Thermal stability engineering of Glomerella cingulata cutinase

Iuan Sheau Chin, Abdul Munir Abd. Murad, Nor Muhammad Mahadi, Sheila Nathan, Farah Diba Abu Bakar

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Cutinase has been ascertained as a biocatalyst for biotechnological and industrial bioprocesses. The Glomerella cingulata cutinase was genetically modified to enhance its enzymatic performance to fulfill industrial requirements. Two sites were selected for mutagenesis with the aim of altering the surface electrostatics as well as removing a potentially deamidation-prone asparagine residue. The N177D cutinase variant was affirmed to be more resilient to temperature increase with a 2.7-fold increase in half-life at 50°C as compared with wild-type enzyme, while, the activity at 25°C is not compromised. Furthermore, the increase in thermal tolerance of this variant is accompanied by an increase in optimal temperature. Another variant, the L172K, however, exhibited higher enzymatic performance towards phenyl ester substrates of longer carbon chain length, yet its thermal stability is inversely affected. In order to restore the thermal stability of L172K, we constructed a L172K/N177D double variant and showed that these two mutations yield an improved variant with enhanced activity towards phenyl ester substrates and enhanced thermal stability. Taken together, our study may provide valuable information for enhancing catalytic performance and thermal stability in future engineering endeavors.

Original languageEnglish
Pages (from-to)369-375
Number of pages7
JournalProtein Engineering, Design and Selection
Volume26
Issue number5
DOIs
Publication statusPublished - May 2013

Fingerprint

Phyllachorales
Thermodynamic stability
Hot Temperature
Esters
Mutagenesis
Biocatalysts
Temperature
Asparagine
Substrates
Enzymes
Static Electricity
Chain length
Half-Life
Electrostatics
Carbon
Mutation
cutinase

Keywords

  • deamidation
  • dynamic light scattering
  • enzyme engineering
  • Glomerella cingulata cutinase
  • thermal stability

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Molecular Biology

Cite this

Thermal stability engineering of Glomerella cingulata cutinase. / Chin, Iuan Sheau; Abd. Murad, Abdul Munir; Mahadi, Nor Muhammad; Nathan, Sheila; Abu Bakar, Farah Diba.

In: Protein Engineering, Design and Selection, Vol. 26, No. 5, 05.2013, p. 369-375.

Research output: Contribution to journalArticle

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