Article

Antibacterial Activity of Cabbage Juice Against Lactic Acid Bacteria

Wiley
Journal of Food Science
Authors:
  • Sejong University, retired
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Abstract

Fresh juice of Cecile cultivar cabbage (Brassica oleracea) was inhibitory to the growth of four species (including seven strains) of lactic acid bacteria. Inhibition was eliminated when the cabbage was heated (steamed 10 min) before juice extraction. When Lcuconostoc mes-enteroides C33 was tested in juices of three other cultivars of cabbage, growth inhibition was invariably noted in fresh juices, but was variable in heated juices. Fresh cabbage juice (CJ) adjusted to higher initial pH (6.5–7.5) showed greater antibacterial activities than at pH 5.0 to 6.0. When pH 4.0 precipitate from fresh CJ was added to noninhi-bitory CJ (cabbage heated before extraction), antibacterial activity was restored. A heat-labile and pH-dependent factor seemed responsible for activating a precursor into an inhibitory compound.

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... Breidt et al. (1995) reported that 700 IU ml À1 of nisin was produced in brined cabbage within 24 h, but after 72 h, nisin activity was no longer detectable. Kyung and Fleming (1994) documented that cabbage juice exhibited antibacterial activity against some lactic acid bacteria strains, but that inhibition was eliminated when the cabbage was heated before juice extraction. Antibacterial activity seemed to be heat labile and pH dependent (Kyung and Fleming, 1994). ...
... Kyung and Fleming (1994) documented that cabbage juice exhibited antibacterial activity against some lactic acid bacteria strains, but that inhibition was eliminated when the cabbage was heated before juice extraction. Antibacterial activity seemed to be heat labile and pH dependent (Kyung and Fleming, 1994). When the antimicrobial activity of different sulphur compounds derived from cabbage was tested, sinigrin did not inhibit the growth of bacteria and yeasts up to 1000 ppm, whereas a breakdown product of sinigrin, allyl-ITC, was very strongly inhibitory to the growth of both oxidative and fermentative yeasts (Kyung and Fleming, 1997). ...
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... Glucosinolate breakdown products also contribute significantly to the typical flavor of Brassica vegetables (6). Further, cabbage juice has been documented in some studies to have antibacterial activity (7,8). The antibacterial activity of fresh cabbage juice, moreover, has been reported to be heat-labile and pH-dependent (7). ...
... Further, cabbage juice has been documented in some studies to have antibacterial activity (7,8). The antibacterial activity of fresh cabbage juice, moreover, has been reported to be heat-labile and pH-dependent (7). ...
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... Natural antimicrobials from the vegetables could be potential future candidates to be used as safe anti infective pharmaceutical products without causing any side effects like chemical medicines. Antimicrobial activity of different vegetables such as spinach, ghuniya, pumpkin, suran, cabbage, has been tested around the world in different times [13,30,35]. Along these lines, we determined the anti-microbial activity of the salad vegetables for the first time in Bangladesh (Table 3). ...
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... Several studies worldwide and in Bangladesh reported antimicrobial activity in different food samples (Kyung et al., 1994;Dubey et al., 2010;Hussain et al., 2010). One of our previous studies was conducted to establish the antimicrobial activity of export quality shrimp samples in Bangladesh (Rahman et al., 2012). ...
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... Though only one sample from this study was found to exhibit such trait against only one type of bacterium, Vibrio spp., yet this result urges an importance of hygienic handling of apple samples after harvesting and during distribution so that the product could be microbiologically safe with a natural medication properties. Several studies suggested that the presence of microorganism-killing agent in different food items could be a potential replacement of chemical medicines which indeed goes a long way to minimize the possible side effects of synthetic medicines, as well as may enhance the shelf life of the food and the consumer safety [15,16,42]. In the context of Bangladesh, this sort of demonstration in case of apple samples have been conducted for the first time, and eventually further experiments with more samples would probably fortify the natural means of disease mitication. ...
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... Natural antimicrobials from the vegetables could be potential future candidates to be used as safe anti infective pharmaceutical products without causing any side effects like chemical medicines. Antimicrobial activity of different vegetables such as spinach, ghuniya, pumpkin, suran, cabbage, has been tested around the world in different times [13,30,35]. Along these lines, we determined the anti-microbial activity of the salad vegetables for the first time in Bangladesh (Table 3). ...
... Several studies worldwide and in Bangladesh reported antimicrobial activity in different food samples (Kyung et al., 1994;Dubey et al., 2010;Hussain et al., 2010). One of our previous studies was conducted to establish the antimicrobial activity of export quality shrimp samples in Bangladesh (Rahman et al., 2012). ...
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Shrimps sold in local markets could be microbiologically spoiled intrinsically or extrinsically. Present study attempted to detect the frequency of such microorganisms in shrimps collected from local markets of Dhaka city. A total of 7 categories of shrimp samples were studied. All of them were found to be contaminated with Staphylococcus spp., Aeromonas spp., Klebsiella spp., Pseudomonas spp. and Shigella spp. ranged from 1.5×104 to.7.9×108 cfu/g with a comparatively higher frequency of Klebsiella spp., Staphylococcus spp., and Aeromonas spp. Study of antibiogram revealed multi-drug resistance of most of the isolates. No antimicrobial activity was detected.
... Plantings after 7 weeks showed no growth reduction or slight growth enhancement (Papavizas 1966). Only fresh cabbage juice inhibited lactic acid (Leuconostoc mesenteroides) bacterial growth (Kyung and Fleming 1994). Inhibition of germination and plant growth by ITCs was most effective when the chemicals were applied to seeds or applied within the first 3 d of the germination period (Leblová-Svobodová and Koštír 1962). ...
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... Similar facts have been reported in case of cabbage as well. Kyung and Fleming (1994) reported that fresh juice of Cecile cultivar cabbage was inhibitory to the growth of LAB due to the presence of antibacterial substances in juice, and the inhibition was eliminated when the cabbage was heated (steamed 10 min) before juice extraction. Preliminary studies showed that heat treated L. digitata and L. saccharina maintained the growth of L. plantarum for 24 h. ...
