Agricultural and Biological Chemistry
Online ISSN : 1881-1280
Print ISSN : 0002-1369
ISSN-L : 0002-1369
Molecular Cloning of the Glucoamylase Gene of Aspergillus shirousami and Its Expression in Aspergillus oryzae
Ichiro SHIBUYAKatsuya GOMIYuzuru IIMURAKojiro TAKAHASHIGakuzo TAMURAShodo HARA
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1990 Volume 54 Issue 8 Pages 1905-1914

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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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