Structure-based stabilization of an enzyme: the case of penicillin acylase from Alcaligenes faecalis

Tianwen Wang; Hu Zhu; Xingyuan Ma; Yushu Ma; Dongzhi Wei

doi:10.2174/092986606775101571

Structure-based stabilization of an enzyme: the case of penicillin acylase from Alcaligenes faecalis

Protein Pept Lett. 2006;13(2):177-83. doi: 10.2174/092986606775101571.

Authors

Tianwen Wang¹, Hu Zhu, Xingyuan Ma, Yushu Ma, Dongzhi Wei

Affiliation

¹ State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, P.R. China.

PMID: 16472081
DOI: 10.2174/092986606775101571

Abstract

The modeled structure of penicillin acylase from Alcaligenes faecali (AFPGA) was constructed by comparative modeling with the Modeller program. Candidate positions that could be replaced with cysteine were estimated by scanning the modeled structure of AFPGA with the program MODIP (modeling disulfide bond in protein). The mutant Q3C/P751C had a higher optimum temperature by three degrees than that of the wild type AFPGA. The half life of the double mutant Q3C/P751C at 55 degrees C was increased by 50%. To our knowledge, this was the first structure-based genetic modification of AFPGA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alcaligenes faecalis / enzymology*
Alcaligenes faecalis / genetics
Cloning, Molecular
Enzyme Stability
Models, Molecular
Mutagenesis, Site-Directed
Penicillin Amidase / chemistry*
Penicillin Amidase / genetics
Penicillin Amidase / metabolism*
Protein Structure, Tertiary
Structural Homology, Protein
Temperature

Substances

Penicillin Amidase