Plasmids for expression of heterologous proteins in Rhizopus oryzae.

Abstract:

:Rhizopus oryzae has long been used for enzyme production (e.g., glucoamylase and lipase), organic acid synthesis, and various fermented food applications. In this work, we describe a set of plasmid-based expression vectors that can be used for the production of heterologous proteins in R. oryzae. Three plasmid vectors have been created using either the glucoamylase A (amyA), pyruvate decarboxylase (pdcA), or phosphoglycerate kinase (pgk1) promoters to drive expression of heterologous proteins. All three plasmids use the pdcA terminator for transcription termination, the pyrG gene for restoration of uracil prototrophy, and an ampicillin resistance gene and origin of replication for maintenance in Escherichia coli. We have expressed green fluorescent protein (GFP) and compared transcription and protein accumulation for each of the expression vectors. Accumulation of GFP transcript and protein was directly correlated with the choice of promoter with pdcA > amyA > pgk1. Transcript level appears to parallel GFP protein accumulation. Plasmid copy number had little impact on transcription or protein accumulation. These vectors should be useful for overexpression of heterologous proteins and potentially, metabolic engineering of Rhizopus strains.

journal_name

Arch Microbiol

journal_title

Archives of microbiology

authors

Mertens JA,Skory CD,Ibrahim AS

doi

10.1007/s00203-006-0121-9

subject

Has Abstract

pub_date

2006-07-01 00:00:00

pages

41-50

issue

1

eissn

0302-8933

issn

1432-072X

journal_volume

186

pub_type

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