Optimization of liquid fermentation conditions of keratinase produced by Bacillus paramycoides Gxun-30
ZHANG Hongyan1, ZHANG Ni1, YANG Mengying1, LIU Cong2, YANG Lifang2, SHEN Naikun1*, JIANG Mingguo1
1(Guangxi Key Laboratory for Polysaccharide Materials and Modifications, Guangxi Key Laboratory of Microbial plant Resources and Utilization, School of Marine Sciences and Biotechnology, Guangxi University for Nationalities, Nanning 530006, China) 2(Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, School of Chemistry and Chemical Engineering, Guangxi University for Nationalities, Nanning 530006, China)
Abstract: In order to enhance the keratinase activity of marine sourced Bacillus paramycoides Gxun-30, the liquid culture medium and conditions were optimized by single factor experiment and response surface method. First, the effects of feathers concentration, carbon source, nitrogen source, inorganic salts, initial pH, fermentation time and inoculation amount on the keratinase production were investigated by single factor experiments. The optimal results corresponding to each single factor were: feathers concentration 15 g/L, fructose 10 g/L, corn steep liquor 4.0 g/L, initial pH 6.5, CaCl2 0.15 g/L, inoculation volume 2.0%, fermentation time 48 h. Secondly, using Plackett-Burman experiment, the main significant factors were identified as corn steep liquor, CaCl2 and feathers amount. Finally, the optimal levels of these three factors were determined with the steepest ascent experiment and response surface method. The optimal conditions for enzyme production were: corn steep liquor 8.17 g/L, CaCl2 0.27 g/L, and feathers amount 13.58 g/L. Under the optimized conditions, the keratinase activity was predicted as 1 866.47 U/mL. The measured keratinase activity reached 1 810.98 U/mL, which was 7.96 times of the initial keratinase activity 227.38 U/mL.
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