Probiotic potential of riboflavin-overproducing Bacillus subtilis ACU-I163MR and ACU-I11MR, isolated from fermented African locust beans
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Riboflavin (vitamin B 2 ) is a water-soluble compound that plays an important role in multiple cellular functions. This study evaluates the probiotic potential of riboflavin-overproducing Bacillus subtilis strains isolated from fermented African locust beans. After strain improvement, B. subtilis ACU-I11MR and ACU-I163MR were selected due to their higher riboflavin production (0.01905±0.0005 mg l −1 to 0.0259±0.0077 mg l −1 and 0.0195±0.0054 mg l −1 to 0.0267±0.0013 mg l −1 , respectively). Their safety was confirmed through haemolytic assay, antibiotic susceptibility tests and the absence of gelatinase and biogenic amine activity. Probiotic potential was assessed via in vitro assays including resistance to low pH, bile salts, phenol, temperature and NaCl; auto-aggregation; cell hydrophobicity; biofilm formation; antibacterial activity; and enzyme and exopolysaccharide production. Both strains were non-haemolytic and negative for gelatinase and biogenic amine activity. They showed significant viability at pH 2 (survival 85.05; 87.09%), 1% bile salts (survival 88.82; 87.64%) and 0.5% phenol (survival 48.80; 59.52%), respectively. ACU-I11MR was susceptible to 9 out of 12 antibiotics, while ACU-I163MR was 100% susceptible. The strains demonstrated strong cell surface adhesion and auto-aggregation and inhibited several pathogenic bacteria. They produce amylase, protease and exopolysaccharide and thrive under various temperature and NaCl conditions. B. subtilis ACU-I163MR, showing superior probiotic potential, could be a promising candidate for developing riboflavin-enriched Bacillus -fermented functional foods.
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I am pleased to tell you that your article has now been accepted for publication in Access Microbiology. This study would be a valuable contribution to the existing literature and will be of interest to the field and community. The necessary corrections requested by reviewers have been made, including addition of statistical rigour in the methods section. I can confirm the sequence data is publicly accessible.
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The paper is well-written and interesting, focusing on the riboflavin biosynthesis potential of a Bacillus subtilis strain. The literature background is relevant and adequate, the methodology is sound and the findings reflect this paper's conclusions. Some minor errors: 1. Line 72: replace 'the overproduce' with 'that overproduce' 2. Methods/Results: Indicate number of biological replicates for each experiment and provide statistical p-values for each comparison. 3. Discussion: what other species produce riboflavin at industrially-relevant level? why is your strain equally good or better compared to known industrial species that produce riboflavin? 4. Discussion: highlight the industrial and/or medical importance of your findings. What new applications and products can be designed?
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Comments to Author
Comments to the Authors Below are my comments: 1. In line 72, the statement "the strains the overproduce riboflavin" is probably a typographical error; it is meant to be 'the strains that overproduce riboflavin.' 2. Across the "Result" sections, the authors did not indicate the number of replicates for their experiments, making the conclusions about statistical significance unclear. For instance, under the "Bile Salts result on lines 295-302, the authors reported statistical significance between survival rates of the two B. subtilis isolates, stating, "However, statistically, B. subtilis ACU-I163MR had a higher survival rate (p
Please rate the manuscript for methodological rigour
Good
Please rate the quality of the presentation and structure of the manuscript
Go…
Comments to Author
Comments to the Authors Below are my comments: 1. In line 72, the statement "the strains the overproduce riboflavin" is probably a typographical error; it is meant to be 'the strains that overproduce riboflavin.' 2. Across the "Result" sections, the authors did not indicate the number of replicates for their experiments, making the conclusions about statistical significance unclear. For instance, under the "Bile Salts result on lines 295-302, the authors reported statistical significance between survival rates of the two B. subtilis isolates, stating, "However, statistically, B. subtilis ACU-I163MR had a higher survival rate (p
Please rate the manuscript for methodological rigour
Good
Please rate the quality of the presentation and structure of the manuscript
Good
To what extent are the conclusions supported by the data?
Strongly support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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