Title :
Heterologous Expression of ksdD Gene Encoding 3-Ketosteroid-Delta1-Dehydrogenase from Arthrobacter simplex in Escherichia coli
Author :
Li, Yu ; Kong, Dong-jun ; Lu, Fu-Ping ; Jia, Hong-hong ; Li, Jiang ; Cao, Yue-ting
Author_Institution :
Key Lab. of Ind. Microbiol., Tianjin Univ. of Sci. & Technol., Tianjin, China
Abstract :
Arthrobacter simplex is extensively used in pharmaceutical industry to introduce C1-C2 double bond into steroid nucleus. This dehydrogenation reaction is known characterized by its 3-ketosteroid-Δ1-dehydrogenase(KSDH), which is encoded by ksdD gene. In this research, we report the expression of KSDH in Escherichia coli. The activity of the enzyme has been determined by spectrophotometric method. We found the activity of purified enzyme was about 5 fold more than the crude enzyme of cell extract. Additionally, compared with A. simplex, the transformation level by broken cell preparations of E. coli recombination cells used in the dehydrogenation reaction of 4-androstene-3, 17-dione was dramatically improved.
Keywords :
biochemistry; cellular biophysics; enzymes; genetics; genomics; microorganisms; proteomics; spectrophotometry; 17-dione; 3-ketosteroid-Δ1-dehydrogenase; 4-androstene-3; Arthrobacter simplex; C1-C2 double bond; Escherichia coli; cell extract; dehydrogenation reaction; enzyme activity; ksdD gene heterologous expression; spectrophotometric method; steroid nucleus; Amino acids; Biochemistry; Biomembranes; Bonding; Business continuity; Chemical industry; Encoding; Microorganisms; Pharmaceuticals; Proteins;
Conference_Titel :
Bioinformatics and Biomedical Engineering (iCBBE), 2010 4th International Conference on
Conference_Location :
Chengdu
Print_ISBN :
978-1-4244-4712-1
Electronic_ISBN :
2151-7614
DOI :
10.1109/ICBBE.2010.5517292
