化工进展Issue(1)：210-215,6.DOI:10.16085/j.issn.1000-6613.2016.01.028
丁酸梭杆菌VPI 3266甘油脱水酶和1,3-丙二醇氧化还原酶的克隆、表达
Cloning and expression of glycerol dehydratase and 1,3-propanediol dehydrogenase fromClostridium butyricum VPI3266
摘要
Abstract
Glycerol dehydratase and 1,3-propanediol dehydrogenase are the key enzymes for 1,3-propanediol metabolism inClostridium butyricum VPI3266. The genedhaB consisted of a 3315bp fragment encoding two proteins subunits,glycerol dehydratase and its activator protein,respectively. The former consisted of 2367bp encoding 788 amino acids,which belonged to family gly-Radical. The latter consisted of 918bp encoding 305 amino acids,which belonged to family Radical-SAM. The activity of glycerol dehydratase was determined by expressingdhaB inE. coli. 1,3-PD dehydrogenase genedhaT consisted of 1166bp encoding 388 amino acids with calculated molecular mass of 4.19×104. The activity of recombinantβ-glucosidase was 5.2U/mL in LB medium by IPTG induction. The optimal activity was achieved at pH=10.0 and 50℃. The purified enzyme was stable over pH range of 8.5—10.0,and had a 2h half-life at 45℃. TheVmax andKm for 1,3-propanediol was 29.2U/mg and 19.8mmol/L,respectively.关键词
丁酸梭杆菌/甘油脱水酶/1,3-丙二醇氧化还原酶/重组表达Key words
Clostridium butyricum/glycerol dehydratase/1,3-propanediol dehydrogenase/recombinant expression分类
生物科学引用本文复制引用
裴建军,屈依然,殷冉,陈安娜,赵林果..丁酸梭杆菌VPI 3266甘油脱水酶和1,3-丙二醇氧化还原酶的克隆、表达[J].化工进展,2016,(1):210-215,6.基金项目
国家自然科学青年基金（30900008），江苏省自然科学基金（BK20131423）和江苏省优势学科建设工程项目（PAPD）。 （）
