Diversity analysis and quantification of lactic acid bacteria in traditionally fermented yaks’ milk products from Tibet

Fermented yaks’ milk is a characteristic fermented dairy product with a rich microflora (especially lactic acid bacteria [LAB]), that has been produced by Tibetan people in China throughout history. Systematic analysis of the LAB composition of fermented yaks’ milk from central Tibet is essential. In the present study, 38 samples of fermented yaks’ milk were collected from three regions in central Tibet. From these, a total of 211 isolates of LAB were generated using traditional pure culture methods. To accurately analyze their biodiversity, 16S rRNA gene sequence analysis and denaturing gradient gel electrophoresis (DGGE) were used. The isolates were identified as belonging to six genera and 22 species/subspecies. Among them, Lactobacillus species predominated (117 isolates) accounting for 55.45% of all isolates. Overall, L. delbrueckii subsp. bulgaricus (60 isolates; 28.43% of the total) was the most commonly isolated species of LAB from fermented yaks’ milk from central Tibet. Seven Lactobacillus species and the Bifidobacterium genus were confirmed by quantitative PCR (q-PCR). The q-PCR results showed that the abundance of these groups in fermented yaks’ milk from central Tibet ranged from 1.12 ± 0.36 log cfu/ml (for L. sakei from Ningzhong) to 7.77 ± 0.84 log cfu/ml (for L. helvetivus from Ningzhong). Lactobacillus acidophilus, L. delbrueckii subsp. bulgaricus, L. fermentum, L. helveticus varied in concentration depending on the region. The results indicated that traditional fermented yaks’ milk from different regions of central Tibet have complex compositions of LAB species. The diversity of LAB might be related to geographical origin.     

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.     

Identification of Lactobacillus from koumiss by conventional and molecular methods

Lactobacilli are considered to be one of the most important potential probiotics in the dairy industry. Twelve strains of Lactobacillus were isolated from home-made koumiss samples, a traditionally fermented mare milk in China. The isolates were identified based on physiological and biochemical characteristics and analysis of 16S RNA sequences. They were proven to be Lactobacillus helveticus, Lactobacillus fermentum, Lactobacillus casei and Lactobacillus plantarum. The results demonstrated that both methods were essential to identify an isolate accurately.     

Safety assessment and antimicrobial properties of the lactic acid bacteria strains isolated from polish raw fermented meat products

Twenty-one strains of the genus Lactobacillus and the genus Pediococcus, isolated from Polish raw fermented meat products, were examined for the potential probiotic properties: resistance to simulated gastric and intestine conditions, safety assessment, and antimicrobial properties. Strains were resistant to gastric enzymes and low pH (3–6 log CFU/mL decrease) and intestinal enzymes and bile salts (1–3 log CFU/mL decrease). Most strains were resistant to gentamycin, streptomycin, vancomycin, tetracycline, ciprofloxacin, and kanamycin. Three of them (Lb. brevis BAL1, BAL10, and KL5) produced β-glucuronidase, which excludes them from qualifying as safe. Seven strains had the ability to produce bacteriocins or bacteriocin-like substances. Overall, strains Lb. brevis SCH6, Pd. pentosaceus BAL6, and KL14 revealed selected superior characteristics (resistance to the gastrointestinal conditions, safety assessment, and antimicrobial properties) as compared to the other LAB strains investigated, which made them a viable bioprotective culture that can be inoculated in raw fermented meat products as starter cultures.     

Invited review: Lactobacillus helveticus—A thermophilic dairy starter related to gut bacteria