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... At 8°C, LAB populations in OPP were lower in shredded cabbage than in shredded carrot by Day 2, but not thereafter. Kyung & Fleming (1994) demonstrated an anti-bacterial activity of cabbage juice against LAB, which was greater at pH values >6.0. In the current work, shredded cabbage in OPP had a pH value >6.0 up to Day 2, but was <5.0 by Day 7. ...
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Sauerkraut fermentation course was observed in 3 cycles and 4 replicates under controlled conditions (2.5% NaCl, 21degreesC) using starter cultures (control; Leuconostoc meseneroides - 700 mil. cfu/ml; Lactococcus lactis ssp. lactis - 500 mil. cfu/ml; preceding fermentation juice). Each of the above mentioned cycles lasted for 28 days. Microbiological and chemical characteristics of the sauerkraut during the 28-day fermentation period were investigated. Lactic acid bacteria content, the concentration of lactic acid, and pH of the medium were monitored daily. The completion of each cycle fermentation was followed by the determination of the final product organoleptic properties which were observed for 6 months with the aim to define the expiration date. The best results relative to sauerkraut quality were obtained by using starter culture L. lactis ssp. lactis, followed by fermentation conducted by natural, spontaneous sauerkraut flora (control variant). Organoleptic properties and expiration date of the final product obtained by the use of lactic acid bacterium L. mesenteroides as a starter culture were better compared to the use of sauerkraut juice obtained from the preceding fermentation cycles and possessing the best organoleptic properties.
Chapter
The fermentation of plant material is an ancient preservation method, the origins of which have been traced to Asia.[1] In Europe, a total of 21 different vegetables are fermented, in addition to an unspecified number of variably composed vegetable blends and fermented vegetable juices (2). The most common products in Europe and the United States are sauerkraut, cucumbers, and olives. Fermented olives are manufactured mainly in the Mediterranean region. The fermentation of these products is carried out in factories or on farms.[2,3] In Korea, kimchi is a traditional fermented vegetable food.[4] According to Buckenhüskes and coworkers,[2] it is generally agreed that fermented plant products are the “food of the future.” Factors supporting this idea include: The high degree of hygienic safety caused by repression of growth of pathogenic bacteria Products can be marketed as “natural” or “biological” Enrichment of desired metabolites such as L-lactic acid or amino acids Build-up of flavor compounds and destruction of negative flavor compounds such as glucosinolates Less energy input than most other methods of preservation Simple handling and storage without cooling Easy method for prehandling of raw material before further processing Most fermented vegetable products including sauerkraut juice are still produced by spontaneous fermentation, which typically involves a succession of microbial populations.
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Crude, chloform and methanol extracts of leaves of two variety of Brassica oleracea e.g., botrytis and capitata were evaluated against Escherichia coli coli Crude and alcoholic extracts of variety botrytis showed high antibacterial activity against E.coli.
Chapter
IntroductionCucumbers for PicklesCabbage for SauerkrautKimchiCulture Development for Vegetable FermentationsReferences
Article
Garlic (Allium sativum L.) contains a specific sulfur compound, the S-allyl derivative of L-cysteine sulfoxide, and has long been known for its antimicrobial activity against various microorganisms, including bacteria, fungi, and protozoa. The principal antimicrobial compound of garlic is S-allyl-L-propenethiosulfinate (allicin) which is generated by an enzyme, alliinase (L-cysteine sulfoxide lyase), from S-allyl-L-cysteine sulfoxide (alliin). This compound exists exclusively in Allium as a major non-protein sulfur-containing amino acid. S-Allyl-L-propenethiosulfinate belongs to the chemical group of thiosulfinates and is a highly potent antimicrobial. The potency of garlic extract is reduced during storage since thiosulfinates are unstable and are degraded to other compounds some of which do not have antimicrobial activity. Diallyl polysulfides and ajoene are sulfur compounds derived from allicin that do possess antimicrobial activity. It was recently found that garlic becomes antimicrobial on heating at cooking temperatures, and that the compound responsible for this is allyl alcohol, which is generated from alliin by thermal degradation.
Chapter
The origin of the preservation of vegetables by fermentation is lost in antiquity. Worldwide, most vegetables are still fermented on a small scale, either in the home, or by entrepreneurs. The current exceptions to this are sauerkraut, cucumbers, and olives. These are of significant commercial importance and will be the focus of this chapter. There is increasing interest in a wide variety of other fermented vegetables and fermented vegetable juices, especially in the European market (Table 2.1); however, most of these are currently produced commercially on a small scale (Buchenhüskes et al.,1990). Korean kimchi, traditionally prepared in the home, has also been recently adapted to commercial production (Cheigh & Park, 1994; Fleming et al., 1995a).
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The fungus Alternaria alternata, is considered one of the major plant pathogens worldwide causing the symptom of leaf spot in vegetables. Glucosinolates are natural products derived from broccoli, containing nitrogen and sulfur, with antifungal activity. Based on the above, the present study aimed to evaluate the effectiveness of broccoli juice in vitro on germination of conidia of A. alternata. To carry out the experiment, leaves of Eruca sativa (arugula) with symptoms of leaf spot were collected, from which the fungus A. alternata was isolated and identified. Conidia of the fungus were placed on PDA medium with different concentrations of glucosinolates found in broccoli juice 0.15, 0.11, 0.07, 0.04, 0.01, and 0 ng/uL. It was observed that the concentrations 0.07, 0.1, 0.15 ug/uL inhibited 100 % spore germination of A. alternata at 6 h evaluation. The average lethal concentration LC was 0.028 ug/uL. In 50 conclusion, it can be mentioned that the glucosinolates present in broccoli juice, have an antifungal potential and can be used as post-harvest treatment, in order to reduce the damage caused by A. alternata in vegetables.
