Effects of ageing time on the properties of polysaccharide in tangerine peel and its bacterial community.

To evaluate the effect of aging time on the quality of tangerine peel (TP) from the perspective of TP polysaccharide (TPP), five polysaccharide samples with different aging times named TPP-0/1/5/10/15 were prepared. Under the conditions of pH 0.5, solid-liquid ratio 1:25 and 80 °C, the TPPs extraction yield ranged from 20.35% to 27.68%. Compared with TPP-0, TPP-1/5/10/15 possesses low molecular weight (Mw) and high methoxy group content. In addition, TPP-15 had the most potent antioxidant activity. And the content of acidic polysaccharides in TPPs was negatively correlated with neutral polysaccharides during aging. Based on the analysis of 16srDNA, the dominant bacteria (Brevundimonas and Pseudomonas) in TP-10 might be critical flora to affect TP quality. This study provided basic information on the relationship between the TPPs and aging time, which could promote a new view to develop TP, and shorten the aging time during TP production.

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.

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.

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.

Effects of monosaccharide composition on quantitative analysis of total sugar content by phenol-sulfuric acid method.

Phenol-sulfuric acid method is one of the most common methods applied to the analysis of total sugar content during polysaccharides study. However, it was found that the results obtained from the phenol-sulfuric acid method was generally lower than the real total sugar content, especially when acidic monosaccharides were contained in the polysaccharides samples. Therefore, the present study focused to unveil the proposed problem. Based on the optimization of colorimetric conditions, such as optimal wave length of absorption, linearity range, color reaction time and temperature, it indicated that the phenol-sulfuric acid method was a convenient and accurate way for the total sugar content analysis. In addition, the color-rendering capabilities of 10 common monosaccharides were systematically analyzed to obtain a relative correction factor for each monosaccharide relative to glucose, which was proved to be the main reason for the deviation in the detection of total sugar content. Moreover, the key points during the application of phenol-sulfuric acid method were suggested. This study provides a scientific theoretical basis and a reliable experimental research method for the accurate determination of total sugar content by the phenol-sulfuric acid method, and which will also promote the application of this convenient method in the polysaccharides study.

Structural features and anticancer mechanisms of pectic polysaccharides: A review.

The anticancer activity of pectic polysaccharides (PPs) was proved by numerous studies, and which also indicated that the bioactivity of PPs was closely related to its complicated structures. Based on the summary and analysis about structure characteristics and corresponding enzymatic process of the reported PPs, the anticancer mechanism and related structural features were systematically clarified. It was found that not only the direct effects on the cancer cells by proliferation inhibition or apoptosis, but also the regulation of immune system, gut microbiota and gut metabolism as indirect effects, jointly played important roles in the anticancer of PPs. Nevertheless, during the study of PPs as promising anticancer components, the exact structure-function relationship, digestion process in vivo, and comprehensive action mechanism are still not well understanding. With the unveiling of the proposed issues, it is believed that PPs are promising to be exploited as effective cancer therapy/adjunctive therapy drugs or functional foods.

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.

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.

Effect of growth phase feeding strategies on succinate production by metabolically engineered Escherichia coli.

Aerobic growth conditions significantly influenced anaerobic succinate production in two-stage fermentation by Escherichia coli AFP111 with knockouts in rpoS, pflAB, ldhA, and ptsG genes. At a low cell growth rate limited by glucose, enzymes involved in the reductive arm of the tricarboxylic acid cycle and the glyoxylate shunt showed elevated activities, providing AFP111 with intracellular redox balance and increased succinic acid yield and productivity.