Determination of Molecular Weight and Antimicrobial Activities of a Purified Bacteriocin from Lactiplantibacillus plantarum MDP 5

Nowadays, unlimited use of antibiotics and preservatives have become a big concern regarding the human health which turn the interest of biotech industries into their research on biologically active molecules from probiotic microbes, since they are nontoxic and suitable for many safer applications. On this background, the present investigation focused on the characterization of a bacteriocin from a Lactobacillus strain. The characterization of a bacteriocin was done using a Lactiplantibacillus plantarum MDP 5 isolated from the local markets of Puducherry and production was performed under standardized cultural conditions. The study on the maximum recovery of bacteriocin using ammonium sulphate precipitation method revealed that the 60% saturation rate evidenced highest activity of 6500 AU/ml with 4.113g/L dry weight followed by the purification was done with RP-HPLC method using C18 column. The purified bacteriocin revealed a novel molecular weight of 22 kDa with the help of SDS-PAGE which has not been reported from Lactobacillus species. Further, the purified bacteriocin evidenced appreciable antimicrobial activities against all the tested human bacterial pathogens of this study. The highest antimicrobial activity was recorded against Escherichia coli MTCC 443 followed by Staphylococcus aureus MTCC 96, Vibrio parahemolyticus MTCC 451, Enterococcus faecalis MTCC 9845, Pseudomonas aeruginosa MTCC 741 and Klebsiella pneumoniae MTCC 109 in the concentrations of 8AU/ml, 16AU/ml, 32AU/ml, 64AU/ml, 256AU/ml and 512AU/ml, respectively. From the overall observation, this study explored a novel bacteriocin purified from a probiotic bacterium represented potential antimicrobial activities against many human pathogens which suggesting its possible use for the safe therapeutic applications.

Microcin MccI47 selectively inhibits enteric bacteria and reduces carbapenem-resistant Klebsiella pneumoniae colonization in vivo when administered via an engineered live biotherapeutic

Background: The gastrointestinal (GI) tract is the reservoir for multidrug-resistant (MDR) pathogens, specifically carbapenem-resistant (CR) Klebsiella pneumoniae and other Enterobacteriaceae, which often lead to the spread of antimicrobial resistance genes, severe extraintestinal infections, and lethal outcomes. Selective GI decolonization has been proposed as a new strategy for preventing transmission to other body sites and minimizing spreading to susceptible individuals. Results: Here, we purify the to-date uncharacterized class IIb microcin I47 (MccI47) and demonstrate potent inhibition of numerous Enterobacteriaceae, including MDR clinical isolates, in vitro at concentrations resembling those of commonly prescribed antibiotics. We then genetically modify the probiotic bacterium Escherichia coli Nissle 1917 (EcN) to produce MccI47 from a stable multicopy plasmid by using MccI47 toxin production in a counterselection mechanism to engineer one of the native EcN plasmids, which renders provisions for inducible expression and plasmid selection unnecessary. We then test the clinical relevance of the MccI47-producing engineered EcN in a murine CR K. pneumoniae colonization model and demonstrate significant MccI47-dependent reduction of CR K. pneumoniae abundance after seven days of daily oral live biotherapeutic administration without disruption of the resident microbiota. Conclusions: This study provides the first demonstration of MccI47 as a potent antimicrobial against certain Enterobacteriaceae, and its ability to significantly reduce the abundance of CR K. pneumoniae in a preclinical animal model, when delivered from an engineered live biotherapeutic product. This study serves as the foundational step towards the use of engineered live biotherapeutic products aimed at the selective removal of MDR pathogens from the GI tract.

Revitalizing the ethanologenic bacterium Zymomonas mobilis for sugar reduction in high-sugar-content fruits and commercial products

