained after prolonged incubation of an excised lymph node in thioglycolate broth. Identification was carried out by sequencing the 16s rRNA. Immunoblots were prepared utilizing protein extracts from the case patient's A. methanolica isolate, an ATCC type strain of A. methanolica and G. bethesdensis.
Results:
Fastidious gram-negative rods grew after prolonged incubation of an excised lymph node in thioglycolate broth. Sequencing of the 16s rRNA identified the organism as A. methanolica. Immunoblot confirmed the pathogen's role in the patient's adenitis by showing the patient's specific immune response to the organism.
Conclusions:
A. methanolica is the second member of the family, Acetobacteaceae to be associated with adenitis in patients with CGD.
Rare earth elements (REEs) are important high-tech materials with widespread industrial applications and a high risk of supply disruption. Current physico-chemical methods for REE recovery are energy intensive, complex, and costly. Thus, it is crucial to undertake measures to secure future REE demand and protect the environment as well. Bioleaching as an alternative technology for the recovery and recycling of REEs from a variety of primary and secondary sources has been increasingly explored. The present review provides an overview on REE content of various primary and secondary sources, applied bioleaching microorganisms and methods, reactor setups, as well as a detailed description of known REE leaching mechanisms. The effects of different process parameters including temperature, pH, medium composition, pulp density, and particle size on efficiency of REE leaching are also evaluated in detail.
Rare earth elements (REEs) are important high-tech materials with widespread industrial applications and a high risk of supply disruption. Current physico-chemical methods for REE recovery are energy intensive, complex, and costly. Thus, it is crucial to undertake measures to secure future REE demand and protect the environment as well. Bioleaching as an alternative technology for the recovery and recycling of REEs from a variety of primary and secondary sources has been increasingly explored. The present review provides an overview on REE content of various primary and secondary sources, applied bioleaching microorganisms and methods, reactor setups, as well as a detailed description of known REE leaching mechanisms. The effects of different process parameters including temperature, pH, medium composition, pulp density, and particle size on efficiency of REE leaching are also evaluated in detail.
This chapter highlights the huge and manifold possibilities of reactions which result from the interactions between microorganisms and the geosphere and which are used for mining, mineral processing, and metal recycling. Besides the introduction (Sect. 1) the contribution is divided into five different sections describing the mobilization (Sect. 2) and immobilization (Sect. 3) of valuable substances, the processes of biosorption and bioaccumulation (Sect. 4), as well as transformation of metals into metal organic compounds (Sect. 5). A special topic (Sect. 6) addresses the application of CO2 as an important component for the formation of energy-rich compounds and chemicals. Each section starts with an overview of the relevant reactions and an explanation of the reaction condition. Afterward information about applications and different technological processes as well as sustainability aspects are provided.
A.ci.do.mo' nas . Gr. adj. acid acid Gr. n. monas unit, monadM .L. fem. n. Acidomonas acidophilic monad.
Proteobacteria / Alphaproteobacteria / Rhodospirillales / Acetobacteraceae / Acidomonas
Cells are Gram negative, nonsporeforming, nonmotile, rod shaped, 0.8–1.0 × 1.5–3.0 µm. Cells occur singly, rarely in pairs. Colonies were white to yellow on PYM medium. Strictly aerobic. Facultative methylotroph. Metabolism is respiratory, never fermentative. Optimal temperature for growth is 30°C. Optimal pH for growth is pH 4.0–4.5. Catalase positive. Nitrate is not reduced to nitrite. Oxidizes ethanol to acetic acid. Overoxidizes acetic acid to CO 2 and H 2 O. Acidomonas contains C 18:1 straight‐chain unsaturated fatty acid as well as C 16:0 3OH and C 16:0 2OH as major components of cellular and hydroxy fatty acids, as do the other genera of the family Acetobacteraceae (Yamada et al., 1981; Urakami and Komagata, 1987a; Urakami et al., 1989a). The ubiquinone system is Q‐10, along with ubiquinone Q‐9 and Q‐11 as minor components.
The mol % G + C of the DNA is : 62.
Type species : Acidomonas methanolica (Uhlig, Karbaum and Steudel 1986) Urakami, Tamaoka, Suzuki and Komagata 1989a, 54 ( Acetobacter methanolicus Uhlig, Karbaum and Steudel 1986, 321.)
A.ce.to.bac.te.ra' ce.ae. M.L. masc. n. Acetobacter type genus of the family; -aceae ending to denote a family; M.L. fem. pl. n. Acetobacteraceae the Acetobacter family. Proteobacteria / Alphaproteobacteria / Rhodospirillales / Acetobacteraceae
This study was carried out to investigate characteristics of acetic acid bacteria for persimmon vinegar fermen-tation. Acetic acid bacteria were isolated from statically fermented vinegar. Microscopically the cells appeared as non-motile and non-flagellated, preferentially occuring in pairs. Isolated strains were able to grow in the presence of acetic acid, ethanol, and glucose. Four of them produced 2-ketogluconic acid but did not produce 5-ketogluconic acid. The four strains isolated from statically fermented vinegar were considered to be grouped into Acetobacter. The strains were inoculated into persimmon juice for persimmon vinegar fermentation. They produced acetic acid in the range of 5.25\?5.68\%.
Acetic acid bacteria are currently accommodated in the acetous group, the family Acetobacteraceae, the class Alphaproteobacteria, based on phylogeny, physiology, and ecology. The acetic acid bacteria are classified at present in 17 genera, of which many species have been reported in the genera Acetobacter, Gluconobacter, Gluconacetobacter, Asaia, and Komagataeibacter. Of the remaining 12 genera, Acidomonas, Kozakia, Swaminathania, Saccharibacter, Neoasaia, Granulibacter, Tanticharoenia, Ameyamaea, Endobacter, Nguyenibacter, and Swingsia are monotypic; the genus Neokomagataea contains two species. In the class Gammaproteobacteria, the genus Frateuria has been mentioned taxonomically as pseudacetic acid bacteria. In addition, isolation and identification of acetic acid bacteria are described.
One-carbon compounds (C1) at all oxidation levels between methane and carbon dioxide occur abundantly throughout nature. Methane is present in fossil deposits and is formed by methanogenic bacteria. Methanol arises by the hydrolysis of methyl esters and ethers such as pectin and lignin, which are present in plants. Methylated amines occur in plants and animals and are produced by microbial degradation of choline derivatives present in plant membrane material and animal tissue. Formate is a major end-product of mixed-acid fermentation and carbon dioxide is present in the atmosphere and, as carbonates, in natural waters and soil. It is not surprising, therefore, that microorganisms are found in nature which are capable of growth on such compounds as carbon and energy sources.
To produce and utilize microbial cellulose being industrially useful, bacterium posessing a high ability to produce cellulose and the uniform structure of cellulose pellicle was isolated from the traditionally fermented vinegar in korea. The isolated strain, GS11 was gram-negative, rod(0.6 x 2.2~3.3 μm) and motile. It also had 58.4% of DNA base composition, posessed straight-chain C(18:l), C(16:0) and C(14:0) fatty acid, and contained ubiquinone Q(10). According to its morphological, physiological and biochemical properties, the isolated strain was identified as Acetobacter xylinum. A. xylinum GS11 was cultured for the study of cellulose productivity during the static culture, 12 days at 30°C. As this result, the amount of glucose was quickly decreased, and then used as substrate for cellulose production. In this culture, the maximal production of cellulose peaked with 2.8 g/l after 8 days.
