Rare ginsenoside Ia synthesized from F1 by cloning and overexpression of the UDP-glycosyltransferase gene from Bacillus subtilis: synthesis, characterization, and in vitro melanogenesis inhibition activity in BL6B16 cells

Background:: Antifungal cyclic lipopeptides, bioactive metabolites produced by many species of the genus Bacillus, are promising alternatives to synthetic fungicides and antibiotics for the biocontrol of human pathogenic fungi. In a previous study, the co- production of five antifungal lipopeptides homologues (designated as AF1, AF2, AF3, AF4 and AF5) by the producer strain Bacillus subtilis RLID 12.1 using unoptimized medium was reported; though the two homologues AF3 and AF5 differed by 14 Da and in fatty acid chain length were found effective in antifungal action, the production/ yield rate of these two lipopeptides determined by High-Performance Liquid Chromatography was less in the unoptimized media. Methods:: In this study, the production/yield enhancement of the two compounds AF3 and AF5 was specifically targeted. Following the statistical optimization (Plackett-Burman and Box-Behnken designs) of media formulation, temperature and growth conditions, the production of AF3 and AF5 was improved by about 25.8- and 7.4-folds, respectively under static conditions. Results:: To boost the production of these two homologous lipopeptides in the optimized media, heat-inactivated Candida albicans cells were used as a supplement resulting in 34- and 14-fold increase of AF3 and AF5, respectively. Four clinical Candida auris isolates had AF3 and AF5 MICs (100 % inhibition) ranging between 4 and 16 μg/ml indicating the lipopeptide’s clinical potential. To determine the in vitro pharmacodynamic potential of AF3 and AF5, time-kill assays were conducted which showed that AF3 (at 4X and 8X concentrations) at 48h exhibited mean log reductions of 2.31 and 3.14 CFU/ml of C. albicans SC 5314, respectively whereas AF5 at 8X concentration showed a mean log reduction of 2.14 CFU/ml. Conclusion:: With the increasing threat of multidrug-resistant yeasts and fungi, these antifungal lipopeptides produced by optimized method promise to aid in the development of novel antifungal that targets disease-causing fungi with improved efficacy.

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Isolation and Identification of a Potent PTP1B Inhibitor, Ursolic Acid, from Carolina Jasmine (Gelsemium sempervirens (L.) J.St.-Hil.)

Letters in Organic Chemistry ◽

10.2174/1570178617666200409101248 ◽

2020 ◽

Vol 17 (12) ◽

pp. 939-943

Author(s):

Toshiro Noshita ◽

Yusuke Kakizoe ◽

Satoshi Tanabe ◽

Hidekazu Ouchi ◽

Akihiro Tai

Keyword(s):

Ursolic Acid ◽

Protein Tyrosine Phosphatase ◽

Tyrosine Phosphatase ◽

Active Agent ◽

Protein Tyrosine Phosphatase 1B ◽

Inhibiting Activity ◽

Inhibition Activity ◽

Isolation And Identification ◽

Gelsemium Sempervirens

Extracts of Carolina jasmine (Gelsemium sempervirens (L.) J.St.-Hil.) petals were evaluated in vitro for inhibition activity against protein tyrosine phosphatase 1B (PTP1B). The principle active agent was also isolated from the extract and identified as ursolic acid (1). This is the first report of ursolic acid from G. sempervirens and of PTP1B-inhibiting activity in the genus Gelsemium.

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Evaluation of Snake Venom’s PhospholipaseA2 Enzyme Inhibition Activity of Cyphostemma adenocoule

Current Chemical Biology ◽

10.2174/2212796814999200917114914 ◽

2020 ◽

Vol 14 (3) ◽

pp. 196-202

Author(s):

Atul Kaushik ◽

Teamrat S. Tesfai ◽

Daniel K. Barkh ◽

Furtuna K. Ghebremeskel ◽

Habtom G. Zerihun ◽

...

