Process Optimization and Antimicrobial Effect of <i>Cytobacillus kochii</i> H Protease

WANG Xuewen; YU Cun; PAN Hongxiang

doi:10.19303/j.issn.1008-0384.2023.01.007

Volume 38 Issue 1

Jan. 2023

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WANG X W, YU C, PAN H X. Process Optimization and Antimicrobial Effect of Cytobacillus kochii H Protease [J]. Fujian Journal of Agricultural Sciences，2023，38（1）：47−57 doi: 10.19303/j.issn.1008-0384.2023.01.007

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WANG X W, YU C, PAN H X. Process Optimization and Antimicrobial Effect of Cytobacillus kochii H Protease [J]. Fujian Journal of Agricultural Sciences，2023，38（1）：47−57 doi: 10.19303/j.issn.1008-0384.2023.01.007

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Process Optimization and Antimicrobial Effect of Cytobacillus kochii H Protease

doi: 10.19303/j.issn.1008-0384.2023.01.007

Forestry College, Guizhou University, Guiyang, Guizhou　550025, China

Received Date: 2022-08-29
Rev Recd Date: 2022-11-21

Available Online: 2023-02-08

Publish Date: 2023-01-28

Abstract

Abstract

Objective Taxonomy of antimicrobial protease-producing Cytobacillus kochii H was studied for the development of a natural disease control agent on plants. Method Taxonomy of C. kochii H was determined by physiological, biochemical, electron microscopy scanning, and 16sRNA sequencing methods. A process of producing the antimicrobial protease secreted by the bacterium was optimized in single-factor and orthogonal experiments. The efficacy of the culture broth on 4 selected pathogenic fungi was applied in an in vitro test as the evaluation criterium. Result The optimized C. kochii H protease-producing process was conducted in a 250 mL triangular flask using 50 mL of pH 8.0 medium, which contained 20.0 g·L⁻¹ of glucose, 8.0 g·L⁻¹ of peptone, 1.0 g·L⁻¹ of MgSO₄, and 0.1g·L⁻¹ of CaSO₄-2H₂O, inoculated with 2.5 mL of 10⁸ CFU·mL⁻¹ C. kochii H fermentation liquid. After 24 h of incubation, the protease activity reached 402.2 U·mL⁻¹, which was 13.92 times greater than it was prior to the optimization. The antimicrobial rates of the broth against Fusarium oxysporum and Phytophthora capsici were 67.32% and 44.87%, respectively, i.e., 9.15% and 12.82%, respectively, increases over those before the optimization. Conclusion The antimicrobial protease-producing C. kochii H was identified taxonomically. The culture process of the enzyme production was optimized to deliver significant in vitro inhibition rates on F. oxysporum and P. capsici.
- Cytobacillus kochii,
- protease,
- optimized fermentation,
- antimicrobial rate on pathogens

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References(27)

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