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Secretion of Bacillus α‐amylase from yeast directed by glucoamylase I signal sequence of Saccharomyces diastaticus
Author(s) -
Kang Dae Ook,
Hwang In Kyu,
Kim Bo Yun,
Ahn Soon Cheol,
Mheen Tae Ick,
Ahn Jong Seog,
Byun Si Myung
Publication year - 1996
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549600201181
Subject(s) - thermostability , amylase , signal peptide , yeast , saccharomyces , saccharomyces cerevisiae , alpha amylase , biochemistry , biology , bacillaceae , bacillales , agar plate , enzyme , microbiology and biotechnology , recombinant dna , gene , bacteria , bacillus subtilis , genetics
For the secretion of Bacillus stearothermophilus α ‐amylase from yeast, a recombinant plasmid pGAT17 was constructed by fusing B. stearothermophilus α‐amylase structural gene in frame to the promoter and signal sequence of Saccharomyces diastaticus glucoamylase I gene (STA1). The secretion of the heterologous α‐amylase from S. diastaticus transformed with pGAT17 was confirmed by the halo formation around colonies on selective starch agar medium. About 80% of the total α‐amylase activity was detected in the extracellular culture medium. The secreted α‐amylase was glycosylated and its molecular weight increased from 61 kDa to 75 kDa. The thermostability of the glycosylated α‐amylase was markedly enhanced, compared with that of the non‐glycosylated enzyme from E. coli.