A comparison of mining methods to extract novel bacteriocins from Lactiplantibacillus plantarum NWAFU-BIO-BS29.

One of the most important challenges in the field of food safety is producing natural and safe substances that act against pathogens in food. Bacteriocins and antimicrobial peptides (AMPs) have an anti-pathogens effect for both Gram-negative and positive bacteria. The aim of this study was to isolate and characterize safe lactic acid bacteria from traditional Chinese fermented milk that can produce anti-bacterial molecule compounds and does not harm for humans and animals. Lactiplantibacillus plantarum NWAFU-BIO-BS29 was found to be safe, lacking 16 genes for virulence factors, biogenic amine production and antibiotic resistance, and no hemolysis activity was observed. In contrast, it has ability to produce a novel potential bacteriocin of Plantaricin Bio-LP1. Precipitation of bacteriocin by Ethyl-acetate proved to be a suitable method for the extraction the bacteriocin. Whilst, the purification steps were performed as follows: the protein purification system (AKTA-Purifier equipped with HiTrap (gel column)), followed by reversed phase high-performance liquid chromatography (RP-HPLC) equipped with C18 column. In addition, LC-MS-MS and MALDI-TOF were used to identify the peptide sequences and estimate the molecular weight, respectively. Notably, among the eight peptide sequences considered, a couple of sequences have been announced as uncharacterized in protein database (FDYYFFDKK and KEIDDNSIAVK) with a molecular mass less than 1.3 kDa. The MIC was 0.552 mg/ml and exhibited high stability under various temperature, pH, and enzymes conditions. The best activity was found at temperature and pH of 4 °C and 6 °C, respectively, which are the optimal conditions for preservation of most foods. We concluded that, the described method can arouse a growing interest in mining novel bacteriocins. Plantaricin Bio-LP1 is a potentially unique bacteriocin that is effective as a bio-preservative and could make a promising contribution in food and animal feed industries or in the medical field with further clinical studies.

Mining, heterologous expression, purification, antibactericidal mechanism, and application of bacteriocins: A review.

Bacteriocins are generally considered as low-molecular-weight ribosomal peptides or proteins synthesized by G+ and G- bacteria that inhibit or kill other related or unrelated microorganisms. However, low yield is an important factor restricting the application of bacteriocins. This paper reviews mining methods, heterologous expression in different systems, the purification technologies applied to bacteriocins, and identification methods, as well as the antibacterial mechanism and applications in three different food systems. Bioinformatics improves the efficiency of bacteriocins mining. Bacteriocins can be heterologously expressed in different expression systems (e.g., Escherichia coli, Lactobacillus, and yeast). Ammonium sulfate precipitation, dialysis membrane, pH-mediated cell adsorption/desorption, solvent extraction, macroporous resin column, and chromatography are always used as purification methods for bacteriocins. The bacteriocins are identified through electrophoresis and mass spectrum. Cell envelope (e.g., cell permeabilization and pore formation) and inhibition of gene expression are common antibacterial mechanisms of bacteriocins. Bacteriocins can be added to protect meat products (e.g., beef and sausages), dairy products (e.g., cheese, milk, and yogurt), and vegetables and fruits (e.g., salad, apple juice, and soybean sprouts). The future research directions are also prospected.

Biodegradation of lignin by Pseudomonas sp. Q18 and the characterization of a novel bacterial DyP-type peroxidase.

Lignin valorization can be obtained through cleavage of selected bonds by microbial enzymes, in which lignin is segregated from cellulose and hemicellulose and abundant phenolic compounds can be provided. In this study, Pseudomonas sp. Q18, previously isolated from rotten wood in China, was used to degrade alkali lignin and raw lignocellulosic material. Gel-permeation chromatography, field-emission scanning electron microscope, and GC-MS were combined to investigate the degradation process. The GC-MS results revealed that the quantities of aromatic compounds with phenol ring from lignin increased significantly after incubation with Pseudomonas sp. Q18, which indicated the degradation of lignin. According to the lignin-derived metabolite analysis, it was proposed that a DyP-type peroxidase (PmDyP) might exist in strain Q18. Thereafter, the gene of PmDyP was cloned and expressed, after which the recombinant PmDyP was purified and the enzymatic kinetics of PmDyP were assayed. According to results, PmDyP showed promising characteristics for lignocellulosic biodegradation in biorefinery.