Article
We have developed a rapid method for the determination of lag times and specific growth rates for bacteria. This method involves the use of a Bio-Tek model EL312 microtiter plate (MP) reader, controlled by KinetiCalc software, and uses a computer program that processes absorbance data from a 96-well MP. The program reads an ASCII file consisting of optical density readings generated by the MP reader, sorts the data into individual growth curves, and determines growth kinetics. No statistically significant difference (P > 0.88) was found between the MP method and standard batch methods. This method allows up to 95 growth curves to be carried out simultaneously. The MP method automates the difficult task of determining growth kinetics for bacterial strains under conditions that result in slow growth rates or long lag times. Using this method we determined the effect of NaCl on the growth kinetics of Listeria monocytogenes and the effects of NaCl and temperature on the growth kinetics of Leuconostoc mesenteroides.
Article
Allium and Brassica vegetables have long been known for their antimicrobial activity against various microorganisms, including Gram-positive and Gram-negative bacteria and fungi. Most microorganisms tested were sensitive to extracts of the Allium and Brassica vegetables and the degree of sensitivity varied depending on the strain under study and test conditions. Among the vegetables, garlic showed the most potent activity, followed by onion. Brassica, including cabbage, showed the least potent activity. The principal antimicrobial compounds of Allium and Brassica have been elucidated as allicin (S-allyl-L-propenethiosulfinate) and methyl methanethiosulfinate, respectively. Both compounds belong to the same chemical group, thiosulfinate, generated from S-allyl and S-methyl derivatives of L-cysteine sulfoxide, respectively, existing in Allium and Brassica as major non-protein sulfur-containing amino acids. There have been only few applications of garlic as a natural food preservative, in spite of numerous studies on antimicrobial activity of the vegetables. Relative instability of the antimicrobial compounds and the strong odor of their mother plants seem to limit the use of them as a practical food preservative.
Article
A paired starter culture system for sauerkraut fermentation was proposed previously by our laboratory, consisting of a nisin-producing strain of Lactococcus lactis and a nisin-resistant strain of Leuconostoc mesenteroides. The objectives of this study were to determine nisin production and stability by a genetically marked culture of L. lactis NCK400 in brined cabbage fermentations and to study the effect of purified nisin on the natural microflora in brined cabbage. We found that NCK400 produced up to 700 IU ml−1 of nisin in brined cabbage within 24 h, but by 72 h nisin activity was no longer detectable. The nisin produced was shown to have an effect on the microflora of the fermentation, however, resulting in a 40% reduction in the population of homofermentative lactic acid bacteria (LAB) at 33 days. We also found that the addition of purified nisin to brined cabbage (up to 12 000 IU ml−1) resulted in a reduction in the cell counts of LAB, as compared to a non-treated cabbage brine, to the extent that fermentation by Gram-negative bacteria occurred. The addition of a nisin-resistant L. mesenteroides strain to the nisin-treated cabbage resulted in a heterolactic fermentation by the added culture, with no evidence of a homolactic fermentation for at least 20 days. We concluded that nisin produced in situ or added to brined cabbage can direct the progression of the species in the resultant fermentations, by preventing the growth of naturally present LAB.
Article
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Film-forming yeasts generate an undesirable yeasty flavor in fermented vegetables such as kimchi in the presence of oxygen. Antimicrobial materials including garlic oil (GO), heated garlic (HG), and allyl alcohol (AA) were investigated for use as alternative natural food preservatives to inhibit the growth of film-forming yeasts in fermented vegetables. Using the fermentation of cucumber pickles as a model system, GO, HG, and AA were effective in preventing film formation at concentrations of 0.006, 3.0, and 0.02%, respectively. The effectiveness of HG in preventing the growth of a film yeast, Hansenula anomala, was not influenced by pH, while that of potassium sorbate, a typical anti-yeast food preservative, was highly dependent on pH. All tested materials were effective when added at the beginning of fermentation due to their negligible inhibitory activity toward lactic acid bacteria.
Article
The anti-listerial effectiveness of selected essential oils (EOs) and shredded fresh herbs (thyme, oregano and rosemary) was examined on a range of modified atmosphere packaged fresh-cut vegetables (lettuce, carrot discs, cabbage and dry coleslaw mix). Anti-listerial effects were in the order: thyme EO > oregano EO > rosemary herb > rosemary EO. While thyme EO demonstrated the best anti-listerial effect, direct application of all the EOs damaged product appearance. Shredded fresh rosemary herb appeared to have a major anti-listerial effect, but shredded fresh thyme and oregano showed no anti-listerial effects. However, fresh rosemary herb was only effective in fresh-cut products when it was stomached with the product prior to microbial analysis. The effectiveness of these antimicrobials varied depending on the product type. Greater anti-listerial effects were recorded on carrot discs and shredded cabbage than on shredded lettuce. Adding shredded carrot to packages enhanced the apparent anti-listerial effects, suggesting a synergistic effect between carrot and rosemary.
Chapter
Many different vegetable fermentation processes are currently carried out on an industrial scale, most of which still rely on selection of the autochthonous microbiota of the raw materials and fermentation plant. The implication of lactic acid bacteria in such processes has been deciphered by classical microbiological techniques in most cases. The application of DNA-based culture-dependent and culture-independent analyses may provide new insights into the microbial successions that take place during the fermentation as well as the microbial diversity. These data can be linked to other issues such as flavor development or regional differences in fermented foods. DNA-based methods can also help to evaluate the fitness of starter cultures used for vegetable fermentations. In this chapter, advances in the understanding of several vegetable fermentations (including caper berries, “Almagro” eggplants, sauerkraut and table olives) by use of molecular techniques are discussed.
Article
Heated (121 °C) garlic extract in which alliinase was inactivated before crushing exhibited complete bacteriostatic activity at 15% against Staphylococcus aureus. Garlic heated for 45 min showed the highest antibacterial activity and the relative peak areas of 4-heptenal, methyl allyl disulfide, diallyl disulfide, 2-vinyl-4H-1,3-dithiin, and diallyl trisulfide (DATS) were highest at 45 min of heating. Other than 4-heptenal, all these compounds are known to possess different degrees of antibacterial activity. DATS was thought to be the primary antibacterial compound in heated garlic extract. It was tentatively concluded that antibacterial compounds were formed from alliin unreacted by alliinase by marked heating. Diallyl compounds with more than three sulfur atoms and the oxidized form of allicin were not detected.
