Expression of functional Candida antarctica lipase B in a cell-free protein synthesis system derived from Escherichia coli.
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.
Park
Chang-Gil
CG
Dept. of Fine Chemical Engineering and Chemistry, Chungnam National University, Daejeon 305-764, Korea.
Kim
Tae-Wan
TW
Oh
In-Seok
IS
Song
Jae Kwang
JK
Kim
Dong-Myung
DM
D002175:Candida / Q000201:enzymology*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins; D015870:Gene Expression; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis; D012995:Solubility
D029968:Escherichia coli Proteins; D005656:Fungal Proteins; D008049:Lipase; C431349:lipase B, Candida antarctica