A galacturonic acid-rich polysaccharide from Diospyros kaki peel: Isolation, characterization, rheological properties and antioxidant activities in vitro.

This research elucidated the structural characteristics and antioxidant activity of a galacturonic acid-rich polysaccharide (PPP-2) isolated from Diospyros kaki peel. PPP-2 was extracted by subcritical water and subsequently purified by DEAE-Sepharose FF column. PPP-2 (12.28 kDa) mainly contained galacturonic acid, arabinose, and galactose with the molar ratios of 87.15: 5.86: 4.31. The structural characteristics of PPP-2 were revealed through FT-IR, UV, XRD, AFM, SEM, Congo red, methylation, GC/MS assay and NMR spectrum. PPP-2 owned the triple helical structure and degradation temperature of 251.09 ℃. The backbone of PPP-2 was formed by â†’4)-α-d-GalpA-6-OMe-(1→ and â†’4)-α-d-GalpA-(1→ with the side chains of â†’5)-α-l-Araf-(1→, →3)-α-l-Araf-(1→, →3,6)-ß-d-Galp-(1→ and α-l-Araf-(1→. Moreover, the inhibitory concentration (IC50) of PPP-2 to ABTS•+, DPPH•, superoxide radical and hydroxyl radical were 1.96, 0.91, 3.63, and 4.08 mg/mL, respectively. Our results suggested that PPP-2 might be a novel candidate of natural antioxidant in pharmaceuticals or functional food.

Protective effect of plantaricin bio-LP1 bacteriocin on multidrug-resistance Escherichia Coli infection by alleviate the inflammation and modulate of gut-microbiota in BALB/c mice model.

The rapid spread of multidrug-resistant pathogens with the low efficacy of common antibiotics for humans and animals in its clinical therapeutics are a global health concern. Therefore, there is a need to develop new treatment strategies to control them clinically. The study aimed to evaluate the effects of Plantaricin Bio-LP1 bacteriocin produced from Lactiplantibacillus plantarum NWAFU-BIO-BS29 to alleviate the inflammation caused by multidrug-resistance Escherichia Coli (MDR-E. coli) infection in BALB/c mice-model. The focus was given on aspects linked to the mechanism of the immune response. Results indicated that Bio-LP1 had highly promising effects on partially ameliorating MDR-E. coli infection by reducing the inflammatory response through inhibiting the overexpression of proinflammatory-cytokines such as secretion of tumor necrosis factor (TNF-α) and interleukin (IL-6 and IL-ß) and strongly regulated theTLR4 signaling-pathway. Additionally, avoided the villous destruct, colon length shortening, loss of intestinal barrier integrity, and increased disease activity index. Furthermore, significantly increased the relative abundance of beneficial-intestinal-bacteria including Ligilactobacillus, Enterorhabdus, Pervotellaceae, etc. Finally, improved the intestinal mucosal barrier to alleviate the pathological damages and promote the production of short-chain fatty acids (SCFAs) a source of energy for the proliferation. In conclusion, plantaricin Bio-LP1 bacteriocin can be considered a safe alternative to antibiotics against MDR-E. coli-induced intestinal inflammation.

Purification, structural characterization and antioxidant activities of two neutral polysaccharides from persimmon peel.

Two neutral polysaccharides (PPP1-1 and PPP1-2) were purified from persimmon peel. PPP1-1 (21.84 kDa) was mainly composed of arabinose (22.92 %), galactose (21.09 %), glucose (35.13 %), and xylose (19.09 %), while PPP1-2 (10.42 kDa) mainly contained arabinose (32.98 %), galactose (20.81 %), glucose (26.86 %), xylose (10.46 %), and mannose (7.63 %). Methylation and NMR spectra analysis demonstrated that the backbone of PPP1-1 appeared to be →6)-α-D-Glcp-(1→, →2,6)-α-D-Glcp-(1→, →5)-α-L-Araf-(1→, and →3,5)-α-L-Araf-(1 â†’ residues with branches consisting of →3)-α-L-Araf-(1→, →4)-α-D-Glcp-(1→, →3)-ß-D-Galp-(1→, →4)-ß-D-Galp-(1→, →4)-ß-D-Xylp-(1→, →6)-ß-D-Galp-(1→, →4)-ß-D-Manp-(1→, and α-L-Araf-(1 â†’ residues. The main chain of PPP1-2 was composed of →6)-α-D-Glcp-(1→, →5)-α-L-Araf-(1→, and →3,5)-α-L-Araf-(1 â†’ residues with branches consisting of →3)-α-L-Araf-(1→, →1,2)-α-D-Glcp-(6→, →4)-α-D-Glcp-(1→, →3)-ß-D-Galp-(1→, →4)-ß-D-Galp-(1→, →6)-ß-D-Galp-(1→, →4)-ß-D-Xylp-(1→, →4,6)-α-D-Glcp-(1→, and →4)-ß-D-Manp-(1 â†’ residues and terminal of α-L-Araf-(1 â†’ residue. PPP1-2 exhibited stronger antioxidant activities and better thermal stability than PPP1-1. Our results provided the foundation for further investigating the structure and biological activities of persimmon peel polysaccharides and highlighted their potential to become potential antioxidants in functional food.

