Application of soy protein isolate-xylose conjugates for improving the viability and stability of probiotics microencapsulated by spray drying.

BACKGROUND: Production and consumption of probiotics need to meet many adverse stresses, which can reduce their health-promoting effects on humans. Microencapsulation is an effective technique to improve the biological activity of probiotics and wall materials are also required during encapsulation. Application of Maillard reaction products (MRPs) in probiotic delivery is increasing. RESULTS: This work aims to study the effects of soy protein isolate (SPI)-xylose conjugates heated at different times on the viability and stability of probiotics. SPI-xylose MRPs formed after heat treatment based on changes in the browning intensity, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Fourier transform infrared spectroscopy. After heat treatment, α-helix and ß-sheet contents of SPI-xylose mixture shifted from 11.3% and 31.3% to 6.4-11.0% and 31.0-36.9%, respectively, and the thermal stability slightly changed. During spray drying, except for MRP240@LAB, probiotic viability was higher in the MRP-based probiotic microcapsules (21.36-25.31%) than in Mix0@LAB (20.17%). MRP-based probiotic microcapsules had smaller particle sizes (431.1-1243.0 nm vs. 7165.0 nm) and greater intestinal digestion tolerance than Mix0@LAB. Moreover, the MRP-based probiotic microcapsules showed better storability than Mix0@LAB and adequate growth and metabolism capacity. CONCLUSION: SPI-xylose Maillard reaction products are a promising wall material for probiotics microencapsulation, which can improve bacterial survivability during spray drying and enhance bacterial gastrointestinal digestion resistance. This study sheds light on preparing probiotic microcapsules with superior properties by spray drying. © 2023 Society of Chemical Industry.

The lactic acid bacteria and yeast community of home-made sauerkraut from three provinces in Southwest China.

The aim of this study was to investigate the lactic acid bacteria (LAB) and yeast community from home-made sauerkraut collected from Southwest China through culture-dependent and culture-independent technology. Forty-eight samples of home-made sauerkraut were collected from households at three different locations in Southwest China. The pH, total acidity and salt contents among these fermented vegetables were 3.69 ± 0.42, 0.86 ± 0.43 g/100 ml, and 3.86 ± 2.55 g/100 ml, respectively. The number of lactic acid bacteria (LAB) and yeasts were 7.25 ± 1.05 log10 colony-forming units (CFU)/ml and 3.74 ± 1.01 log CFU/ml, respectively. A total of 182 LAB and 81 yeast isolates were identified. The dominant isolates were Lactobacillus plantarum, L. brevis, Pediococcus ethanolidurans, Pichia membranifaciens, P. fermentans and Kazachstania bulderi. Denaturing gradient gel electrophoresis (DGGE) showed that L. plantarum, uncultured Lactobacillus sp, P. ethanolidurans, and K. exigua were the predominant microflora. Our studies demonstrated that the DGGE technique combined with a culture-dependent method is very effective for studying the LAB and yeast community in Chinese traditional fermentation vegetables. The results will give us an understanding of LAB and yeast community of Chinese sauerkraut and improve the knowledge of LAB and yeast community of Chinese sauerkraut.

Metatranscriptomics reveals the gene functions and metabolic properties of the major microbial community during Chinese Sichuan Paocai fermentation.

Chinese Sichuan Paocai (CSP) is one of the world's best-known fermented vegetables with a large presence in the Chinese market. The dynamic microbial community is the main contributor to Paocai fermentation. However, little is known about the ecological distribution and functional importance of these community members. In this study, metatranscriptomics was used to comprehensively explore the active microbial community members and key transcripts with significant functions in the Paocai fermentation process. Enterobacter, Leuconostoc, and Lactobacillus dominated the three-fermentation stages (Pre-, Mid- and Lat-), respectively. Carbon metabolism was the most abundant pathway. GH (glycoside hydrolase) and GT (lycosyl transferase) were the two most highly expressed carbohydrate-active enzymes. The most highly differentially expressed genes were grouped in the biosynthesis of amino acids, followed by glycolysis. Meta-pathways in the Sichuan Paocai fermentation ecosystem were reconstructed, Lactobacillaceae and Enterobacteriaceae were the two most important metabolic contributors. In addition, the nrfA and nirB were two genes referred to distinct nitrite reductase enzymes and 9 specialized genes, such as eclo, ron and ent were expressed to produce autoinducer 2 (AI-2) kinase in response to population density. The present study revealed functional enzymes and meta-pathways of the active microbial communities, which provide a deeper understanding of their contribution to CSP products.