Introduction and Historical ViewBiological FundamentalsIndustrial Production ProceduresProduct Recovery and ProcessingUtilization of Gluconic Acid and GluconatesProduction FiguresReferences
Ten types of farm-made brewing vinegars were collected and four high acetic acid-producing strains (CV1, CV3, CV5, and CV6) were isolated. Among them strain CV1, exhibiting highly alcohol-resistant and acetic acid-producing properties, was selected and its taxonomic properties were investigated by phenotypic (particularly chemotaxonomic) characterization and phy- logenetic inference based on 16S rRNA gene sequence analysis. On SM broth agar, cells of strain CV1 were gram-staining- negative and formed pale white colonies with smooth to rough surfaces. Strain CV1 produced acetate from ethanoi and was resistant to up to 8% (v/v) ethanoi in LM broth. Strain CV1 had a G+C content of 61.0 mol%, contained meso-DAP as the cell wall amino acid, and possessed Q-10 as the major ubiquinone. A comparison of 16S rRNA gene sequences showed that strain CV1 was most closely related to Gluconacetobacter saccharivorans (299.0% identity). In liquid media, the optimum growth conditions for acetic acid production were 30°C and pH >3.0 and strain CV1 produced 9.3% and 8.4% acetic acids from 10% and 9% alcohol concentrations, respectively.
Part I: Acetic, Lactic, Gluconic, Succinic and Polyhydroxyalkanoic AcidsPalmer Rogers, Jiann-Shin Chen Mary Jo ZidwickIn MemoriumShortly before the completion of this chapter, Palmer Rogers passed away suddenly. Palmer was the one who motivated and shepherded us to the completion of this work. He had a lifelong dedication to education of students and helped them attain the satisfaction that achieving in-depth understanding of science through hard work can bring. Palmer was kind, creative, energetic and uncompromising in his scientific integrity and will be remembered with fondness by the many people whose lives he touched.General IntroductionThe objective of this chapter is to present the ways bacteria are effectively harnessed as biocatalysts to perform the synthesis of bulk organic acids and solvents. Prior to the development of the petroleum-based chemical industry, microbial fermentations of agricultural biomass were ...
The objective of this chapter is to present the ways bacteria are effectively harnessed as biocatalysts to perform the synthesis of bulk organic acids and solvents. Prior to the development of the petroleum-based chemical industry, microbial fermentations of agricultural biomass were a major source of a number of useful bulk organic chemicals. Commercial chemical production often emerged from a much earlier food-processing technology where grains, corns, milks, and fruits were fermented to wines, beers, cheeses, and vinegars. Beginning at the end of the nineteenth century and continuing to the present, specific bacterial strains were selected from nature to produce commercially needed bulk chemicals such as lactic acid, acetic acid, acetone and butanol, and more recently gluconic acid and polyhydroxyalkanoates. Lactic acid currently is produced at very large volumes for a multitude of food and industrial uses. Using the tools of metabolic engineering, bacterial strains are being altered for production of propanediols, butanediol, and succinic acid at higher yields and productivity than are possible using natural strains.
The family Acetobacteraceae is taxonomically included in the order Rhodospirillales of the class Alphaproteobacteria, and 32 genera are validly described. The genera are basically classified into two groups, an acetous group and an acidophilic group, in the light of application, ecology, and phylogeny. The acetous group comprises genera in acetic acid bacteria like Acetobacter, Gluconacetobacter, Gluconobacter, Asaia, Granulibacter, and Komagataeibacter. The acidophilic group consists of acidophilic and neutrophilic genera like Acidiphilium and Roseomonas. In the 1960s, taxonomy of acetic acid bacteria was significantly affected by the chemotaxonomic study with G+C content of DNA, quinone systems, cellular fatty acid composition, and DNADNA similarity. Further, data of phylogenetic analysis based on 16S rRNA gene sequences have had a profound impact on the systematics of acetic acid bacteria and other bacteria over all. Membrane-bound dehydrogenases are responsible for the oxidation of alcohols and sugars in acetic acid bacteria. The dehydrogenases are located in the periplasmic side of the cytoplasmic membrane of the bacteria. The direct electron acceptor of the dehydrogenases is ubiquinone in the respiratory chain of the acetic acid bacteria. Production of acetic acid from ethanol and of D-gluconate, 2-keto-D-gluconate, 5-keto-D-gluconate, and 2,5-diketo-D-gluconate from D-glucose is due to the membrane-bound dehydrogenases. Acetic acid bacteria are widely distributed in alcoholic and acidic environments, and they are isolated from vinegar, wine, beer, sake, cider, fermented foods, fruits, flowers, and other alcoholic materials. Asaia strains are isolated from flowers and even from mosquitoes and other insects. Granulibacter is known for its pathogenicity for humans. Acetic acid bacteria are widely used for production of vinegar. D-Gluconic acid and keto-D-gluconic acids are produced from D-glucose by Gluconobacter strains, which are used in the food, pharmaceutical, and chemical industries. L-Sorbose is produced from sorbitol by Gluconobacter strains, which is further converted to 2-keto-L-gulonic acid as a penultimate intermediate in the industrial production of vitamin C.
Ribosomal 5SRNA (rRNA) was isolated from 12 strains belonging to the genera Acidomonas, Acetobacter and Gluconobacter and sequenced. A dendrogram constructed from the data indicated that methylotrophic and nonmethylotrophic strains of the genus Acetobacter formed two separate clusters. The non-methylotrophic members of the genus Acetobacter were phylogenetically closer to Gluconobacter than to the methylotrophic strains of Acetobacter. The methylotrophic strains of Acetobacter were recovered as a clade with the type strain of Acidomonas methanolica. These data support an earlier proposal which reclassified methylotrophic strains of Acetobacter into the genus Acidomonas.
The facultative methylotrophic bacterium Acidomonas methanolica MB58 can utilize C1 compounds via the ribulose monophosphate pathway. A large gene cluster comprising three components related to C1 metabolism was found in the genome. From upstream, the first was an mxa cluster encoding proteins for oxidation of methanol to formaldehyde; the second was the rmp cluster encoding enzymes for formaldehyde fixation; and the third was the cbb gene cluster encoding proteins for carbon dioxide (CO2) fixation. Examination of CO2 requirements for growth of A. methanolica MB58 cells demonstrated that it did not grow on any carbon source under CO2-free conditions. Measurement of ribulose-1,5-bisphosphate carboxylase activity and RT-PCR analysis demonstrated enzymatic activity was detected in A. methanolica MB58 at growth phase, regardless of carbon sources. However, methanol dehydrogenase and 3-hexlose-6-phosphate synthase expression was regulated by methanol or formaldehyde; it were detected during growth and apparently differed from ribulose-1, 5-bisphosphate carboxylase expression. These results suggested that A. methanolica MB58 may be initially dependent on autotrophic growth and that carbon assimilation was subsequently coupled with the ribulose monophosphate pathway at early- to mid-log phases during methylotrophic growth.
Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.
The characteristics of a strain that contaminates the manufacturing of rice vinegar by Acetobacter pasteruianus was invetigated. Conditions for inhibiting pellicle formation and growth of the contaminant, which occurs during static culture and storage, were also observed. Examining the morphological, cultural, and physiological characteristics and measuring the amount of cellulose production during static culture for 14 day, we found that the strain was known to be Acetobacter xylinum. No growth was observed below as well as over . Also, the extent of growth was limited when the concentrations of ethanol, NaCl, and acetic acid were more than 10%, 1.5%, and 7%, respectively.
This chapter explains the manifold geobiotechnological possibilities to separate industrial valuable metals from various industrial residues and stored waste products of the past. In addition to an overview of the different microbially catalyzed chemical reactions applicable for a separation of metals and details of published studies, results of many individual investigations from various research projects are described. These concern the separation of rare earth elements from phosphorous production slags, the attempts of tin leaching from mining flotation residues, the separation of metals from spent catalysts, or the treatment of ashes as valuable metal-containing material. The residues of environmental technologies are integrated into this overview as well. The description of the different known microbial processes offers starting points for suitable and new technologies. In addition to the application of chemolithoautotrophic microorganisms the use of heterotrophic microorganisms is explained.
The 16S rRNA gene (rDNA) sequence from Acetobacter methanolicus MB58 was determined and aligned with known sequences of Acetobacter and Gluconobacter species from a previous study (Sievers et al. 1994). Acetobacter methanolicus forms a single branch between Acetobacter pasteurianus / Acetobacter aceti and the cluster comprising Acetobacter europaeus, Acetobacter xylinum, Acetobacter hansenii, Acetobacter diazotrophicus and Acetobacter liquefaciens. The phylogenetic data provide sufficient and clear evidence for the revival of Acetobacter methanolicus from “Acidomonas methanolica”.
