Amino acid residues involved in organic solvent-stability of the LST-03 lipase.
LST-03 lipase from Pseudomonas aeruginosa LST-03 is highly active and stable in the presence of various organic solvents. To further characterize and improve the organic solvent-stability of the LST-03 lipase, residues that potentially provide this stability were identified and mutated to other amino acids in an effort to increase the organic solvent-stability of the protein. S155L, G157R, S164K, S194R, and D209N mutations were found to improve the organic solvent-stability of the wild-type LST-03 lipase. Such mutations were found to induce structural changes, including the formation of a salt bridge, hydrogen bonds, lead to an improved packing of the hydrophobic core, and pI shift of side chain. These changes increased the stability of the protein, thereby improving the organic solvent-stability of the wild-type LST-03 lipase. In addition, a single mutation was found to stabilize the lipase by single or multiple factors.
Kawata
Takuya
T
Department of Chemical Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan.
Ogino
Hiroyasu
H
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000596:Amino Acids / Q000737:chemistry* / Q000235:genetics; D006358:Hot Temperature; D008049:Lipase / Q000737:chemistry* / Q000235:genetics; D016296:Mutagenesis; D011487:Protein Conformation; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D012995:Solubility; D012997:Solvents / Q000737:chemistry*
D000596:Amino Acids; D012997:Solvents; D008049:Lipase