Genomic analysis revealed adaptive mechanism to plant-related fermentation of Lactobacillus plantarum NCU116 and Lactobacillus spp.

Lactobacillus plantarum NCU116 is the first sequenced strain derived from traditional Chinese sauerkraut (TCS). Since NCU116 manifested outstanding probiotic effects in vitro and in vivo, it is crucial to comprehend a clear genetic background for NCU116. Functional re-annotation and comparative analysis were performed to excavate the unique and representative genes in NCU116, in order to investigate its metabolic preference and adaptive mechanism. Horizontal gene transfer (HGT) seemed to occur frequently, which endows NCU116 with a strong ability to transport carbohydrates, as a strain-specific fructose/mannose-PTS was identified, and opu and osmC coding genes were retrieved as NCU116-specific. In addition, a strain-specific type I R/M system and several prophage loci were found in NCU116, which could play vital roles in self-defense mechanism. Pathways of bacterial metabolism on plant-related substrates fermentation were then generated by reconstruction of associated pathways. Moreover, a unique potential plantaricin-producing locus with high homology to that of JDM1 was defined in the genome of NCU116, which could be very important for the preservation of fermented-food. Our results would provide critical basis for the application of NCU116 in food and pharmaceuticals industries.

Interactions between Lactobacillus plantarum NCU116 and its environments based on extracellular proteins and polysaccharides prediction by comparative analysis.

Lactic acid bacteria (LAB) play a significant role in food industry and artisan fermented-food. Most of the applicable LABs were commonly obtained from natural fermented food or human gut. And Lactobacillus plantarum NCU116 was screened from a LAB-dominated traditional Chinese sauerkraut (TCS). In order to comprehend the interaction between NCU116 and its environments, comparative genomics were performed to identify genes involved in extracellular protein biosynthesis and secretion. Four secretory pathways were identified, including Sec and FPE pathways, holins and efflux ABC transporter system. Then 348 potential secretory proteins were identified, including 11 alpha-amylases responsible for degradation of macromolecules, and 8 mucus binding proteins which attribute to adherence to intestine epithelium. Besides, EPS clusters of NCU116 (EPS116) were identified and analyzed by comparing to other strains, which suggested a novel genotype of EPS clusters. These findings could be critical to extend the application of NCU116 in food and pharmaceuticals industries.

Composition and functional diversity of fecal bacterial community of wild boar, commercial pig and domestic native pig as revealed by 16S rRNA gene sequencing.

The bacterial community in mammalian gastrointestinal tract is abundant and complex. To date, little is known about the gut microbiota of wild boar. This study aimed to investigate the fecal bacterial diversity of wild boar and compare with commercial pig and domestic native pig. The diet composition showed that the diets of wild boar, commercial pig and domestic native pig were different from each other. More than 1,760,000 quality-filtered sequences were obtained, and the results revealed distinct compositions and diversity of fecal microbiota in three groups. PCoA and NMDS analyses showed that fecal bacterial communities of wild boar, commercial pig and domestic native pig formed distinctly different clusters. Although the three groups shared a large size of OTUs comprising a core microbiota community, a strong distinction existed at family and genus levels. Ruminococcaceae, Prevotellaceae and Christensenellaceae were more abundant in the feces of wild boar than in domestic native pig and commercial pig. At the genus level, the proportion of unidentified Christensenellaceae was remarkably higher in wild boar group, while commercial pig and domestic native pig group had a higher abundance of Streptococcus and Lactobacillus. Tax4Fun predictions of metagenome function showed statistically significant differences in the functions of fecal microbiota in three groups. There were more bacteria genes with amino acid metabolism, cell growth and death, cell motility, energy metabolism, immune system and environmental adaptation observed in wild boar feces, while commercial pig feces contained more bacteria genes with carbohydrate metabolism, drug resistance, aging, infectious diseases, lipid metabolism, endocrine and metabolic diseases. These results indicated that the fecal microbial ecosystem of the wild boar is significantly different from that of domestic native pig and commercial pig, suggesting that diet is an important factor leading to differences in bacterial abundance and diversity in feces.

