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Titolo:
Establishment of a hyper-protein production system in submerged Aspergillus oryzae culture under tyrosinase-encoding gene (mel0) promoter control
Autore:
Ishida, H; Matsumura, K; Hata, Y; Kawato, A; Suginami, K; Abe, Y; Imayasu, S; Ishishima, E;
Indirizzi:
Gekkeikan Sake Co Ltd, Res Inst, Fushimi Ku, Kyoto 6128361, Japan Gekkeikan Sake Co Ltd Kyoto Japan 6128361 shimi Ku, Kyoto 6128361, Japan Soka Univ, Grad Sch Engn, Dept Bioengn, Hachioji, Tokyo 1928577, Japan Soka Univ Hachioji Tokyo Japan 1928577 gn, Hachioji, Tokyo 1928577, Japan
Titolo Testata:
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
fascicolo: 1-2, volume: 57, anno: 2001,
pagine: 131 - 137
SICI:
0175-7598(200110)57:1-2<131:EOAHPS>2.0.ZU;2-H
Fonte:
ISI
Lingua:
ENG
Soggetto:
SOLID-STATE CULTURE; HOMOLOGOUS TRANSFORMATION SYSTEM; AMYLASE-A GENE; NUCLEOTIDE-SEQUENCE; ESCHERICHIA-COLI; BETA-GLUCURONIDASE; FUSION PROTEIN; GLUCOAMYLASE; EXPRESSION; ENZYME;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
35
Recensione:
Indirizzi per estratti:
Indirizzo: Ishida, H Gekkeikan Sake Co Ltd, Res Inst, Fushimi Ku, 300 Katahara Chou, Kyoto 6128361, Japan Gekkeikan Sake Co Ltd 300 Katahara Chou Kyoto Japan 6128361 pan
Citazione:
H. Ishida et al., "Establishment of a hyper-protein production system in submerged Aspergillus oryzae culture under tyrosinase-encoding gene (mel0) promoter control", APPL MICR B, 57(1-2), 2001, pp. 131-137

Abstract

UV-mediated mutagenesis generated a high glucoamylase-producing mutant of Aspergillus oryzae exhibiting strong melanization in solid-state culture. Expression of the glucoamylase-encoding gene (glaB), which is specifically expressed in solid-state culture, and the tyrosinase-encoding gene (melO), was analyzed using an E. coli P-glucuronidase (GUS) reporter assay to investigate this phenomenon. Although no common regulation was found for melO andglaB expression, the former was greatly enhanced in submerged culture. Interestingly, the melO promoter was about four times stronger for GUS production than the powerful promoters amyB, glaA, and modified agdA, previously isolated for industrial heterologous gene expression in A. oryzae. These findings indicated that the melO promoter would be suitable for hyper-production of heterologous protein in Aspergillus. The glaB-type glucoamylase selected as the target protein was produced in a submerged culture of A. oryzae under the control of the melO promoter. The maximum yield was 0.8 g/l broth, and the total extracellular protein purity was 99%. Repeated batch culture, to improve productivity, gave a maximum yield of 3.3 g/l broth. The importance of this work is in the establishment of a both high-level and high-purity protein overproduction system in A. oryzae by use of the melO promoter.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 06/04/20 alle ore 07:50:42