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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Optimization of extracellular alkaline protease production from species of Bacillus
1Laboratory of Microbiology, School of Life Science and Technology, China Pharmaceutical University, No. 24 Tongjiaxiang, Nanjing 210009, People's Republic of China. chuweihua2002@yahoo.com.cn
Abstract:
Thirty-five strains capable of secreting extracellular alkaline proteases were isolated from the soil and waste water near the milk processing plant, slaughterhouse. Strain APP1 with the highest-yield alkaline proteases was identified as Bacillus sp. The cultural conditions were optimized for maximum enzyme production. When the initial pH of the medium was 9.0, the culture maintained maximum proteolytic activity for 2,560 U ml(-1) at 50 degrees C for 48 h under the optimized conditions containing (g(-1)): soyabean meal, 15; wheat flour, 30; K(2)HPO(4), 4; Na(2)HPO(4), 1; MgSO(4) x 7H(2)O, 0.1; Na(2)CO(3), 6. The alkaline protease showed extreme stability toward SDS and oxidizing agents, which retained its activity above 73 and 110% on treatment for 72 h with 5% SDS and 5% H(2)O(2), respectively.
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