The excessive consumption of sugars can cause health issues. Different strategies have been developed to reduce sugars in the diets. However, sugars in fruits and commercial products may be difficult to reduce, limiting their usage among certain populations of people. Zymomonas mobilis is a generally recognized as safe (GRAS) probiotic bacterium with the capability to produce levan-type prebiotics, and thrives in high-sugar environments with unique characteristics to be developed for lignocellulosic biofuel and biochemical production. In this study, the sugar reduction capabilities of Z. mobilis ZM4 were examined using two fruits of pear and persimmon and three high-sugar-content commercial products of two pear pastes (PPs) and one Chinese traditional wine (CTW). Our results demonstrated that Z. mobilis ZM4 can utilize sugars in fruits with about 20 g/L ethanol and less than 5 g/L sorbitol produced within 22 h using pears, and about 45 g/L ethanol and 30 g/L sorbitol produced within 34 h using persimmons. When PPs made from pears were used, Z. mobilis can utilize nearly all glucose (ca. 60 g/L) and most fructose (110 g/L) within 100 h with 40 ~ 60 g/L ethanol and more than 20 g/L sorbitol produced resulting in a final sorbitol concentration above 80 g/L. In the high-sugar-content alcoholic Chinese traditional wine, which contains mostly glucose and ethanol, Z. mobilis can reduce nearly all sugars with about 30 g/L ethanol produced, resulting in a final ethanol above 90 g/L. The ethanol yield and percentage yield of Z. mobilis in 50 ~ 60% CTW were 0.44 ~ 0.50 g/g and 86 ~ 97%, respectively, which are close to its theoretical yields—especially in 60% CTW. Although the ethanol yield and percentage yield in PPs were lower than those in CTW, they were similar to those in fruits of pears and persimmons with an ethanol yield around 0.30 ~ 0.37 g/g and ethanol percentage yield around 60 ~ 72%, which could be due to the formation of sorbitol and/or levan in the presence of both glucose and fructose. Our study also compared the fermentation performance of the classical ethanologenic yeast Saccharomyces cerevisiae BY4743 to Z. mobilis, with results suggesting that Z. mobilis ZM4 had better performance than that of yeast S. cerevisiae BY4743 given a higher sugar conversion rate and ethanol yield for sugar reduction. This work thus laid a foundation for utilizing the advantages of Z. mobilis in the food industry to reduce sugar concentrations or potentially produce alcoholic prebiotic beverages. Graphical Abstract

Siderophore-mediated zinc acquisition enhances enterobacterial colonization of the inflamed gut

Zinc is an essential cofactor for bacterial metabolism, and many Enterobacteriaceae express the zinc transporters ZnuABC and ZupT to acquire this metal in the host. However, the probiotic bacterium Escherichia coli Nissle 1917 (or “Nissle”) exhibits appreciable growth in zinc-limited media even when these transporters are deleted. Here, we show that Nissle utilizes the siderophore yersiniabactin as a zincophore, enabling Nissle to grow in zinc-limited media, to tolerate calprotectin-mediated zinc sequestration, and to thrive in the inflamed gut. We also show that yersiniabactin’s affinity for iron or zinc changes in a pH-dependent manner, with increased relative zinc binding as the pH increases. Thus, our results indicate that siderophore metal affinity can be influenced by the local environment and reveal a mechanism of zinc acquisition available to commensal and pathogenic Enterobacteriaceae.

Chickpea-Derived Prebiotic Substances Trigger Biofilm Formation by Bacillus subtilis

Chickpea-based foods are known for their low allergenicity and rich nutritional package. As an essential dietary legume, chickpea is often processed into milk or hummus or as an industrial source of protein and starch. The current study explores the feasibility of using the chickpea-derived prebiotic substances as a scaffold for growing Bacillus subtilis (a prospective probiotic bacterium) to develop synbiotic chickpea-based functional food. We report that the chickpea-derived fibers enhance the formation of the B. subtilis biofilms and the production of the antimicrobial pigment pulcherrimin. Furthermore, electron micrograph imaging confirms the bacterial embedding onto the chickpea fibers, which may provide a survival tactic to shield and protect the bacterial population from environmental insults. Overall, it is believed that chickpea-derived prebiotic substances provide a staple basis for developing functional probiotics and synbiotic food.

Potential of Inulin-Fructooligosaccharides Extract Produced from Red Onion (Allium cepa var. viviparum (Metz) Mansf.) as an Alternative Prebiotic Product

Red onion is a popular ingredient in many Thai dishes and has recently been promoted for commercial cultivation. In this study, inulin-fructooligosaccharides (inulin-FOSs) were extracted from red onions in a simplified extraction method. The extract contained 24.00 ± 0.38 g/L free glucose, fructose and sucrose, while the level of FOSs was recorded at 74.0 ± 2.80 g/L with a degree of polymerization of 4.1. The extract was resistant to simulated gastrointestinal conditions, while selectively promoting probiotic lactobacilli. These outcomes resulted in inhibitory effects against various pathogenic bacteria. The in vitro batch culture fermentation of the extract by natural mixed culture indicated that an unknown sugar identified as neokestose was more rapidly fermented than 1-kestose and other longer-chain inulin-FOSs. Notably, neokestose selectively encouraged a bifidogenic effect, specifically in terms of the growth of Bifidobacteirum breve, which is an infant-type probiotic bacterium. This is the first report to state that neokestose could selectively enhance the bifidogenic effect. In summary, inulin-FOSs extract should be recognized as a multifunctional ingredient that can offer benefits in food and pharmaceutical applications.

