Molecular Cloning of the Glucoamylase Gene of Aspergillus shirousami and Its Expression in Aspergillus Oryzae

Ichiro Shibuya; Gakuzo Tamura; Katsuya Gomi; Yuzuru Iimura; Kojiro Takahashi; Shodo Hara

doi:10.1271/bbb1961.54.1905

Molecular Cloning of the Glucoamylase Gene of Aspergillus shirousami and Its Expression in Aspergillus Oryzae

Ichiro Shibuya, Gakuzo Tamura, Katsuya Gomi, Yuzuru Iimura, Kojiro Takahashi, Shodo Hara

AGR - Bioscience and Biotechnology for Future Bioindustries

Research output: Contribution to journal › Article › peer-review

35 Citations (Scopus)

Abstract

The glucoamylase enzyme (GAase) gene from Aspergillus shirousami was cloned and sequenced from genomic and cDNA libraries. The genomic gene was located in the 5.4 kb EcoRI fragment. The deduced amino acid sequence of GAase contained 639 amino acid residues with a relative molecular mass of approximately 68,000 daltons (non-glycosylated form). The genomic gene of A. shirousami GAase was introduced into Aspergillus oryzae. These transformants had increased GAase and raw starch degradation (RSD) activity in culture media and in rice-koji extracts. Analysis by Southern, Northern, SDS-PAGE, and Western blot techniques confirmed the foreign gene was correctly transcribed, translated, and expressed in A. oryzae.

Original language	English
Pages (from-to)	1905-1914
Number of pages	10
Journal	Agricultural and Biological Chemistry
Volume	54
Issue number	8
DOIs	https://doi.org/10.1271/bbb1961.54.1905
Publication status	Published - 1990

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Agricultural and Biological Sciences(all)

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title = "Molecular Cloning of the Glucoamylase Gene of Aspergillus shirousami and Its Expression in Aspergillus Oryzae",

abstract = "The glucoamylase enzyme (GAase) gene from Aspergillus shirousami was cloned and sequenced from genomic and cDNA libraries. The genomic gene was located in the 5.4 kb EcoRI fragment. The deduced amino acid sequence of GAase contained 639 amino acid residues with a relative molecular mass of approximately 68,000 daltons (non-glycosylated form). The genomic gene of A. shirousami GAase was introduced into Aspergillus oryzae. These transformants had increased GAase and raw starch degradation (RSD) activity in culture media and in rice-koji extracts. Analysis by Southern, Northern, SDS-PAGE, and Western blot techniques confirmed the foreign gene was correctly transcribed, translated, and expressed in A. oryzae.",

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AU - Gomi, Katsuya

AU - Iimura, Yuzuru

AU - Takahashi, Kojiro

AU - Hara, Shodo

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