Cloning and expression of an active aspartic proteinase from Mucor circinelloides in Pichia pastoris.

ABSTRACT:

Background

Extracellular aspartic proteinase (MCAP) produced by Mucor circinelloides in solid state fermentations has been shown to possess milk clotting activity and represents a potential replacement for bovine chymosin in cheese manufacturing. Despite its prospects in the dairy industry, the molecular characteristics of this enzyme remain unknown. This work focuses on MCAP cloning and optimization of heterologous expression in Pichia pastoris, and characterization of the enzyme.

Results

The cloning of cDNA sequence encoding MCAP from M. circinelloides was performed using a fragment of approximately 1 kbp as a probe. The fragment was amplified using non-specific primers designed from the NDIEYYG and KNNYVVFN consensus motifs from aspartic proteinases of different fungi. Ge

SUBMITTER: Gama Salgado JA 

PROVIDER: S-EPMC3828624 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature