CONSTRUCTION OF AN ALPHA-AMYLASE GLUCOAMYLASE FUSION GENE AND ITS EXPRESSION IN SACCHAROMYCES-CEREVISIAE

被引:25
作者
SHIBUYA, I
TAMURA, G
SHIMA, H
ISHIKAWA, T
HARA, S
机构
[1] BREWING RESOURCES CO LTD, RES INST, KITA, TOKYO 114, JAPAN
[2] NATL RES INST BREWING, KITA, TOKYO 114, JAPAN
关键词
D O I
10.1271/bbb.56.884
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A fusion gene which encoded a polypeptide comprised of 1116 amino acids was constructed using the alpha-amylase and glucoamylase cDNAs of Aspergillus shirousamii. When the fusion gene was expressed in Saccharomyces cerevisiae using a yeast expression plasmid under the control of the yeast ADH1 promoter, a bifunctional fusion protein (145 kDa) having both alpha-amylase and glucoamylase activities was secreted into the culture medium. The fusion protein had higher raw-starch-digesting activity than those of the original alpha-amylase and glucoamylase, and adsorbed onto raw starch like the glucoamylase. It was suggested that the characteristics are a result of the raw-starch-affinity site in the glucoamylase domain of the fusion protein.
引用
收藏
页码:884 / 889
页数:6
相关论文
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