The strain Lactobacillus helveticus DPC4571 has emerged as a promising flavor adjunct culture for Cheddar cheese given that it is consistently associated with improved flavor. The availability of the complete genome sequence of Lb. helveticus DPC4571 has enabled the search for the presence or absence of specific genes on the genome, in particular those of technological interest. Indeed, this analysis has facilitated a greater understanding into the functioning of lactic acid bacteria as a whole. The biochemical pathways of Lb. helveticus responsible for producing flavor compounds during cheese ripening are poorly understood but now with the availability of a complete genomic sequence are ripe for exploitation. Bioinformatic analysis of the genome of Lb. helveticus DPC4571 has revealed a plethora of genes with industrial potential including those responsible for key metabolic functions that contribute to cheese flavor development such as proteolysis, lipolysis, and cell lysis. In addition, it has been demonstrated that Lb. helveticus has the potential to produce bioactive peptides such as angiotensin converting enzyme inhibitory activity in fermented dairy products, demonstrating the therapeutic value of this species. A most intriguing feature of the genome of Lb. helveticus DPC4571 is the remarkable similarity in gene content with many intestinal lactobacilli, although originating from considerably different environments. Bioinformatic analysis demonstrated that 65 to 75% of genes were conserved between the commensal and dairy lactobacilli, which allowed key niche-specific gene sets to be described. This review focuses on the isolation, characterization, and exploitation of the Lb. helveticus species with particular emphesis on taking into consideration recent genome sequence data for Lb. helveticus and other Lactobacillus species.     

Diversity analysis of lactic acid bacteria in Korean rice wines by culture-independent method using PCR-denaturing gradient gel electrophoresis

A polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method was used to determine the presence and diversity of lactic acid bacteria (LAB) in takju, a traditional Korean rice wine. Bacterial DNAs were extracted from 15 commercial rice wines and amplicons of partial 16S rRNA genes were separated by DGGE and intense bands were sequenced. Lactobacillus (Lb.) paracasei, Lb. plantarum, and Leuconostoc pseudomesenteroides were detected in all samples and Lb. harbinensis and Lb. parabuchneri were found with above 80% frequency of occurrence. Unknown species of Lactobacillus were also widely detected. This result revealed that, regardless of products and raw materials, the distribution profiles of LAB in takju products have a common pattern comprising of above predominant species and, furthermore, takju can be regarded as a LAB-rich fermented food providing various probiotics.     

益生乳酸菌Lactobacillus casei Zhang从基础研究到产业化开发

2001年-2009年,内蒙古农业大学乳品生物技术与工程教育部重点实验室从采集自中国西部7个省市及蒙古国14个省市不同少数民族地区951份自然发酵乳制品等样品中分离鉴定出3 388株乳酸菌,建立了中国首个原创性乳酸菌菌种资源库;从菌种库中分离自传统发酵酸马奶(koumiss)的243株乳杆菌中筛选获得1株性能优良的乳酸菌—Lactobacillus casei Zhang(L.casei Zhang)。采用5种动物模型和人体试验进行了功能评价,并利用基因组学和蛋白质组学研究技术对L.casei Zhang益生分子机制进行了深入剖析。经过多年系统的基础研究,自主解决了其产业化的关键技术问题,包括其直投式发酵剂、发酵乳制品和发酵豆乳益生菌饮料的研发并实现了产业化。     

High γ-aminobutyric acid production from lactic acid bacteria: Emphasis on Lactobacillus brevis as a functional dairy starter

γ-Aminobutyric acid (GABA) and GABA-rich foods have shown anti-hypertensive and anti-depressant activities as the major functions in humans and animals. Hence, high GABA-producing lactic acid bacteria (LAB) could be used as functional starters for manufacturing novel fermented dairy foods. Glutamic acid decarboxylases (GADs) from LAB are highly conserved at the species level based on the phylogenetic tree of GADs from LAB. Moreover, two functionally distinct GADs and one intact gad operon were observed in all the completely sequenced Lactobacillus brevis strains suggesting its common capability to synthesize GABA. Difficulties and strategies for the manufacture of GABA-rich fermented dairy foods have been discussed and proposed, respectively. In addition, a genetic survey on the sequenced LAB strains demonstrated the absence of cell envelope proteinases in the majority of LAB including Lb. brevis, which diminishes their cell viabilities in milk environments due to their non-proteolytic nature. Thus, several strategies have been proposed to overcome the non-proteolytic nature of Lb. brevis in order to produce GABA-rich dairy foods.     

