Isolation and characterisation of lactic acid bacteria from yan-jiang (fermented ginger), a traditional fermented food in Taiwan

BACKGROUND: Yan‐jiang (fermented ginger) is a popular traditional fermented food in Taiwan. The lactic acid bacteria (LAB) microflora in yan‐jiang has not been studied in detail. In this study, LAB from yan‐jiang were isolated, characterised and identified. RESULTS: A total of 176 LAB were isolated; 160 cultures were isolated from yan‐jiang samples and 16 cultures were isolated from raw ginger. These isolates were characterised phenotypically and then divided into nine groups (A to I) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Lactobacillus sakei and Lactococcus lactis subsp. lactis were the major LAB found in the initial 2 days of fermentation without pickled plums; these species were mostly replaced by Weissella cibaria and L. plantarum after 3 days of fermentation. In the fermentation bucket with added pickled plums, W. cibaria was the most abundant LAB found during fermentation. The antibacterial activities of the isolates were determined. Twenty‐four Lc. lactis subsp. lactis and 19 W. cibaria strains showed inhibitory activity against the indicator strain L. sakei JCM 1157^T. CONCLUSION: Results demonstrate that various LAB species were more numerous when fermentation was carried out without pickled plums. LAB also had effects on the aroma and flavour of yan‐jiang. Copyright © 2011 Society of Chemical Industry     

Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia

Spontaneous milk fermentation has a long history in Mongolia, and beneficial microorganisms have been handed down from one generation to the next for use in fermented dairy products. The objective of this study was to investigate the diversity of lactic acid bacteria (LAB) communities in fermented yak, mare, goat, and cow milk products by analyzing 189 samples collected from 13 different regions in Mongolia. The LAB counts in these samples varied from 3.41 to 9.03 log cfu/mL. Fermented yak and mare milks had almost identical mean numbers of LAB, which were significantly higher than those in fermented goat milk but slightly lower than those in fermented cow milk. In total, 668 isolates were obtained from these samples using de Man, Rogosa, and Sharpe agar and M17 agar. Each isolate was considered to be presumptive LAB based on gram-positive and catalase-negative properties, and was identified at the species level by 16S rRNA gene sequencing, multiplex PCR assay, and restriction fragment length polymorphism analysis. All isolates from Mongolian dairy products were accurately identified as Enterococcus faecalis (1 strain), Enterococcus durans (3 strains), Lactobacillus brevis (3 strains), Lactobacillus buchneri (2 strains), Lactobacillus casei (16 strains), Lactobacillus delbrueckii ssp. bulgaricus (142 strains), Lactobacillus diolivorans (17 strains), Lactobacillus fermentum (42 strains), Lactobacillus helveticus (183 strains), Lactobacillus kefiri (6 strains), Lactobacillus plantarum ssp. plantarum (7 strains), Lactococcus lactis ssp. lactis (7 strains), Leuconostoc lactis (22 strains), Leuconostoc mesenteroides (21 strains), Streptococcus thermophilus (195 strains), and Weissella cibaria (1 strain). The predominant LAB were Strep. thermophilus and Lb. helveticus, which were isolated from all sampling sites. The results demonstrate that traditional fermented dairy products from different regions of Mongolia have complex compositions of LAB species. Such diversity of LAB provides useful information for further studies of probiotic strain selection and starter culture design, with regard to the industrial production of traditional fermented milk.     

Diversity of lactic acid bacteria in sian-sianzih (fermented clams), a traditional fermented food in Taiwan

BACKGROUND: Sian‐sianzih (fermented clams) is a popular traditional fermented food in Taiwan. The lactic acid bacteria (LAB) microflora in sian‐sianzih have not been studied in detail. In this study, LAB from sian‐sianzih were isolated, characterized and identified. RESULTS: A total of 186 cultures of LAB were isolated from seven sian‐sianzih samples and 29 cultures were isolated from its main raw substrate: clams. The identification results revealed up to 11 distinct bacterial species belonging to five genera in sian‐sianzih, and three species belonging to two genera in clams. The most common bacterial genera in sian‐sianzih were Lactobacillus and Weissella, followed by Leuconostoc, Pediococcus and Lactococcus. A regional similarity in LAB, with differences in diversity, was observed in the current study. On the other hand, Lactococcus lactis subsp. lactis was the most common species found in raw clam samples. The results also suggested that greater LAB diversity could be observed in wild clams than in cultured ones. Furthermore, antibacterial activities of the isolates were determined, and one Weisella hellenica strain showed inhibitory activity against the indicator strain Lactobacilluas sakei JCM 1157^T. A sensory assessment of seven sian‐sianzih samples was also performed and the results indicated that diversity of LAB has a great effect on its aroma and taste formation. CONCLUSION: The results demonstrate that various LAB species are distributed in sian‐sianzih and have a great effect on the flavor of sian‐sianzih. Copyright © 2011 Society of Chemical Industry     

