Crystal structure and enhanced activity of a cutinase-like enzyme from Cryptococcus sp. strain S-2.
The structural and enzymatic characteristics of a cutinase-like enzyme (CLE) from Cryptococcus sp. strain S-2, which exhibits remote homology to a lipolytic enzyme and a cutinase from the fungus Fusarium solani (FS cutinase), were compared to investigate the unique substrate specificity of CLE. The crystal structure of CLE was solved to a 1.05 A resolution. Moreover, hydrolysis assays demonstrated the broad specificity of CLE for short and long-chain substrates, as well as the preferred specificity of FS cutinase for short-chain substrates. In addition, site-directed mutagenesis was performed to increase the hydrolysis activity on long-chain substrates, indicating that the hydrophobic aromatic residues are important for the specificity to the long-chain substrate. These results indicate that hydrophobic residues, especially the aromatic ones exposed to solvent, are important for retaining lipase activity.
Kodama
Yuya
Y
Institute of Life Sciences, Ajinomoto Co., Inc., Kawasaki-ku, Kawasaki-shi 210-8681, Japan.
Masaki
Kazuo
K
Kondo
Hidemasa
H
Suzuki
Mamoru
M
Tsuda
Sakae
S
Nagura
Tomohiro
T
Shimba
Nobuhisa
N
Suzuki
Ei-ichiro
E
Iefuji
Haruyuki
H
D001665:Binding Sites; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry*; D002384:Catalysis; D003454:Cryptococcus / Q000378:metabolism*; D018360:Crystallography, X-Ray / Q000379:methods; D004220:Disulfides; D004926:Escherichia coli / Q000201:enzymology; D005670:Fusarium / Q000201:enzymology; D006868:Hydrolysis; D008958:Models, Molecular; D008968:Molecular Conformation; D016297:Mutagenesis, Site-Directed; D012997:Solvents / Q000737:chemistry; D013379:Substrate Specificity
D004220:Disulfides; D012997:Solvents; D002265:Carboxylic Ester Hydrolases; C019952:cutinase