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Mutagenesis of cutinase-producing strain Thermobifida fusca WSH03-11 and optimization of fermentation conditions of mutant with high cutinase production

ZHANG Shouliang,CHEN Jian,HUA Zhaozhe,DU Guocheng   

  1. Key Lab of Industrial Biotechnology,Ministry of Education; Environmental Biotechnology laboratory,
    School of Biotechnology,Southern Yangtze University
  • Online:2006-05-25 Published:2006-05-25

角质酶产生菌Thermobifida fusca WSH03-11
诱变及高产突变株发酵条件优化

张守亮,陈 坚,华兆哲,堵国成   

  1. 江南大学,工业生物技术教育部重点实验室;生物工程学院环境生物技术室

Abstract: Thermobifida fusca WSH03-11 was mutagenized by using DES and a mutant was isolated after three runs of screening. The cutinase activity was improved from 2.3 U/mL to 8.9 U/mL. After sodium acetate was selected as fermentation carbon source,batch microbial cutinase fermentation by a mutant of Thermobifida fusca WSH03-11 at various pH values ranging from 7.0 to 7.9 was studied. Based on time courses of the specific cell growth rate and specific cutinase formation rate at different pH values,a two-stage pH value-shift strategy in which pH value was controlled at 7.3 in the first 20 h,and then switched to 7.6 after 20 h was developed. By applying this pH value-shift strategy in cutinase fermentation,the maximal cutinase activity reached 19.8 U/mL and was increased by 125%,64%,37% and 47%,compared to those with constant pH values at 7.0,7.3,7.6 and 7.9,respectively.

摘要: 以Thermobifida fusca WSH03-11为出发菌株,经硫酸二乙酯诱变,获得一株遗传稳定性好的高产角质酶的突变菌株(酶活由2.3 U/mL提高为8.9 U/mL)。确定了以乙酸钠为碳源,并在2.5 L发酵罐中研究了不同pH值对突变株角质酶分批发酵的影响。根据不同pH值下发酵过程中细胞比生长速率及产物比生长速率的变化,确定了分阶段控制pH值的策略,即在发酵前20 h控制pH值7.3、20 h后控制pH值7.6,这种条件下角质酶活达到19.8 U/mL,比采用单一pH值7.0、7.3、7.6、7.9下的最大值分别提高了125%,64%,37%,47%。

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