18 Mutanten von Acetobacter methanolicus MB 58, die nach MNNG- oder UV-Licht-Behandlung nicht mehr auf Methanol als Kohlenstoff- und Energiequelle wachsen konnten, wurden auf Defekte bezüglich der linearen und zyklischen dissimilatorischen Sequenz von Formaldehyd sowie der Phosphofructokinase getestet. Die meisten Mutanten waren pleiotroph und defekt in der C1-regulierten Methanol-Dehydrogenase und/oder der konstitutiven Formaldehyd-Dehydrogenase. Cytochrom c-Verlust war immer gekoppelt mit dem Ausfall an Methanol-Dehydrogenase-Aktivität. Formiat-Dehydrogenase-Defekt war nur in einer pleiotrophen Mutante nachweisbar. Keine der heterotroph vermehrten Mutanten wies Defekte im dissimilatorischen RuMP-Zyklus sowie in der Phosphofructokinase auf. In 3 zur Oxydation von Methanol noch befähigten Mutanten konnte durch Methanol die Hexulose-6-phosphat-Isomerase nicht induziert werden, was als Hinweis auf Isoenzyme angesehen werden kann. In 2 Mutanten konnten an den untersuchten Stellen keine Enzymdefekte gefunden werden, was wahrscheinlich macht, daß assimilatorische Enzyme getroffen worden sind.
Die Produkte der Essigsäurebakterien sind überwiegend das Ergebnis unvollständiger Oxydationen. Sie sind seit langem bekannt und im Gebrauch. Bakteriencellulose ist eine Ausnahme. Bisher ungenutzt, scheinen sich jetzt industrielle Anwendungsmöglichkeiten für sie zu erschießen. Eine neue beschriebene Art, Acetobacter methanolicus, eröffenet neue Möglichkeiten für Prozesse und Produkte: SCP auf der Basis von Methanol, Abwasserreinigung, aerobe Säuerung als Bestandteil der Silierung, Gluconsäure. Aufgrund ihrer physiologischen Spezifik und technologischen Vielseitigkeit ist die Art gleichzeitig ein interessantes Untersuchungsobjekt im Hinblick auf Bakteriophagen und neue Wirt-Vector-Systeme.
We report here the construction of a promoter-probe vector, pRS2, which can be utilized in either Acetobacter methanolicus MB 58 or Escherichia coli due to the presence of broad-host-range replicon RSF 1010. The vector provides several unique restriction sites for promoter cloning as well as resistance markers for the selection of transformants. The promoter-probe vector was constructed by inserting an EcoRI-SalI-polylinker fragment of pUC 19 into EcoRI/SalI digested pMK 16. The resulting plasmid, pRS1, was cloned into the unique EcoRI site of the broad-host-range plasmid RSF 1010.
The vector was used to clone promoter-containing sequences derived from the A. methanolicus MB 58 chromosome as well as the E. coli lac-promoter.
Using a methanol utilizing acidophilic MB 58 bacteria strain (IMET 10945 T ), a stable fermentation process was performed in a pilot plant pressure fermenter of 250 l total volume for more than four months.
Under the fermentation conditions (system pressure and retention time) used, productivities ranged between 2.3 and 9.4 BDM kg ⁻¹ . h ⁻¹ and growth yields between 0.31 and 0.42.
Models describe the correlations between yield and retention time. The mean value of growth yield amounts to 0.41 at a retention time of 6.5 h.
Electrophoretic analysis of the proteins bound to poly(3-hydroxybutyric acid), PHB-, granules in Methylobacterium extorquens, M. rhodesianum as well as the PHB-leaky mutants Mu 1 and Mu 11, which were isolated from the latter, resulted in two dominant low-molecular weight proteins, which were referred to as GA 11 and GA20. After purification of these proteins antibodies against the GA11 and GA20 protein of M. extorquens were obtained. Both proteins bound to the surface of PHB granules as revealed by immunoelectron microscopy of whole cells of M. extorquens and M. rhodesianum. With cells of the PHB-leaky mutants Mu 1 and Mu 11 no specific labeling was observed. The N-terminal amino acid sequences of the GA11 and the GA20 protein were determined. We found significant homologies between the sequences of the investigated strains. The use of oligonucleotide probes based on the N-terminal sequences of the GA20 protein from M. rhodesianum to identify the corresponding structural genes in various genomic libraries failed.
Hitherto, lower growth yields were found with acidophilic methylotrophic bacteria, than those found with neutrophilic methylotrophic ones. Since differences with respect to the oxidative phosphorylation might be responsible for this phenomenon, P/0 quotients were determined for the oxidation by Acetobacter methanolicus sp. MB 70 of endogenous substrates, methanol and other substrates. P/0 quotients for endogenous substrates were calculated from the formation of “energy-rich” phosphate bonds and the concomitant oxygen uptake on transition of the cells from anaerobiosis to aerobiosis in the range between 0.07 to 0.17 nmol P/nmol 0. P/0 quotients for methanol between 0.1 to 0.25 were determined under aerobic conditions by intracellular energization. For this purpose, the energy charge was decreased to values lower than 0.5 by changing the extracellular pH from 4.0 to 7.0, and increased again to values from 0.7 to nearly 1.0, by adding methanol. The addition of FCCP and DCCD2) totally prevented the increase of the ATP concentration. The energization of the same cell preparation produced similar P/0 quotients for methanol, ethanol and glucose. But in a certain cell preparation, the P/0 quotient for glucose (0.01) was considerably lower than that for methanol (0.18); this was the only quotient reduced from 0.12 to 0.05 by aerating the former cell preparation for 4 h. The results are discussed in terms of the coupling of dehydrogenase reactions, for endogenous and exogenous substrates, to the cytochromes of the respiratory chain. Its composition and structure before cytochrome c appears to be responsible for the fact that the growth yields of these bacteria are smaller than expected for such a type of methylotrophs.
Methanol oxidation in Mefhyfophuga marina appears to be organized in a way similar to that in other Gram- negative methylotrophs, involving a quinoprotein methanol dehydrogenase (MDH; EC 1.1.99.8) and two soluble cytochromes. The MDH is composed of two different subunits, most probably arranged in an a2P2 structure. The two cytochromes are of the e-type and differ in size (molecular mass 19.5 and 12.5 kDa) and isoelectric point (PI 4.6 and 9.2). The one with the lowest isoelectric point, commonly designated as cytochrome eL, is able to oxidize reduced MDH. Taking advantage of the ability of M. marina, a restricted facultative methylotroph, to utilize fructose as a growth substrate, mutants impaired in methanol utilization were isolated after application of optimal concentrations of ethylmethane sulphonate. Three classes of methanol oxidation mutants were obtained. Class I mutants were affected in a global regulation of the synthesis of both apo-methanol-dehydrogenase and cytochrome cL as well as PQQ (pyrroloquinoline quinone). Class I1 mutants did not produce an active MDH, but instead a comparable amount of a 65 kDa protein was found in the cell-free extract upon SDS-PAGE. This mutant protein was purified and compared to wild-type MDH. It was located in the periplasm, but unlike MDH it was composed of only two identical large subunits. Each of these subunits was able to bind one molecule of PQQ. An antiserum raised against wild-type MDH did not react with the mutant protein. Conversely, an antiserum raised against mutant protein weakly cross-reacted with wild-type MDH, suggesting that the presence of the P-subunit in MDH dramatically changes its immunochemical behaviour. Three of the class I1 mutants did not produce PQQ. In the presence of PQQ, partial revertants able to grow on methanol medium were obtained (class 111). Class 111 mutants produced a stable apo-MDH consisting of a- and P-subunits and showing a normal reaction with the anti-MDH serum. Although this apo-MDH can bind PQQ, enzymic activity was not restored in nitro. This suggests that, in addition to apo-MDH and PQQ, other factors are required for the assembly of an enzymically active MDH.