Roles of intestinal Parabacteroides in human health and diseases.

The stability of gut microbiota is essential for the host's health. Parabacteroides spp., core members of the human gut microbiota, have an average abundance of 1.27% in humans of 12 populations. Parabacteroides have recently been reported to have a close relationship with host health (e.g. metabolic syndrome, inflammatory bowel disease and obesity). Parabacteroides have the physiological characteristics of carbohydrate metabolism and secreting short chain fatty acids. However, antimicrobial resistance of Parabacteroides to antibiotics (such as clindamycin, moxifloxacin and cefoxitin) should not be ignored. In this review, we primarily focus on Parabacteroides distasonis, Parabacteroides goldsteinii, Parabacteroides johnsonii and Parabacteroides merdae and discuss their relationships with host disease, diet and the prevention or induction of diseases. Pa. distasonis and Pa. goldsteinii may be viewed as potential next generation probiotic candidates due to their protective effects on inflammation and obesity in mice. We also discuss the potential therapeutic application of Parabacteroides spp. in maintaining host-intestine homeostasis.

Lactic acid bacteria isolated from Chinese traditional fermented milk as novel probiotic strains and their potential therapeutic applications.

Lactic acid bacteria (LAB) are believed to have health-promoting properties to the host and can be used in therapeutics interventions; intriguingly, they have the property to produce bio-preservatives substances. Therefore, this study aimed to mine probiotics and evaluate their safety, functional properties, and cholesterol-lowering capability. Seven potential probiotic strains were compared from 56 LAB strains isolated from traditional Chinese fermented milk. The results showed that all tested strains are tolerant to gastric acidity (45.5-83.26) and bile salts (11.92-92.91%) and have antibacterial activity against Staphylococcus aureus ATCC25923 and Escherichia coli ATCC25922. Likewise, it lowered the cholesterol levels in vitro by live cells (26.57-45.76%) and dead cells (29.53-50.97%) with remarkable aggregation ability (13.8-43.71%). Antioxidant properties and produce short chain fatty acids (SCFAs) were strain-dependent features. Upon assessment of the safety, Enterococcus faecium NWAFU-BIO-AS14 exhibited virulence factors genes (VFs) of (mur-2ed, odc, and tet(K)) and + hemolysis activity. While Enterococcus faecium NWAFU-BIO-A-B24 and Limosilactobacillus fermentum NWAFU-BIO-B-S6 have VFs of (odc, vanC2, and ant(6)-Ia). Limosilactobacillus fermentum NWAFU-BIO-D-B2 has only (odc). Thus, they are not considered as safe probiotics. In contrast, Lactiplantibacillus plantarum NWAFU-BIO-BS29, Companilactobacillus crustorum NWAFU-BIO-AS16, and Lactobacillus gallinarum NWAFU-BIO-D-S7 are the safest and best strains, respectively, due to the absence of 16 VFs and their sensitivity to antibiotics such as kanamycin, erythromycin, tetracycline, gentamycin, vancomycin, streptomycin, chloramphenicol, and ampicillin. Accordingly, these strains have a high potentiality to be used as starter cultures or safely applied as perfect probiotics in functionals food and feed. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03403-z.

Probiotic of Lactiplantibacillus plantarum NWAFU-BIO-BS29 Isolated from Chinese Traditional Fermented Milk and Its Potential Therapeutic Applications Based on Gut Microbiota Regulation.