Keyword(s):

Enzyme Inhibition ◽

Snake Venom ◽

Ethanol Extract ◽

Egg Yolk ◽

Chloroform Extract ◽

Inhibition Activity ◽

Traditional Use ◽

Snake Bites ◽

Ethanol Extracts

Background: A snake bite is fundamentally an injury often resulting in puncture wounds meted out by the animal's fangs and occasionally resulting in envenomation. Rate of snake bites around 5,400,000 bites per year leads to over 2,500,000 envenomings and around 125,000 fatal cases annually. Snake venom enzymes are rich in metalloproteinases, phospholipaseA2, proteinases, acetylcholinesterases and hyaluronidases. Objective: Cyphostemma adenocoule is traditionally being used for the treatment of snake bites in Eritrea. The present research was aimed at evaluating the snake venom enzyme inhibition activity of C. adenocoule against puff adder venom and developing a base for the traditional use of the plant against snakebites in Eritrea. Methods: The anti-venom activity of C. adenocoule was assessed in-vitro through phospholipaseA2 enzyme inhibition assay using egg yolk as a cell. The ethanol and chloroform extracts of C. adenocoule showed in vitro anti phospholipase A2 activity, whereas the water extracts of the plant showed no activity. Results: Among the extracts of C. adenocoule, the highest percentage of inhibition was obtained from chloroform extract (95.55% at 100mg/ml). The extract showed prominent activity at different concentrations (34.7% at10mg/ml, 48.8% at 20mg/ml, 54.8% at 40mg/ml, 60.9% at 60mg/ml, 80.5% at 80mg /ml). The ethanol extract also showed certain activity at various concentrations (25.22% at10mg/ml, 14.78% at 20mg/ml, 2.6% at40mg/ml). The activity of the chloroform extracts increases as concentration increases, whereas the activity of the ethanol extracts decreases as concentration increases. The aqueous extract of C. adenocoule did not show any activity at all concentrations. Conclusion: In this study, the chloroform and ethanol extracts of the plant inhibited the enzyme of interest and thus proved the efficacy of anti-snake venom activity of the plant.

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Evaluating the potential of Bacillus subtilis fermented okara as a functional food ingredient through in vitro digestion and fermentation

Food Biotechnology ◽

10.1080/08905436.2021.1909615 ◽

2021 ◽

pp. 1-22

Author(s):

Wai Kit Mok ◽

Yong Xing Tan ◽

Wei Ning Chen

Keyword(s):

Bacillus Subtilis ◽

Functional Food ◽

In Vitro Digestion ◽

Food Ingredient

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Purification and in vitro DNA-binding Specificity of the Bacillus subtilis Phage ϕ105 Repressor

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)63768-8 ◽

1989 ◽

Vol 264 (25) ◽

pp. 14784-14791

Author(s):

L Van Kaer ◽

M Van Montagu ◽

P Dhaese

Keyword(s):

Bacillus Subtilis ◽

Dna Binding ◽

Binding Specificity ◽

Subtilis Phage ◽

Dna Binding Specificity ◽

Bacillus Subtilis Phage

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Safety Assessment of Bacillus subtilis MB40 for Use in Foods and Dietary Supplements

Nutrients ◽

10.3390/nu13030733 ◽

2021 ◽

Vol 13 (3) ◽

pp. 733

Author(s):

Jessica L. Spears ◽

Richard Kramer ◽

Andrey I. Nikiforov ◽

Marisa O. Rihner ◽

Elizabeth A. Lambert

Keyword(s):

Antibiotic Resistance ◽

Bacillus Subtilis ◽

Dietary Supplements ◽

Genome Sequencing ◽

In Silico ◽

Safety Assessment ◽

In Vivo Studies ◽

Strain Identification ◽

Consumer Safety

With the growing popularity of probiotics in dietary supplements, foods, and beverages, it is important to substantiate not only the health benefits and efficacy of unique strains but also safety. In the interest of consumer safety and product transparency, strain identification should include whole-genome sequencing and safety assessment should include genotypic and phenotypic studies. Bacillus subtilis MB40, a unique strain marketed for use in dietary supplements, and food and beverage, was assessed for safety and tolerability across in silico, in vitro, and in vivo studies. MB40 was assessed for the absence of undesirable genetic elements encoding toxins and mobile antibiotic resistance. Tolerability was assessed in both rats and healthy human volunteers. In silico and in vitro testing confirmed the absence of enterotoxin and mobile antibiotic resistance genes of safety concern to humans. In rats, the no-observed-adverse-effect level (NOAEL) for MB40 after repeated oral administration for 14 days was determined to be 2000 mg/kg bw/day (equivalent to 3.7 × 1011 CFU/kg bw/day). In a 28 day human tolerability trial, 10 × 109 CFU/day of MB40 was well tolerated. Based on genome sequencing, strain characterization, screening for undesirable attributes and evidence of safety by appropriately designed safety evaluation studies in rats and humans, Bacillus subtilis MB40 does not pose any human health concerns under the conditions tested.

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