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Characteristics of the Leuconostoc mesenteroides subsp. mesenteroides strains from fresh vegetables

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Strains synthesizing extracellular polysaccharide dextran on a medium with 10% sucrose were isolated from different kind of vegetables (cabbage, cucumber, cauliflower, kohlrabi, carrot, green beans, red beet, pepper, eggplant, radish). Carbohydrate fermentation was examined using a bioMerieux API 50 CHL test system. Among micropopulations with characteristic spherical cell morphology, 94.9% belonged to Leuconostoc mesenteroides subsp. mesenteroides and 5.1% were identified as Leuconostoc mesenteroides subsp. dextranicum. According to fermentation of pentoses L. mesenteroides strains were divided into three groups with a certain number of biotypes; 10 strains were tested on acid production. .
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UDC 635.1/.6:579.64:579.8
APTEFF, 37, 1-192 (2006) BIBLID: 1450–7188 (2006) 37, 3-11
Original scientic paper
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CHARACTERISTICS OF THE Leuconostoc mesenteroides
subsp. mesenteroides STRAINS FROM FRESH VEGETABLES
*RUGDQD5'LPLü
Strains synthesizing extracellular polysaccharide dextran on a medium with 10%
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kohlrabi, carrot, green beans, red beet, pepper, eggplant, radish). Carbohydratefermentationbohydratefermentationfermentation
was examined using a bioMerieux API 50 CHL test system. Among micropopulations with
characteristic spherical cell morphology, 94.9% belonged to Leuconostoc mesenteroides
subsp. mesenteroides and 5.1% were identifíed as Leuconostoc mesenteroides subsp.
dextranicum. According to fermentation of pentoses L. mesenteroides strains were divi-
ded into three groups with a certain number of biotypes; 10 strains were tested on acid
production.
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'5D¿QRVH –+–
*URXS%LVFKDUDFWHUL]HGE\DVPDOOHUQXPEHURIVWUDLQVEXWE\DKLJKHUYDULDELO-
LW\FRQFHUQLQJIHUPHQWDWLRQFKDUDFWHULVWLFV7DEOH$OORIWKHPEHVLGH/DUDELQRVHXVH
