Expression of functional Candida antarctica lipase B in a cell‐free protein synthesis system derived from Escherichia coli |
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| Authors: | Chang‐Gil Park Tae‐Wan Kim In‐Seok Oh Jae Kwang Song Dong‐Myung Kim |
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| Affiliation: | 1. Dept. of Fine Chemical Engineering and Chemistry, Chungnam National University, Daejeon 305‐764, Korea;2. School of Chemical and Biological Engineering, College of Engineering, Seoul National University, Seoul 151‐742, Korea;3. Korea Research Institute of Chemical Technology, Daejeon 305‐600, Korea |
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| Abstract: | 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 |
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| Keywords: | candida antartica lipase B protein synthesis Pichia pastoris |
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