Evaluation and comparison of the microbial communities and volatile profiles in homemade suansun from Guangdong and Yunnan provinces in China.

BACKGROUND: Suansun is a traditional salt-free fermented bamboo shoot product that has been widely consumed as a cooking ingredient in south China for centuries. The aim of this study was to evaluate and compare the microbial and metabolic diversity in samples of two kinds of suansun, namely Guangdong suansun (GD) and Yunnan suansun (YN), using high-throughput sequencing (HTS) and headspace solid-phase microextraction-gas chromatograph-mass spectrometry (HS-SPME/GC-MS), respectively, and then to assess the influence of environmental factors on the microbial communities. RESULTS: The results showed that Lactobacillus and Serratia were the most abundant bacterial genera in both the GD and YN groups. For the fungi, Pichia, Candida, and Debaryomyces were the major genera in the GD group, whereas Pichia and Zygosaccharomyces were the dominant genera in the YN group. The canonical correlation analysis (CCA) results demonstrated that three environmental factors - temperature, longitude, and altitude - play a more important role in affecting the microbial community composition of suansun than physical and chemical factors. The fugal community composition was more influenced by environmental factors than the bacterial community. The volatile profile of the GD group differed from that of the YN group, and the difference was mainly reflected in the relative alcohol, aldehyde, ester, and aromatic compound content. CONCLUSIONS: This study provided insights into the microbial and metabolic profiles of suansun products. The findings might be useful for the improvement and standardization of suansun production. © 2020 Society of Chemical Industry.

Comparison of bacterial diversity in traditionally homemade paocai and Chinese spicy cabbage.

This study aimed to investigate bacterial diversity in paocai and Chinese spicy cabbage and compare the microbial communities using high-throughput sequencing. Bacteria representing 26 phyla, 480 genera and 338 species were observed in these Chinese fermented vegetables. Firmicutes and Proteobacteria were the main phyla observed in both paocai and Chinese spicy cabbage. Additionally, Lactobacillus, Pediococcus, Serratia, Stenotrophomonas and Weissella were the major genera observed in both paocai and Chinese spicy cabbage. Overall, the relative abundances of Lactobacillus, Pediococcus and Weissella in Chinese spicy cabbage were much higher than those in paocai, but the proportions of Stenotrophomonas and Serratia in Chinese spicy cabbage were less than those in paocai. The results showed that the composition of the microbial community in Chinese spicy cabbage was positively correlated with total titratable acidity (TA), lactic acid and acetic acid contents but was negatively correlated with salinity. In contrast, the composition of the microbial community in paocai was negatively correlated with TA, lactic acid and acetic acid contents but was positively correlated with salinity. This study provides insights into the relationship between bacterial profiles and environmental factors in Chinese spicy cabbage and paocai, and its findings will aid in guiding future research on fermented vegetables.

Bacterial community dynamics and physicochemical characteristics in natural fermentation of jiang-shui, a traditional food made in northwest China.

BACKGROUND: Chinese jiang-shui is fermented food without salt. In consideration of the few studies of the bacterial community and dynamics of jiang-shui fermentation, the aim of this study was to investigate the microbial diversity and dynamics of jiang-shui fermentation using high-throughput sequencing (HTS), denaturing gradient gel electrophoresis (DGGE) and the culture-dependent method. RESULTS: The culture-dependent method showed that pathogenic bacteria (Staphylococcus aureus, Salmonella and Escherichia coli) and yeast mainly existed in the early stage and lactic acid bacteria dominated until the end stage. HTS and DGGE revealed that Serratia marcescens, Serratia sp., Lactobacillus curvatus, Lactococcus lactis, uncultured bacterium and Bacillus thuringiensis started the fermentation, followed by the middle stage with Lactococcus lactis, Weissella sp. and Bacillus arthracis as the predominant species. The end stage was characterized with Lactobacillus pentosus, Weissella cibaria and Weissella sp. as the major bacteria. CONCLUSIONS: The results showed that genera Serratia, Lactococcus, Weissella and Lactobacillus dominated the whole process of jiang-shui fermentation. This study provided a good analysis of the bacterial changes of jiang-shui fermentation, and future studies should consider the relationships among the sensory characteristics, microbial communities and metabolites. © 2019 Society of Chemical Industry.

Isolation and characterization of potential Lactobacillus acidophilus strains isolated from pig feces.