Natural and anthropogenic acidic environments are dominated by bacteria and Archaea. As many as 86 genera or species have been identified or isolated from pH Keywords: Sulfolobus; Thiobacillus; acid mine drainage; solfataras; springs Document Type: Regular Paper Affiliations: U.S. Geological Survey, National Center MS 956, Reston, VA 20192, U.S.A. Publication date: August 1, 2000 $(document).ready(function() { var shortdescription = $(".originaldescription").text().replace(/\\&/g, '&').replace(/\\, '<').replace(/\\>/g, '>').replace(/\\t/g, ' ').replace(/\\n/g, ''); if (shortdescription.length > 350){ shortdescription = "" + shortdescription.substring(0,250) + "... more"; } $(".descriptionitem").prepend(shortdescription); $(".shortdescription a").click(function() { $(".shortdescription").hide(); $(".originaldescription").slideDown(); return false; }); }); Related content In this: publication By this: publisher By this author: Robbins, E.I. GA_googleFillSlot("Horizontal_banner_bottom");
The poly(3-hydroxybutyric acid) (PHB) granules from eight methylotrophic bacteria that use the serine pathway were isolated in a sucrose gradient (1–2 M); these bacteria included members of the genus Methylobacterium, Mycoplana rubra, and PHB-leaky mutants of Methylobacterium rhodesianum. As shown by sodium dodecyl sulfate – polyacrylamide gel electrophoresis, the granules from all investigated methylotrophic strains revealed two major bands representing small proteins. An efficient purification procedure for these two low molecular weight proteins associated with the PHB granules was developed by solubilization of the proteins with Triton X-114 and affinity chromatography on Procion Blue-H-ERD.Key words: poly(3-hydroxybutyric acid), granule-associated proteins, methylotrophic bacteria.
Laboratorium voor Microbiologie en microbiele Genetica, Rijksuniversiteit, B-9000 Ghent, Belgium'; Deutsche Sammlung von Mikroorganismen und Zellkulturen, 0-3300 Braunschweig, Federal Republic of Germany,; and EMBRAPA, Programa Nncional de Pesquisa em Biologia do Solo, Seropkdica, 23851 Rio de Janeiro, Brazil3 Results of deoxyribonucleic acid (DNA)-ribosomal ribonucleic acid and DNA-DNA hybridizations, together with a phenotypic and chemotaxonomic analysis, revealed that nitrogen-fixing bacteria isolated from roots and stems of sugarcane belong to a new species in the genus Acetobacter, for which the name Acetobacter diazotrophicus sp. nov. is proposed. Strain LMG 7603 (= Dobereiner PA1 5 = ATCC 49037) is the type strain. New microaerobic, gram-negative, N,-fixing bacteria were isolated from roots and stems of sugarcane in Brazil (1). Because of their ability to grow at low pH values and their ability to form acetic acid from ethanol, these organ-isms could belong to the acetic acid bacteria (the genera Acetobacter and Gluconobacter) (3, 4) or to the genus Frateuria (14). However, Cavalcante and Dobereiner (1) believed that the differences between these genera and the N,-fixing sugarcane isolates were sufficiently great to pro-pose the name "Saccharobacter nitrocaptans" (not validly published). The original description (1) was based on more than 20 strains isolated in four different regions of Brazil. In this paper we present genomic, phenotypic, and chemo-taxonomic evidence that these isolates constitute a new species in the genus Acetobacter, for which we propose the name Acetobacter diazotrophicus. Details of the strains and their sources are given in Table 1. Cells for genomic studies, protein electrophoresis, and determination of ubiquinones were grown on GYC medium (5% glucose, 1% yeast extract, 3% CaCO,, 2.5% agar). The methods which we used have been described previously (2, 4, 6-12). Differentiating phenotypic tests were
Methylotrophic bacteria which use hexulose-6-phosphate synthase for fixing formaldehyde are able to oxidize it cyclically via 6-phosphogluconate. This also holds true for Acetobacter methanolicus MB58. Generally methylotrophic bacteria oxidize formaldehyde via formate to CO2. Up to now only the presence of a membrane-bound, dye-mediated formate dehydrogenase has been demonstrated in Acetobacter methanolicus MB58. In this paper evidence for the presence of an aldehyde dehydrogenase is provided which catalyzes the oxidation of formaldehyde by means of toluylene blue as an electron acceptor. Hence Acetobacter methanolicus MB58 possesses at least two routes for the dissimilation of formaldehyde to CO2.
The lysine content of the biomass of the acidophilic facultatively methylotrophic bacterium Acetobacter methanolicus MB 58 was increased by genetic manipulations.
A homoserine auxotroph, MB 58.196, and a threonine auxotroph, MB 58.195, were obtained from Acetobacter methanolicus MB 58 by N-methyl-N′-nitro-N-nitrosoguanidine treatment.
Investigations of enzyme activities revealed that the homoserine auxotroph lacks homoserine dehydrogenase activity, and the threonine auxotroph lacks homoserine kinase activity.
Concerning the lysine-producing ability, only the homoserine auxotrophic mutant accumulates lysine in the intracellular pool. The intracellular lysine content of this mutant was increased 40-fold.
An excretion of amino acids into the medium was not detected. A homoserine resistant mutant, MB 58.196.10, isolated from MB 58.196 by UV-irradiation, was able to excrete lysine. About 95% of free lysine were found in the culture medium. Altogether, the free lysine concentration was increased 800-fold in comparison to the wild-type strain.
By these genetic manipulations the total lysine concentration of MB 58.196 was increased to 2.7% and of MB 58.196.10 to 56% in comparison to the wild-type strain.
The optimization task was performed using the gluconic acid synthesis by the Acetobacter methanolicusMB 58 strain. The microorganisms were grown continuously on methanol as the growth substrate. After finishing the growth process by the deficiency of N and P, the gluconic acid synthesis was started by adding glucose. The synthesis process was performed continuously. The oxygen transfer rate depended on the gluconic acid concentration. During the growth process, the oxygen transfer rate reached a value of about 13 g O2 · kg−1 · h−1using a 30-l glass fermenter equipped with a 6 blade stirrer and fully baffled. This rate declined to a value of between 2 and 5 g O2 · kg−1 · h−1 in the presence of gluconic acid concentrations above 150 g gluconic acid · kg−1medium. The yield (g gluconic acid · g−1glucose) depended on the gluconic acid concentration and amounted to y = 0.7 in relation to 150 g gluconic acid · kg−1medium and y = 0.8 in relation to 200 g · kg−1medium, respectively. The fermenters were coupled with ultrafiltration moduls (Fa. ROMICON and Fa. SARTORIUS). The biomass concentrations amounted from 5 to 40 g dry mass kg−1medium. The ultrafiltration modules retained the biomass within the fermentation system. A glucose solution (30 to 50 weight percent glucose) was continuously dosed into the fermenter. The retention time was chosen between 2 and 30 h. The gluconic acid synthesis rate reached values of up to 32 g gluconic acid · kg−1 · h−1. Within a range of up to 250 g gluconic acid · kg−1medium, the acid concentration had no influence on the enzyme activity.
Ninety-eight Gluconobacter strains of various origins were examined by numerical analysis of 177 phenotypic features and by gel electrophoresis of soluble proteins. Gluconobacter was phenotypically quite different from Aceto- bacter and Frateuria. An extensive phenotypic description and a minimal description of the genus Gluconobacter are given. The genus Gluconobacter contained two groups, A and B, by both techniques. Phenons A and B could be differentiated only by the requirement for nicotinic acid and by their electropho- retic patterns. Protein electrophoresis showed clearly that Gluconobacter strains are genetically stable over several decades. The strains of all five subspecies of Gluconobacter oxydans cited in the Approved Lists of Bacterial Names (Skerman et al., Int. J. Syst. Bacteriol. 30:225-420, 1980) were distributed randomly over the phenotypic and electrophoretic clusters and subclusters, and the type strains of the subspecies all fell in phenon B. We conclude that the single species Gluconobacter oxydans should not be further divided into subspecies.