Diversity and screening of lactic acid bacteria in la-baicai made by Korean-Chinese in northeastern China

This study investigated the lactic acid bacteria (LAB) population in la-baicai (spicy cabbage), a traditional fermented food made by the Korean-Chinese community in northeastern China and screened for functional LAB. LAB diversity was analyzed using denaturing gradient gel electrophoresis (DGGE), and 81 LAB strains were isolated and identified based on 16S rDNA sequencing analysis. Polymerase chain reaction DGGE detected 21 LAB species, belonging to the genera Lactobacillus (Lb.), Leuconostoc (Leu.), Pediococcus and Weissella, in 45 la-baicai samples. Lb. plantarum and Lb. sakei were considered to be dominant in the bacterial community. Among 81 LAB isolated by traditional pure culture methods were Lb. plantarum (25 strains), Lb. brevis (two strains), Lb. casei, (four strains), Lb. pentosus (three strains), Enterococcus faecium (45 strains), and E. durans (two strains). The tolerances of these LAB were investigated. Six LAB with high salt (NaCl)-tolerance were screened from the isolates, and 16% (w/v) was the highest salt-tolerance reached. Among the isolates, strain N1, identified as Lb. pentosus, survived well in a simulated digestive environment. Traditional fermented la-baicai from northeastern China is a rich LAB resource. Further study is needed and would be worthwhile to advance the benefits of these LAB.     

Probiotic monocultures in fermented goat milk beverages – sensory quality of final product

The aim of our study was to conduct a selection of the monocultures capable of providing the most attractive sensory features of the final product. Four fermented goat's milk beverages were produced with probiotic monocultures containing Lactobacillus (Lb. acidophilus La‐5, Lb. rhamnosus K3 and Lb. plantarum O20) and Bifidobacterium (Bif. animalis subsp. lactisBB‐12). A sensory analysis and microbiological assessment of fermented goat's milk beverages were made at the beginning of the study and after 3, 7, 10 and 14 days of refrigerated storage (5 ± 1 °C). We found that samples including monocultures Lb. plantarum O20 and Bif. animalis subsp. lactisBB‐12 were differentiated from other goat's milk beverages.     

Optimization of sour milk fermentation for the production of ACE-inhibitory peptides and purification of a novel peptide from whey protein hydrolysate

The effect of fermentation conditions on the production of angiotensin-I converting enzyme (ACE) inhibitory peptide in sour milk fermented by Lactobacillus helveticus LB10 was investigated using response-surface methodology. Optimal conditions to produce the maximum production of ACE-inhibitory peptides were found to be 4% (v/w) inoculum, 7.5 initial pH of medium and 39.0 °C. The fermented milk resulted in 75.46% inhibition in ACE activity. The cell-envelope proteinase, assisted by X-prolyldipeptidyl aminopeptidase of Lb. helveticus LB10 produced the ACE-inhibitory peptides. A novel ACE-inhibitory peptide from whey protein hydrolysate produced by crude proteinases of Lb. helveticus LB10 was purified. The separations were performed with Sephadex^® G-75 and Sephadex G-15 gel filtration chromatography and reversed-phase, high-performance liquid chromatography. The peptide with the RLSFNP sequence was isolated from β-lactoglobulin hydrolysate and its IC₅₀ while inhibiting ACE activity was 177.39 μm.     

Employment of autochthonous microflora in Pecorino Sardo cheese manufacturing and evolution of physicochemical parameters during ripening

The isolation and identification of lactic acid bacteria (LAB) from raw ewes’ milk and traditional Pecorino Sardo cheese made from this milk without the addition of starter culture was carried out to define the autochthonous lactic microflora present in milk and the evolution of LAB during cheese ripening. Isolation of 275 strains belonging to different Lactococcus, Lactobacillus, Streptococcus and Enterococcus species was achieved. Coccal-shaped LAB were found to predominate during cheese fermentation, while lactobacilli were preponderate during the latter phase of ripening. The technological selection of a total of 174 LAB strains belonging to the species Lactococcus lactis, Streptococcus thermophilus, Lactobacillus helveticus and Lb. casei allowed an experimental starter to be prepared, in which a potentially probiotic species, Lb. casei was used. The suitability of the autochthonous starter culture was tested in cheese-making trials, using thermised ewes’ milk, by comparing experimental Pecorino Sardo cheese with a control cheese produced at industrial scale using a whey starter culture from previous batches of manufacture. In particular, microbiological and physicochemical parameters were determined over 210 days of cheese ripening. Although sensory evaluation did not show any significant difference between experimental and control Pecorino Sardo cheeses, the use of the selected autochthonous starter allowed the production of experimental cheese with a significantly higher level of free amino acids, in particular essential amino acids, in comparison with the Pecorino Sardo control cheeses.     

