Essential role of Gly33 in a novel organic solvent-tolerant lipase from Serratia marcescens ECU1010 as determined by site-directed mutagenesis.
A novel lipase lipB from Serratia marcescens ECU1010 is highly stable in the presence of organic solvents. By sequence and structure comparison with homologous lipase lipA, three amino acid residues were found to be different between them. To identify the residues which increase the organic solvent stability of lipB, residues that potentially provide this stability were mutated to the ones of lipA at equivalent positions. The replacement of Gly at position 33 by Asp obviously decreased its stability in organic solvents. Molecular modeling and structural analysis also suggested that the Gly33 residue is important for the organic solvent stability of lipB.
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, People's Republic of China, suxia5664@163.com.
Su-Xia
SX
Li
Qiang
Q
Ma
Kang
K
Lin
Jiao-Jiao
JJ
Wu
Yi-Xin
YX
Wu
Jian-He
JH
Xu
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002087:Butyrates / Q000737:chemistry; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005998:Glycine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012706:Serratia marcescens / Q000737:chemistry* / Q000201:enzymology; D012997:Solvents
D001426:Bacterial Proteins; D002087:Butyrates; D011994:Recombinant Proteins; D012997:Solvents; C033592:4-nitrophenyl butyrate; D008049:Lipase; D005998:Glycine