Article
ABSTRACTS-Methyl-L-cysteine sulfoxide (SMCSO) was present in several cultivars of cabbage (Brussicn oleracea) at 690–1,120 ppm. SMCSO was not inhibitory to Leuconostoc mesenreroides, but its hydrolysis product, methyl methanethiolsulfinate (MMTSO), was inhibitory. MMTSO was formed in fresh cabbage juice (CJ) and in a model system containing SMCSO and fresh cabbage (pH 4.0) precipitate, perhaps by cysteine sulfoxide lyase. Growth inhibition by 1 mM MMTSO was reversed by 5 mM dithiothreitol, cysteine, or reduced glutathione, but not by oxidized glutathione or thiamine.
Article
Dichloromethane extracts of juices from fresh cabbages, including four known (Brutus, Galaxy, Bentley, Structon) and two unknown cultivars, were analyzed by X-MS for the presence of sinigrin degradation products. Allyl isothiocyanate (AITC), which has been reported to be the important aroma compound of freshly disrupted cabbage was not detected in any of the dichloromethane extracts of cabbage juice. Instead, 1-cyano-2,3-epithiopropane, which is one of the isomers of AITC, was the primary volatile compound in all cabbage extracts. AITC was detected in relative trace concentration only when cabbage juice was injected into the GC, using a wide bore, packed column. Thus, the relative importance of AITC and 1-cyano-2,3-epithiopropane to the aroma of freshly disrupted cabbage should be considered.
Article
Autoclaved cabbage juice was inhibitory to growth of Staphylococcus aureus. S-Methyl-L-cysteine sulfoxide (SMCSO), autoclaved either together with or separately from nutrient broth, also inhibited the growth of S. aureus, but inhibition was greater when SMCSO was autoclaved separately. Methyl methanethiosulfonate (MMTSO2), a thermal breakdown product of SMCSO, completely inhibited growth of S. aureus at 10 ppm. MMTSO2 was formed in both autoclaved samples of cabbage juice and aqueous solution of SMCSO. Thus, evidence indicates that the bacterial inhibitory activity in autoclaved cabbage juice was due to heatinduced formation of MMTSO2 from SMCSO.
Article
The bulky precipitate formed in the washwaters from the processing of Spanish-style green olives, as a consequence of HCl acidification, presented inhibitory activity against lactic acid bacteria growth, the intensity of the effect being influenced by the type of washwater, pH and inoculum size. The delay or complete inhibition of microorganism growth was confirmed for three strains of lactobacilli (Lactobacillus pentosus LP99, L. pentosus ATCC 8041 and L. plantarum ATCC 14917), and this phenomenon can allow growth of undesirable microorganisms during washwater fermentation. The inhibitory activity against lactobacilli in acidified washwaters disappeared when the bulky precipitate was eliminated by centrifugation or simply by sonicating the solutions. It seems that sonication modified the macromolecular structure of the precipitate and the biosorption of microorganisms in it did not occur. Also, the inhibitory activity of this precipitate was confirmed in brines of Spanish-style green olives. Therefore, this new lactobacillus growth inhibitor will have to be taken into account in the future for improving olive fermentation.
Article
Functional foods are claimed to have several health-specific advantages. In addition to their basic nutritive value, they contain a proper balance of ingredients which help in the prevention and treatment of illnesses and diseases. Within this category, products containing lactic acid bacteria or probiotics are increasingly gaining importance. The recognition of the beneficial effects of dairy products containing probiotics has been well established. The allergy to dairy products, lactose intolerance, and cholesterol content are the major drawbacks related to the use of fermented dairy products for a large percentage of consumers. Modern consumers are increasingly interested in their personal health, and expect the food that they eat to be healthy or even capable of preventing illness. Because of this, probiotic food products made out of fermentation of cereals and fruits and vegetables is receiving attention from the scientific world as well as consumers and constitutes the major part of this review. The use of mathematical models for the probiotic fermentation will help in reducing the time and effort involved in the optimization of the probiotic fermentation process. We have tried to summarize the developments in the use of mathematical models for probiotic fermentation. Future technological prospects exist in innovations which represent solutions for the stability and viability problems of probiotics in new food environments. Current research on novel probiotic formulations and microencapsulation technologies exploiting biological carrier and barrier materials has also been discussed.
Article
The aim of this study is to develop a growth inhibitory material against some pathogenic microorganisms, using beneficial bacteria such as Bifidobacterium species and certain types of vegetables which can be good substrates for the growth of the beneficial bacteria. At first, various vegetable juices were screened for the growth promotion of Bifidobacterium longum etc. Among the vegetables tested, broccoli (Brassica oleracea L. var. botrytis L.) and cabbage (Brassica oleracea L. var. capitata L.) showed excellent growth promoting activities for B. longum. Secondly, the B. longum-fermented broccoli (BFB) and Lactobacillus pentosus-fermented broccoli (LFB) supernatants were prepared and the growth inhibitory activities against Candida albicans were determined. Both of them showed dose-dependent, growth inhibitory effects, and the effect of BFB was superior to LFB. It was thought that the superior effect of BFB could be mainly attributed to the acids, especially acetic acid, produced by B. longum. BFB also inhibited some pathogenic bacteria such as Streptococcus mutans and Porphylomonas gingivalis. In conclusion, broccoli was found to be a good growth-promoting substance for B. longum. The fermented product, BFB, appears to be a usable material that inhibits the growth of C. albicans and some pathogenic bacteria.
Article
In the present study, the effects of concentration, pH, growth phases and heat treatment on the inhibition growth against several strains of food pathogenic bacteria by azelaic acid (Aza) was investigated to evaluate its possibility as food preservatives. The result showed that 300 ppm of Aza almost completely inhibited the growth of tested bacteria. Aza added at the beginning of the incubation had the high inhibitory effect. In the range of pH 5-9, Aza had the highest inhibitory at pH 9. The degree of antibacterial activity of Aza was higher than those of sodium benzoate and potassium sorbate and similar to those of dimethylfumarate and fumaric acid. Antibacterial activity of Aza was stable even in the heating treatment.
