Site-saturation mutagenesis of Glomerella cingulata cutinase gene for enhanced enzyme thermostability

Wan Nurhidayah Wan Hanapi, Chin Iuan-Sheau, Nor Muhammad Mahadi, Abdul Munir Abd. Murad, Farah Diba Abu Bakar

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Citations (Scopus)

Abstract

Cutinase is an important biocatalyst for various industrial applications. This enzyme which has dual functionality comparable to esterases and lipases, is efficient in the hydrolysis of soluble esters and emulsified triacylglycerols. Naturally-occurring enzymes usually have disadvantages when applied in non-natural catalysis due to Glomerella cingulata cutinase enzyme thermostability. It is postulated that by increasing the rigidity at certain amino acid positions showing high mobility based on the three-dimensional structure of G. cingulata cutinase, the improvement in thermostability will be achieved. The amino acid N82 of G. cingulata cutinase was selected based on its high B-factor value determined via the B-FITTER program. Megaprimer PCR was employed to introduce mutations at the chosen site by randomization using NNK degenerate primers. About 300 transformants were selected for screening of positive cutinase variants. The N82-V14 cutinase variant was observed to be more thermostable at an almost 2-fold increase when exposed at 50°C for 1 hr as compared to the wild-type enzyme. This study may provide valuable information regarding thermal stability of cutinases denaturation at high temperatures.

Original languageEnglish
Title of host publication2015 UKM FST Postgraduate Colloquium: Proceedings of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology 2015 Postgraduate Colloquium
PublisherAmerican Institute of Physics Inc.
Volume1678
ISBN (Electronic)9780735413252
DOIs
Publication statusPublished - 25 Sep 2015
Event2015 Postgraduate Colloquium of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology, UKM FST 2015 - Selangor, Malaysia
Duration: 15 Apr 201516 Apr 2015

Other

Other2015 Postgraduate Colloquium of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology, UKM FST 2015
CountryMalaysia
CitySelangor
Period15/4/1516/4/15

Fingerprint

mutagenesis
genes
enzymes
thermal stability
saturation
amino acids
primers
biopolymer denaturation
mutations
rigidity
catalysis
hydrolysis
esters
screening

ASJC Scopus subject areas

  • Physics and Astronomy(all)

Cite this

Hanapi, W. N. W., Iuan-Sheau, C., Mahadi, N. M., Abd. Murad, A. M., & Abu Bakar, F. D. (2015). Site-saturation mutagenesis of Glomerella cingulata cutinase gene for enhanced enzyme thermostability. In 2015 UKM FST Postgraduate Colloquium: Proceedings of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology 2015 Postgraduate Colloquium (Vol. 1678). [030021] American Institute of Physics Inc.. https://doi.org/10.1063/1.4931242

Site-saturation mutagenesis of Glomerella cingulata cutinase gene for enhanced enzyme thermostability. / Hanapi, Wan Nurhidayah Wan; Iuan-Sheau, Chin; Mahadi, Nor Muhammad; Abd. Murad, Abdul Munir; Abu Bakar, Farah Diba.

2015 UKM FST Postgraduate Colloquium: Proceedings of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology 2015 Postgraduate Colloquium. Vol. 1678 American Institute of Physics Inc., 2015. 030021.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Hanapi, WNW, Iuan-Sheau, C, Mahadi, NM, Abd. Murad, AM & Abu Bakar, FD 2015, Site-saturation mutagenesis of Glomerella cingulata cutinase gene for enhanced enzyme thermostability. in 2015 UKM FST Postgraduate Colloquium: Proceedings of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology 2015 Postgraduate Colloquium. vol. 1678, 030021, American Institute of Physics Inc., 2015 Postgraduate Colloquium of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology, UKM FST 2015, Selangor, Malaysia, 15/4/15. https://doi.org/10.1063/1.4931242
Hanapi WNW, Iuan-Sheau C, Mahadi NM, Abd. Murad AM, Abu Bakar FD. Site-saturation mutagenesis of Glomerella cingulata cutinase gene for enhanced enzyme thermostability. In 2015 UKM FST Postgraduate Colloquium: Proceedings of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology 2015 Postgraduate Colloquium. Vol. 1678. American Institute of Physics Inc. 2015. 030021 https://doi.org/10.1063/1.4931242
Hanapi, Wan Nurhidayah Wan ; Iuan-Sheau, Chin ; Mahadi, Nor Muhammad ; Abd. Murad, Abdul Munir ; Abu Bakar, Farah Diba. / Site-saturation mutagenesis of Glomerella cingulata cutinase gene for enhanced enzyme thermostability. 2015 UKM FST Postgraduate Colloquium: Proceedings of the Universiti Kebangsaan Malaysia, Faculty of Science and Technology 2015 Postgraduate Colloquium. Vol. 1678 American Institute of Physics Inc., 2015.
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