'[\ORVHDQGULERVH5HJDUGLQJ WKHIHUPHQWDWLYHDFWLYLW\WKHELRW\SH%ZDVGRPLQDQW
DQGWKHORZHVWDFWLYLW\VKRZHG%ZLWKWKHDELOLW\WRXVHRQO\VDOLFLQH,QFRPSDULVRQWKH
JURXS$WKHSDUWLFLSDWLRQRIUHSUHVHQWDWLYHVIHUPHQWLQJODFWRVHZDVVRPHZKDWKLJKHU
7
Table 2. 'LIIHUHQWLDWLQJFKDUDFWHULVWLFVRIL. mesenteroidesVWUDLQVJURXS%
Characteristic B1 B2 B3 B4 B5 B6
0DQQLWRO ––+
$P\JGDOLQ ++–
$UEXWLQ ++–+
6DOLFLQ +++++
&HOORELRVH +++
/DFWRVH +–++
ȕ*HQWLELRVH ++++ –
*OXFRQDWH –++
NHWR
JOXFRQDWH ––+
7KHODVWJURXS & LQFOXGHG WKH VWUDLQV IHUPHQWLQJ/DUDELQRVHDQG ' [\ORVH )LYH
VXFKVWUDLQVZHUHLGHQWL¿HG7DEOH$FFRUGLQJWRFKDUDFWHULVWLFVWKH\ZHUH JURXSHGLQ
WKUHHELRW\SHVDQGWKHPRVWDFWLYHRQHZDV&
Table 3. 'LIIHUHQWLDWLQJFKDUDFWHULVWLFVRIL. mesenteroidesVWUDLQVJURXS&
Characteristic C1 C2 C3
$P\JGDOLQ –+–
$UEXWLQ ++–
6DOLFLQ –+–
&HOORELRVH ++–
0HORELRVH –++
'5DI¿QRVH –++
*OXFRQDWH +––
$P\JGDOLQVKRZHGWREHPRVWO\IHUPHQWHGE\DOOWKUHHJURXSVL. mesenteroides7D-
EOH5HDFWLRQVLGHQWLFDO IRUDOOWKHVWUDLQVDUH QRWSUHVHQWHG7KHH[DPLQDWLRQVKRZHG
WKDWWKHVWUDLQVRIJURXS$ZHUHWKHPRVWIUHTXHQWO\SUHVHQWRQJUHHQEHDQVDQG
WKHVWUDLQVRIJURXS% RQFDXOLÀRZHUDQG FDUURW7DEOH6LPLODUUHVXOWVIRU ODFWLFDFLG
EDFWHULDZHUHUHSRUWHGE\9DOGH]HWDO
8
Table 4. 1XPEHURISRVLWLYHVWUDLQVZLWKIHUPHQWDWLRQDELOLW\RILVRODWHGL. mesenteroidesJURXSV
Source of
fermentation
Group
ABC
$P\JGDOLQ   3
$UEXWLQ  4
&HOORELRVH  4
'5DI¿QRVH   4
*DODFWRVH 
*OXFRQDWH 8
/DFWRVH   
0DQQLWRO   
0HORELRVH 4
6DOLFLQ  3
E*HQWLELRVH 
NHWRJOXFRQDWH  
Table 5. )UHTXHQFHRIL. mesenteroidesJURXSVLQYHJHWDEOHV
Vegetable Group [%]
ABC
3HSSHU   
&DXOLÀRZHU  
&DEEDJH  
*UHHQ%HDQ   
&XFXPEHU  
(JJSODQW 
5DGLVK  
5HG%HHW  
.RKOUDEL   
&DUURW 
7KHDFLGSURGXFWLRQ E\  L. mesenteroidesVWUDLQVRIJURXSV$DQG%LVSUHVHQWHG
LQ7DEOH7KHDQDO\VLVRIUHVXOWVVKRZVWKDWWKHKLJKHVWSHUFHQWDFLGLW\H[SUHVVHGDVWKH
FRQWHQWRISURGXFHGODFWLFDFLGZDVREWDLQHGDIWHUWKH¿UVWGD\RIIHUPHQWDWLRQZKLOHWKH
SURGXFWLRQZDV DIWHUZDUGVVORZHU7KHELJJHVWGHFUHDVH RIS+YDOXHZDV UHFRUGHGDIWHU
WKH¿UVWGD\FRPSDUHGWRWKHLQLWLDOYDOXHRI
Table 6. $FLGSURGXFWLRQDQGS+YDOXHVRIL. mesenteroides
DWWDLQHGDIWHUJURZWKIRUDQGGD\V
Strain 1. day 7. day
Acid (%) pH Acid (%) pH
$    
$    
$    
$    
$    
$    
%    
%    
%    
%    
,QLWLDOS+ 
5HJDUGLQJWKHIHUPHQWDWLYHDFWLYLW\¿YHVWUDLQVDUHVWDQGLQJRXWZLWKKLJKHVWSURGXF-
