Cloning, expression and biochemical characterization of the cholesterol oxidase CgChoA from Chryseobacterium gleum.

ABSTRACT:

Background

Cholesterol oxidases are important enzymes for applications such as the analysis of cholesterol in clinical samples, the synthesis of steroid derived drugs, and are considered as potential antibacterial drug targets.

Results

The gene choA encoding a cholesterol oxidase from Chryseobacterium gleum DSM 16776 was cloned into the pQE-30 expression vector and heterologously expressed in Escherichia coli JM109 co-transformed with pRARE2. The N-terminally His-tagged cholesterol oxidase (CgChoA) was assigned to be a monomer in solution by size exclusion chromatography, showed a temperature optimum of 35°C, and a pH optimum at 6.75 using 0.011 M MOPS buffer under the tested conditions. The purified protein showed a maximum activity of 15.5 U/mg. CgChoA showed a Michaelis-M

SUBMITTER: Reiss R 

PROVIDER: S-EPMC4053396 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature