Antifungal Activity of Lactobacillus plantarum ZZUA493 and Its Application to Extend the Shelf Life of Chinese Steamed Buns

Lactic acid bacteria (LAB) can produce many kinds of antifungal substances, which have been widely proven to have antifungal activity. In this study, 359 strains of LAB were screened for antifungal activity against Aspergillus niger (A. niger) using the 96-well microtiter plate method, and three showed strong activity. Of these, ZZUA493 showed a broad-spectrum antifungal ability against A. niger, Aspergillus oryzae, Trichoderma longibrachiatum, Aspergillus flavus and Fusarium graminearum. ZZUA493 was identified as Lactobacillus plantarum. Protease treatment, the removal of hydrogen peroxide with catalase and heat treatment had no effect on the antifungal activity of the cell-free supernatant (CFS) of ZZUA493; organic acids produced by ZZUA493 appeared to have an important role in fungal growth inhibition. The contents of lactic acid, acetic acid and phenyllactic acid in the CFS tended to be stable at 48 h, and amounted to 28.5, 15.5 and 0.075 mg/mL, respectively. In addition, adding ZZUA493, as an ingredient during their preparation, prolonged the shelf life of Chinese steamed buns. Overall, ZZUA493 appears to have good potential as a fungal inhibitor for food preservation.

Effect of Brine fermented Pickling to Physicochemical, Anti-nutritional, and Microbiological Attributes of Pickled gboma Eggplant (Solanum macrocarpon)

Gboma eggplant (Solanum macrocarpon) has low calorie, high phenolic and antioxidant, excellent dietary fibre and mineral content. Besides healthy effects, gboma eggplant also contained numerous anti-nutrients unbeneficial for human health. Due to high moisture content, gboma eggplant was highly perishable during post-harvest. This research observed the changes of nutritional proximate, mineral contents, acidification indexes, anti-nutrients, phytochemical and antioxidant properties, texture profiles, microbiological characteristics of both raw and pickled gboma eggplant. Gboma eggplant fruits were soaked in clean water for 2 minutes before cutting their calyx lobes. The pre-treated fruits were submerged in sterilized brine (5% salt) for 8 days. Periodically, pickled samples were taken to examine physicochemical, anti-nutritional, and microbiological attributes of pickled gboma eggplant. Results showed that nutritional proximate of moisture, ash, protein, fat, fibre, carbohydrate was varied slightly during fermentation. There was a decreasing trend of moisture and carbohydrate; meanwhile, there was an increasing trend of ash, protein, fibre during 8 days of fermentation. There was no significant difference of fat during pickling. There was a minor increment of mineral contents in all samples. There was accumulation of phenyllactic acid, ascorbic acid content and titratable acidity with exception of pH. Anti-nutrient contents like tannin, phytate, oxalate, steroidal glycoalkaloid greatly decreased in raw and pickled eggplant. Remarkable increments of total phenolic, flavonoid, DPPH free radical scavenging and FRAP ferric reducing antioxidant of raw and pickled eggplant was presented. Texture profiles of hardness, crispness, fracturability, crunchiness revealed a minor reduction of sensory scores during 8 days of fermentation. Lactic acid bacteria, Bacillus significantly proliferated; meanwhile, Micrococcus and S. aureus were absolutely retarded in pickled eggplant. Yeast and fungi increased in the first 4 days and decreased afterwards. There was no significant difference of proximate compositions; mineral contents; phenyllactic acid, ascorbic acid content, pH and titratable acidity; anti-nutritional contents; phytochemical and antioxidant properties; texture profile (sensory score); microbiological load (with exception of S. aureus) between pickled gloma eggplants at day 6th and day 8th of fermentation. At a quick glance, the length of fermentation could be shorted to 6 days instead of 8 days. However, at the 6th day, S. aureus load was still presented at 0.33±0.01 log CFU/g. The fermentation should be lasted to 8th day so that S. aureus load could be dropped down to zero to ensure microbial food safety. Raw gboma eggplant should be fermented in 8% brine solution for 8 days to obtain the best physicochemical, anti-nutritional, and microbiological properties of pickle. Findings of this research suggested that fermentation pickling would be an appropriate approach to improve nutritional, physicochemical and functional criteria while controlling toxic chemical residues, retarding the growth and proliferation of spoilage and pathogen microorganisms.