The properties and taxonomic positions of 11 strains previously identified as members of "Acetobacter aurantius Kondb and Ameyama" (this name is not on the Approved Lists) were reexamined. For each we determined about 100 phe- notypic features, the protein gel electropherograms, and the parameters of deoxy- ribonucleic acid: "C-ribosomal ribonucleic acid (DNA:( "ClrRNA) hybrids. The 11 strains fell into three taxonomically distinct groups. Strain IF0 3248 was the only one which belonged in Acetobacter. Strain IF0 3246 was an atypical Gluconobacter. The remaining nine strains formed a tight cluster, with very similar phenotypic features and protein gel electropherograms. Taxonomically, this cluster is quite removed from Gluconobacter and Acetobacter, and the properties of its DNA:rRNA hybrids suggest that it is closer to Pseudomonas Section I (R. E. Buchanan and N. E. Gibbons (ed.), BergeyS Manual of Deter- minative Bacteriology, 8th ed.) and Xanthomonas. We propose the name Fra- teuria gen. nov. for $his cluster, with Frateuria aurantia sp. nov. as the type species and IF0 3245 as the type strain. An extensive phenotypic description and minimal standards of the new genus are given, as is the phenotypic differentiation from Glucono bacter and Aceto bacter. The name "Acetobacter aurantium" (sic) was proposed by Kondb and Ameyama (17) to refer to four strains of acetic acid bacteria isolated from Lilium auratum Lindl. Because these strains share diagnostic features with Glucono- bacter as well as with Acetobacter, Asai (3) and Asai et al. (4) judged that it was difficult to assign them to either of these two genera and therefore referred to them as "intermediate" strains. Additional characteristics of these strains were given by Ameyama and Kondb (1, 2). Yamada et al. (24) showed that these strains produce ubiquinone Q8, whereas Glucono bacter possesses ubiquinone QlO and Acetobacter pos- sesses ubiquinone Q9. Yamada et al. (25) isolated and characterized six additional polarly flagel- lated intermediate strains from Rubus parvifol- ius.
Von 6 methanutilisierenden und 11 methanolutilisierenden Bakterienspecies wurden spektralphotometrische Analysen durchgeführt. Damit liegen einschließlich der Befunde von DAVEY u. MITTON (1973), TONGE et al. (1974) sowie ANTHONY (1975) die Cytochromspektren von 9 methan‐ und 14 methanolutilisierenden Bakterien vor. Obwohl zwischen obligat und fakultativ methylotrophen Bakterien unterschieden werden kann, die Assimilation der C1‐Verbindungen über zwei prinzipiell verschiedene Sequenzen erfolgt und sich die einzelnen Species in morphologischen, ultrastrukturellen und physiologischen Merkmalen erheblich unterscheiden, besitzen alle methanutilisierenden Bakterien ein qualitativ nahezu identisches Cytochromkomplement, das sich von den Spektren der methylotrophen, aber Methan nicht utilisierenden Bakterien, die bis auf Ausnahmen fakultativ methylotroph sind, nicht signifikant unterscheidet. Charakteristisch ist der hohe Gehalt an Cytochrom c und o sowie die sehr geringe Konzentration an b. Alle methanutilisierenden Bakterienstämme sowie die Hyphomicrobium‐Species verfügen über den Cytochrom aa3‐Komplex; in den methanolutilisierenden Bakterien konnte dieser nicht in allen Fällen sicher nachgewiesen werden. Das Cytochromspektrum ist weder zur Differenzierung methanoxydierender Bakterien mit verschiedenen internen Membransystemen geeignet noch kann es zur Unterscheidung von obligat und fakultativ methylotrophen Species dienen. Das prinzipiell identische Cytochromkomplement sowie die Tatsache, daß die Cytochrome der „methanotrophen” Bakterien mit Methan wie mit Methanol reduzierbar sind, werden als Hinweis auf die generelle funktionelle Bedeutung für die Methylotrophie diskutiert.
At the meeting of the Judicial Commission
of the ICSB held in Jerusalem on the 29th
March, 1973 an Ad Hoc Committee was
appointed (Minute 22) to organize a review of
the currently valid names of bacteria with the
object of retaining only names for those taxa
which were adequately described and, if
cultivable, for which there was a Type, Neotype
or Reference strain available; to compile these
names under the title of Approved Lists of
Bacterial Names and to publish the lists in the
International Journal of Systematic Bacteriology,
to become effective on January 1, 1980...
~~~~ ~~~~ ~~ ~ pared with some of its neighbors. In view of these results, we expressed the opinion that to classify M. organophilum in a genus separate from all of the other PPFMs solely on the basis of methane utilization was unjustified. This opinion was reinforced by the finding that nei- ther the National Collection of Industrial Bacte- ria strain nor the American Type Culture Collec- tion strain of M. organophilrtm was any longer able to grow on methane, thus rendering them virtually indistinguishable (phenotypically) from many other PPFMs. We also suggested that the PPFMs constituted a distinct taxon which could be excluded from most of the genera to which they had been assigned. We had previously reviewed in detail the somewhat checkered taxonomic history of the PPFMs (5); at different times these orga- nisms had been assigned individually or collec- tively to the genera Bacillus, Flavobncterium, Ch rom o ha c t e rium , Vi brio, Pro t a m ino ba ct e r , My cop la n u , Ps e udom on as, and Methylobacterium. We pointed out that current concepts of the first five of these genera by definition could not accommodate the PPFMs. 'The genus Protaininobacter was no longer recognized, and our argument for the exclusion of the PPFMs from the genera Myco- plann and Pserrdomonas was based largely on our own numerical taxonomic study (6). The two Cory n e b a c' t r rill m ,
A new genus and a new species of methane-oxidizing bacteria are described. The colonies produced by these bacteria are pink, circular, and convex with entire margins, Cells are gram negative and are normally found singularly with some rosettes. Negative stains indicate polar flagellation. In thin sections, intracytoplasmic membranes, similar to those described as type I1 in other methylotrophs, were present when the cells were grown with methane. No such membranes were apparent when the cells were grown with the other carbon and energy sources tested. The serine pathway for formaldehyde incorporation is the pathway of C, metabolism. The deoxyribonucleic acid base composition is 66 mol% guanine plus cytosine. Methylobacterium is proposed as t,he name for this new genus of rod-shaped, methane-oxidizing bacteria. The specific epithet in the name of the type species, Methylobacterium organophilum sp. nov., denotes the preference of this organism for organic carbon and energy sources more complex than methane. The type strain of M. organophilum is XX (= ATCC 27886). This bacterium differs from all previously described genera and species of methane- oxidizing bacteria in its ability to utilize a variety of organic substrates with carbon-carbon bonds as sources of carbon and energy. The pathways for methane oxidation and the assimilation of one-carbon units are repressed during growth on complex organic substrates.
The cellular fatty acid composition was determined for 53 strains of acetic acid bacteria by gas-liquid chromatography. The representatives of the genera Gluconobacter and Acetobacter contained the straight-chain unsaturated acid of C18:1 as a major constituent. Other acids were the straight-chain saturated acids of C16:0 and C18:0 and unidentified acids. The genus Acetobacter was discriminated from the genus Gluconobacter by the former having a detectable amount of the straight-chain saturated acid of C14:0 under experimental conditions. In contrast, the polarly flagellated intermediate strains represented the cellular fatty acid profiles composed of the iso-branched chain acid of C15:0. These results are discussed in connection with the ubiquinone system. The cellular fatty acid composition is proposed here as a novel criterion for characterizing acetic acid bacteria, especially the polarly flagellated intermediate, or A. aurantius.
Fifty strains of acetic acid bacteria including closely related strains were taxonomically studied.
Except for five brown pigment-producing strains, they were appropriately divided into two separate genera Gluconobacter and Acetobacter mainly in accordance with the flagellation and the oxidative behavior towards acetate and lactate. Thus the differentiation of acetic acid bacteria into two separate genera Gluconobacter and Acetobacter which was firstly proposed by ASAI was again verified. Species characterization within each genus of acetic acid bacteria was discussed and considered to be unreliable.
