耐热过氧化氢酶基因工程菌的构建及其发酵条件  

Construction of Recombinant Thermo-Stable Catalase Engineering Strain and Optimization of Its Fermentation Condition

作　　者：段绪果[1] 沈微[1] 李艳丽[1] 饶志明[1] 唐雪明[1] 方慧英[1] 刘吉泉[1] 诸葛健[1] 

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214036

出　　处：《食品与生物技术学报》2006年第2期74-78,共5页Journal of Food Science and Biotechnology

摘　　要：利用PCR扩增技术以嗜热脂肪芽孢杆菌IAM11001染色体DNA为模板,扩增得到嗜热脂肪芽孢杆菌过氧化氢酶编码基因perA。再与经EcoRⅠ酶切的温控表达载体pBV220连接,构建重组质粒,并转化宿主大肠杆菌JM109,得到耐热过氧化氢酶基因工程菌,然后对该工程菌在LB培养基和半合成培养基中的发酵条件进行了优化。结果表明:在LB培养基中,诱导时期和酶合成最佳诱导时间均为5 h,最大产酶量达到72.9 U/mL;在半合成培养基中,于30℃培养4 h,再于42℃诱导培养6 h,产酶量最高达到131 U/mL。The gene encoding a thermo-stable catalase was amplified from the chromosomal DNA of Bacillus stearothermophilus IAM11001 by PCR. It was then inserted into the temperature control expression vector pBV220 and expressed in E. coli JM109. The fermentation conditions were optimized both in the LB medium and semi-synthetic medium. The results indicated that both of the inducing time and the length of inducing time were 5 hours and the maximal enzyme activity reached 72.9 U/mL in LB medium; The maximal enzyme activity of 131 U/mL in semisynthetic medium could be obtained when keeping temperature at 30 ℃ for 4 hours in the growth phase and then shifting to 42 ℃ for 6 hours.

关 键 词：耐热过氧化氢酶 基因工程菌 温控表达载体 发酵条件 

分 类 号：TQ920[轻工技术与工程—发酵工程]