WLRQRIDFLG$%DQG$$$6WUDLQ$SUR-
GXFHGWKHORZHVWDPRXQWRIDFLGGXULQJWKHVHYHQGD\VRIIHUPHQWDWLRQ±
&21&/86,21
5HVHDUFKRIIHUPHQWDWLRQIHDWXUHVRIODFWLFDFLGEDFWHULDLVWKH¿UVWVWHSLQWKHVHOHFWLRQ
RIVWUDLQV FDSDEOHWRSURGXFH VXI¿FLHQWDPRXQWRI DFLG7KH DLPRIIXUWKHU LQYHVWLJDWLRQ
ZDVWRFKRRVHWKHPRVWVXLWDEOHVWUDLQVRIL. mesenteroides DVVWDUWHUFXOWXUHVIRUWKHFRQ-
WUROOHGIHUPHQWDWLRQRIYHJHWDEOHV
5()(5(1&(6
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Leuconostoc mesenteroidesKH[DGHFDQH6FLHQFHVGHV$OLPHQWV17 
 7DOOJUHQ$+8$LUDNVLQHQ5YRQ:HLVVHQEHUJ+2MDPR -.XXVLVWRDQG0
/HLVROD([SRO\VDFFKDULGHSURGXFLQJ EDFWHULD IURP VXJDU EHHWV$SSOLHG DQG (QYL-
URQPHQWDO0LFURELRORJ\65 
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KHWHURODFWLFDFLGEDFWHULDLQJUHHQEHDQMXLFH-RXUQDORI)RRG6FLHQFH48 
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ɄȺɊȺɄɌȿɊɂɋɌɂɄȿɋɈȳȿȼȺLeuconostoc mesenteroides
subsp. mesenteroides ɂɁɈɅɈȼȺɇɂɏɂɁɋȼȿɀȿȽɉɈȼɊȶȺ
ȽɨɪɞɚɧɚɊȾɢɦɢʄ
ɂɡɪɚɡɥɢɱɢɬɨɝɩɨɜɪʄɚ ɤɭɩɭɫ ɤɪɚɫɬɚɜɚɰ ɤɚɪɮɢɨɥ ɤɟɥɟɪɚɛɚɦɪɤɜɚɡɟɥɟɧɚɛɨ
ɪɚɧɢʁɚ ɰɜɟɤɥɚ ɩɚɩɪɢɤɚ ɩɥɚɜɢ ɩɚɬɥɢʇɚɧ ɪɨɬɤɜɚ ɢɡɨɥɨɜɚɧɢɫɭ ɫɨʁɟɜɢ ɤɨʁɢ ɧɚ ɩɨ
ɞɥɨɡɢ ɫɚ  ɫɚɯɚɪɨɡɟ ɫɢɧɬɟɬɢɲɭ ɟɤɫɬɪɚɰɟɥɭɥɚɪɧɢ ɩɨɥɢɫɚɯɚɪɢɞ ɞɟɤɫɬɪɚɧ Ɏɟɪ
ɦɟɧɬɚɰɢʁɚɭɝʂɟɧɢɯɯɢɞɪɚɬɚɢɫɩɢɬɚɧɚ ʁɟ ɤɨɪɢɲʄɟʃɟɦELRɆɟULHX[Ⱥ3,  &+/ɬɟɫɬ
ɫɢɫɬɟɦɚ Ɉɞ ɦɢɤɪɨɩɨɩɭɥɚɰɢʁɚɤɨʁɟ ɫɭ ɫɟ ɤɚɪɚɤɬɟɪɢɫɚɥɟ ɫɮɟɪɢɱɧɨɦ ɦɨɪɮɨɥɨɝɢʁɨɦ
ʄɟɥɢʁɚ  ɩɪɢɩɚɞɚɥɨ ʁɟ ɜɪɫɬɢ Leuconostoc mesenteroides VXEVS mesenteroides
ɞɨɤʁɟɢɞɟɧɬɢɮɢɤɨɜɚɧɨɤɚɨLeuconostoc mesenteroidesVXEVSdextranicumɉɪɟ

ɦɚɮɟɪɦɟɧɬɚɰɢʁɢɩɟɧɬɨɡɚL.mesenteroidesɫɨʁɟɜɢ ɫɭɫɜɪɫɬɚɧɢ ɭ ɬɪɢ ɝɪɭɩɟ ɫɚ ɨɞɝɨ
ɜɚɪɚʁɭʄɢɦɛɪɨʁɟɦɛɢɨɬɢɩɨɜɚɫɨʁɟɜɚL. mesenteroidesʁɟɬɟɫɬɢɪɚɧɨɧɚɩɪɨɢɡɜɨɞʃɭ
ɤɢɫɟɥɢɧɟ
Received 5 May 2006
Accepted 29 September 2006
... These heterolactic organisms produce carbon dioxide and organic acids which rapidly lower the pH of the beverage to 4.0 or 4.5 and inhibit the development of undesirable microorganisms. The carbon dioxide produced replaces the oxygen, making the environment anaerobic [34] and suitable for the growth of subsequent organisms such as Lactobacillus. Besides, the anaerobic environment created by the CO2 has a preservative effect on the beverage since it inhibits the growth of unwanted bacteria contaminants [35]. ...
