Optimization of Bovine Cathepsin C Production by Cultivation of Recombinant Methylotrophic Yeast Candida boidinii.

Abstract

Bovine cathepsin C (CTC) production from a recombinant methylotrophic yeast Candida boidinii CT714 was investigated, in which CTC gene was driven by formate dehydrogenase promoter and strongly induced in the presence of methanol. Two step fermentation using DYS medium containing 1.0% (w/v) glucose for cell growth and MYS medium containing 2.5% (v/v) methanol for inducible production of CTC was suitable for efficient production. Methanol concentration within 1.5 to 3.5% (v/v) was preferable for CTC induction. After 24 h of cell growth and 72 h of induction, CTC production reached approximately 12 U/I (this corresponded to 2.15 U/g-dry cells). When vitamins were enriched in growth medium, CTC productivity was improved more than 3 times higher, and it was found that the effective vitamins were biotin, p-aminobenzoic acid and thioctic acid.