Expression of functional Candida antarctica lipase B in a cell‐free protein synthesis system derived from Escherichia coli

• Published in: Biotechnology Progress
• Main Authors: Park, Chang‐Gil, Kim, Tae‐Wan, Oh, In‐Seok, Song, Jae Kwang, Kim, Dong‐Myung
• Format: Article in Journal/Newspaper
• Language: English
• Published: Wiley 2009
• Subjects: Antarc*; Antarctica
• Online Access: http://dx.doi.org/10.1002/btpr.109; https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fbtpr.109; https://onlinelibrary.wiley.com/doi/full/10.1002/btpr.109

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