A culture-dependent and -independent approach for the identification of lactic acid bacteria associated with the production of nem chua,a Vietnamese fermented meat product

In this study, the diversity of the native lactic acid bacteria (LAB) population in nem chua, a popular traditional Vietnamese uncooked fermented meat, was described using a combination of culture-dependent and culture-independent methods. A total of two hundred seventy-three LAB isolates were subjected to a polyphasic identification approach combining (GTG)₅-PCR fingerprinting and phenylalanyl-tRNA synthase α subunit (pheS) and RNA polymerase α subunit (rpoA) gene sequence analysis. LAB associated with nem chua were identified as Lactobacillus pentosus (21%), Lactobacillus plantarum (29.7%), Lactobacillus brevis (5%), Lactobacillus paracasei (0.4%), Lactobacillus fermentum (0.7%), Lactobacillus acidipiscis (0.4%), Lactobacillus farciminis (23%), Lactobacillus rossiae (0.4%), Lactobacillus fuchuensis (0.7%), Lactobacillus namurensis (0.4%), Lactococcus lactis (0.4%), Leuconostoc citreum (9.5%), Leuconostoc fallax (1%), Pediococcus acidilactici (1%), Pediococcus pentosaceus (4%), Pediococcus stilesii (1%), Weissella cibaria (0.7%) and Weissella paramesenteroides (0.7%). Furthermore, PCR-DGGE was also applied as a culture-independent method in this study. Results indicated the presence of species of which no isolates were recovered, i.e. Lactobacillus helveticus/crispatus, Lactococcus garvieae and Vagococcus sp. Conversely, not all isolated bacteria were detected by PCR-DGGE. Principal component and discriminant analysis disclosed correlations between the different production locations and certain isolated LAB species and strains and/or DGGE bands suggesting possible influences of locally prevailing production practices on the nem chua LAB microbiota.     

Diversity of lactic acid bacteria in suan-tsai and fu-tsai, traditional fermented mustard products of Taiwan

Fu-tsai and suan-tsai are spontaneously fermented mustard products traditionally prepared by the Hakka tribe of Taiwan. We chose 5 different processing stages of these products for analysis of the microbial community of lactic acid bacteria (LAB) by 16S rRNA gene sequencing. From 500 LAB isolates we identified 119 representative strains belonging to 5 genera and 18 species, including Enterococcus (1 species), Lactobacillus (11 species), Leuconostoc (3 species), Pediococcus (1 species), and Weissella (2 species). The LAB composition of mustard fermented for 3 days, known as the Mu sample, was the most diverse, with 11 different LAB species being isolated. We used sequence analysis of the 16S rRNA gene to identify the LAB strains and analysis of the dnaA, pheS, and rpoA genes to identify 13 LAB strains for which identification by 16S rRNA gene sequences was not possible. These 13 strains were found to belong to 5 validated known species: Lactobacillus farciminis, Leuconostoc mesenteroides, Leuconostoc pseudomesenteroides, Weissella cibaria, and Weissella paramesenteroides, and 5 possibly novel Lactobacillus species. These results revealed that there is a high level of diversity in LAB at the different stages of fermentation in the production of suan-tsai and fu-tsai.     