An amendment was added to the previous publication concerning the flagellation of brown pigment-producing three strains, i, e., G. liquefaeiens G-1 (IAM 1834), G. melanogenus AC-8 (IAM 1835) and G. melanogenus U-4 (IAM 1836). They were confirmed to be peritrichous, not polar, and therefore were excluded from genus Gluconobacter.
A remarkable mutation transforming from non-acetate oxidizing strain into acetate oxidizing one during the long periods of preservation in the laboratory was presented in the case with G. liquefaciens G-1 (IAM 1834).
A new finding concerning the existence of polarly flagellated, . acetate-and lactate-oxidizing brown pigment-producing strains was also presented. Accordingly we have at present two different types of brown pigment-producing and acetate- and lactate-oxidizing strains besides G. melanogenus (Beijerinck) Asai (Syn. A, melanogenus Beijerinck); one is characterized by peritrichous flagellation and the other is characterized by polar flagellation.
The generic situation of those organisms was discussed and was decided temporarily as "intermediate" strains neither assignable to genus Gluconobacter nor to genus Acetobacter.
Phylogenetic relationship of genera Gluconobacter and Acetobacter with reference to the "intermediate" strains was also discussed.
One hundred and fifty pink-pigmented facultatively methylotrophic bacteria (PPFMs) of the 'Pseudomonas extorquens' type, 28 other facultative methylotrophs and 16 non-methylo- trophic marker strains of the genera Pseudomonas, A lcaligenes, Mycoplana and Microcyclus were compared in a numerical phenetic study using 140 unit characters. Data were analysed using the simple matching (SsM) and Jaccard (S,) coefficients and single, complete and average linkage algorithms. Cluster composition was largely the same with each similarity coefficient and linkage method. Four major and seven minor clusters ,containing 187 of the 194 strains were defined above the 80% similarity level. The non-pink methylotrophs were recovered in two major, four minor and two single-membered clusters. One of the major clusters could be equated with Microcyclus aquaticus but strains in the other were not identified. Strains in two of the minor clusters were identified as Pseudomonas species but members of the other two minor and one of the single-strain clusters were not positively identified. They had pseudomonad properties but were not closely associated with any marker strains. The other single-membered methylotroph cluster was thought to be a cytophaga or flexibacter. The remaining minor and single-membered clusters contained only marker strains. All the PPFMs were recovered in the two remaining major clusters which were closely related to each other but not to the rest of the organisms studied. The generic assignment of the PPFMs is discussed and the suggestion is made that the genus Methylobacterium may be the most appropriate place for them, despite their apparent- inability to utilize methane.
Mit Hilfe einer polarographischen O2-Meßanordnung im geschlossenen System wurde die Fähigkeit von nicht-wachsenden Zellen von Ac. calcoaceticus für die Veratmung von n-Alkanen und deren Derivaten untersucht. Die Bakterien waren auf Hexadecan kultiviert, in der logarithmischen Phase geerntet und 2 Std. lang zur Substratverarmung in C-freiem Medium inkubiert worden. Die initialen Atmungsraten mit n-Alkanen als Substrat sinken vom Ende der log-Phase an kontinuierlich ab. Mit steigender Tetradecan-Konzentration folgen die Atmungsraten Sättigungskurven (bis etwa 2 · 10−5 M), bei weiterer Erhöhung gibt es es ein Minimum (bei etwa 10−4 M) und dann wieder einen erneuten Anstieg ohne erkennbare Sättigung. Bis zum Ende der Oxydation werden pro Mol Tetradecan etwa 10 Mol O2 verbraucht; das entspricht einer Oxydation über das Acetat hinaus, aber nicht bis zu Kohlendioxid/Wasser. Die Atmungsraten steigen bei gleichen molaren Konzentrationen mit der Kettenlänge der Alkane an, und zwar stärker als es dem erhöhtem O2-Bedarf, der durch die Kettenverlängerung bedingt ist, entspricht. Es werden auch mittelkettige n-Alkane (bis herunter zum Hexan) veratmet, obwohl diese nicht als C-Quelle genutzt werden können. Derivate der n-Alkane werden ebenfalls veratmet, nur l,ω-Dibrom-Alkane werden nicht umgesetzt. Für die Alkane sowie für die meisten der Derivate wird die Fähigkeit zur Oxydation durch Kultivierung auf n-Alkanen erst induziert. Alkanole und Alkanone können in geringerem Umfange allerdings auch nach Wachstum auf Acetat veratmet werden. Die Oxydation der Alkansäuren ist konstitutiv.
Das stäbchenförmige, gram-negative und Katalase-positive Bakterium MB 58 ist fakultativ methylotroph. Es unterscheidet sich z. B. von Acetobacter suboxydans ATCC 621 H, das ebenfalls Methanol assimilieren kann (Hexulosephosphat-Weg), kaum, weder in der Fähigkeit zur Oxydation verschiedener Substrate (Methanol, Äthanol, Glucose, Acetaldehyd u. a.) noch in den Cytochromprofilen oder im Enzym-Muster nach Wachstum auf Methanol bzw. Glucose.
Beim Wachstum auf Glucose zeigt Bakterium MB 58 wie Gluconobacter oxydans diauxisches Verhalten, was darauf zurückgeführt wird, daß Glucose assimiliert und ein Teil davon gleichzeitig zu Gluconsäure oxydiert wird.
Außerdem können Phagen von Bakterium MB 58 verschiedene Acetobacter- und Gluconobacter-Species lysieren, und auch mit Hilfe der Immunofluoreszenz sowie der Objektträgeragglutination läßt sich Bakterium MB 58 von den genannten „Essigsäurebakterien” nicht unterscheiden.
Auf Grund dieser Befunde, die auf enge verwandtschaftliche Beziehungen hinweisen, wird vorgeschlagen, Bakterium MB 58 in die Gruppe der „Essigsäurebakterien” einzuordnen.
Von 6 methanutilisierenden und 11 methanolutilisierenden Bakterienspecies wurden spektralphotometrische Analysen durchgeführt. Damit liegen einschließlich der Befunde von DAVEY u. MITTON (1973), TONGE et al. (1974) sowie ANTHONY (1975) die Cytochromspektren von 9 methan- und 14 methanolutilisierenden Bakterien vor. Obwohl zwischen obligat und fakultativ methylotrophen Bakterien unterschieden werden kann, die Assimilation der C1-Verbindungen über zwei prinzipiell verschiedene Sequenzen erfolgt und sich die einzelnen Species in morphologischen, ultrastrukturellen und physiologischen Merkmalen erheblich unterscheiden, besitzen alle methanutilisierenden Bakterien ein qualitativ nahezu identisches Cytochromkomplement, das sich von den Spektren der methylotrophen, aber Methan nicht utilisierenden Bakterien, die bis auf Ausnahmen fakultativ methylotroph sind, nicht signifikant unterscheidet. Charakteristisch ist der hohe Gehalt an Cytochrom c und o sowie die sehr geringe Konzentration an b. Alle methanutilisierenden Bakterienstämme sowie die Hyphomicrobium-Species verfügen über den Cytochrom aa3-Komplex; in den methanolutilisierenden Bakterien konnte dieser nicht in allen Fällen sicher nachgewiesen werden.
Das Cytochromspektrum ist weder zur Differenzierung methanoxydierender Bakterien mit verschiedenen internen Membransystemen geeignet noch kann es zur Unterscheidung von obligat und fakultativ methylotrophen Species dienen.
Das prinzipiell identische Cytochromkomplement sowie die Tatsache, daß die Cytochrome der „methanotrophen” Bakterien mit Methan wie mit Methanol reduzierbar sind, werden als Hinweis auf die generelle funktionelle Bedeutung für die Methylotrophie diskutiert.
Acetobacter sp. MB 58 assimilates methanol via the fructose-1,6-bisphosphate variant of the hexulose phosphate pathway. Glyceraldehyde-3-phosphate originates as net product of an assimilation loop involving the regeneration of the C1-acceptor ribulose-5-phosphate and must be available for the de novo synthesis of the C1-acceptor as well as for the oxidative glycolysis.
