RRC ID 27488
著者 Suzuki H, Yoshida K, Ohshima T.
タイトル Polysaccharide-degrading thermophiles generated by heterologous gene expression in Geobacillus kaustophilus HTA426.
ジャーナル Appl Environ Microbiol
Abstract Thermophiles have important advantages over mesophiles as host organisms for high-temperature bioprocesses, functional production of thermostable enzymes, and efficient expression of enzymatic activities in vivo. To capitalize on these advantages of thermophiles, we describe here a new inducible gene expression system in the thermophile Geobacillus kaustophilus HTA426. Six promoter regions in the HTA426 genome were identified and analyzed for expression profiles using β-galactosidase reporter assay. This analysis identified a promoter region upstream of a putative amylose-metabolizing gene cluster that directed high-level expression of the reporter gene. The expression was >280-fold that without a promoter and was further enhanced 12-fold by maltose addition. In association with a multicopy plasmid, this promoter region was used to express heterologous genes. Several genes, including a gene whose product was insoluble when expressed in Escherichia coli, were successfully expressed as soluble proteins, with yields of 0.16 to 59 mg/liter, and conferred new functions to G. kaustophilus strains. Remarkably, cellulase and α-amylase genes conferred the ability to degrade cellulose paper and insoluble starch at high temperatures, respectively, generating thermophiles with the potential to degrade plant biomass. Our results demonstrate that this novel expression system expands the potential applications of G. kaustophilus.
巻・号 79(17)
ページ 5151-8
公開日 2013-9-1
DOI 10.1128/AEM.01506-13
PII AEM.01506-13
PMID 23793634
PMC PMC3753961
MeSH Cellulase / biosynthesis* Cellulase / genetics Gene Dosage Gene Expression Geobacillus / genetics* Hot Temperature Hydrolysis Metabolic Engineering / methods* Metabolic Networks and Pathways / genetics* Plasmids Polysaccharides / metabolism* Promoter Regions, Genetic Recombinant Proteins / biosynthesis Recombinant Proteins / genetics alpha-Amylases / biosynthesis* alpha-Amylases / genetics
IF 4.016
引用数 19
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY
リソース情報
一般微生物 JCM 9974 JCM 10545 JCM 11548