Isolation and characterization of lactic acid bacteria from jiang‐sun (fermented bamboo shoots), a traditional fermented food in Taiwan

BACKGROUND: In this study, lactic acid bacteria (LAB) were isolated, characterized and identified from jiang‐sun (fermented bamboo shoots; a traditional fermented food in Taiwan). Samples were collected at seven time intervals from a fixed fermenting bucket during the fermentation process of jiang‐sun and its initial ingredients, dochi (fermented soybeans) and bamboo shoots. RESULTS: A total of 57 LAB cultures were isolated; 42 cultures were isolated from jiang‐sun samples and 15 cultures were isolated from dochi and bamboo shoots. These isolates were characterized phenotypically and then divided into three groups (A–C) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Alteration of microbial populations during the fermentation process was observed. While various LAB were found in the dochi and bamboo shoots, it was mostly replaced by Lactobacillus plantarum after 1 day of fermentation. Furthermore, the antibacterial activities of the isolates were determined, and one Enterococcus faecium strain showed inhibitory activity against all the indicator strains. CONCLUSION: Results suggest that L. plantarum is the main LAB present during the fermentation of jiang‐sun. To our knowledge, this is the first report describing the distribution and varieties of LAB that exist in the jiang‐sun fermentation process. Copyright © 2010 Society of Chemical Industry     

The microbiology of South African traditional fermented milks

A total of 15 samples of traditional fermented milk were collected from individual households in South Africa and Namibia. Lactic acid bacteria dominated the microflora of these samples, especially the genera Leuconostoc, Lactococcus and Lactobacillus. Other groups identified included pyogenic streptococci and enterococci. The dominant lactococci species was Lactococcus lactis subsp. lactis. Eighty-three percent of the leuconostoc isolates were identified as Leuconostoc mesenteroides subsp. dextranicum. Other species identified included Leuconostoc citreum, Leuconostoc lactis, Lactobacillus delbrueckii subsp. lactis and Lactobacillus plantarum.     

利用16S rDNA序列及tuf-RFLP鉴定蒙古国发酵乳中的乳酸菌

运用16S rDNA序列分析和tuf-RFLP技术对采于蒙古国扎布汗省的25份发酵乳样中分离出的110株乳酸菌进行鉴定。首先将分离的110株乳酸菌的16S rRNA基因进行扩增,测序并构建系统发育树,初步鉴定为41株嗜热链球菌,40株瑞士乳杆菌,11株德氏乳杆菌保加利亚亚种,2株发酵乳杆菌,1株乳明串珠菌,2株肠膜明串肠膜亚种,1株乳酸乳球乳酸亚种和12株属于干酪族的菌株。由于干酪乳杆菌族的16S rDNA序列差异很小,故采用tuf-RFLP技术对这12株进行了进一步的验证,通过分离菌株与模式菌株tuf-RFLP图谱的比较分析,结果表明这12株菌均为干酪乳杆菌。     

Microbial characterisation and stability of a farmhouse natural fermented milk from Spain

This work reports the microbial characterisation of a farmhouse natural fermented milk (NFM) with good sensorial properties produced in Spain. Culturing and denaturing gradient gel electrophoresis (DGGE) analyses showed thatLactococcus lactissubsp.lactis and L. lactissubsp.cremoris(approximate levels of 10⁹ cfu/mL) were dominant in this NFM, whileLactobacillus plantarumappeared at a lower level (10⁶–10⁷ cfu/mL). Repetitive extragenic palindromic (REP)‐PCR typing of the isolates identified single strains each ofLc. lactissubsp.lactis, Lc. lactissubsp.cremorisandLb. plantarum. These three strains formed a stable microbial association which has been maintained for at least some decades.     

Isolation of probiotic bacteria from turmeric (Curcuma longa Linn.) and its application in enriched beverages

The purpose of this study is to obtain lactic acid bacteria (LAB) from fresh turmeric rhizomes that could be used as starter culture with the ability to ferment turmeric and to investigate its influence on total phenolic content (TPC) and antioxidant activity. Three LAB strains, namely Enterococcus faecium, Lactococcus lactis subsp. lactis, and Lactobacillus plantarum, were isolated from turmeric rhizomes. These microorganisms showed similar characteristics as probiotic organisms. The TPC of fermented turmeric beverages was measured using the Folin–Denis reagent method, while the antioxidant activity assays were quantified by 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) and ferric reducing antioxidant power assay. This study confirms that bacteria isolated from turmeric can be used as starter. Moreover, fermentation time and LAB lead to dominant amount of TPC and that they have high levels of antioxidant activities.     

