Effect of mutation at oxyanion hole residu (H110F) on activity of Lk4 lipase.

Autor:	Ma'ruf IF; Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Indonesia.; Genetic and Molecular Biotechnology Research Group, School of Life Sciences and Technology, Institut Teknologi Bandung, Indonesia., Widhiastuty MP; Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Indonesia., Suharti; Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Indonesia.; Department of Chemistry, Faculty of Science and Computer, Universitas Pertamina, Indonesia., Moeis MR; Genetic and Molecular Biotechnology Research Group, School of Life Sciences and Technology, Institut Teknologi Bandung, Indonesia., Akhmaloka; Biochemistry Research Group, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Indonesia.; Department of Chemistry, Faculty of Science and Computer, Universitas Pertamina, Indonesia.
Jazyk:	angličtina
Zdroj:	Biotechnology reports (Amsterdam, Netherlands) [Biotechnol Rep (Amst)] 2021 Jan 16; Vol. 29, pp. e00590. Date of Electronic Publication: 2021 Jan 16 (Print Publication: 2021).
DOI:	10.1016/j.btre.2021.e00590
Abstrakt:	Mutant of lipase at oxyanion hole (H110 F) was constructed. The gene was highly expressed in Eschericia coli BL21 (DE3) and the recombinant protein was purified using Ni-NTA affinity chromatography. The activity of mutant enzyme was significantly increased compared to that the wild type. Further comparison showed that both of the enzymes exhibited same optimum pH and temperature, and showed highest lipolytic activity on pNP-decanoate (C10). The wild type appeared lost of activity on C14 and C16 substrates meanwhile the mutant still showed activity up to 20 %. In the presence of non polar organic solvent such as n-hexane, the wild type became inactive enzyme meanwhile the mutant still remained 50 % of its activity. The results suggested that mutation at oxyanion hole (H110 F) caused enzyme-substrate interaction change resulting on elevation of activity, better activity toward longer carbon chain substrate and improving the activity in the present of non polar organic solvent. Competing Interests: The authors report no declarations of interest. (© 2021 The Authors. Published by Elsevier B.V.)
Databáze:	MEDLINE
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