Lactic acid bacteria are one of the bioresources that can promote the host's health and have potential therapeutic applications. This study aimed to evaluate the probiotic properties of novel Lactiplantibacillus plantarum NWAFU-BISO-BS29 isolated in vitro from traditional Chinese fermented milk, assess its safety, and study its interaction with the gut microbiota using a BALB/c mouse model. The findings reveal that this strain had a high tolerance to gastric acidity (64.4%) and bile salts (19.83-87.92%) with remarkable auto-aggregation and co-aggregation abilities (33.01-83.96%), respectively. Furthermore, it lowered the cholesterol levels in dead cells (44.02%) and live cells (34.95%) and produced short-chain fatty acids (SCFAs). Likewise, it showed good antioxidant properties and strong antipathogen activity against Escherichia coli and Staphylococcus aureus with inhibition zones at 21 and 25 mm, respectively. The safety assessment results indicate that all of the virulence factor genes were not detected in the whole DNA; additionally, no hemolysis or resistance to antibiotics commonly used in food and feed was observed. Interestingly, the 16S rRNA gene sequencing of the mouse gut microbiota showed a marked alteration in the microbial composition of the administrated group, with a noticeable increase in Firmicutes, Patescibacteria, Campylobacterota, Deferribacterota, Proteobacteria, and Cyanobacteria at the phylum level. The modulation of gut microbial diversity significantly improved the production of SCFCs due to the abundance of lactobacillus genera, which was consistent with the functional gene predictive analysis and is believed to have health-promoting properties. Based on these results, our novel strain is considered a safe and good probiotic and could hold high potential to be used as a starter culture or to safely supplement functional foods as a probiotic and may provide new insights into therapeutic interventions.

Extraction optimization and characterization of persimmon peel pectin extracted by subcritical water.

Persimmon peel pectin (PPP) was extracted by subcritical water via the response surface methodology. The optimal crude PPP extraction yield of 7.62 ± 0.7 % was found at 138 °C, 2.84 min, and liquid-solid ratio of 1:10.02. After treatment of deproteinization and decolorization with papain and hydrogen peroxide, 83.19 % of protein and 78.56 % of the colour in crude PPP were removed, respectively. PPP owned the Mw of 21.79 kDa and its uronic acids content was 64.03 %. PPP was further affirmed by fourier transform infrared, X-ray diffractometer and 1H NMR analysis. Moreover, the degradation temperature (228.05 â„ƒ) of PPP was verified via differential scanning calorimetry. Then, the IC50 of PPP to ABTS•+ was 9.8 times that of commercial citrus pectin. Moreover, PPP could change microbial communities and selectively enrich Bacteroides, Cetobacterium, Erysipelatoclostridium, Parabacteroides and Phocaeicola sartorii. This study demonstrated that subcritical water was practicable for extraction of persimmon peel pectin.

Purification, characterization and bioactivity of exopolysaccharides produced by Lactobacillus plantarum KX041.

A novel crude exopolysaccharide (EPS) produced by Lactobacillus plantarum KX041 possessed prominent antioxidant activity which was proved in our previous study. In our present study, the further purifications were conducted to obtain EPS fractions, which were called as EPS-1-1, EPS-2-1 and EPS-3. The structures and conformational characterizations were determined through FT-IR, UV, GC, HPLC, NMR, SEM and Congo red test analysis. The Mw of EPS-1-1, EPS-2-1 and EPS-3 were estimated to be 57,201, 70,734, and 26,387 Da, respectively. EPS-1-1 and EPS-2-1 had the similar structure, composed of arabinose, mannose, glucose and galactose with a ratio of 1.09:88.53:3.99:6.39 and 0.58:94.11:3.55:1.76, and both were α-type configurations. Whereas EPS-3 contained rhamnose, fucose, arabinose, xylose, mannose, glucose, galactose and galacturonic acid in a molar ratio of 2.01:2.65:10.95:4.62:4.07:27.81:44.16:3.73 and possessed a triple helical structure. SEM results indicated EPS-1-1 and EPS-2-1 appeared flake shapes piling up into compact structures with a rough surface. In addition, these purified EPS fractions all possessed the immune activity, DPPH/ABTS radicals scavenging activities and DNA damage protective effect.