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Comparison of three different promoter systems for secretory α‐amylase production in fed‐batch cultures of recombinant Saccharomyces cerevisiae
Author(s) -
Park Yong Soo,
Shiba Sumihisa,
lijima Shinji,
Kobayashi Takeshi,
Hishinuma Fumio
Publication year - 1993
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260410904
Subject(s) - plasmid , amylase , biology , saccharomyces cerevisiae , extracellular , recombinant dna , gene , secretion , microbiology and biotechnology , low copy number , promoter , cell growth , chemistry , enzyme , gene expression , biochemistry
Cloned gene expression in recombinant Saccharomyces cerevisiae 20B‐12 containing three different plasmids was compared in batch and fed‐batch cultures. The plasmids pNA3, pNA7, and pNA9 contain the α‐amylase gene under the control of SUC2, PGK , and GAL7 Promoters, respectively. The synthesis of α‐amylase was therefore induced by low glucose concentration for the SUC2 and PGK promoters, and by galactose for GAL7 promoter. The specific cell growth rates were similar among cells harboring the three different plasmids; they decreased from 0.35 to 0.38 h −1 during the cell growth phase to 0.03 to 0.06h −1 during the production phase. The secretory α‐amylase activity of cells harboring plasmid pNA7 was 129 U/mL in fed‐batch culture, which was 1.4 and 2 times as high as the activities of cells harboring plasmids pNA3 and pNA9, respectively. The secretion ratios (amount of extracellular α‐amylase activity/amounts of total α‐amylase activity) of cells harboring plasmids pNA3, pNA7, and pNA9 were 91.4%, 94.5%, and 95.3%, respectively. © 1993 Wiley & Sons, Inc.