Article
ABSTRACT Brassica tissues are potentially useful in the control of Aphanomyces root rot of peas (Pisum sativum), but identity of the responsible compounds and specific impacts of those compounds on the pathogen's infection potential remain uncertain. Brassica napus seed meals and water extracts from these meals were used to determine the effect of glucosinolate hydrolysis products on Aphanomyces euteiches f. sp. pisi. B. napus meal ('Dwarf Essex') containing glucosinolates and intact myrosinase, the enzyme responsible for glucosinolate hydrolysis, completely inhibited infection by A. euteiches f. sp. pisi oospores. Water extracts from this meal, likewise, severely inhibited infection by oospores, as well as mycelial growth. Extracts from autoclaved 'Dwarf Essex' meal, in which myrosinase was denatured, and a low glucosinolate B. napus variety ('Stonewall') produced little disease reduction and had less impact on mycelial growth. Gas chromatographic analysis of Brassica tissues and water extracts confirmed that glucosinolates remained in autoclaved 'Dwarf Essex' meal and that 'Stonewall' meal contained low glucosinolate concentrations. 5-Vinyloxazolidine-2-thione was identified by mass spectrometry as a dominant glucosinolate hydrolysis product in aqueous extracts of the inhibitory meal. Bioassays conducted with aqueous solutions of this compound reduced mycelial growth, but not to the extent of those from intact 'Dwarf Essex' meal. Water-soluble compounds produced from the hydrolysis of glucosinolates in B. napus tissues reduced A. euteiches oospore infection and inhibited mycelial growth, thus, demonstrating potential utility of Brassica species in the control of A. euteiches.
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Seventy-nine cultivars and lines of cabbage Brassica oleracea L. (Capitata group) were analyzed for 11 glucosinolates to provide a data base of the levels of these potential toxicants. Aglucon hydrolytic products of glucosinolates from fresh cabbage (mean of 79 cultivars) include 24 ppm allyl isothiocyanate, 45 ppm 3-methylsulfinylpropyl isothiocyanate, 18 ppm SCN ion, 17 ppm 4-methylsulfinylbutyl isothiocyanate, and 4 ppm goitrin. Composition of the cultivars are summarized by type (red, white, savoy) and by end use (market, storage, kraut). Glucosinolates with a 3-carbon aglucon (excluding the sinolate carbon) predominate over 4-carbon glucosinolates in white and savoy types. Four-carbon glucosinolates (including goitrin precursor) predominate in red cabbages. Savoy cabbages are high in glucosinolates yielding SCN ion. Distinctions between market, storage, and kraut cultivars are less well defined. No differences could be seen between open pollinated and hybrid cultivars. Year-to-year variation for 12 cultivars is discussed.
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Nisin-resistant Leuconostoc mesenteroides NCK293 and nisin-producing Lactococcus lactis subsp. lactis NCK401 were evaluated separately and in combination for growth and nisin production in a model sauerkraut fermentation. Strains were genetically marked and selectively enumerated by using antibiotic-containing media. The growth and survival of L. mesenteroides were similar in the presence and absence of Lactococcus lactis subsp. lactis. The growth of Lactococcus lactis subsp. lactis was not inhibited, although the maximum cell density was reduced and the population decline was more pronounced in the presence of L. mesenteroides. Nisin was detected within 24 h, and levels were relatively constant over the 12-day test period. The maximum cell populations and nisin level achieved could be altered by changing the initial cell ratios of L. mesenteroides and lactococcus lactis subsp. lactis. Isogenic nisin-producing and nisin-negative Lactococcus lactis subsp. lactis derivatives were used in combination with nisin-resistant L. mesenteroides to demonstrate that nisin levels produced in mixed culture were sufficient to retard the onset of the growth of nisin-sensitive, homofermentative Lactobacillus plantarum ATCC 14917.
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Two Lactococcus lactis subsp. lactis strains, NCK400 and LJH80, isolated from a commercial sauerkraut fermentation were shown to produce nisin. LJH80 was morphologically unstable and gave rise to two stable, nisin-producing (Nip+) derivatives, NCK318-2 and NCK318-3. NCK400 and derivatives of LJH80 exhibited identical morphological and metabolic characteristics, but could be distinguished on the basis of plasmid profiles and genomic hybridization patterns to a DNA probe specific for the iso-ISS1 element, IS946. NCK318-2 and NCK318-3 harbored two and three plasmids, respectively, which hybridized with IS946. Plasmid DNA was not detected in NCK400, and DNA from this strain failed to hybridize with IS946. Despite the absence of detectable plasmid DNA in NCK400, nisin-negative derivatives (NCK402 and NCK403) were isolated after repeated transfer in broth at 37 degrees C. Nisin-negative derivatives concurrently lost the ability to ferment sucrose and became sensitive to nisin. A 4-kbp HindIII fragment containing the structural gene for nisin (spaN), cloned from L. lactis subsp. lactis ATCC 11454, was used to probe genomic DNA of NCK318-2, NCK318-3, NCK400, and NCK402 digested with EcoRI or HindIII. The spaN probe hybridized to an 8.8-kbp EcoRI fragment and a 10-kbp HindIII fragment in the Nip+ sauerkraut isolates, but did not hybridize to the Nip- derivative, NCK402. A different hybridization pattern was observed when the same probe was used against Nip+ L. lactis subsp. lactis ATCC 11454 and ATCC 7962. These phenotypic and genetic data confirmed that unique Nip+ L. lactis subsp. lactis strains were isolated from fermenting sauerkraut.