Characterization of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation

Lactic acid bacteria (LAB) isolated from raw materials (fish, rice, garlic and banana leaves) and processed som-fak (a Thai low-salt fermented fish product) were characterized by API 50-CH and other phenotypic criteria. Lactococcus lactis subsp. lactis and Leuconostoc citreum were specifically associated with fish fillet and minced fish, Lactobacillus paracasei subsp. paracasei with boiled rice and Weisella confusa with garlic mix and banana leaves. In addition, Lactobacillus plantarum, Lactobacillus pentosus and Pediococcus pentosaceus were isolated from raw materials. A succession of aciduric, homofermentative lactobacillus species, dominated by Lb. plantarum/pentosus, was found during fermentation. In total, 9% of the strains fermented starch and 19% fermented garlic, the two main carbohydrate components in som-fak. The ability to ferment garlic was paralleled by a capacity to ferment inulin. An increased percentage of garlic fermenting strains was found during fermentation of som-fak, from 8% at day 1 to 40% at day 5. No starch fermenting strains were isolated during fermentation. Three mixed LAB cultures, composed of either starch fermenting Lc. lactis subsp. lactis and Lb. paracasei subsp. paracasei, or garlic fermenting Lb. plantarum and Pd. pentosaceus, or a combination of these strains were inoculated into laboratory prepared som-fak with or without garlic. In som-fak without garlic, pH was above 4.8 after three days, irrespective of addition of mixed LAB cultures. The starch fermenting LAB were unable to ferment som-fak and sensory spoilage occurred after three days. Fermentation with the combined mix of starch and garlic fermenting strains led to production of 2.5% acid and a decrease in pH to 4.5 in two days. The fermentation was slightly slower with the garlic fermenting strains alone. This is the first report describing the role of garlic as carbohydrate source for LAB in fermented fish products.     

Isolation of novel homopolysaccharide-producing lactic acid bacteria from Romanian raw milk and fermented dairy products

Traditionally fermented foods can be a rich source of diverse lactic acid bacteria (LAB) with interesting functional properties, such as exopolysaccharide (EPS) production. The objectives of this study were to map the mucoid and/or ropy LAB isolated from raw milk and traditionally fermented dairy products, collected in different regions of Romania, to study the species diversity within the samples and to further explore the EPS-producing capacity of the isolates. Seventy-three LAB strains were isolated and identified through (GTG)₅-PCR genomic fingerprinting and SDS-PAGE of whole-cell proteins. Lactococcus lactis was the most frequently encountered species, followed by Lactobacillus plantarum, Leuconostoc spp., and Enterococcus spp. Nine strains produced homopolysaccharides (HoPS, glucose monomers), namely L. lactis 1.8; Leuc. citreum 1.10, 1.11, 1.12, 2.8, and 4.11; Leuc. mesenteroides 21.2; Leuc. pseudomesenteroides 20.6; and Weisella confusa/cibaria 38.2, six of them in the concentrations above 10 g/L, both in milk and MRS medium supplemented with sucrose. In all EPS, the glucose constituents were connected by different α-linkages, among which α-1,6-linkages were the most prevalent.     

Phenotypic and genotypic identification of lactic acid bacteria isolated from ethnic fermented bamboo tender shoots of North East India

Mesu, soidon, soibum and soijim are ethnic fermented bamboo tender shoot products prepared by the people in North East India. Microbiological analysis of mesu, soidon, soibum and soijim showed the population dominated by lactic acid bacteria (LAB) ranging up to 10(8) cfu g(-1). The phenotypic characterisation of predominant LAB isolated from the fermented bamboo shoot products was based on general morphology, physiological tests, API and Biolog systems. The genotypic characterisation of LAB was based on RAPD-PCR, rep PCR, species-specific PCR techniques, 16S rRNA gene sequencing and DNA-DNA hybridisation. Predominant functional LAB strains associated with the fermented bamboo shoot products were identified as Lactobacillus brevis, Lb. plantarum, Lb. curvatus, Pediococcus pentosaceus, Leuconostoc mesenteroides subsp. mesenteroides, Leuc. fallax, Leuc. lactis, Leuc. citreum and Enterococcus durans.     

