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

fYear

2010

fDate

18-20 June 2010

Firstpage

1

Lastpage

5

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;

fLanguage

English

Publisher

ieee

Conference_Titel

Bioinformatics and Biomedical Engineering (iCBBE), 2010 4th International Conference on

Conference_Location

Chengdu

ISSN

2151-7614

Print_ISBN

978-1-4244-4712-1

Electronic_ISBN

2151-7614

Type

conf

DOI

10.1109/ICBBE.2010.5517292

Filename

5517292