... Besides, the anaerobic environment created by the CO2 has a preservative effect on the beverage since it inhibits the growth of unwanted bacteria contaminants [35]. As reported by [36], L. pseudomesenteroides is widely present in many fermented foods such as dairy, wine and beans while L. mesenteroides is associated with sauerkraut and pickled fermented products [34]. The organism produces dextrans and aromatic compounds (diacetyl, acetaldehyde, and acetoin) which could contribute to the taste and aromatic profile. ...
Article
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The appropriate solution to the problem of quality variability and microbial stability of traditional non-alcoholic pearl millet fermented beverages (NAPMFB) is the use of starter cultures. However, potential starter cultures need to be tested in the production process. We aimed to identify and purify bioburden lactic acid bacteria from naturally fermented pearl millet slurry (PMS) and assess their effectiveness as cultures for the production of NAPMFB. Following the traditional Kunun-zaki process, the PMS was naturally fermented at 37 °C for 36 h. The pH, total titratable acidity (TTA), lactic acid bacteria (LAB), total viable count (TVC) and the soluble sugar were determined at 3 h interval. The presumptive LAB bacteria were characterized using a scanning electron microscope , biochemical tests and identified using the VITEK 2 Advanced Expert System for microbial identification. The changes in pH and TTA followed a non-linear exponential model with the rate of significant pH decrease of 0.071 h −1 , and TTA was inversely proportional to the pH at the rate of 0.042 h −1. The Gompertz model with the mean relative deviation modulus, 0.7% for LAB and 2.01% for TVC explained the variability in microbial growth during fermentation. The LAB increased significantly from 6.97 to 7.68 log cfu/mL being dominated by Leuconostoc, Pediococcus, Streptococcus and Enterococcus with an optimum fermentation time of 18 h at 37 °C and 4.06 pH. L. mesenteroides and P. pentosaceus created an acidic environment while E. gallinarum increased the pH of the pearl millet extract (PME). Innovative NAPMFB was produced through assessment of LAB from PMS to PME fermented with L. mesentoroides (0.05%) and P. pentosaceus (0.025%) for 18 h, thereby reducing the production time from the traditional 24 h.
... Leuconostoс mesenteroides является преобладающим микроорганизмом на ранних стадиях ферментации капусты и оказывает большое влияние на вкус и качество продукта ферментации, поэтому важной задачей являлось изучение характеристик генома L. mesenretoides, а также разнообразие геномов молочнокислых бактерий (Lactobacillus gasseri, L. casei, L. bulgaricus, and L. brevis, L. mesenteroides, Oenococcus oeni, Lactococcus cremoris, P. pentosaceus, Streptococcus thermophilus, Brevibacterium linens) и формирование их в 1 геном консорциум. Данные показали, что бактериофаги могут играть важную роль в естественной смене молочнокислых бактерий [6]. Исследование последовательности бактериофага Leuconostoc mesenteroides Φ1-A4, выделенного в процессе промышленной ферментации овощей, представляет собой первую полную геномную и молекулярную характеристику фага Leuconostoc, и результаты исследования могут положить начало развитию технологии управления фагами. ...
... Темпы нарастания концентрации микроорганизмов вида υ=f_v (τ) определяли дифференцированием функций (2) -(4) по их аргументу: (5) где υ -темп нарастания концентрации микроорганизмов, [lg (КОЕ/г)]/ч. Для удобства графического представления логики дальнейших рассуждений, функции темпов были преобразованы следующим образом: (6) При выраженной lg N, значения темпа нарастания концентрации микроорганизмов для разных моментов времени будут логически неэквивалентны друг другу даже в случае их численного равенства. Поэтому дальнейшие расчёты выполняли приведёнными значениями темпа: ...
