Improvement of Bacillus clausii cell HH1 density via culture medium optimization and pH-stat fed batch fermentation

Tuan Anh Pham; Thi Phuong Nguyen; Kim Anh To

doi:10.15625/2525-2518/17250

Author affiliations

Authors

Tuan Anh Pham School of Biotechnology and Food Technology, Hanoi University of Science and Technology, No. 1, Dai Co Viet Road, Hai Ba Trung, Ha Noi, Viet Nam;
Laboratory of Applied Miro-organisms, Hanoi University of Science and Technology, No. 1, Dai Co Viet Road, Hai Ba Trung, Ha Noi, Viet Nam
Thi Phuong Nguyen School of Biotechnology and Food Technology, Hanoi University of Science and Technology, No. 1, Dai Co Viet Road, Hai Ba Trung, Ha Noi, Viet Nam
Kim Anh To School of Biotechnology and Food Technology, Hanoi University of Science and Technology, No. 1, Dai Co Viet Road, Hai Ba Trung, Ha Noi, Viet Nam

DOI:

https://doi.org/10.15625/2525-2518/17250

Keywords:

Bacillus clausii, culture medium optimization, pH-stat fed batch fermentation

Abstract

In this study, the culture medium and fermentation modes were studied aiming to improve the cell density of Bacillus clausii. Firstly, the factorial design method using Minimum Run Resolution IV design was used to evaluate the relative importance of culture medium components to the growth of Bacillus clausii. The results showed that three components peptone, yeast extract and malt extract were the components significantly affecting the bacterial biomass. Then, the optimization of these three ingredient concentrations using a response surface methodology with the Box-Behnken design resulted that the maximal biomass had been achieved using the medium containing 7.64 g/L peptone, 10 g/L yeast extract and 6.36 g/L malt extract. Finally, the pH-stat fed batch fermentation was conducted in a 2-liter bioreactor where the 9X concentrated optimal culture medium was feed into the bioreactor based on the pH signal. As a result, the microbial cell density increased by 2.9-fold compared to that achieved through batch fermentation.

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References

Suresh K., Krishna S., Govender P. - The use of probiotics and safety concerns: A review. Afr. J. Microbiol. Res., 6 (2012) 6871-6877. DOI: https://doi.org/10.5897/AJMR12.1281

Urdaci M. C., Bressollier P., Pinchuk I. - Bacillus clausii probiotic strains: antimicrobial and immunomodulatory activities. J. Clin. Gastroenterol, 38 (2004) 86-90. DOI: https://doi.org/10.1097/01.mcg.0000128925.06662.69

Abbrescia A., Palese L. L., Papa S., Gaballo A., Alifano P., Sardanelli A. - Antibiotic sensitivity of Bacillus clausii strains in sommercial preparation. Clin. Immunol. Endocr. Metab. Drugs, 1 (2015) 102-110. DOI: https://doi.org/10.2174/2212707002666150128195631

de Castro J.-A. A., Guno M. J. V.-R., Perez M. O. - Bacillus clausii as adjunctive treatment for acute community-acquired diarrhea among Filipino children: a large-scale, multicenter, open-label study (CODDLE). Trop. Dis. Travel Med. Vaccines., 5 (2019) 14-14. DOI: https://doi.org/10.1186/s40794-019-0089-5

Fakhry S., Sorrentini I., Ricca E., De Felice M., Baccigalupi L. - Characterization of spore forming Bacilli isolated from the human gastrointestinal tract. J. Appl. Microbiol., 105 (2008) 2178-2186. DOI: https://doi.org/10.1111/j.1365-2672.2008.03934.x

Senesi S., Celandroni F., Tavanti A., Ghelardi E. - Molecular characterization and identification of Bacillus clausii strains marketed for use in oral bacteriotherapy. Appl. Environ. Microbiol., 67 (2001) 834-839. DOI: https://doi.org/10.1128/AEM.67.2.834-839.2001

Singh V., Haque S., Niwas R., Srivastava A., Pasupuleti M., Tripathi C. K. M. - Strategies for fermentation medium optimization: an in-depth review. Front. Microbiol., 7 (2017) DOI: https://doi.org/10.3389/fmicb.2016.02087

