RRC ID 2210
Author Nakagawa T, Inagaki A, Ito T, Fujimura S, Miyaji T, Yurimoto H, Kato N, Sakai Y, Tomizuka N.
Title Regulation of two distinct alcohol oxidase promoters in the methylotrophic yeast Pichia methanolica.
Journal Yeast
Abstract In this study, two Pichia methanolica alcohol oxidase (AOD) promoters, P(MOD1) and P(MOD2), were evaluated in a promoter assay system utilizing the acid phosphatase (AP) gene from Saccharomyces cerevisiae (ScPHO5) as a reporter. Heterologous gene expression driven by the P(MOD1) and P(MOD2) promoters was found to be strong and tightly regulated by carbon source at the transcriptional level. P(MOD1) was induced not only by methanol but also by glycerol. P(MOD2) was induced only by methanol, although it was not repressed on the addition of glycerol to a methanol medium, suggesting that P(MOD2) is regulated in a manner distinct from that of other AOD-gene promoters. On the other hand, methanol and oxygen level-influenced gene expression mediated by P(MOD1) and P(MOD2). P(MOD1) expression was optimal at low methanol concentrations, whereas P(MOD2) was predominantly expressed at high methanol and high oxygen concentrations. Based on these results, both P(MOD2) and P(MOD1) should be useful tools for controlling heterologous gene expression in P. methanolica. In particular, it should be possible to differentially control the production phases of two heterologous proteins, using P(MOD1) and P(MOD2) in the same host cell and in the same flask.
Volume 23(1)
Pages 15-22
Published 2006-1-15
DOI 10.1002/yea.1334
PMID 16411161
MeSH Acid Phosphatase / genetics Acid Phosphatase / metabolism Alcohol Oxidoreductases / genetics* Alcohol Oxidoreductases / metabolism Gene Expression Regulation, Enzymologic Gene Expression Regulation, Fungal Histocytochemistry Methanol / metabolism Mutagenesis, Insertional Oxygen / metabolism Pichia / genetics* Pichia / metabolism* Promoter Regions, Genetic / genetics*
IF 3.143
Times Cited 17
WOS Category MYCOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Yeast ?ScPHO5 gene