Article
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Relevance . Cabbage is one of the most popular products, which is mainly fermented with the addition of various vegetables. When fermentation is not only the original nutrients such as vitamin C, amino acids, dietary fibers, etc., but also develop functional microorganisms such as lactic acid bacteria. Fermentation has an important effect on the quality and taste of cabbage, so it is important to study the fermentation process, microbial diversity and changes in nutrients and chemical elements in the fermentation process. L. mesenteroides is considered to be the dominant species on heterofermentative early stages of fermentation. However, there is little information on the diversity of species and strains of Leuconostoc involved in fermentation of sauerkraut. Studies that used traditional biochemical methods to study fermentation of sauerkraut showed that four main types of lactic acid bacteria were involved in the fermentation process: Leuconostoc mesenteroides, Lactobacillus plantarum, Pediococcus pentosaceus and Lactobacillus brevis. Taking into account the importance of two-stage fermentation of vegetable raw materials in order to create optimal conditions for the development of the "main" pool of lactic acid microorganisms at the first stage, it becomes urgent to conduct a complex of studies aimed at reproducing the "natural" process in which the main role is played by bacteria of the genus Leuconostoc mesenteroides at the second stage – monocultures of lactic acid microorganisms and their consortia. Methods . The paper studies the dynamics of the type of interaction of lactic acid microorganisms in paired consortiums on model media pretreated by the culture of the species Leuconostoc mesenteroides, at the main stage of step fermentation of white cabbage of the "Parus" variety. Results . It is established that the sum of the criteria, the consortium "L. mesenteroides \ L. casei + L. plantarum" demonstrates the most pronounced advantage compared with monoculture cultivation of appropriate format of pseudotensorial; despite the pronounced synergy in the cultivation of the consortium "L. mesenteroides \ L. brevis + L. plantarum", the dynamics of the comparison index on the rate of increase in the concentration of microorganisms indicates the need for additional research.
... Most of bacteria were grown in single and isolated cells underneath the dextran film. This is similar to the observation of McCleskey et al. (1947) and Dimic (2006). Interestingly, Fig. 6a also shows random white spots which are foreign colonies distributed on the film and throughout the soil grains. ...
Article
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The bio-clogging using bacteria can be an eco-friendly and sustainable alternative to conventional grouting methods for seepage control. However, it remains unclear to date how the dilute concentration of bacterium and medium during field installation can affect the setting time of bacterium and its correlation with permeability reduction. In this study, the setting time of bacterium and its effectiveness in permeability reduction were addressed through experimental and theoretical investigations. A series of sand column was cultivated using different concentrations of Leuconostoc mesenteroides and culture medium. The distribution and composition of the bacterial product (i.e. dextran) were observed by refractometer, scanning electron microscope (SEM), and energy dispersive X-ray spectroscopy (EDS). Soil permeability was recorded using a constant head test. The results revealed that bacterium was effective to produce dextran at the setting time of about 5 d after installation. This dextran can reduce the permeability of bio-mediated soil by two orders of magnitude, even without culture medium supply. In general, the dextran production decreased proportionally with increase of bacterium and medium concentration. However, at 50% bacterium and medium concentration by weight, it still has a significant influence on permeability reduction with similar setting time, compared to 100% concentration.
... Morphological characteristics were confirmed by visual and microscopic observations as listed in Table 1. S. macedonicus MBF 10-2 colonies grown in MRS medium and modified colonies showed similar characteristics. L. mesenteroides and Bacillus subtilis, as indicator bacteria, showed typical colony formation [27] as presented in Fig. 1. Molecular identification using 16S rDNA polymerase chain reaction (PCR) and sequencing was used to confirm the starter culture as S. macedonicus MBF10-2 at 99% homology. ...
... One of them, L. mesenteroides produces water-soluble and -insoluble dextran, this compound is a biodegradable glucose polymer which has applications in pharmaceutical, cosmetics, oil, and food industries because its biological activities such as immunomodulatory and antitumor effects (Shukla and Goyal, 2014). In addition, this compound can be used as a cryoprotective agent for plant, animal, and human cells (Dimic, 2006). In addition, different LAB have been identified in this beverage; these bacteria have the ability to produce antimicrobial compounds called bacteriocin, which are antimicrobial agents of protein origin which are considered as safe for human health. ...