Monitoring the lactic acid bacterial diversity during shochu fermentation by PCR-denaturing gradient gel electrophoresis

The presence of lactic acid bacteria (LAB) during shochu fermentation was monitored by PCR-denaturing gradient gel electrophoresis (DGGE) and by bacteriological culturing. No LAB were detected from fermented mashes by PCR-DGGE using a universal bacterial PCR primer set. However, PCR-DGGE using a new primer specific for the 16S rDNA of Lactococcus, Streptococcus, Tetragenococcus, Enterococcus, and Vagococcus and two primers specific for the 16S rDNA of Lactobacillus, Pediococcus, Leuconostoc, and Weissella revealed that Enterococcus faecium, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus nagelii, Lactobacillus plantarum, Lactococcus lactis, Leuconostoc citreum, Leuconostoc mesenteroides, and Weissella cibaria inhabited in shochu mashes. It was also found that the LAB community composition during shochu fermentation changed after the main ingredient and water were added during the fermentation process. Therefore, we confirmed that PCR-DGGE using all three primers specific for groups of LAB together was well suited to the study of the LAB diversity in shochu mashes. The results of DGGE profiles were similar to the results of bacteriological culturing. In conclusion, LAB are present during shochu fermentation but not dominant.     

Diversity and probiotic potentials of lactic acid bacteria isolated from gilaburu,a traditional Turkish fermented European cranberrybush (Viburnum opulus L.) fruit drink

The aim of the present study was to characterize lactic acid bacteria (LAB) strains isolated from traditional fermented gilaburu fruit juice and their probiotic potential. The LAB counts of the fermented gilaburu fruit juice were in the range of 3.92–8.30 log cfu/g. Total of 332 isolates belonging to Lactobacillus and Leuconostoc species were characterized from traditional fermented gilaburu juice by genotypic methods. It was also determined that the major LAB strains belong to Lactobacillus plantarum (173 isolates), Lactobacillus casei (52 isolates) and Lactobacillus brevis (24 isolates), while Lactobacillus buchneri, Lactobacillus parabuchneri, Lactobacillus pantheris, Leuconostoc pseudomesenteroides and Lactobacillus harbinensis were the least in isolated LAB strains. In terms of the probiotic potentials, Lb. plantarum strains were able to grow at pH 2.5, but 3 of Lb. casei strains, one of each Lb. brevis and Lb. buchneri strains could not grow at the same pH. All selected LAB stains were resistant to bile salt at ≤ 0.3% concentration. While all the LAB species grew at 15 °C, two Lactobacillus hordei strains could also grow at 45 °C. The highest cell hydrophobicity degrees were for Lb. casei (G20a) and Lb. plantarum (G19e) as 87.5 and 86.0%, respectively. Listeria monocytogenes and Bacillus cereus were the most sensitive bacteria against the selected LAB strains, while Escherichia coli and Staphylococcus aureus were the most resistant. Again all the isolated LAB species were resistant to three antibiotics; kanamycin, streptomycin and vancomycin. Characterization and probiotic potentials of the LAB isolated from fermented gilaburu (Viburnum opulus) juice were studied first time, and further research needs to be done on their behaviors in similar food formulations as a probiotic.     

Microbial diversity and dynamics of Pecorino di Filiano PDO,a traditional cheese of Basilicata region (Southern Italy)

The evolution of microbial populations of ‘Pecorino di Filiano’ (PF) cheese was investigated during ripening in natural cave and storeroom. 62.5% of isolates grow at 45 and 15 °C and 77.7% showed high salt concentrations tolerance. Brevibacterium linens was dominant in surface samples. Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus paracasei subsp. paracasei were more frequently isolated both surface and core samples, while Leuconostoc lactis and Leuconostoc mesenteroides subsp. mesenteroides prevailed among Leuconostoc isolates. Our results suggest the importance of the ripening environment of cheeses and how a biological ecosystem affects and produces the typical features of artisanal products.     

Natural populations of lactic acid bacteria isolated from vegetable residues and silage fermentation

Natural populations of lactic acid bacteria (LAB) and silage fermentation of vegetable residues were studied. Fifty-two strains of LAB isolated from cabbage, Chinese cabbage, and lettuce residues were identified and characterized. The LAB strains were gram-positive and catalase-negative bacteria, which were divided into 6 groups (A to F) according to morphological and biochemical characteristics. The strains in group A were rods that did not produce gas from glucose and formed the d and l isomers of lactate. Groups B and C were homofermentative cocci that formed l-lactic acid. Groups D, E, and F were heterofermentative cocci that formed d-lactic acid. Based on 16S rDNA gene sequence analysis, group A to F strains were identified as Lactobacillus plantarum, Lactococcus piscium, Lactococcus lactis, Leuconostoc citreum, Weissella soli and Leuconostoc gelidum, respectively. The prevalent LAB, predominantly homofermentative lactobacilli, consisted of Lactobacillus plantarum (34.6%), Weissella soli (19.2%), Leuconostoc gelidum (15.4%), Leuconostoc citreum (13.5%), Lactococcus lactis (9.6%), and Lactococcus piscium (7.7%). Lactobacillus plantarum was the dominant member of the LAB population in 3 types of vegetable residues. These vegetable residues contained a high level of crude protein (20.2 to 28.4% of dry matter). These silages prepared by using a small-scale fermentation system were well preserved, with low pH and a relatively high content of lactate. This study suggests that the vegetable residues contain abundant LAB species and nutrients, and that they could be well preserved by making silage, which is a potentially good vegetable protein source for livestock diets.     