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Cabbage (Brassica oleracea var capitata) leaves were used as a source of cystine lyase. Diethylaminoethyl-cellulose chromatography resolved two peaks of activity, designated I and II.Cystine lyase I (molecular weight 145,000) and O-acetylserine sulfhydrylase (molecular weight 70,000) were resolved by Bio-Gel A-0.5M chromatography. This isozyme catalyzed an alpha,beta-elimination reaction with cystine, cysteine, O-acetylserine, and several S-substituted cysteines. The substrate specificity was similar to previously reported S-alkylcysteine lyases. The elution profiles during purification, and heat inactivation studies indicated that the above reactions were catalyzed by a single protein. The pH optimun with cystine and cysteine as substrate was 8.5 to 9.0, and the K(m) values were: cystine (0.3 mm), cysteine (0.3 mm), O-acetylserine (6 mm), and S-methylcysteine sulfoxide (1.8 mm).Cystine lyase II was resolved into three peaks (molecular weight greater than 500,000, 240,000, and 145,000) using Bio-Gel A-0.5M chromatography. This enzyme degraded l-cystine, l-cysteine, O-acetylserine, S-methylcysteine sulfoxide, and djenkolic acid. The pH optimum with cystine and cysteine was 8.5 to 9.0, and the K(m) values were: cystine (0.3 mm), cysteine (0.3 mm), O-acetylserine (12.5 mm), and S-methylcysteine sulfoxide (3.7 mm).
Article
Thirteen varieties of cabbage, including five commonly used commercial varieties and eight newly developed hybrids, were analyzed for their ability to undergo lactic acid fermentations. The rates of fermentation, total quantities of acids produced, and chemical analyses indicate that 11 of the 13 selections could support adequate fermentation at any of the three stages of maturation. However, two varieties G27 × G51 (105) and G60 × W-1 (106), harvested during the earlier phases of the 1967 and 1968 growing seasons, failed to undergo proper fermentations and consistently resulted in producing putrid products. These hybrids when harvested at the late stage of maturation were capable of producing total titratable acidities comparable to the acceptable varieties. However the resulting sauerkrauts were judged to be of poor quality. The inability of these varieties to support adequate fermentations may be due to growth inhibitory substances or the lack of inherent maturation-dependent growth factor(s) essential for the growth of lactic acid bacteria.
Article
Green olives were found to contain an inhibitor(s) of several species of lactic acid bacteria usually associated with the Spanish-type brined olive fermentation. The inhibitor was demonstrated by the presence of inhibition zones surrounding tissue which had been cut from frozen olives and implanted in a seeded nutritive agar medium. Relative potencies of aqueous extracts of frozen olives were determined by a paper disc assay method. The Mission variety of olive contained the most inhibitor, and the Manzanillo and Ascolano, about 50 and 40% as much as the Mission variety, respectively. Sevillano and Barouni varieties contained comparatively little inhibitor. Effects of the inhibitor on growth rates of lactic acid bacteria were determined by adding various amounts of a concentrated aqueous extract of olives to a nutritive broth medium contained in screw-capped tubes. Of the four species of lactic acid bacteria tested, Leuconostoc mesenteroides was the most sensitive, and Lactobacillus plantarum was the least sensitive; Pediococcus cerevisiae and Lactobacillus brevis were intermediate in sensitivity. Extracts possessed a bactericidal property, as evidenced by their effect on L. mesenteroides . Sodium chloride, especially at concentrations of about 5% and higher, greatly increased the effectiveness of the inhibitor. The inhibitor was ethyl alcohol-soluble and was stable when heated at 100 C in aqueous solution. Potencies of extracts were reduced greatly by adjustment to p H 10, but no appreciable effect was noted by adjustment to p H 0.8. Images Fig. 1
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キャベツの重要細菌病である黒腐病に対する抵抗性の機作を解明する目的で,黒腐病細菌Xanthomonas campestris pv. campestris (Xcc),キャベツに非病原性のインゲン葉焼病菌X. campestris pv. phaseoli (Xcp),及び腐生細菌の霊菌Serratia marcescens (Sm)を用い,キャベツ葉組織におけるこれら細菌の増殖と接種組織における抗菌活性の変化との関係を比較検討した。 1. 3種類の細菌をキャベツ葉に針接種及び注入接種した場合, Xccは対数的によく増殖して発病に至ったが, XcpとSmの2細菌は,増殖が強く抑制されて徐々に減少した。しかし,健全キャベツ葉から採取した細胞間汁液中では3細菌とも同等によく増殖し,非病原細菌がとくに増殖阻害を受けるということはなかった。 2. Xcpを注入接種後3日目の接種葉から得た水抽出液及び酢酸エチル粗抽出物は,健全葉からのものよりXccとXcpに対して高い抗菌活性を示し,その抗菌物質は低分子性の物質であろうと推定された。 3. 供試3細菌を接種後,経時的に細菌の増殖経過と酢酸エチルで抽出される抗菌物質の生成量とを比較検討した結果, Xcp接種葉では,抗菌物質が1日後にすでに多量に生成され, 7日目まで高濃度で推移することが明らかとなった。組織内の細菌の増殖は抗菌物質の生成量と対応して接種当初から著しく阻害された。これとほぼ同様のパターンが腐生細菌Smの接種葉でもみられたが,抗菌物質の生成量はかなり低かった。一方, Xcc接種葉では,抗菌物質の生成より先行して細菌が急速に増殖し,病徴を発現した。抗菌物質は感染初期には全く増加せず,発病期ごろからわずかに増え始め,発病末期にはかなりの生成量に達した。なお,健全葉からも微量の抗菌物質が検出された。
Article
The effects of pH values and NaCl concentrations on the growth rates of five species of lactic acid bacteria commonly associated with the sauerkraut fermentation were determined in filter-sterilized cabbage juice. Growth rates of all cultures, with the exception of Pediococcus cerevisiae, were retarded by addition of salt, lower pH, or interaction of both pH and salt. Based upon lag and generation times, P. cerevisiae was the culture most tolerant to the pH and salt concentration employed, whereas Streptococcus faecalis was the most sensitive species. Of the heterofermentative cultures, Lactobacillus brevis was less subject to growth inhibition than Leuconostoc mesenteroides. Under conditions simulating those found during the initial phases of the sauerkraut fermentation (2.25% salt, pH 6.2), L. mesenteroides displayed the shortest lag and generation times of all cultures examined. This rapid growth rate coupled with a marked accelerated death rate may explain, in part, the reason this species is both the first to dominate and the first to die during the early phases of the sauerkraut fermentation. Although cabbage juice previously fermented by L. mesenteroides appears to inhibit growth of P. cerevisiae, it had no apparent inhibitory or stimulatory effects on the other cultures. Under conditions simulating those found during the initial phases of the sauerkraut fermentation (2.25% salt, pH 6.2), L. mesenteroides displayed the shortest lag and generation times of all cultures examined. This rapid growth rate coupled with a marked accelerated death rate may explain, in part, the reason this species is both the first to dominate and the first to die during the early phases of the sauerkraut fermentation. Although cabbage juice previously fermented by L. mesenteroides appears to inhibit growth of P. cerevisiae, it had no apparent inhibitory or stimulatory effects on the other cultures.
