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

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

doi:10.1063/1.4931242

Cited by 4 publications

(1 citation statement)

References 19 publications

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“…Protein engineering techniques have also been applied in order to increase the thermostability of cutinases. G. cingulata single mutants aiming at the removal of an asparagine prone to deamination [71] and in increasing the rigidity of the structure [75], led to enhanced thermostability. TfCut2 from T. fusca has been shown to increase its T m by 13 • C, through the addition of 10 mM Ca 2+ [76].…”

Section: Host Selection and Protein Engineering Techniquesmentioning

confidence: 99%

A Middle-Aged Enzyme Still in Its Prime: Recent Advances in the Field of Cutinases

et al. 2018

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Cutinases are α/β hydrolases, and their role in nature is the degradation of cutin. Such enzymes are usually produced by phytopathogenic microorganisms in order to penetrate their hosts. The first focused studies on cutinases started around 50 years ago. Since then, numerous cutinases have been isolated and characterized, aiming at the elucidation of their structure–function relations. Our deeper understanding of cutinases determines the applications by which they could be utilized; from food processing and detergents, to ester synthesis and polymerizations. However, cutinases are mainly efficient in the degradation of polyesters, a natural function. Therefore, these enzymes have been successfully applied for the biodegradation of plastics, as well as for the delicate superficial hydrolysis of polymeric materials prior to their functionalization. Even though research on this family of enzymes essentially began five decades ago, they are still involved in many reports; novel enzymes are being discovered, and new fields of applications arise, leading to numerous related publications per year. Perhaps the future of cutinases lies in their evolved descendants, such as polyesterases, and particularly PETases. The present article reviews the biochemical and structural characteristics of cutinases and cutinase-like hydrolases, and their applications in the field of bioremediation and biocatalysis.

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Section: Host Selection and Protein Engineering Techniquesmentioning

confidence: 99%

A Middle-Aged Enzyme Still in Its Prime: Recent Advances in the Field of Cutinases

et al. 2018

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Protein Engineering of Enzyme Robustness Relevant to Organic and Pharmaceutical Chemistry and Applications in Biotechnology

2023

Enzyme Engineering

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Utility of B-Factors in Protein Science: Interpreting Rigidity, Flexibility, and Internal Motion and Engineering Thermostability

et al. 2019

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The term B-factor, sometimes called the Debye–Waller factor, temperature factor, or atomic displacement parameter, is used in protein crystallography to describe the attenuation of X-ray or neutron scattering caused by thermal motion. This review begins with analyses of early protein studies which suggested that B-factors, available from the Protein Data Bank, can be used to identify the flexibility of atoms, side chains, or even whole regions. This requires a technique for obtaining normalized B-factors. Since then the exploitation of B-factors has been extensively elaborated and applied in a variety of studies with quite different goals, all having in common the identification and interpretation of rigidity, flexibility, and/or internal motion which are crucial in enzymes and in proteins in general. Importantly, this review includes a discussion of limitations and possible pitfalls when using B-factors. A second research area, which likewise exploits B-factors, is also reviewed, namely, the development of the so-called B-FIT-directed evolution method for increasing the thermostability of enzymes as catalysts in organic chemistry and biotechnology. In both research areas, a maximum of structural and mechanistic insights is gained when B-factor analyses are combined with other experimental and computational techniques.

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

Cited by 4 publications

References 19 publications

A Middle-Aged Enzyme Still in Its Prime: Recent Advances in the Field of Cutinases

A Middle-Aged Enzyme Still in Its Prime: Recent Advances in the Field of Cutinases

Protein Engineering of Enzyme Robustness Relevant to Organic and Pharmaceutical Chemistry and Applications in Biotechnology

Utility of B-Factors in Protein Science: Interpreting Rigidity, Flexibility, and Internal Motion and Engineering Thermostability

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