Comparison of the effect of exopolysaccharide-producing lactic acid bacteria from sourdough on dough characteristics and steamed bread quality

The use of exopolysaccharide (EPS)-producing lactic acid bacteria (LAB) is promising in sourdough fermentation. However, the knowledge of the effects of various species of LAB on steamed bread making remains limited. In this study, the effects of two LAB with high EPS-producing capacity, namely Weissella cibaria L32 and Lactobacillus brevis L17 on dough fermentation and steamed bread quality were estimated. The addition of these two LAB strains significantly increased the titratable acidity and protease activity during the dough fermentation, especially L. brevis L17. Although the in situ EPS synthesised by LAB could improve the steamed bread quality, excessive acidification of L. brevis L17 would still increase the protease activity and thus destroy its gluten network structure. As a result, the steamed bread fermented with L. brevis L17 had the lowest specific volume and hardest texture in comparison with the steamed bread fermented with W. cibaria L32 and with added EPS produced by W. cibaria L32 and L. brevis L17. These results indicated that different EPS-producing LAB exhibited distinctive dough fermentation characteristics, and the in situ EPS-producing W. cibaria L32 could improve steamed bread quality, which confirmed its potential application in steamed bread making.     

Isolation and applied potential of lactic acid bacteria from Chinese traditional fermented food in specific ecological localities

The isolation and screening of lactic acid bacteria (LAB) from natural sources have been one of the powerful means to obtain strains for the food industry. A total of 275 indigenous isolates were obtained from 43 samples of traditional fermented foods in specific ecological niches throughout the northwestern China, and among which 13 strains of LAB were selected for their potential in food preservation and production. Among the 13 isolates, Lactobacillus (10) was dominant over Lactococcus (3). The distribution of the isolates was as follows: Lactobacillus paracasei ssp. paracasei (J23, M10, M20, M22), Lactobacillus rhamnosus (J20, M18), Lactococcus lactis ssp. lactis (X20, Q7), Lactobacillus casei (Q1, Q12), Lactobacillus plantarum (J11), Lactococcus lactis ssp. cremoris (X8), Lactobacillus delbrueckii ssp. bulgaricus (Q5). All 13 isolates produced bacteriocin with a broad inhibitory spectrum against selected Gram-positive as well as Gram-negative pathogenic and spoilage species. Biochemical analysis revealed that they possessed high acidification and coagulation activity. Several strains possessed the high activity of 2 or 3 technological characteristics, related to novel starters and food preservatives.     

Diversity of lactic acid bacteria in fermented brines used to make stinky tofu

Stinky tofu is a kind of fermented tofu with a strong odor. Although stinky tofu is a very popular snack in the Asian region, the community of microbes, and especially lactic acid bacteria (LAB), indigenous to the fermented brine from which it is made remains poorly described. We examined 168 isolates obtained from the original fermented brine (brine A) and two brines in which the hard tofu (brine B) and soft tofu (brine C) had been soaked. Through random amplified polymorphic DNA (RAPD) analysis for typing and 16S rDNA sequencing, 136 representative strains were identified as belonging to 7 genera and 32 species: Enterococcus (2 species), Lactobacillus (14 species), Lactococcus (3 species), Leuconostoc (6 species), Pediococcus (1 species), Streptococcus (2 species), and Weissella (4 species). The LAB composition of brine A was the most diverse: 19 different species were isolated, and 9 of them were classified as Lactobacillus species. The 16S rDNA sequences of 9 strains (6 from brine A and 3 from brine C) showed low values of similarity (below 98%) with currently known species by analysis using the FASTA software. Thus, a wide variety of LAB strains were associated with the fermentation of stinky tofu brines.