Using Unnatural Protein Fusions to Engineer a Coenzyme Self-Sufficiency System for D-Phenyllactic Acid Biosynthesis in Escherichia coli

The biosynthetic production of D-penyllactic acid (D-PLA) is often affected by insufficient supply and regeneration of cofactors, leading to high production cost, and difficulty in industrialization. In this study, a D-lactate dehydrogenase (D-LDH) and glycerol dehydrogenase (GlyDH) co-expression system was constructed to achieve coenzyme NADH self-sufficiency and sustainable production of D-PLA. Using glycerol and sodium phenylpyruvate (PPA) as co-substrate, the E. coli BL21 (DE3) harboring a plasmid to co-express LfD-LDH and BmGlyDH produced 3.95 g/L D-PLA with a yield of 0.78 g/g PPA, similar to previous studies. Then, flexible linkers were used to construct fusion proteins composing of D-LDH and GlyDH. Under the optimal conditions, 5.87 g/L D-PLA was produced by expressing LfD-LDH-l3-BmGlyDH with a yield of 0.97 g/g PPA, which was 59.3% increased compared to expression of LfD-LDH. In a scaled-up reaction, a productivity of 5.83 g/L/h was reached. In this study, improving the bio-catalytic efficiency by artificial redox self-equilibrium system with a bifunctional fusion protein could reduce the bio-production cost of D-PLA, making this bio-production of D-PLA a more promising industrial technology.

Effects of Phenyllactic Acid, Lactic Acid Bacteria, and Their Mixture on Fermentation Characteristics and Microbial Community Composition of Timothy Silage

This study investigated the effects of phenyllactic acid (PL), lactic acid bacteria (LAB), and their mixture on fermentation characteristics and microbial community composition of timothy silage. Timothy silages were treated without (CK) or with PL [10 mg/kg fresh matter (FM) basis], LAB inoculant (IN; a mixture of Lactobacillus plantarum and L.buchneri, 105 cfu/g FM), and their mixture (PI) and stored at ambient temperature (5°C∼15°C) in a dark room for 60 days. Compared with CK, all treated silages showed lower (P < 0.05) levels of butyric acid and ammonia-N. Treatment with PL enhanced (P < 0.05) the crude protein preservation of silage by favoring the growth of L. curvatus and Saccharomyces cerevisiae and inhibition of lactic acid–assimilating yeast belonging to Issatchenkia during ensiling. In particular, treatment with PL advanced (P < 0.05) the productions of lactic acid and volatile fatty acid in IN-treated silage. Therefore, PL used as a new additive exhibited potential for improving silage fermentation when it is combined with LAB IN during ensiling.

Bacterial vaginosis and health-associated bacteria modulate the immunometabolic landscape in 3D model of human cervix

Bacterial vaginosis (BV) is an enigmatic polymicrobial condition characterized by a depletion of health-associated Lactobacillus and an overgrowth of anaerobes. Importantly, BV is linked to adverse gynecologic and obstetric outcomes: an increased risk of sexually transmitted infections, preterm birth, and cancer. We hypothesized that members of the cervicovaginal microbiota distinctly contribute to immunometabolic changes in the human cervix, leading to these sequelae. Our 3D epithelial cell model that recapitulates the human cervical epithelium was infected with clinical isolates of cervicovaginal bacteria, alone or as a polymicrobial community. We used Lactobacillus crispatus as a representative health-associated commensal and four common BV-associated species: Gardnerella vaginalis, Prevotella bivia, Atopobium vaginae, and Sneathia amnii. The immunometabolic profiles of these microenvironments were analyzed using multiplex immunoassays and untargeted global metabolomics. A. vaginae and S. amnii exhibited the highest proinflammatory potential through induction of cytokines, iNOS, and oxidative stress-associated compounds. G. vaginalis, P. bivia, and S. amnii distinctly altered physicochemical barrier-related proteins and metabolites (mucins, sialic acid, polyamines), whereas L. crispatus produced an antimicrobial compound, phenyllactic acid. Alterations to the immunometabolic landscape correlate with symptoms and hallmarks of BV and connected BV with adverse women’s health outcomes. Overall, this study demonstrated that 3D cervical epithelial cell colonized with cervicovaginal microbiota faithfully reproduce the immunometabolic microenvironment previously observed in clinical studies and can successfully be used as a robust tool to evaluate host responses to commensal and pathogenic bacteria in the female reproductive tract.