Key Laboratory of Microbial Metabolism, Ministry of Education, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 800 Dong-Chuan Road, Shanghai, 200240, China.
Bioprocess Biosyst Eng. 2010 Aug;33(6):657-63. doi: 10.1007/s00449-009-0330-0. Epub 2009 May 23.
Bacillus cereus ZH14 was previously found to produce a new type of antiviral ribonuclease, which was secreted into medium and active against tobacco mosaic virus. In order to enhance the ribonuclease production, in this study the optimization of culture conditions using response surface methodology was done. The fermentation variables including culture temperature, initial pH, inoculum size, sucrose, yeast extract, MgSO(4).7H(2)O, and KNO(3) were considered for selection of significant ones by using the Plackett-Burman design, and four significant variables (sucrose, yeast extract, MgSO(4).7H(2)O, and KNO(3)) were further optimized by a 2(4) factorial central composite design. The optimal combination of the medium constituents for maximum ribonuclease production was determined as 8.50 g/l sucrose, 9.30 g/l yeast extract, 2.00 g/l MgSO(4).7H(2)O, and 0.62 g/l KNO(3). The enzyme activity was increased by 60%. This study will be helpful to the future commercial development of the new bacteria-based antiviral ribonuclease fermentation process.
先前发现蜡状芽孢杆菌 ZH14 能够产生一种新型抗病毒核糖核酸酶,这种酶分泌到培养基中后能对烟草花叶病毒产生作用。为了提高核糖核酸酶的产量,本研究采用响应面法对培养条件进行了优化。通过 Plackett-Burman 设计,从包括培养温度、初始 pH 值、接种量、蔗糖、酵母提取物、MgSO4·7H2O 和 KNO3 在内的发酵变量中筛选出显著变量,然后通过 2(4) 因子中心组合设计对四个显著变量(蔗糖、酵母提取物、MgSO4·7H2O 和 KNO3)进行进一步优化。确定了获得最大核糖核酸酶产量的最佳培养基成分组合为:8.50 g/L 蔗糖、9.30 g/L 酵母提取物、2.00 g/L MgSO4·7H2O 和 0.62 g/L KNO3。酶活提高了 60%。本研究将有助于未来基于新型细菌的抗病毒核糖核酸酶发酵工艺的商业化开发。