Heterologous expression of williamsia marianensis induced cholesterol oxidase in E. coli

Most common treatment protocols have limitations and drawbacks. Thus, finding effective treatments is the goal of many researchers. Therapeutic peptides are a promising class of drugs that offer numerous benefits over conventional pharmaceuticals including proteins and antibodies. Cholesterol oxidase, produced from Williamson marianensis, has been shown to have therapeutic benefits. Burn and ear samples, wound swabs, sputum and urine were obtained to confirm the identification of previously identified isolates by conventional biochemical assays. In this study, we were able to clone the cholestrol oxidase gene using PCR with primers specific to an expression vector (pET28b) and express it in E. coli (BL-21/DE3) Rosetta after identification with IPTG. Gyncholestrol oxidase (~500 bp) was sequenced to provide a cox sequence and submitted for synthesis by Genscript Corporation, USA. A double restriction digestion pattern was demonstrated pET-28a(+) cox-william. The obtained pattern consisted of two strands: one of them was a carrier plasmid (4200 bp) and a strand of 2800 base pairs carried the cholesterol oxidase gene. As a result, the cholesterol oxidase gene was successfully cloned and expressed. Cholestrol oxidase derived from Williamson marianensis will be used in future therapeutic outcomes.