1 School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia.
2 Centre for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke Street West, Montréal, Québec, Canada H4B 1R6 ; Department of Biology, Concordia University, 7141 Sherbrooke Street West, Montréal, Québec, Canada H4B 1R6.
3 Centre for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke Street West, Montréal, Québec, Canada H4B 1R6.
4 Malaysia Genome Institute, Jalan Bangi, 43000 Kajang, Selangor, Malaysia.
5 Department of Bioprocess Engineering, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia.

Development of a pyrG mutant of Aspergillus oryzae strain S1 as a host for the production of heterologous proteins.

ScientificWorldJournal. 2013;2013:634317

Authors: Ling SO, Storms R, Zheng Y, Rodzi MR, Mahadi NM, Illias RM, Abdul Murad AM, Abu Bakar FD

Abstract

The ease with which auxotrophic strains and genes that complement them can be manipulated, as well as the stability of auxotrophic selection systems, are amongst the advantages of using auxotrophic markers to produce heterologous proteins. Most auxotrophic markers in Aspergillus oryzae originate from chemical or physical mutagenesis that may yield undesirable mutations along with the mutation of interest. An auxotrophic A. oryzae strain S1 was generated by deleting the orotidine-5'-monophosphate decarboxylase gene (pyrG) by targeted gene replacement. The uridine requirement of the resulting strain GR6 pyrG?0 was complemented by plasmids carrying a pyrG gene from either Aspergillus nidulans or A. oryzae. ß -Galactosidase expression by strain GR6 pyrG?0 transformed with an A. niger plasmid encoding a heterologous ß -galactosidase was at least 150 times more than that obtained with the untransformed strain. Targeted gene replacement is thus an efficient way of developing auxotrophic mutants in A. oryzae and the auxotrophic strain GR6 pyrG?0 facilitated the production of a heterologous protein in this fungus.

PMID: 24381522 [PubMed - indexed for MEDLINE]