Search for Influence Factors in Lactobacilli with Probiotic Properties Isolated from Traditional Kazakh Foods.

The aim of this study was to investigate the factors influencing the probiotic potential of lactobacilli isolated from traditional Kazakh foods, focusing on the identification and characterisation of multifunctional proteins. The basis of the methodological approach in this scientific study is an empirical, experimental study of the factors of influence produced by lactobacilli, which were obtained from traditional Kazakh foods and have pronounced probiotic properties. In this scientific work, results were obtained indicating that the expressed factors of probiotic action can perform adhesive and signalling functions by analogy with homologous proteins of pathogenic and commensal/probiotic bacteria. This largely determines the usefulness of these factors for studying the mechanisms of their probiotic action. In addition, it was found that potential probiotic strains of lactobacilli, which were isolated from food products traditional for Kazakhstan, contain adhesion proteins to the components of the mammalian organism, namely human plasminogen and porcine mucin. It is going about ENO, GAPDH, and p66/DnaK, which, along with P40 and P75 muramidases, are classified as probiotic factors. They are also called multifunctional proteins and are designated as factors of probiotic action. The practical significance of the results obtained during the implementation of this study lies in the possibility of their application in the planning and implementation of activities in the field of ensuring food security in Kazakhstan.

Characterization of a new bacteriocin-like inhibitory peptide produced by Lactobacillus sakei B-RKM 0559.

The biopreservation strategy allows extending the shelf life and food safety through the use of indigenous or controlled microbiota and their antimicrobial compounds. The aim of this work was to characterize an inhibitory substance with bacteriocin-like activity (Sak-59) produced by the potentially probiotic L. sakei strain from artisanal traditional Kazakh horse meat product Kazy. The maximum production of Sak-59 occurred at the stationary phase of the L. sakei growth. Sak-59 showed inhibitory activity against gram-positive meat spoilage bacteria strains of Listeria monocytogenes, Staphylococcus aureus, and pathogenic gram-negative bacteria strains of Serratia marcescens and Escherichia coli, but not against the tested Lactobacilli strains. Sak-59 activity, as measured by diffusion assay in agar wells, was completely suppressed after treatment with proteolytic enzymes and remained stable after treatment with α-amylase and lipase, indicating that Sak-59 is a peptide and most likely not glycosylated or lipidated. It was concluded that Sak-59 is a potential new bacteriocin with a characteristic activity spectrum, which can be useful in the food and feed industries.

Metagenomic analysis of gut microbial communities from a Central Asian population.

OBJECTIVE: Changes in the gut microbiota are increasingly recognised to be involved in many diseases. This ecosystem is known to be shaped by many factors, including climate, geography, host nutrition, lifestyle and medication. Thus, knowledge of varying populations with different habits is important for a better understanding of the microbiome. DESIGN: We therefore conducted a metagenomic analysis of intestinal microbiota from Kazakh donors, recruiting 84 subjects, including male and female healthy subjects and metabolic syndrome (MetS) patients aged 25-75 years, from the Kazakh administrative centre, Astana. We characterise and describe these microbiomes, the first deep-sequencing cohort from Central Asia, in comparison with a global dataset (832 individuals from five countries on three continents), and explore correlations between microbiota, clinical and laboratory parameters as well as with nutritional data from Food Frequency Questionnaires. RESULTS: We observe that Kazakh microbiomes are relatively different from both European and East Asian counterparts, though similar to other Central Asian microbiomes, with the most striking difference being significantly more samples falling within the Prevotella-rich enterotype, potentially reflecting regional diet and lifestyle. We show that this enterotype designation remains stable within an individual over time in 82% of cases. We further observe gut microbiome features that distinguish MetS patients from controls (eg, significantly reduced Firmicutes to Bacteroidetes ratio, Bifidobacteria and Subdoligranulum, alongside increased Prevotella), though these overlap little with previously published reports and thus may reflect idiosyncrasies of the present cohort. CONCLUSION: Taken together, this exploratory study describes gut microbiome data from an understudied population, providing a starting point for further comparative work on biogeography and research on widespread diseases. TRIAL REGISTRATION NUMBER: ISRCTN37346212; Post-results.