The aim of this study was to isolate and characterize Lactic Acid Bacteria (LAB) from 16 feces samples of pig farm, and to evaluate the probiotic potential of these isolates as potential oral probiotic candidates. The selection process was based on the isolation, identification, and a series of experiments for the selection of appropriate candidates with beneficial properties. The results demonstrated that most of LAB showed relatively strong resistance to pH 2.5 and high bile salts (1%), and had good survival in simulated gastric and intestinal juice. 9 isolates displayed antimicrobial activities against Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa and Enterobacter sakazakii. Almost all isolates were sensitive to ampicillin, chloramphenicol, vancomycin and amoxicillin, and most of isolates exhibited resistance against tetracycline and vancomycin. The adhesion rates of LAB varied greatly. The results of the study suggested that the Lactobacillus acidophilus NCUA065001 have the important functional property of probiotic candidates to enhance gut integrity and could considered to be the potential antibiotic alternatives in the pig feed industry.

The microbial succession and their correlation with the dynamics of flavor compounds involved in the natural fermentation of suansun, a traditional Chinese fermented bamboo shoots.

Suansun is a typical fermented bamboo shoot product widely consumed in southern China. Complex microorganisms are involved during spontaneous fermentation of suansun. However, little is known about the succession and roles of these microbiota throughout the fermentation process, especially for the functional microbiota involved in aroma formation. Thus, in this study, the dynamic changes in physicochemical parameters, flavor compounds, and microbiota community during the natural production of suansun were investigated. The principal component analysis (PCA) results showed that physicochemical parameters and flavor compounds varied substantially during the 30-day fermentation. Moreover, the microbial flora exhibited a dynamic change from the acid-sensitive genera of Lactococcus, Leuconostoc, Weissella, and Enterobacter to an acid-resistant genus of Lactobacillus due to lactic acid and acetic acid accumulation. Furthermore, bidirectional orthogonal partial least squares (O2PLS) indicated that four bacterial genera, namely, Lactobacillus, Clostridium_sensu_stricto_1, Enterobacter, and Leuconostoc, showed strong connections with various characteristic flavors, thus playing crucial roles in the unique flavor formation of suansun. These results may shed light on the development of effective starter cultures for the industrial manufacture of high-quality suansun products.

Screening, Safety Evaluation, and Mechanism of Two Lactobacillus fermentum Strains in Reducing the Translocation of Staphylococcus aureus in the Caco-2 Monolayer Model.

Staphylococcus aureus is a common commensal of humans, and its translocation from gastrointestine to peripheral organs and tissues could cause severe diseases and complications. This study focuses on the screening and characterization of Lactobacillus strains with significant inhibitory effect on the translocation of S. aureus through Caco-2 monolayers. First, strains with strong affinity for mucin and Caco-2 cells were obtained, via microtiter plate assay and adhesion assay, respectively. Obtained bacteria were further tested for their inhibitory effects on the growth of S. aureus by well diffusion assay. Subsequently, two strains preincubated with Caco-2 monolayers were found to inhibit the translocation of S. aureus CMCC26003 by 80.95 and 43.96%, respectively, via the transcellular translocation assay. These two strains were then identified to be Lactobacillus fermentum NCU3087 and L. fermentum NCU3088. Second, the mechanism of inhibition was investigated by analyzing the relative concentration of tight junction proteins and proinflammatory cytokines of Caco-2 cells, by Western blot and enzyme-linked immunosorbent assay, respectively. Results showed that both NCU3087 and NCU3088 significantly attenuated the degradation of occludin, claudin-1, ZO-1, and JAM-1 and suppressed the secretion of interleukin 6 and tumor necrosis factor-α induced by S. aureus, to different extent. Moreover, two Lactobacillus strains could barely translocate the Caco-2 monolayers, had no hemolytic activity, displayed strong resistance to gastrointestinal fluids, and were sensitive or moderate sensitive to nine clinically relevant antibiotics. Collectively, this study identified two Lactobacillus strains with significant inhibitory effect on the translocation of S. aureus, and their safeness for application was evaluated, thereby providing potential solutions for prevention of S. aureus and prophylaxis of related diseases.

Comparison of microbial communities and physiochemical characteristics of two traditionally fermented vegetables.