It is made probable in a regulatory model that this is accomplished via alternating anabolic and catabolic phases which are controlled by concerted action of PEP-carboxylase and pyruvate kinase. Whereas Ac-CoA is a crucial effector and ä-ketoglutarate and aspartate are inhibitors for the PEP-carboxylase, the pyruvate kinase is assumed to be regulated by energy charge.
Six strains were obtained from raspberries of Rubus parvifolius, which were characterized by a gram-negative short rod with polar flagellation, oxidation of acetate and lactate, and ubiquinone with 8 isoprene units. The properties of these organisms were very similar to those of "polarly flagellated intermediate strains" in acetic acid bacteria, except for the formation of dihydroxyacetone from glycerol. Some discussions are made on the taxonomic position of these organisms.
Die Hexulose-6-phosphat-Synthase aus dem fakultativ methylotrophen Bakterium MB 58, dem obligat Methanol utilisierenden Bakterium Pseudomonas W 6 und dem obligat Methan utilisierenden Bakterium GB 3 wurde nach Permeabilisierung der Zellen in situ bzw. nach Ultraschall-Desintegration in vitro untersucht.
Die kinetischen Kennlinien (v gegen Ribose-5-phosphat- bzw. Formaldehyd-Konzentration) sind bei Bakterium MB 58 und Pseudomonas W 6 durch einen komplexen Verlauf charakterisiert. Es tritt ein intermediäres Plateau auf, das bei Pseudomonas W 6 in vitro nicht mehr zu beobachten ist. Bakterium GB 3 zeigt in vitro und in situ MICHAELIS-MENTEN-Kinetik. Eine Beeinflussung der Hexulosephosphat-Synthase aus Pseudomonas W 6 durch Intermediate des Kohlenstoff- und Energiestoffwechsels wurde nicht festgestellt.
Die Bedeutung einer komplexen Kinetik der Hexulosephosphat-Synthase für die Regulation des methylotrophen Stoffwechsels wird diskutiert.
A new genus and species of obligately methylotrophic bacteria are described. These bacteria are nonmotile, gram-negative rods occurring singly and in pairs. Only methanol and methylamine can support growth. Formaldehyde fixation occurs mainly via the 3-hexulose phosphate pathway, and cell extracts contain a glutathione-independent, nicotinamide adenine dinucleotide-linked formaldehyde dehydrogenase. The deoxyribonucleic acid base composition is 54.1 mol% guanine plus cytosine. Nitrogen-limited cells accumulate over 5% of their dry weight as a glycogen-like reserve material. This polysaccharide is a homoglucan which is similar to glycogen in its iodine-staining properties and its degree of degradation by phosphorylase a. Some of the glucose molecules are a- 1,4 linked, and the presence of other types of bonds in the glucan is implied. The name of the genus proposed for these bacteria is Methylobacillus gen. nov. The name of the type species, Methylobacillus glycogenes sp. nov., refers to the ability of this species to form a glycogen-like reserve material. The type strain of M. glycogenes is T-11 (= ATCC 29475). Methylotrophy has been defined as the "abil- ity to grow nonautotrophically at the expense of carbon compounds containing one or more car-
A rapid and simple test for the simultaneous detection of 2-keto-, 5-keto- and 2.5-diketo-gluconic acids by thin-layer chromatography and their revelation with o-phenylene-diamine. HCl is described. 32 Acetobacter and 83 Gluconobacter strains were scanned with this method. The formation of keto-gluconic acids appears to be a stable and reliable feature for the classification of acetic accid bacteria.
Ninety-eight strains, representing all Acetobacter species and subspecies from the Approved Lists of Bacterial Names (Skerman et al., 1980), were examined in a numerical analysis of 177 phenotypic features and compared to ninety-eight Gluconobacter and seven Frateuria strains. Four phenons could be delineated, corresponding to Frateuria (phenon 1), A. aceti subsp. liquefaciens (phenon 2), Gluconobacter (phenon 3) and Acetobacter minus A. aceti subsp. liquefaciens (phenon 4). Acetobacter, Frateuria and Gluconobacter are well- could be distinguished. Comparison of the protein electrophoregrams of Acetobacter strains revealed a fairly high internal homogeneity within phenon 2, subphenons C and D. Strains of the subphenon E gave very divergent protein patterns. The following classificatory changes are proposed within the genus Acetobacter: (1) Acetobacter liquefaciens sp. nov. is proposed for the homogeneous phenon 2, containing all 12 A. aceti subsp. liquefaciens strains (% G + C range of 62.3 to 64.6; IAM 1834 as type strain); (2) for the homogeneous subphenon D containing 8 A. aceti subsp. aceti strains, the name Acetobacter aceti emend, should be retained (% G + C range of 55.9 to 59.5; NCIB 8621 as type strain); (3) for subphenon E, a heterogeneous group, containing a variety of Acetobacter subspecies (all with their type strain) the species name Acetobacter pasteurianus emend, is preserved with LMD 22.1 as type strain; this species has the broad % G + C range of 52.8 to 62.5; (4) for subphenon C, a new species, Acetobacter hansenii sp. nov. is proposed (% G + C range of 58.1 to 62.6, NCIB 8746 as type strain). Minimal descriptions and differentiating keys are provided.
A method has been described for the isolation of DNA from micro-organisms which yields stable, biologically active, highly polymerized preparations relatively free from protein and RNA. Alternative methods of cell disruption and DNA isolation have been described and compared. DNA capable of transforming homologous strains has been used to test various steps in the procedure and preparations have been obtained possessing high specific activities. Representative samples have been characterized for their thermal stability and sedimentation behaviour.
Generally, methylotrophic bacteria grow optimally in a pH range between 6 and 7.2. The assimilation of methanol can take place via several pathways. Acetobacter methanolicus preiers an acidic pH range for growth, the pH optimum is about 4, and it uses the FBP variant for methanol assimilation.
The latter is interesting from a regulatory point of view because phosphofructokinase disappears during growth on glucose, which is assimilated via the hexosemonophosphate pathway. Since Entner-Doudoroff enzymes and phosphoketolase are absent in A. `ethanolicus as well as in non-methylotrophic Acetobacter and Gluconobacter species phosphofructokinase becomes a key enzyme of the assimilation of methanol.
Although A. methanolicus uses the hexulosephosphate pathway the growth yield on methanol is smaller than with other “hexulosephosphate pathway bacteria” e. g. with obligate methanol assimilating bacteria.
At first sight it may appear that the acidic optimum pH is responsible for the smaller growth yield and the discrepancy between the experimental and predicted values.
The relationship between the dependence on and the protection from, high external proton concentration on the one hand and the causes of the low growth yield on the other are discussed. Accordingly, A. methanolicus and another heterotrophic acidophiles seem to be acidoresistant above all, their machinery guaranteeing the protection from the high proton concentration is responsible for the acidophily and the low growth efficiency is caused by a simple respiratory chain.
Eine neue, spezifische Farbreaktion ermglicht den Nachweis der Ameisensure und des Formiats ohne vorherige Reduktion zu Formaldehyd. Der Nachweis beruht auf der roten Frbung, die man durch Reaktion von Citronensure bzw. Aconitsuren mit Ameisensure oder Formiat in Gegenwart von Essigsureanhydrid, i-Propanol oder einer geeigneten Acetamid-i-Propanol-Lsung und einer kleinen Menge Alkali erhlt. Die bei Verwendung von Citronensure erzeugte rote Farbe zeigt ein Lichtabsorptionsmaximum bei 515 nm. Weniger als 20 g Formiat/ml knnen nachgewiesen werden.A new, specific colour reaction permits the identification of formic acid and formate without proceeding reduction to formaldehyde. It is based on the red colour obtained by reaction of citric or aconite acids with formic acid or formate in presence of acetanhydride, i-Propanol or a suitable acetamide-i-Propanol-solution and a small quantity of alkali. The red colour produced with citric acid shows an absorption maximum at 515 nm. Less than 20 g of formate/ml can be detected.