Chapter
Coyol wine, better known as “tavern” is a traditional beverage produced from the fermented sap of coyol palm (Acrocomia aculeata); in México, this beverage is only produced in the State of Chiapas specifically in the regions of the Frailesca, Centro, and Soconusco. This beverage has been consumed for a long time, the palm name derives from the Nahuatl “coyoli,” which means “rattlesnake” and was formerly known as “cuauchcoyolli” or rattlesnake tree. The sap obtained from this palm is white, slightly thick, and refreshing. This sap is obtained in the months of March and April and without fermentation this sap is clean, colorless, and sweet containing about 10%–12% of sugar (sucrose), proteins, vitamins, and a low fat content which makes it attractive for various microorganisms such as bacteria and yeasts which are responsible for tavern fermentation. Where the level of sugars rapidly decreases as they are converted into alcohol making the sap turn milky white, thick and with a certain degree of alcohol, fermentation takes place in a short period (24 h) having in the end an alcohol content of approximately 13%. Fermentation is performed with a native consortium of several microorganisms, where it has been reported that the presence of yeasts such as Saccharomyces cerevisiae and bacteria of the Leuconostoc and Lactobacillus genera, the last one being considered as lactic acid bacteria. In this chapter, the chemical and physical properties of tavern and the microorganisms involved in fermentation as well as its potential applications have been discussed.
... L. mesenteroides forms an alphaglucan polysaccharide and dextran in the sucrose-rich medium by the action of dextran sucrose enzyme which in result changes the viscosity of the sugarcane juice during the storage. (Dimic, 2006;Singh, Gaikwad, & Omre, 2014). The fermentation by yeasts like Saccharomyces cerevisiae, under anaerobic conditions, will produce unwanted products. ...
Article
In this work, sugarcane juice was preserved using non-thermal hurdles consisting of microfluidisation and natural polypeptides (nisin and polylysine). The effects were compared with thermal hurdles which comprised of pasteurization and Potassium Metabisulfite (KMS) and the untreated control sample. All the samples were stored in glass bottles at 5±2 °C and evaluated for physico-chemical, microbial and sensory parameters till 63 days. In the untreated sample, pH and total soluble solids (TSS) decreased, whereas, titrable acidity and color (L* and a*) values increased significantly (P ≤ 0.01). Both thermal and non-thermal hurdles were able to cause a complete reduction in polyphenoloxidase activity which was 23 units in the control sample. In the sample treated with thermal hurdles small changes in pH, TSS, titrable acidity, color (L* and a*) occurred during storage. On the contrary, the physico-chemical properties of juice samples treated with non-thermal hurdles were stable throughout the storage period and were more acceptable to the sensory panel than the pasteurized sample. Using non-thermal hurdles, 100% reduction in initial microbial load of 3.19 × 10⁶ CFU/ml was achieved and remained within 50 CFU/ml ensuring the shelf life of 56 days.
Article
We studied the possibility of using RAPD-PCR with primers: ERIC1R-1, ERIC2-1, BOXA1R, BOXA2R and Rep-PCR with primers P15, P16, XD8, XD9, RAPD-mes, (GTG)5 to identify genetic heterogeneity of 9 strains and 8 isolates of Leuconostoc mesenteroides. Three clusters of cultures with a high level of bootstrap support were identified as a result of phylogenetic analysis obtained when typing Leuconostoc. The obtained results indicate the possibility of revealing genetic differences in the profile of the generated amplicons among Leuconostoc mesenteroides strains using the combined methods of Rep-PCR and RAPD-PCR.