Peptidoglycan Hydrolases of Local Lactic Acid Bacteria from Kazakh Traditional Food.

INTRODUCTION: Peptidoglycan (PG) is a major component of the cell wall of Gram-positive bacteria and is essential for maintaining the integrity of the bacterial cell and its shape. The bacteria synthesize PG hydrolases, which are capable of cleaving the covalent bonds of PG. They also play an important role in modeling PG, which is required for bacterial growth and division. In an era of increasing antibiotic-resistant pathogens, PG hydrolases that destroy these important structures of the cell wall act as a potential source of new antimicrobials. The aim of this study is to identify the main PG hydrolases of local lactic acid bacteria isolated from traditional foods that enhance probiotic activity of a biological preparation. METHODS: Lactococcus lactis 17A and Lactococcus garvieae 19A were isolated from the traditional sausage-like meat product called kazy. They were isolated according to standards methods of microbiology. Genetic identification of the isolates were tested by determining the nucleotide sequences of 16S rDNA. The Republican collection of microorganisms took strains of Lactobacillus casei subsp. Rhamnosus 13-P, L. delbrueckii subsp. lactis CG-1 B-RKM 0044 from cheese, Lactobacillus casei subsp. casei B-RKM 0202 from homemade butter. They used the standard technique of renaturating polyacrylamide gel electrophoresis to detect PG hydrolases activity. RESULTS: According to the profiles of PG hydrolase activity on zymograms, the enzymes of Lactococci 17A and 19A in kazy are similar in electrophoretic mobility to major autolysin AcmA, while the lactobacilli of industrial and home-made dairy products have enzymes similar to extracellular proteins p40 and p75, which have probiotic activity. CONCLUSIONS: Use of peptidoglycan hydrolases seems to be an interesting approach in the fight against multi-drug resistant strains of bacteria and could be a valuable tool for the treatment of diseases caused by these microorganisms in Kazakhstan.

Examination of Lactic Acid Bacteria to Secretion of Bacteriocins.

INTRODUCTION: Bacteriocins produced by lactic acid bacteria (LAB) have the potential to cover a very broad field of applications, including the food industry and the medical sector. In the food industry, bacteriocinogenic LAB strains can be used as starter cultures, co-cultures, and bioprotective cultures, which would be used to improve food quality and safety. In the medical sector, bacteriocins of probiotic LAB might play a role in interactions, which take place in human gastrointestinal tract, and contribute to gut health. The aim of this study was the examine the effect of LAB antimicrobial activity. METHODS: LAB were isolated from different commercial and home made products, such as kazy and sour cream. To screen for bacteriocin producing LAB, we used an agar diffusion bioassay, described in a previous study by Dr. Yang, with three modifications in cell-free supernatant (CFS). First we had a clear supernatant, second we adjusted the CFS to pH 6.0 to eliminate acids antimicrobial effects, and third the CFS pH 6.0 was treated with catalase to exclude the action of H2O2 and confirm action of bacteriocin-like substances. Pathogenic S.marcescens, E. coli, S.aureus cultures were used as indicators. RESULTS: Screening of 95 strains of LAB through deferred antagonism to six indicator cultures showed that all of the selected strains had a high value of antibacterial activity. However, CFS of only 50 strains retained their antimicrobial activity, and 10 of them lost this activity in the second modification of CFS with pH 6.0 to test culture S.marcescens, which confirmed the acidic nature of antimicrobial activity of CFS. Lb.rhamnosus (P-1), Lb.fermentum (N-6), and Lc.lactis (7M) lost antibacterial activity in the presence of the catalase. All modifications of CFS of three strains: Lb.pentosus (16al), Lb.pentosus (P-2), and Pediococcusacidilactici (8) retained inhibitory activity to E.coli and S. aureus. Supernatants of only Lactococcusgarvieae (10a) and Pediococcusacidilactici (25) extracted from homemade meat food kazy (Karaganda) and sour cream (Astana), respectively retained antibacterial activity to all three indicator cultures. CONCLUSION: The antibacterial activity (pH 6.0, added catalase) of Lactococcusgarvieae (10a) and Pediococcusacidilactici (25) to S. marcescens, E. coli, and S.aureus indicates these strains as promising strains for further use in the preparation of bacteriocins.