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Many plants that serve for human nutritional purposes contain compounds from which physiologically active substances are formed by enzymic reactions. Usually such compounds, and the enzymes which react with them, are located in different cells, so that the active substances are formed only on crushing the plant: Examples are the lachrymatory factor and the antimicrobial substances formed from the different cysteine-S-oxides of the onion and garlic, as well as the goitrogenic compounds formed from the thioglucosides occuring in the Brassica species.
Article
Mechanisms for the formation of methanethiol, dimethyl disulfide, and dimethyl trisulfide in disrupted cabbage tissues were investigated. Dimethyl disulfide was produced in both air- and nitrogen-saturated disrupted cabbage tissues without significant differences (p ≤ 0.05), which indicated that air oxidation of methanethiol is not the predominant mechanism for the formation of dimethyl disulfide. These results favored the mechanism in which the formation of dimethyl disulfide occurs from chemical disproportionation of methyl methanethiosulfinate. Methanethiol and dimethyl trisulfide were formed rapidly in model systems containing either methyl methanethiosulfinate or methyl methanethiosulfonate and hydrogen sulfide. This indicated that the reactions of the thiosulfinate and thiosulfonate compounds with hydrogen sulfide are prominent mechanisms for the formation of methanethiol and dimethyl trisulfide following the action of cysteine sulfoxide lyases. Methyl methanethiosulfinate and methyl methanethiosulfonates were found to possess characterizing sauerkraut aroma notes.
Article
The naturally occurring sulfur compound S-methylcysteine sulfoxide (SMCSO) was determined for five common Brassica vegetables, broccoflower, broccoli, Brussels sprouts, cabbage, and cauliflower. This natural chemical has a role in the aroma and flavor of these vegetables and, perhaps more importantly, appears to be involved with the inhibition of experimental carcinogenesis. Brussels sprouts contained the highest level of SMCSO. This vegetable was then used to identify the cystine lyase-mediated enzymatic conversion products including methyl methanethiosulfinate (MMTSO). After it was demonstrated that MMTSO, dimethyl trisulfide, and pyruvate can be formed facilely in a simplified enzymatic model system, generation of MMTSO was confirmed in a water extract of macerated Brussels sprouts. Formation of MMTSO was shown to be strongly dependent on pH of the vegetable juice. After 24 h, only trace amounts of MMTSO were observed at the developed acidic pH of Brussels sprouts, while considerable amounts were formed at an adjusted basic pH. This is the first evidence that MMTSO is enzymatically derived from SMCSO under natural conditions.
Article
A quantitative study of the hydrolysis of S-methyl-L-cysteine sulfoxide by boiling 1 N hydrochloric acid shows that most of the material is converted to pyruvic acid, ammonia, dimethyl disulfide and methyl methanethiolsulfonate. The sulfur-containing products are probably formed from methyl methanethiolsulfinate, which has been shown to disproportionate to them under the conditions of the experiment. The thiolsulfinate may be formed from methanesulfenic acid. Alanine, S-methylcysteine and a carbonyl compound are formed in small amounts. Hydrolysis of the sulfoxide in neutral solution follows the same course as in acid solution. Two quantitative procedures for the determination of methyl methanethiolsulfonate have been developed.
Article
Combinations of Leuconostoc mesenteroides, Lactobacillus brevis, Pediococcus cerevisiae, and Lactobacillus plantarum were grown in sterile cabbage juice broth. The effects of culture interactions were determined by lag times, generation times, pH change, and total acidity measurements. Fresh filter-sterilized cabbage juice was a better medium than autoclaved cabbage juice, or refrigerated filter-sterilized cabbage juice. The addition of NaCl (2.25%) had a minimal effect on the cultures, except for L. mesenteroides, which was inhibited. In paired association studies, L. mesenteroides was inhibited by all the other cultures, conversely, L. plantarum and L. brevis benefited from most of the culture interactions.
Article
An enzyme which degrades cysteine sulfoxides has been found in members of the genus Brassica of the family Cruciferae. The name 3-alkylsulphinylalanine alkyl sulphenate lyase (deaminating) (cysteine sulfoxide lyase) is proposed for this enzyme.1 The enzyme has been purified about eleven-fold from broccoli buds. The broccoli lyase is similar in its mode of action to alliinase which has been described in Allium species. The pH optimum of the broccoli enzyme is between 8·4 and 8·6 in borate buffer. The products of the reaction are pyruvate, ammonia, and alkyl alkane thiosulfinates. The Km is 2·7 × 10−3M with l-methylsulphinylalanine as the substrate. Studies with acetone powders and aged preparations, and inhibition studies indicate that pyridoxal-5′-phosphate is a coenzyme. s-methyl-l-cysteine was not a substrate, nor did it inhibit the reaction when present in equimolar concentration with the sulfoxide. Equimolar amounts of l-cysteic acid and l-cysteine sulfinic acid resulted in inhibitions of 30–40 per cent in the utilization of l-methylsulphinylalanine.
