Heterologous gene expression and functional analysis of a type III polyketide synthase from Aspergillus niger NRRL 328

Kotaro Kirimura, Shotaro Watanabe, Keiichi Kobayashi

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Type III polyketide synthases (PKSs) catalyze the formation of pyrone- and resorcinol-types aromatic polyketides. The genomic analysis of the filamentous fungus Aspergillus niger NRRL 328 revealed that this strain has a putative gene (chr-8-2: 2978617-2979847) encoding a type III PKS, although its functions are unknown. In this study, for functional analysis of this putative type III PKS designated as An-CsyA, cloning and heterologous expression of the An-CsyA gene (An-csyA) in Escherichia coli were performed. Recombinant His-tagged An-CsyA was successfully expressed in E. coli BL21 (DE3), purified by Ni2+-affinity chromatography, and used for in vitro assay. Tests on the substrate specificity of the His-tagged An-CsyA with myriad acyl-CoAs as starter substrates and malonyl-CoA as extender substrate showed that His-tagged An-CsyA accepted fatty acyl-CoAs (C2-C14) and produced triketide pyrones (C2-C14), tetraketide pyrones (C2-C10), and pentaketide resorcinols (C10-C14). Furthermore, acetoacetyl-CoA, malonyl-CoA, isobutyryl-CoA, and benzoyl-CoA were also accepted as starter substrates, and both of triketide pyrones and tetraketide pyrones were produced. It is noteworthy that the His-tagged An-CsyA produced polyketides from malonyl-CoA as starter and extender substrates and produced tetraketide pyrones from short-chain fatty acyl-CoAs as starter substrates. Therefore, this is the first report showing the functional properties of An-CsyA different from those of other fungal type III PKSs.

Original languageEnglish
Pages (from-to)1106-1110
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume473
Issue number4
DOIs
Publication statusPublished - 2016 May 13
Externally publishedYes

Fingerprint

Pyrones
Functional analysis
Aspergillus niger
Aspergillus
Gene expression
Starters
Malonyl Coenzyme A
Gene Expression
Substrates
Polyketides
Escherichia coli
Resorcinols
Genes
Affinity chromatography
Cloning
Substrate Specificity
Fungi
Affinity Chromatography
flavanone synthetase
Organism Cloning

Keywords

  • Aspergillus niger NRRL 328
  • Pyrone-type polyketide
  • Resorcinol-type polyketide
  • Triacetic acid lactone
  • Type III polyketide synthase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Heterologous gene expression and functional analysis of a type III polyketide synthase from Aspergillus niger NRRL 328. / Kirimura, Kotaro; Watanabe, Shotaro; Kobayashi, Keiichi.

In: Biochemical and Biophysical Research Communications, Vol. 473, No. 4, 13.05.2016, p. 1106-1110.

Research output: Contribution to journalArticle

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