嗜热厌氧乙醇菌JW200中乙醛脱氢酶的纯化  

Purification of Acetaldehyde Dehydrogenase From Thermoanaerobacter ethanolicus JW200

作　　者：彭惠[1] 毛忠贵[1] 武国干[2] 邵蔚蓝[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 [2]南京师范大学微生物工程重点实验室,江苏南京210097

出　　处：《南京师大学报（自然科学版）》2007年第1期78-81,共4页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：中国973项目子课题基金(2004CB719600);国家自然科学基金(30170511)资助项目

摘　　要：研究了嗜热厌氧乙醇菌(Thermoanaerobacter ethanolicus)JW200中乙醇代谢途径的关键酶之一乙酰CoA依赖型的乙醛脱氢酶(acetaldehyde dehydrogenase,ALDH,EC 1.2.1.10)的纯化.结果表明,经DEAE Sepha-rose Fast Flow阴离子交换层析、两次为30%与80%硫酸铵盐析和相应盐浓度的Butyl-HIC疏水层析、TOYOPE-ARL HW-55F分子筛层析等提纯步骤,可得到电泳纯的ALDH,其提纯倍数为910倍,得率为7%.由SDS-PAGE和梯度PAGE测得全酶由4个亚基组成,全酶相对分子质量为360 000,亚基相对分子质量为100 000.A acetaldehyde dehydrogenase （CoA-acetylating） hol metabolic pathway was purified from which is one of the key enzymes of the alcoethanolicus JW200 through following steps： （1） ion exchange chromatography on DEAE-Sepharose Fast Flow, （2） ammonium sulfate fractionation with saturation 30% and hydrophobic interaction chromatography on Butyl - HIC, （3） ammonium sulfate frac- tionation with saturation 80% and hydrophobic interaction chromatography on Butyl-HIC, （4） gel filtration on TOYOPEARL HW-55F. The purified enzyme showed a single band on SDS-polyacrylamide gel electrophoresis with a purification of 910 fold, and a yield of 7%. The molecular weight of the subunit and the whole enzyme were estimated by SDS-PAGE and gradation PAGE as 100 000 and 360 000, respectively.

关 键 词：乙醛脱氢酶 嗜热厌氧乙醇菌 纯化 

分 类 号：Q556[生物学—生物化学]