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Introduction: Fermentation is a biotechnological process of preserving the biological potential of raw materials and transforming them in order to impart new organoleptic properties and to increase nutritional value of the product allowing diversification of daily meals; thus, in some countries fermented products make up a significant part of the human diet. Despite the fact that fermented products are very useful for humans, the fermentation process itself remained rather complicated for reproduction during a long time. Currently, starter cultures are used in industrial production of fermented food products enabling the production of foodstuffs with a guaranteed range of consumer properties. Such species of lactic acid bacteria as Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Oenococcus, Pediococcus, Streptococcus, Tetragenococcus, Vagococcus, and Weissella play the main role in production of fermented food and drinks while L. mesenteroides plays the primary role in starting fermentation of many types of plant materials including cabbage, beet, turnip, cauliflower, green beans, chopped green tomatoes, cucumbers, olives, etc. Objective: To control and manage the industrial fermentation process, it is important to determine the main processes occurring at different stages and the types of lactic acid microorganisms responsible for initiation, continuation and completion of the process. Results: This review shows that, despite the variety of fermentable vegetables, L. mesenteroides species of lactic acid bacteria are of particular importance at the primary heteroenzymatic stage since during this very period the processed raw materials form conditions for inhibiting pathogenic and facultative pathogenic microflora and create optimal environment for subsequent development of targeted microorganisms determining the quality of finished products. Conclusions: When developing food technology, L. mesenteroides species of lactic acid bacteria must be an indispensable component of industrial starter cultures for obtaining final products of consistently high quality.6
Article
Bacteria p. Leuconostoc is a technologically important group of lactic acid bacteria that is part of starter cultures for production of various dairy products. Two species are most important in the dairy industry: Leuconostoclactis and Leuconostocmesenteroides , which includes three subspecies: dextranicum , mesenteroides and cremoris . The main problem of identifying representatives of the p. Leuconostoc that these microorganisms can often be misidentified as enterococci or lactobacilli. In comparison with traditional methods of species detection, the establishment of species identity using PCR is characterized by universality, a deeper level of species differentiation, high reproducibility and reliability. The article presents the results of designing specific primers for Leuconostocmesenteroides ssp. mesenteroides and Leuconostocmesenteroides ssp. dextranicum . The specificity of developed primers was confirmed by in silico testing using available Leuconostocmesenteroides genomic sequences, and experimentally using DNA samples of Leuconostoc mesenteroides clear cultures. The taxonomic affiliation of 5 isolates of leuconostocci isolated from natural samples was established using the developed primers. Methodological Instructions have been developed that regulate the procedure for determining the taxonomic position of bacteria of genus Leuconostoc to a subspecies. Methodological guidelines for identification of leuconostocs will be used in collections of industrial microorganisms for the accurate identification of deposited strains.
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Relevance Fermentation of vegetables is usually carried out in the traditional way (spontaneous fermentation using native microflora), but the quality of the finished product is difficult to predict. Very often, due to the low initial amount of lactic acid bacteria or their low activity, the result of the process remains unpredictable, which can lead to the loss of a significant amount of product. In the fermentation of vegetables involved several types of facultatively anaerobic lactic acid bacteria. In order to control the fermentation process and make it directed, it is necessary to study which lactic acid bacteria are involved in the fermentation process, the period in which their growth and death, and how it affects the organoleptic properties of the finished product, as well as to study the activity of lactic acid microorganisms in the fermentation process. When fermentation of vegetables are not only the original nutrients such as vitamin C, amino acids, dietary fibers, etc., but also develop functional microorganisms such as lactic acid bacteria. Fermentation has an important effect on the quality and taste, so it is very important to study the fermentation process, microbial diversity and changes in nutrients and chemical elements in the fermentation process. Reducing the rate or preventing microbial spoilage of food is based on four main principles: minimization of product contamination by microorganisms; suppression of growth and reproduction of microorganisms-contaminants; destruction of microorganisms-contaminants; removal of microorganisms-contaminants. Fermentation is based on a combination of the first three principles and is achieved by creating conditions for the growth of specific microorganisms that can give food the desired taste, aroma, texture and appearance. Results This review is devoted to the scientific aspects of vegetable fermentation, including crops that contribute to the creation of optimal conditions for the development of the main pool of lactic acid microorganisms, the production of finished products of high quality and the prevention of microbial spoilage. It is shown that at the first stage of fermentation lactobacilli of the genus L. mesenteroides play a determining role. It is their "work" to create optimal conditions for the development of the target lactic microflora depends on the quality of the finished product. This fact should be taken into account when creating industrial bacterial starter cultures – "starter cultures" for the directed process of fermentation of vegetables.
