Development of a pyrG mutant of Aspergillus oryzae strain s1 as a host for the production of heterologous proteins

Selina Oh Siew Ling, Reginald Storms, Yun Zheng, Mohd Rohaizad Mohd Rodzi, Nor Muhammad Mahadi, Rosli Md Illias, Abdul Munir Abd. Murad, Farah Diba Abu Bakar

Research output: Contribution to journalArticle

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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 pyrGΔ0 was complemented by plasmids carrying a pyrG gene from either Aspergillus nidulans or A. oryzae. β-Galactosidase expression by strain GR6 pyrGΔ0 transformed with an A. niger plasmid encoding a heterologous β-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 pyrGΔ0 facilitated the production of a heterologous protein in this fungus.

Original languageEnglish
Article number634317
JournalThe Scientific World Journal
Volume2013
DOIs
Publication statusPublished - 2013

Fingerprint

Aspergillus oryzae
Aspergillus
Genes
protein
Galactosidases
gene
Proteins
plasmid
mutation
Plasmids
replacement
Aspergillus nidulans
Mutation
Carboxy-Lyases
Uridine
Mutagenesis
Fungi
mutant
fungus

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Environmental Science(all)
  • Medicine(all)

Cite this

Development of a pyrG mutant of Aspergillus oryzae strain s1 as a host for the production of heterologous proteins. / Ling, Selina Oh Siew; Storms, Reginald; Zheng, Yun; Rodzi, Mohd Rohaizad Mohd; Mahadi, Nor Muhammad; Illias, Rosli Md; Abd. Murad, Abdul Munir; Abu Bakar, Farah Diba.

In: The Scientific World Journal, Vol. 2013, 634317, 2013.

Research output: Contribution to journalArticle

Ling, Selina Oh Siew ; Storms, Reginald ; Zheng, Yun ; Rodzi, Mohd Rohaizad Mohd ; Mahadi, Nor Muhammad ; Illias, Rosli Md ; Abd. Murad, Abdul Munir ; Abu Bakar, Farah Diba. / Development of a pyrG mutant of Aspergillus oryzae strain s1 as a host for the production of heterologous proteins. In: The Scientific World Journal. 2013 ; Vol. 2013.
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