Upregulation of promoter activity of the Aspergillus oryzae xylanase gene by site-directed mutagenesis.
In Aspergillus oryzae, inductive expression of the xynF1 gene is mediated by a transcriptional activator, AoXlnR. Promoter activity of the xynF1 gene, monitored by beta-galactosidase activity, was successfully upregulated by mutating two non-canonical AoXlnR binding sequences to what is thought to be the strongest sequence. Transformants carrying three canonical binding sequences in the promoter region produced enzyme 2.8 times more active (33,000 units mg(-1) protein) than that of transformants carrying the authentic promoter (12,000 units mg(-1) protein).
Marui
Junichiro
J
Department of Biological Mechanisms and Functions, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
Kato
Masashi
M
Kobayashi
Tetsuo
T
Tsukagoshi
Norihiro
N
D001236:Aspergillus oryzae / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000235:genetics; D024101:Mitochondrial Proteins; D016297:Mutagenesis, Site-Directed; D010088:Oxidoreductases / Q000235:genetics; D010940:Plant Proteins; D011401:Promoter Regions, Genetic / Q000235:genetics; D015202:Protein Engineering / Q000379:methods; D013045:Species Specificity; D043325:Xylan Endo-1,3-beta-Xylosidase; D014995:Xylosidases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics; D000099188:Alternative Oxidase
D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D043325:Xylan Endo-1,3-beta-Xylosidase; D014995:Xylosidases; D001616:beta-Galactosidase; D000099188:Alternative Oxidase