Genetic diversity of Lactobacillus delbrueckii isolated from raw milk in Hokkaido,Japan

Lactic acid bacteria (LAB) play important roles in acid production and flavor formation in fermented dairy products. Lactic acid bacteria strains with distinct characteristics confer unique features to products. Diverse LAB have been identified in raw milk and traditional fermented milk prepared from raw milk. However, little is known about LAB in raw milk in Japan. To preserve diverse LAB as potential starters or probiotics for future use, we have isolated and identified various kinds of LAB from raw milk produced in Japan. In this study, we focused on Lactobacillus delbrueckii, one of the most important species in the dairy industry. We identified L. delbrueckii subspecies isolated from raw milk in Hokkaido, Japan, by analyzing intraspecific diversity using 4 distinct methods, hsp60 cluster analysis, multilocus sequence analysis, core-genome analysis, and whole-genome analysis based on average nucleotide identity. The subspecies distribution and a new dominant subset of L. delbrueckii from raw milk in Japan were revealed. The discovery of new strains with different genotypes is important for understanding the geographic distribution and characteristics of the bacteria and further their use as a microbial resource with the potential to express unconventional flavors and functionalities. The strains identified in this study may have practical applications in the development of fermented dairy products.     

Probiotic potential of lactic acid bacteria from fermented Malaysian food or milk products

This study investigated the probiotic characteristics of lactic acid bacteria (LAB) isolated from 14 Malaysian fermented food or milk products. In total, 22.3% (121 of 542) of the LAB isolated from the local fermented products exhibited antibacterial activity against Micrococcus luteus. Twelve LAB that demonstrated better antibacterial activity against M. luteus as compared to a commercial strain Lactobacillus casei strain Shirota (LABPC) were selected for further characterisation. Based on 16S rRNA gene sequence, the LAB were identified as pediococci (seven) and lactobacilli (five). All 12 LAB showed bile tolerance, but only eight were acid tolerant at pH ≥ 3.0. The highest level of adhesion to HT‐29 cells was observed among the Lactobacillus sp. LAB 1 and 10. The LAB also showed antimicrobial activity against Escherichia coli and Staphylococcus aureus through the production of organic acids. LAB isolated from Malaysian fermented food and milk products, especially fermented tapioca, contains potential probiotic candidates.     

Investigating the bacterial microbiota of traditional fermented dairy products using propidium monoazide with single-molecule real-time sequencing

Traditional fermented dairy foods have been the major components of the Mongolian diet for millennia. In this study, we used propidium monoazide (PMA; binds to DNA of nonviable cells so that only viable cells are enumerated) and single-molecule real-time sequencing (SMRT) technology to investigate the total and viable bacterial compositions of 19 traditional fermented dairy foods, including koumiss from Inner Mongolia (KIM), koumiss from Mongolia (KM), and fermented cow milk from Mongolia (CM); sample groups treated with PMA were designated PKIM, PKM, and PCM. Full-length 16S rRNA sequencing identified 195 bacterial species in 121 genera and 13 phyla in PMA-treated and untreated samples. The PMA-treated and untreated samples differed significantly in their bacterial community composition and α-diversity values. The predominant species in KM, KIM, and CM were Lactobacillus helveticus, Streptococcus parauberis, and Lactobacillus delbrueckii, whereas the predominant species in PKM, PKIM, and PCM were Enterobacter xiangfangensis, Lactobacillus helveticus, and E. xiangfangensis, respectively. Weighted and unweighted principal coordinate analyses showed a clear clustering pattern with good separation and only minor overlapping. In addition, a pure culture method was performed to obtain lactic acid bacteria resources in dairy samples according to the results of SMRT sequencing. A total of 102 LAB strains were identified and Lb. helveticus (68.63%) was the most abundant, in agreement with SMRT sequencing results. Our results revealed that the bacterial communities of traditional dairy foods are complex and vary by type of fermented dairy product. The PMA treatment induced significant changes in bacterial community structure.     