Sreekumar G., Krishnan S. - Enhanced biomass production study on probiotic Bacillus subtilis SK09 by medium optimization using response surface methodology. Afr. J. Biotechnol., 9 (2010) 8078-8084. DOI: https://doi.org/10.5897/AJB10.1283

Zhong J., Zhang X., Ren Y., Yang J., Tan H., Zhou J. - Optimization of Bacillus subtilis cell growth effecting jiean-peptide production in fed batch fermentation using central composite design. Electron. J. Biotechnol., 17 (2014) 132-136. DOI: https://doi.org/10.1016/j.ejbt.2014.04.010

Levisauskas D., Galvanauskas V., Simutis R., Zilinskas A. - Optimization of biomass production in fed-batch culture by feed and dilution control actions. Inf. Technol. Control., 35 (2006) 383-390.

Suzuki T., Yamane T., Shimizu S. - Phenomenological background and some preliminary trials of automated substrate supply in pH-Stat modal fed-batch culture using a setpoint of high limit. J. Fermentat. Bioeng., 69 (1990) 292-297.

Cho Y. H., Song J. Y., Kim K. M., Kim M. K., Lee I. Y., Kim S. B., Kim H. S., Han N. S., Lee B. H., Kim B. S. - Production of nattokinase by batch and fed-batch culture of Bacillus subtilis. N Biotechnol, 27 (2010) 341-346. DOI: https://doi.org/10.1016/j.nbt.2010.06.003

Kim B. S., Lee S. C., Lee S. Y., Chang Y. K., Chang H. N. - High cell density fed-batch cultivation of Escherichia coli using exponential feeding combined with pH-stat. Bioprocess Biosys. Eng., 26 (2004) 147-150. DOI: https://doi.org/10.1007/s00449-003-0347-8

Sousa C., Klainer B., Lima K., Pinto G. - Biomass production from Bacillus sp. RAB9 using several carbon sources. BMC Proc. 2014 Oct 1;8(Suppl 4):P172. doi: 10.1186/1753-6561-8-S4-P172., - P172. DOI: https://doi.org/10.1186/1753-6561-8-S4-P172

Koim-Puchowska B., Kłosowski G., Dróżdż-Afelt J. M., Mikulski D., Zielińska A. - Influence of the medium composition and the culture conditions on surfactin biosynthesis by a native Bacillus subtilis natto BS19 strain. Molecules, 26 (2021) DOI: https://doi.org/10.3390/molecules26102985

Mosquera S., González-Jaramillo L. M., Orduz S., Villegas-Escobar V. - Multiple response optimization of Bacillus subtilis EA-CB0015 culture and identification of antifungal metabolites. Biocatal. Agric. Biotechnol., 3 (2014) 378-385. DOI: https://doi.org/10.1016/j.bcab.2014.09.004

Ojha S. K., Singh P. K., Mishra S., Pattnaik R., Dixit S., Verma S. K. - Response surface methodology based optimization and scale-up production of amylase from a novel bacterial strain Bacillus aryabhattai KIIT BE-1. Biotechnol. Rep., 27 (2020) e00506. DOI: https://doi.org/10.1016/j.btre.2020.e00506

Tabandeh F., Hosseinian Moghaddam H. R., Yakhchali B., Shariati P., Hamed Mousavian M. T., Ghasemi F. - Fed-batch fermentation of Bacillus clausii for efficient production of alkaline protease using different feeding strategies. Chem. Eng. Commun., 198 (2011) 1063-1074. DOI: https://doi.org/10.1080/00986445.2011.552024

Wang C. K., Duan K. J., Yeh K. W., Chen W. C. - Production of a fusion protein of sweet potato sporamin from recombinant E. coli XL1 Blue by fed-batch fermentations. Biotechnol. Lett., 23 (2001) 475-479.

Suzuki T., Yamane T., Shimizu S. - Phenomenological background and some preliminary trials of automated substrate supply in pH-stat modal fed-batch culture using a setpoint of high limit. J. Ferment. Bioeng., 69 (1990) 292-297. DOI: https://doi.org/10.1016/0922-338X(90)90108-9

Monteiro S., Clemente J., Cunha A. - Enhanced spore production of Bacillus subtilis grown in a chemically defined medium. Adv. Microbiol., 04 (2014) 444-454. DOI: https://doi.org/10.4236/aim.2014.48049