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Expression of functional Candida antarctica lipase B in a cell‐free protein synthesis system derived from Escherichia coli
Author(s) -
Park ChangGil,
Kim TaeWan,
Oh InSeok,
Song Jae Kwang,
Kim DongMyung
Publication year - 2009
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.109
Subject(s) - candida antarctica , escherichia coli , lipase , pichia pastoris , biochemistry , enzyme , cell free system , cell free protein synthesis , cell , chemistry , pichia , yeast , biology , protein biosynthesis , recombinant dna , gene
This article reports the cell‐free expression of functional Lipase B from Candida antarctica (CalB) in an Escherichia coli extract. Although most of the cell‐free synthesized CalB was insoluble under conventional reaction conditions, the combined use of molecular chaperones led to the soluble expression of CalB. In addition, the functional enzyme was generated by applying the optimal redox potential. When examined using p ‐nitrophenyl palmitate as a substrate, the specific activity of the cell‐free synthesized CalB was higher than that of the reference protein produced in Pichia pastoris . These results highlight the potential of cell‐free protein synthesis technology as a powerful platform for the rapid expression, screening and analysis of industrially important enzymes. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009