Crystal structure and enhanced activity of a cutinase-like enzyme from Cryptococcus sp. strain S-2

Proteins. 2009 Nov 15;77(3):710-7. doi: 10.1002/prot.22484.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Carboxylic Ester Hydrolases / chemistry*
  • Catalysis
  • Cryptococcus / metabolism*
  • Crystallography, X-Ray / methods
  • Disulfides
  • Escherichia coli / enzymology
  • Fusarium / enzymology
  • Hydrolysis
  • Models, Molecular
  • Molecular Conformation
  • Mutagenesis, Site-Directed
  • Solvents / chemistry
  • Substrate Specificity

Substances

  • Disulfides
  • Solvents
  • Carboxylic Ester Hydrolases
  • cutinase