Die zu einer nahezu zuckerfreien, asparaginhaltigen Nhrlsung zugesetzten Salze entfalten einen berraschend starken Einflu auf die Oxydation des beigegebenen thylalkohols durch Bacterium acetigenoideum ebenso wie auf das Wachstum, die Bildung von Involutions-formen und die Beweglichkeit dieses von uns aus einer Obstessigmaische reingezchteten haplotrophen Essigbakteriums. Lediglich durch Variierung der Salzzustze gelingt es, die Bildung der Essigsure entweder zu unterbinden oder bis auf das hchste (etwa 90% der theoretisch zu erwartenden Menge) zu steigern oder eine Weiterbrennung der entstandenen Essigsure zu CO2 (beroxydation) bis zu ihrem fast gnzlichen Verschwinden zu erreichen.Die geprften Salzionen — die in Form von Einzelsalzen bzw. Salzgemischen der Nhrlsung zugesetzt wurden — lassen sich nach ihrem Verhalten zu Suerung und Wachstum unserer Essigbakterie unter den gegebenen Bedingungen in Reihen anordnen, die an die lyotropen Salzeinflsse bzw. an Ionenquilibrierungen im Sinne J. Loebs erinnern, wobei im allgemeinen von den Kationen bzw. Anionen Ca, Mg bzw. H2PO4 sowei Cl frdern, die alkalien K, Na bzw. SO4 hemmen. Hierbei geht der Einflu auf Suerung und Wachstum nicht immer parallel. Bezglich der zahlreichen Einzelergebnisse mu auf die Arbeit selbst verwiesen werden.Die Konzentrationswirkung der Einzelsalzgaben ergab Optimumkurven mit einem beraus steilen Ansteig in den niedrigen Konzentrationsstufen von etwa 0,00001 n bis 0,005 n, einem von der Art der Salze ziemlich unabhngigen Optimum von 0,025 n bis 0,05 n und einem Wendepunkt im absteigenden Ast, also ein Verhalten nach Art der bekannten Ertragskurven, wie sie z. B. die Abhngigkeit der produzierten Trockensubstanz von der Nhrstoffmenge darstellt.
The previously discovered linear relation between the base composition of DNA, expressed in terms of percentage of guanine plus cytosine bases, and the denaturation temperature, Tm, has been further investigated. By means of measurements on 41 samples of known base composition the previously observed relation has been confirmed. It can be summarized thus : for a solvent containing 0·2 M-Na+, Tm = 69·3 + 0·41 (G-C) where Tm is in degrees Centigrade and G-C refers to the mole percentage of guanine plus cytosine. The deviations of experimental points from this relation are no more than that expected from the uncertainties of base analysis and the variations of a half degree in the reproducibility of determining the Tm. Consequently it appears that the measurement of the Tm is a satisfactory means of determining base composition in DNA. The Tm values are most simply measured by following the absorbance at 260 mμ as a function of temperature of the DNA solution and noting the midpoint of the hyperchromic rise. Only 10 to 50 μg of DNA are required.A number of other DNA samples of unknown base composition have been examined in this manner and their base compositions recorded.
The nitrogen requirements of 96 Gluconobacter, 55 Acetobacter and 7 Frateuria strains were examined. Only some Frateuria strains were able to grow on 0.5% yeast extract broth or 0.5% peptone broth. In the presence of D-glucose or D-mannitol as a carbon source, ammonium was used as the sole source of nitrogen by all three genera. With ethanol, only a few Acetobacter strains grew on ammonium as a sole nitrogen source. Single L-amino acids cannot serve as a sole source of carbon and nitrogen for growth of Gluconobacter, Acetobacter or Frateuria. The single L-amino acids which were used by most strains as a sole nitrogen source for growth are: asparagine, aspartic acid, glutamine, glutamic acid, proline and alanine. Some Acetobacter and Gluconobacter strains deaminated alanine, asparagine, glutamic acid, threonine, serine and proline. No Frateuria strain was able to develop on cysteine, glycine, threonine or tryptophan as a sole source of nitrogen for growth. An inhibitory effect of valine may explain the absence of growth on this amino acid. No amino acid is "essential" for Gluconobacter, Acetobacter or Frateuria.
Summary TheAcetobacter genus as presently constituted include two distinctly different morphological types, one having peritrichous flagella with
a wavelength averaging 2.9 μ and the other having polar multitrichous flagella with a wavelength averaging 1.4 μ. All of the
peritrichously flagellated types actively oxidized acetic and lactic acids to CO2 and H2O, while none of the polar flagellated types showed this property. It is proposed that the presentAcetobacter genus be divided into two genera:Acetobacter andAcetomonas gen.nov. The redefinedAcetobacter genus should include only peritrichously flagellated species and nonflagellated species with similar physiology.Acetomonas gen. nov. should include only polar multitrichous species and nonflagellated species of similar physiology. Typical species
of the redefinedAcetobacter genus should oxidize acetic or lactic acid to CO2 and H2O while typical species ofAcetomonas gen. nov. should not have this property.
Acetic acid bacteria. Classification and biochemical activities
- T Asai
Asai, T. 1968. Acetic acid bacteria. Classification and biochemical activities. University of Tokyo Press, Tokyo.
Bergey's manual of systematic bacteriology
- Acetobacteraceae Gillis
- De Ley
Acetobacteraceae Gillis and De Ley 1980, 23, p. 267-278. In
N. R. Krieg and J. G. Holt (ed.), Bergey's manual of systematic
bacteriology, vol. 1. The Williams & Wilkins Co., Baltimore.
Drews, G. 1974. Mikrobiologisches Praktikum, 2nd ed. Springer-Verlag, Berlin.
A taxonomic study of some Gram-negative facultatively methylotrophic bacteria Emendation of Methylobacterium Patt Methylobacterium rhodinum (Heumann 1962) comb. nov. corrig., Methylobacterium radiotolerans (Ito and Iizuka 1971) comb. nov. corrig., and Methylobacterium mesophilicum
- P N Green
- I J Bousfield
- P N Green
- I J Bousfield
Green, P. N., and I. J. Bousfield. 1982. A taxonomic study of
some Gram-negative facultatively methylotrophic bacteria. J.
Gen. Microbiol. 128:623-638.
Green, P. N., and I. J. Bousfield. 1983. Emendation of
Methylobacterium Patt, Cole, and Hanson 1976, Methylobacterium rhodinum (Heumann 1962) comb. nov. corrig.,
Methylobacterium radiotolerans (Ito and Iizuka 1971) comb.
nov. corrig., and Methylobacterium mesophilicum (Austin and
Goodfellow 1979) comb. nov. Int. J. Syst. Bacteriol.
The problems on the fermentation engineering of methanol assimilating microorganisms
- A Mimura
- M Wada
Mimura, A,, and M. Wada. 1978. The problems on the fermentation engineering of methanol assimilating microorganisms.
Hakko Kogaku Kaishi 56:662-675.
Der Abbau von Methylalkohol, Formaldehyd und Ameisensaure durch lebende und getotete Essigbakterien
- D Muller
Muller, D. 1932. Der Abbau von Methylalkohol, Formaldehyd
und Ameisensaure durch lebende und getotete Essigbakterien.
Biochem. Z. 254:97-111.
Process for producing bacterial single cell protein from methanol
- T Urakami
- I Terao
- I Nagai
Urakami, T., I. Terao, and I. Nagai. 1981. Process for producing
bacterial single cell protein from methanol, p. 349-359. In H.
Dalton (ed.), Microbial growth on C1 compounds. Heyden,
London.
Method for identifying acetic acid bacteria
- J G Carr
- London
- J De Ley
- M Gillis
- J Swings
Carr, J. G. 1968. Method for identifying acetic acid bacteria, p.
1-8. In B. M. Gibbs and D. A. Shapton (ed.), Identification
methods for microbiologists, part B. Academic Press, Inc.,
London.
De Ley, J., M. Gillis, and J. Swings. 1984. Family VI.