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Optimization of Spore Production in Bacillus coagulans Using Response Surface Methodology Approach

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Abstract

Due to its spore-forming ability, Bacillus coagulans has advantages over the other non-spore-forming probiotics. Among them, survival and stability during food processing and storage, resistance to acid pH, and digestive enzymes are important. However, there are few studies on the quality and amount of sporulation in B. coagulans. This study investigated the spore densities and formation efficiency of B. coagulans. The optimal medium formulation consisted of yeast extract (1.00 g L−1), potassium acetate (20.00 g L−1), and MnSO4 (0.01 g L−1 and 0.03 g L−1). After reaching the optimal medium, a response surface regression equation was established based on the results of central composite design (CCD) experimental designs to optimize time, temperature, and pH parameters. The predicted results thus obtained were in good agreement (R2 = 95.19%) with the results obtained by performing experiments. Multiple regression analysis and analysis of variance (ANOVA) showed that pH is negative, and temperature and time dose are positive factors. The maximum spore cell densities by optimization plots have obtained 9.80 log at temperature 83.77 °C, pH 3.05, and time 111.19 h, considering that B. coagulans needs special environmental and cellular conditions to enter the sporulation stage. In this study, the composition of the culture medium and factors such as temperature, time, and pH were considered influencing factors in B. coagulans sporulation.

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All generated or analyzed data as well as the utilized software and materials have been included in the published article. The generated results during the study are available from the corresponding authors upon reasonable request.

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Acknowledgements

This study was a part of Seyedeh Habibeh Mirmajidi’s PhD dissertation proposed and approved in the Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran.

Funding

This work was supported by the Vice-Chancellor for Research and Technology of Shiraz University of Medical Sciences, Shiraz, Iran (grant No. 22866).

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Authors

Contributions

Seyedeh Habibeh Mirmajidi has performed the experimental analysis study. Seyedeh Habibeh Mirmajidi and Mohammad Hossein Morowvat have carried out the analysis and interpretation of the results. Cambyz Irajie, Mohammad Hossein Morowvat, and Seyedeh Habibeh Mirmajidi have designed the study concept and interpreted the data. Navid Nezafat, Younes Ghasemi, and Amir Savardashtaki read and revised the manuscript and approved the final version.

Corresponding authors

Correspondence to Mohammad Hossein Morowvat or Younes Ghasemi.

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The study was approved by the ethics committee of Shiraz University of Medical Sciences, Shiraz, Iran (IR.SUMS.REC.1401.153). This article does not contain any studies with human participants or animals performed by any of the authors.

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Not applicable.

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The authors declare no competing interests.

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Mirmajidi, S.H., Irajie, C., Savardashtaki, A. et al. Optimization of Spore Production in Bacillus coagulans Using Response Surface Methodology Approach. Appl Biochem Biotechnol (2024). https://doi.org/10.1007/s12010-024-04934-2

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