基于16S rDNA高通量测序方法比较新疆西北部地区乳品中微生物的多样性

为更加全面地了解新疆特色乳品中微生物多样性,比较不同动物来源的原奶和酸奶的细菌群落结构,运用高通量测序技术,对乳品中细菌16S r DNA V4-V5区测序,进而对新疆克州和塔城地区牛奶、驼奶、马奶、羊奶、酸牛奶、酸驼奶和酸马奶7种乳品中细菌群落组成和多样性进行分析。研究共获得539 557条有效序列,379个OTU。多样性分析表明,原奶样品中细菌Shannon-Wiener指数明显高于酸奶样品。微生物群落组成分析发现,不同乳品之间菌群组成差异较大。7种乳品中的菌群均以厚壁菌门和变形菌门为主,但原奶样品主要以变形菌门为主,而酸奶样品主要以厚壁菌门为主。在属水平上,牛奶主要以假单胞菌属(Pseudomonas)为主,驼奶主要以埃希菌属-志贺菌属(Escherichia-Shigella),马奶主要以明串珠菌属(Leuconostoc)为主,羊奶中的优势菌属为乳球菌属(Lactococcus),而酸牛奶、酸驼奶和酸马奶都是以乳杆菌属(Lactobacillus)为优势菌属。不同动物来源的原奶和酸奶样品中的微生物多样性存在显著差异,并且原奶中检测到的环境污染菌和致病菌(或条件致病菌)的丰度也相对较高。本研究结果将为准确评估乳品中的微生物群落对新疆地区少数民族健康的影响提供一定的数据基础。     

Effect of adzuki bean sprout fermented milk enriched in γ-aminobutyric acid on mild depression in a mouse model

This study focused on the ability of adzuki bean (Vigna angularis) sprout fermented milk, which is rich in γ-aminobutyric acid (GABA), to relieve anxiety and mild depression. A high-yield GABA-producing strain, Lactobacillus brevis J1, from a healthy cow was screened, and its physiological and probiotic properties were evaluated. The effect of adzuki bean sprout fermented milk was investigated in vivo in a chronic depression mouse model. The results showed that Lb. brevis J1 had excellent probiotic properties, grew well at low pH and 3% NaCl, and adhered to the surface of HT-29 cells. The GABA-enriched (241.30 ± 1.62 µg/mL) adzuki bean sprout fermented milk prepared with Streptococcus thermophilus, Lactobacillus bulgaricus, and Lactobacillus plantarum, and Lb. brevis J1 can reduce and possibly prevent mild depression-like symptoms in mice (C57/B6) by increasing social interaction and enhancing the pleasure derived from movement. The research revealed that the GABA_B-cyclic AMP-protein kinase A-cAMP-response element binding protein (GABA_B-cAMP-PKA-CREB) signaling pathway was related to the depression-like symptoms and that levels of 5-hydroxytryptamine, norepinephrine, and dopamine in the hippocampus of mice increased after treatment with the adzuki bean sprout fermented milk. Our results suggest that GABA-enriched dairy products have the potential to prevent or treat mild depression-like symptoms in mice, which suggests a new approach for a dietary therapy to treat chronic social stress.     

