【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。更多还原

【Abstract】 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 semi-synthetic medium could be obtained when keeping temperature at 30 ℃ for 4 hours in the growth phase and then shifting to 42 ℃ for 6 hours.更多还原