【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.更多还原

【Abstract】 A acetaldehyde dehydrogenase(CoA-acetylating) which is one of the key enzymes of the alcohol metabolic pathway was purified from Thermoanaerobacter ethanolicus 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 fractionation 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.更多还原