Article
Knowledge of glucosinotate (GS) content in cabbage (Brassica oleracea) is needed to establish levels at which these compounds or their enzymically released products are consumed. New methods of analysis for individual and total GS were applied to 12 open-pollinated and 10 hybrid varieties. Total glucosinolates in cabbage as harvested ranged from 299 to 1288 ppm. Of the 12 GS's determined, those found in the greatest amount were: two 3-indotylmethyl GS's (18 to 63 ppm as thiocyanate ion); allyl GS [4 to 146 ppm as allyl isothiocyanate (ITC)]; 3-methyisulfinylpropyl GS (30 to 164 ppm as ITC); 4methylsulfinylbutyl GS (0.2 to 119 ppm as ITC). The last two have not been quantitated previously in edible cabbage. Goitrin content ranged from 1.2 to 26 ppm. Significant differences among varieties were found and are attributed to genetic variability. Variation in GS content from head to head within a variety was greater in the open-pollinated than in the hybrid varieties.
Article
Under controlled conditions, extracts of garlic, onion, turnip, green peppers and radishes were used to inhibit Escherichia coli, Salmonella typhosa, Shigella dysentriae and Staphylococcus aureus, which are all pathogenic bacteria. It was found that 1–4% by vol. of garlic extract completely inhibited the growth of all the bacteria used. 4% by vol. of onion extract completely inhibited the growth of both Shigella dysentriae and Staphylococcus aureus at 10–6 dilution. Salmonella typhosa and E. coli were not completely inhibited; the inhibition was 48·3% for E. coli and 95·3% for Salmonella typhosa. At 10–4 bacterial dilution, onion extract decreased the colony number substantially in all four bacteria. 4% extracts from turnip, green peppers and radishes did not show a definite antibacterial action against any bacterium at the given dilutions. On the contrary some growth stimulating activity of these extracts on some bacteria was observed.
Article
Thirteen varieties of cabbage, including five commonly used commercial varieties and eight newly developed hybrids, were analyzed for their ability to undergo lactic acid fermentations. The rates of fermentation, total quantities of acids produced, and chemical analyses indicate that 11 of the 13 selections could support adequate fermentation at any of the three stages of maturation. However, two varieties G27 x G51 (105) and G60 x W-1 (106), harvested during the earlier phases of the 1967 and 1968 growing seasons, failed to undergo proper fermentations and consistently resulted in producing putrid products. These hybrids when harvested at the late stage of maturation were capable of producing total titratable acidities comparable to the acceptable varieties. However the resulting sauerkrauts were judged to be of poor quality. The inability of these varieties to support adequate fermentations may be due to growth inhibitory substances or the lack of inherent maturation-dependent growth factor(s) essential for the growth of lactic acid bacteria.
Article
Green olives were found to contain an inhibitor(s) of several species of lactic acid bacteria usually associated with the Spanish-type brined olive fermentation. The inhibitor was demonstrated by the presence of inhibition zones surrounding tissue which had been cut from frozen olives and implanted in a seeded nutritive agar medium. Relative potencies of aqueous extracts of frozen olives were determined by a paper disc assay method. The Mission variety of olive contained the most inhibitor, and the Manzanillo and Ascolano, about 50 and 40% as much as the Mission variety, respectively. Sevillano and Barouni varieties contained comparatively little inhibitor. Effects of the inhibitor on growth rates of lactic acid bacteria were determined by adding various amounts of a concentrated aqueous extract of olives to a nutritive broth medium contained in screw-capped tubes. Of the four species of lactic acid bacteria tested, Leuconostoc mesenteroides was the most sensitive, and Lactobacillus plantarum was the least sensitive; Pediococcus cerevisiae and Lactobacillus brevis were intermediate in sensitivity. Extracts possessed a bactericidal property, as evidenced by their effect on L. mesenteroides. Sodium chloride, especially at concentrations of about 5% and higher, greatly increased the effectiveness of the inhibitor. The inhibitor was ethyl alcohol-soluble and was stable when heated at 100 C in aqueous solution. Potencies of extracts were reduced greatly by adjustment to pH 10, but no appreciable effect was noted by adjustment to pH 0.8.
Article
Cystine lyase degrades l-cystine by a beta-elimination to form cysteine persulfide, pyruvate, and ammonia. This enzyme is common in Brassica sp. and has been purified to homogeneity from extracts of broccoli (Brassica oleracea var botrytis) buds. Two isozymes were separated on DEAE-Fractogel columns and the first peak, cystine lyase I further purified to homogeneity. The purified enzyme had a narrow range of substrate specificity with l-cystine and S-alkyl-l-cysteine sulfoxides being the primary substrates. The K(m) for l-cystine was 1.9 millimolar and for S-ethyl-l-cysteine sulfoxide was 15.6 millimolar, suggesting that l-cystine would be preferred in vivo. Using gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis the molecular weight of the holoenzyme was estimated as 152,000 composed of subunits of approximately 49,000. This strongly suggests the native enzyme is a trimer. The presence of carbohydrate in the native enzyme was detected at the level of 5.8% on a weight basis. Except for the ability to utilize l-cystine as a substrate there are many similarities between cystine lyase I and the alliin lyase of onion (Allium cepa).
Article
A fermentor was designed and constructed for study of the physical, microbiological, and chemical changes that occur during the sauerkraut fermentation. The fermentor has some essential features that include restriction in volume of the sauerkraut bed, construction of clear plastic to permit visual determination of liquid-level changes as a result of gas entrapment within the sauerkraut bed, and a gas-lift device for use in nitrogen purging of the fermenting brine. Fermentations exhibited two distinct stages, the first one gaseous and the second non-gaseous. The gaseous stage was characterized by rapid CO(2) and acid production due to growth by hetero-fermentative lactic acid bacteria with resultant gas entrapment within the sauerkraut bed and a rise in liquid level. Also, rapid disappearance of fructose and rapid appearance of mannitol occurred during this stage. The nongaseous stage was characterized by growth of homo-fermentative lactic acid bacteria with little or no CO(2) production and a gradual increase in lactic acid until all fermentable sugars were metabolized. Nitrogen purging appeared to offer several potential advantages, including a means for brine circulation, removal of CO(2) from the brine, and anaerobiosis to ensure retention of ascorbic acid, desirable color, and other oxygen-sensitive traits in sauerkraut.
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Die antimikrobielle Wirkung van Thiocyanaten und Isothiocyanaten
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