重组氨基酰化酶1与N-乙酰鸟氨酸脱乙酰酶酶学性质比较

Comparisons of Enzymatic properties of Aminoacylase1 and N-acetyl-ornithine Deacetylase

摘要:

利用基因工程手段重组表达了牛肾来源的氨基酰化酶1（ACY1），并与实验室之前构建的N-乙酰鸟氨酸脱乙酰酶（NAO）进行比较，以N-乙酰-DL-蛋氨酸为底物研究了其酶学性质，考察了pH、温度、反应时间、表面活性剂、金属离子等因素对拆分DL-蛋氨酸的影响。结果表明：重组NAO和ACY1拆分300 mmol/L的N-乙酰-DL-蛋氨酸所需时间分别是2 h和6 h；重组NAO的酶活及N-乙酰-L-蛋氨酸的转化率分别为1139.41 U/g 和98%，高于重组ACY1的 671.24 U/g和58.78%，约是重组ACY1的1.7倍。

Abstract:

Aminoacylase 1 (ACY1) from bovine kidney was recombinant expressed in E.coli BL21 (DE3) with the vector pETDuet. The enzymatic properties comparison between ACY1 and N-acetyl-ornithine deacetylase (NAO) were studied. Several influence factors of enzyme reaction such as pH, temperature, enzyme reaction time, surfactant and metal irons were investigated. The results indicated that the resolution of 300 mmol/L N-Ac-DL-Methionine with ACY1 needs 6 h, while NAO needs 2 h. The enzyme activity and conversion rate of NAO (1139.41 U/g and 98%) are about 1.7 times higher than ACY1 (671.24 U/g and 58.78%).