Guangxi Suansun (Guangxi SS) and Guangxi Suancai (Guangxi SC) are two kinds of traditionally fermented vegetables consumed as cooking ingredients in Guangxi Province, China, for thousands of years. However, little is known about their microbial communities as well as the differences between them. Thus, this study was aimed to investigate and compare the physicochemical indexes as well as the bacterial and fungal profiles of Guangxi SS and Guangxi SC. Results showed that the titratable acidity, lactic acid and acetic acid content in SS were significantly higher than those in SC, while the salinity of SS was significantly lower than that in SC, and the nitrite contents in all samples were are far lower than the limit of nitrite contents in fermented vegetables. Firmicutes, Proteobacteria, Ascomycota and Basidiomycota were the main phyla observed in both SS and SC samples. Lactobacillus, Serratia, Stenotrophomonas and Lactococcus were the major bacterial genera in both SS and SC samples, the predominant fungal genera in SS group were Kazachstania, Debaryomyces and Pichia, while the major genera in SC group were Kazachstania, Debaryomyces and Nakaseomyces. At the species level, Lactobacillus acetotolerans, Pichia kudriavzevii, Debaryomyces hansenii, Pichia norvegensis, Kazachstania exigua and Kazachstania humilis were the major species observed in SS, while L. delbrueckii, L. fermentum, L. aviarius, and Pichia kudriavzevii and Debaryomyces hansenii were the predominant species in SC. Salinity was found to be more strongly correlated to the bacterial and fungal communities of both SS and SC than other physicochemical factors (pH, the titratable acidity, lactic acid and acetic acid content). This study provided detailed insight into the microbial communities of Guangxi SS and Guangxi SC, and the findings may help understand the microbial structures of Chinese traditional fermented vegetables.

Comparison of the bacterial communities in home-made Nanfeng yancai with and without salt.

The differences in bacterial communities and physicochemical characteristics between salted and salt-free yancai samples from Nanfeng County were compared in this study. Total acid, salinity and lactic acid content in salted group were significantly higher than those in salt-free group, whereas pH value as well as the contents of acetic acid, sucrose, glucose and fructose showed no significant difference between the two groups. 20 phyla, 339 genera and 203 species were observed in all yancai samples. Firmicutes and Proteobacteria were the major phyla. Lactobacillus was the predominant genus in both the salted and the salt-free samples, whereas Pediococcus, as the second most abundant genus in the salted samples, was in fairly low abundance in the salt-free samples. Culture-dependent and -independent methods showed that the major lactic acid bacteria in the salted samples were Lactobacillus plantarum, L. pentosus, Pediococcus pentosus and L. brevis, while the salt-free samples were predominated by L. fermentum, L. coryniformis and L. sp. The result indicated that salinity had a significant effect on the bacterial communities in Nanfeng yancai, which will improve our understanding of the bacterial ecology of yancai, as well as the relationships between physicochemical characteristics and the microbiota of yancai.

Bacterial community and composition in Jiang-shui and Suan-cai revealed by high-throughput sequencing of 16S rRNA.

Fermented vegetables have a long history in many cultures. Jiang-shui and Suan-cai are two of the most well-known instances in North China. They are made by a process of natural lactic acid fermentation. However, they have the different characteristics, i.e. acidity, taste and flavor, which are influenced by the specific bacterial community. Therefore, we used high-throughput sequencing methods to identify the bacterial community structure of Jiang-shui and Suan-cai in this study. Subsequently, we figured out the bacterial differences of these two products using the statistical analysis. Firmicutes and Proteobacteria were the dominant phyla in both Jiang-shui and Suan-cai. However, Lactobacillus was the main genus in Jiang-shui samples, whereas both Lactobacillus and Pediococcus were the major genera in the Suan-cai samples. At the species level, Lactobacillus amylolyticus, Lactobacillus fermentum and Lactobacillus pontis were the major species in Jiang-shui samples, while Pediococcus parvulus, Lactobacillus coryniformis, Lactobacillus pentosus and Lactobacillus parabrevis were the dominant species in the Suan-cai samples. These results suggested that Jiang-shui and Suan-cai had their own unique bacterial community, leading to the specific characteristics. Furthermore, the bacterial communities of both fermented vegetables varied at different locations. This study revealed the flora present in the Jiang-shui and the Suan-cai, providing a deep insight of the microbial species of Chinese fermented vegetables and guidance for the production of the Jiang-shui and the Suan-cai.