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Nearly 500 strains isolated from different media used to study the aerobic mesophilic and lactic acid flora of Valdeón cheese (a Spanish hand-made blue cheese) have been identified. Nearly 95% of aerobic mesophiles were lactic acid bacteria (LAB). From these, Enterococcus (40·4%) and Lactococcus (42·2%) were the dominant genera, with Lactobacillus (4·1%) and Leuconostoc (5·0%) being also found. The selectivity of the other media used was variable and this is discussed. Several species of Enterococcus were isolated from our samples (Ent. avium, Ent. faecium and Ent. durans), although one was outstanding (Ent. faecalis, 24·7% of the total of strains identified). The dominating LAB species found was Lact. lactis subsp. lactis (31·1%). Other LAB identified were Lact. raffinolactis, Lb. plantarum, Lb. casei, Leuc. mesenteroides subsp. dextranicum, Leuc. mesenteroides subsp. mesenteroides and Leuc. paramesenteroides. The evolution of the lactic-acid flora found during the manufacture and ripening of this variety of cheese showed a pattern marked by the dominance of lactococci and enterococci during the first stages and the substitution of lactococci by lactobacilli and leuconostoc from drying onwards which would be, along with enterococci, the major genera found in the cheese at the stage of consumption. From a technological point of view, the quantitative importance of certain species such as Lact. lactis and Ent. faecalis, as well as the presence of others, i.e. Lb. plantarum and Leuc. mesenteroides, suggest their possible use as starters in the industrial manufacture of this variety.
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Green bean juice was fermented with 10 species (14 strains) of heterofermentative and two homofermentative lactic acid bacteria to select organisms which might be used to carry out a complete fermentation. Lactobacillus cellobiosus was the only organism to remove all fermentable sugars from bean juice with or without 2.5% NaCI. Nine other cultures used from 75–95% of the sugars. Lactobacillus cellobiosus also produced the lowest final pH among the 14 strains. A complete analysis of the major fermentation substrates and products was done for each of the organisms. Fermentation balance calculations showed a range from 74–132% carbon recovery. These bacteria showed considerable variation in the ability to degrade malic acid and to form mannitol and acetic acid.
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Leuconostoc mesenteroides B-1299 dextrans are separated into two kinds: fraction L, which is precipitated by an ethanol concentration of 38%, and fraction S, which is precipitated at an ethanol concentration of 40%. Fraction S dextran contained 35% of α-1,2 branch linkages, and fraction L contained 27% α-1,2 branch linkage with 1% α-1,3 branch linkages. We have isolated mutants constitutive for dextransucrase from L. mesenteroides NRRL B-1299 using ethyl methane sulfonate. The mutants produced extracellular as well as cell-associated dextransucrases on glucose media with higher activities (2.5–4.5 times) than what the parental strain produced on sucrose. Based on Penicillium endo-dextranase hydrolysis, mutant B-1299C dextransucrases produced slightly different dextrans when they were elaborated on a glucose medium and on a sucrose medium. Mutant B-1299CA dextransucrase elaborated on a glucose medium and on a sucrose medium synthesized the same dextran, although the dextran was different from those of other mutants and the parental strain. Mutant B-1299CB dextransucrase, elaborated on a glucose medium and on a sucrose medium, formed different dextrans. Differences in water solubility, susceptibility to endo-dextranase hydrolysis, and the physical appearance of the ethanol precipitated dextrans elaborated by different mutants grown on glucose media and sucrose media were found. All mutant dextransucrases elaborated on a glucose medium bound to Sephadex G-200. After activity staining of nondenaturing sodium dodecyl sulfate—polyacrylamide gel electrophoresis activity bands, 184 and 240 Kd for each enzyme preparation, although each dextransucrase formed different dextran(s).
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The development of the dominant bacterial populations during traditional Mozzarella cheese production was investigated using physiological analyses and molecular techniques for strain typing and taxonomic identification. Analysis of RAPD fingerprints revealed that the dominant bacterial community was composed of 25 different biotypes, and the sequence analysis of 16S rDNA demonstrated that the isolated strains belonged to Leuconostoc mesenteroides subsp. mesenteroides, Leuc. lactis, Streptococcus thermophilus, Strep. bovis, Strep. uberis, Lactococcus lactis subsp. lactis, L. garviae, Carnobacterium divergens, C. piscicola, Aerococcus viridans, Staphylococcus carnosus, Staph. epidermidis, Enterococcus faecalis, Ent. sulphureus and Enterococcus spp. The bacterial populations were characterized for their physiological properties. Two strains, belonging to Strep. thermophilus and L. lactis subsp. lactis, were the most acidifying; theL. lactis subsp. lactis strain was also proteolytic and eight strains were positive to citrate fermentation. Moreover, the molecular techniques allowed the identification of potential pathogens in a non-ripened cheese produced from raw milk.