Effect of surface charge alteration on stability of L-asparaginase II from Escherichia sp

Enzyme Microb Technol. 2014 Mar 5;56:15-9. doi: 10.1016/j.enzmictec.2013.12.012. Epub 2013 Dec 22.

Abstract

Escherichia coli L-asparaginases have great significance in the treatment of leukemia. Consequently, there is considerable interest in engineering this enzyme for improving its stability. In this work, the effect of surface charge on the stability of the enzyme l-asparaginase II was studied by site-directed mutagenesis of the cloned ansB gene from Escherichia sp. Replacement of two positively charged residues (K139 and K207) on the surface loops with neutral and reverse charges resulted in altered thermo stability in designed variants. Neutral charge substitutions (K139A and K207A) retained greater tolerance and stability followed by negative charge substitutions (K139D and K207D) compared to control mutant K139R and wild enzyme. From the results, it was concluded that the optimization of surface charge contributed much to the thermal properties of proteins without affecting the structure.

Keywords: Site-directed mutagenesis; Surface charge alteration; Thermal tolerance; l-Asparaginase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Asparaginase / chemistry*
  • Asparaginase / genetics
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Base Sequence
  • Cattle
  • Cloning, Molecular
  • Escherichia / enzymology*
  • Escherichia coli
  • Feces / microbiology
  • Hot Temperature
  • Hydrogen-Ion Concentration
  • Models, Molecular
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Protein Conformation
  • Protein Denaturation
  • Protein Stability
  • Static Electricity
  • Surface Properties

Substances

  • Bacterial Proteins
  • Asparaginase