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Secretory Overexpression of Bacillus thermocatenulatus Lipase in Saccharomyces cerevisiae Using Combinatorial Library Strategy
Author(s) -
Kajiwara Shota,
Yamada Ryosuke,
Ogino Hiroyasu
Publication year - 2018
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201700409
Subject(s) - saccharomyces cerevisiae , pichia pastoris , yeast , lipase , terminator (solar) , signal peptide , biology , plasmid , secretion , promoter , recombinant dna , pichia , biochemistry , gene , gene expression , enzyme , ionosphere , physics , astronomy
Simple and cost‐effective lipase expression host microorganisms are highly desirable. A combinatorial library strategy is used to improve the secretory expression of lipase from Bacillus thermocatenulatus (BTL2) in the culture supernatant of Saccharomyces cerevisiae . A plasmid library including expression cassettes composed of sequences encoding one of each 15 promoters, 15 secretion signals, and 15 terminators derived from yeast species, S. cerevisiae , Pichia pastoris , and Hansenula polymorpha , is constructed. The S. cerevisiae transformant YPH499/D4, comprising H. polymorpha GAP promoter, S. cerevisiae SAG1 secretion signal, and P. pastoris AOX1 terminator, is selected by high‐throughput screening. This transformant expresses BTL2 extra‐cellularly with a 130‐fold higher than the control strain, comprising S. cerevisiae PGK1 promoter, S. cerevisiae α‐factor secretion signal, and S. cerevisiae PGK1 terminator, after cultivation for 72 h. This combinatorial library strategy holds promising potential for application in the optimization of the secretory expression of proteins in yeast.