重组精氨酸脱亚胺酶制备L-瓜氨酸的工艺条件优化  

Optimization of Preparing L-citrulline by Recombinant Arginine Deiminase

作　　者：马越[1,2] 宿玲恰[1,2] 吴丹[1,2] 吴敬[1,2] 

机构地区：[1]江南大学食品科学与技术国家重点实验室,无锡214122 [2]江南大学生物工程学院工业生物技术教育部重点实验室,无锡214122

出　　处：《生物技术通报》2015年第8期180-185,共6页Biotechnology Bulletin

摘　　要：将来源于Pseudomonas putida ACCC 10185的ADI编码基因克隆到表达载体p ET-24a(+)中,转化Escherichia coli BL21(DE3),通过超声波破碎得到粗酶液,酶活检测ADI酶活为26 U/m L发酵液。对酶转化L-精氨酸盐酸盐生成L-瓜氨酸的反应条件进行了优化,结果表明,当底物L-精氨酸盐酸盐浓度650 g/L,反应初始p H6.0,温度37℃,加酶量24 U/g底物,转速100-200 r/min,转化时间7 h,L-瓜氨酸转化率达到100%,是目前国内外报道的酶法制备L-瓜氨酸的最高水平。The arcA gene encoding ADT from Pseudomonas putida ACCC 10185 was cloned into the expression vector pET-24a （ ＋ ） . The vector was then transformed into Escherichia coli BL21 （ DE3 ） for intracellular production of ADI. The crude enzyme was obtained by ultrasonic treatment, and activity in the fermentation broth of recombinant E. coli BL21 （ DE3 ） was 26 U/mL. Furthermore, the condition for enzymatic conversion of L-arginine monohydrochloride to L-citrulline by the recombinant ADI was optimized. At 650 g/L of L-arginine monohydrochloride, pH6.0, 37℃, 100-200 r/min, and 24 U ADI per gram substrate incubated for 7 hours, 100% of the L-arginine monohydrochloride was transformed into L-citrulline, which was the highest level of preparing L-citrulline by enzyme method in home and abroad presently.

关 键 词：精氨酸脱亚胺酶 L-瓜氨酸 酶转化 重组表达 

分 类 号：TQ922[轻工技术与工程—发酵工程] Q814[生物学—生物工程]