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Directed evolution of recombinant c-terminal truncated staphylococcus epidermidis lipase AT2 for the enhancement of thermostability


Citation

Veno, Jiivittha and Ahmad Kamarudin, Nor Hafizah and Mohamad Ali, Mohd Shukuri and Masomian, Malihe and Raja Abd. Rahman, Raja Noor Zaliha (2017) Directed evolution of recombinant c-terminal truncated staphylococcus epidermidis lipase AT2 for the enhancement of thermostability. International Journal of Molecular Sciences, 18 (11). art. no. 2202. pp. 1-17. ISSN 1661-6596; ESSN: 1422-0067

Abstract

In the industrial processes, lipases are expected to operate at temperatures above 45 °C and could retain activity in organic solvents. Hence, a C-terminal truncated lipase from Staphylococcus epidermis AT2 (rT-M386) was engineered by directed evolution. A mutant with glycine-to-cysteine substitution (G210C) demonstrated a remarkable improvement of thermostability, whereby the mutation enhanced the activity five-fold when compared to the rT-M386 at 50 °C. The rT-M386 and G210C lipases were purified concurrently using GST-affinity chromatography. The biochemical and biophysical properties of both enzymes were investigated. The G210C lipase showed a higher optimum temperature (45 °C) and displayed a more prolonged half-life in the range of 40-60 °C as compared to rT-M386. Both lipases exhibited optimal activity and stability at pH 8. The G210C showed the highest stability in the presence of polar organic solvents at 50 °C compared to the rT-M386. Denatured protein analysis presented a significant change in the molecular ellipticity value above 60 °C, which verified the experimental result on the temperature and thermostability profile of G210C.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
DOI Number: https://doi.org/10.3390/ijms18112202
Publisher: Multidisciplinary Digital Publishing Institute
Keywords: Directed evolution; Staphylococcal lipases; Thermostability; Characterization; Circular dichroism
Depositing User: Ms. Ainur Aqidah Hamzah
Date Deposited: 04 Apr 2022 08:13
Last Modified: 04 Apr 2022 08:13
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.3390/ijms18112202
URI: http://psasir.upm.edu.my/id/eprint/61489
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