Polyclonal Aptamers for Specific Fluorescence Labeling and Quantification of the Health Relevant Human Gut Bacterium Parabacteroides distasonis

Single-stranded DNA aptamers as affinity molecules for the rapid, reliable detection of intestinal bacteria are of particular interest to equip health systems with novel robust and cheap diagnostic tools for monitoring the success of supplementation strategies with selected probiotic gut bacteria in the fight against major widespread threats, such as obesity and neurodegenerative diseases. The human gut bacterium Parabacteroides distasonis (P. distasonis) is positively associated with diseases such as obesity, non-alcoholic fatty liver disease and multiple sclerosis with reduced cell counts in these diseases and is thus a promising potential probiotic bacterium for future microbial supplementation. In this paper we report on the evolution of a specific polyclonal aptamer library by the fluorescence based FluCell-SELEX directed against whole cells of P. distasonis that specifically and efficiently binds and labels P. distasonis. The aptamer library showed high binding affinity and was suited to quantitatively discriminate P. distasonis from other prominent gut bacteria also in mixtures. We believe that this library against a promising probiotic bacterium as a prototype may open new routes towards the development of novel biosensors for the easy and efficient quantitative monitoring of microbial abundance in human microbiomes in general.

Hypocholesterolemic Operating Parameters of Novel Probiotics In vitro

The Hypocholesterolemic is an effective and friendly process for the removal of cholesterol and an important aspect of human health. Removal of cholesterol by probiotic bacteria was investigated in this study. The potential probiotic bacterium was preliminarily identified as Lactobacillus casei. In addition, the various operating parameters like cholesterol concentration, incubation time, bile salts concentration, probiotic dose and initial pH was studied. The optimum parameters for most probiotics studied are cholesterol concentration is 100 μg /ml, incubation time is 48 h, bile salts concentration is 0.5%, probiotic dose is 200μl and initial pH is 6. The maximum percentage removal of cholesterol using Lactobacillus casei is 93.6. The probiotics bacteria are favorable for Hypocholesterolemic.

High dimensional geometry of fitness landscapes identifies master regulators of evolution and the microbiome

A longstanding goal of biology is to identify the key genes and species that critically impact evolution, ecology, and health. Yet biological interactions between genes (1, 2), species (3–6), and different environmental contexts (7–9) change the individual effects due to non-additive interactions, known as epistasis. In the fitness landscape concept, each gene/organism/environment is modeled as a separate biological dimension (10), yielding a high dimensional landscape, with epistasis adding local peaks and valleys to the landscape. Massive efforts have defined dense epistasis networks on a genome-wide scale (2), but these have mostly been limited to pairwise, or two-dimensional, interactions (11). Here we develop a new mathematical formalism that allows us to quantify interactions at high dimensionality in genetics and the microbiome. We then generate and also reanalyze combinatorically complete datasets (two genetic, two microbiome). In higher dimensions, we find that key genes (e.g. pykF) and species (e.g. Lactobacillus plantarum) distort the fitness landscape, changing the interactions for many other genes/species. These distortions can fracture a “smooth” landscape with one optimal fitness peak into a landscape with many local optima, regulating evolutionary or ecological diversification (12), which may explain how a probiotic bacterium can stabilize the gut microbiome.

Nutritionally Enhanced Probioticated Whole Pineapple Juice

Nutritionally enhanced probioticated whole pineapple juice (WPJ, comprising juice of pineapple pulp and peel) beverages were produced by fermentation of WPJ with the probiotic bacterium Lactobacillus plantarum WU-P19. The 12 h fermented juice contained between 2.1 × 109 and 3.7 × 109 live cells of the probiotic per milliliter, depending on the beverage formulation. The beverage had a pH of around 4.1 and a lactic acid content of ~12.8 g L−1. It had a total sugar (glucose, sucrose, fructose, maltose) content of ~100.2 g L−1. During fermentation, some of the initial glucose and fructose were consumed by the probiotic, but sucrose and maltose were not consumed. The original WPJ was free of vitamin B12, but fermentation enhanced vitamin B12 content (~19.5 mg L−1). In addition, fermentation enhanced the concentrations of vitamins B2, B3, and B6, but the bacterium consumed some of the vitamin B1 originally present. From a nutritional perspective, the final probioticated beverage was a good source of vitamin B12, vitamin C and vitamin B6. In addition, it contained nutritionally useful levels of vitamins B1, B2, and B3. The calorific value of the final beverage was 56.94 kcal per 100 mL. The product was stable during 21-day refrigerated (4 °C) storage.