Isolation and identification of high viscosity-producing lactic acid bacteria from a traditional fermented milk in Xinjiang and its role in fermentation process

Forty-four lactic acid bacteria (LAB) isolated from traditional Sayram ropy fermented milk (SRFM) in southern Xinjiang of China. Further two strains were selected based on their high viscosity-producing activity. Based on the API 50 CHL strip and 16S rDNA sequence analysis, strain MB 2-1 was a Gram-positive, rod-shaped Lactobacillus helveticus and strain MB 5-1 was identified as Streptococcus thermophilus. Both the two LAB strains were grown in the milk fermentations for pure and mixed cultures and were evaluated for their growth, acidification properties, EPS production, and ability to increase the apparent viscosity of fermented milk, respectively. L. helveticus MB 2-1 displayed a high increasing in viable cell counts and the acidifying capacity in pure cultures, whereas the relatively high EPS production and viscosity-producing capacity detected in pure culture with S. thermophilus MB 5-1 as starter culture. In addition, the mixed culture of the two strains showed a higher cell growth, EPS production, and high viscosity-producing capacity at 37?°C. Values of apparent viscosity were 4.03- and 2.41-fold higher in mixed culture than for pure cultures of L. helveticus MB 2-1 and S. thermophilus MB 5-1, respectively. There was a positive correlation between the viscosity and high molecular weight EPS production with pure and mixed cultures. Our data showed two strains combination, with high viscosity-producing and acidifying capacity, can be used in mixed cultures for the manufacture of fermented milk with improved functional properties.     

一种新型开菲尔风味复合发酵剂的研制

为研制一款新型的风味突出、理化性质稳定的开菲尔风味复合发酵剂,该研究从源自中国新疆的10种开菲粒中分离获得51株乳酸菌、22株酵母菌。51株乳酸菌中,3株乳酸菌的产香能力和蛋白分解能力突出,单菌发酵乳的全质构特性最稳定。其中,开菲尔乳杆菌MLK5的乙醛质量浓度为15.82 mg/L、双乙酰质量浓度为3.99 mg/L、氨基氮质量浓度为597.09 mg/L,干酪乳杆菌SLC1的乙醛质量浓度为20.02 mg/L、双乙酰质量浓度为4.69 mg/L、氨基氮质量浓度为684.92 mg/L,肠膜明串珠菌NLM2的乙醛质量浓度为18.01 mg/L、双乙酰质量浓度为4.44 mg/L、氨基氮质量浓度为600.58 mg/L。22株酵母菌中,马克思克鲁维酵母菌菌株FY1的乙醇产量适宜,遗传稳定性好,乳糖利用率最高为56.56%。最终确定,发酵剂中最佳比例为乳酸菌:酵母菌=5:1,3株乳酸菌间最优例为肠膜明串珠菌:开菲尔乳杆菌:干酪乳杆菌为1:2:1。复合发酵乳的凝乳时间为6.0 h、酸度为84.65 ºT、持水力为62.31%、乙醇体积分数为0.53%、乳酸菌活菌数为3.89×109 CFU/mL、酵母菌活菌数为4.61×106 CFU/mL、感官评分为89.21,与开菲尔粒发酵乳的各项指标类似。而复合发酵乳中的挥发性风味物质间构成和谐,更易于消费者接受。上述研究结果,有助于稳定和简化开菲尔的工业生产,为开发不同风味酸乳产品提供参考。     

Analysis of volatile compounds produced by different species of lactobacilli in rye sourdough using multiple headspace extraction

Profiles of volatile organic compound (VOC) produced by nine individual lactic acid bacteria (LAB) during rye sourdough fermentation were compared by automated SPME and GC/MS‐Tof. The dough samples were inoculated with individual strains, placed inside the headspace vials and incubated during next 24 h. The production or loss of VOC‐s was followed by adsorbing volatiles onto 85‐m Car/PDMS fibre in every 4 h. Volatile profiles differed among LAB species and divided LAB into two main groups – hetero‐ and homofermentative. Hetrofermentative LAB (Lactobacillus brevis; Leuconostoc citreum; Lactobacillus vaginalis, Lactobacillus panis) showed high production of acetic acid, CO₂, ethanol, ethylacetate, producing also hexyl acetate, ethyl hexanoate and isopentyl acetate. Whereas homofermentative LAB species (Lactobacillus helveticus; Lactobacillus casei; Lactobacillus sakei; Lactobacillus curvatus) produced a considerable amount of 2,3‐butanedione. Production of l ‐leucine methyl ester was unique for Lb. sakei, Lb. casei and Lb. curvatus strains. Lb. helveticus was the only LAB that produced benzaldehyde.