普通脱硫弧菌产氢化酶条件及提取贮存因素研究

FACTORS AFFECTING PRODUCTION, EXTRACT AND STORAGE OF HYDROGENASE FROM DESULFOVIBRIO VULGARIS

普通脱硫弧菌D-2氧化酶在培养基N上产量较高。金属螯合剂EDTA能增加菌体酶含量,其最适浓度为0.02g/L。碱性缓冲液(含有EDTA)在30℃、15分钟内能抽提约90%的周质氢化酶。该酶在低温、充氮条件下能保持很高的起始酶活,且牛血清白蛋白(BSA)能明显增加酶在空气中的稳定性,但还原的氢化酶纯样品氧失活比较明显。

An examination of conditions for the extract and storage of hydrogenase from D. vulgaris D-2, with the aim of optimizing hydrogenase production, was reported. The medium N gave 6-7 times as much hydrogenase activity as the medium B. The metal-chelating agents gave a reproducible increasing in hydrogenase activity. The optimum concentration was about 0.02g/1. Nearly 90% of hydrogenase activity was extracted by using the slightly alkaline buffer(containing 50m mol/L EDTA) within 15 min at 30 ℃. The enzyme could be stored frozen in air for two weeks with only small losses in activity. The oxidized form of the enzyme was more stable than the reduced form and the addition of bovine serum albumin(BSA) conferred even greater stabilitythan the use of a nitrogen gas phase.