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In silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability

Alkaline-stable lipases are highly valuable biocatalysts that catalyze reactions under highly basic conditions. Herein, computational predictions of lipase from Acinetobacter haemolyticus and its mutant, Mut-LipKV1 was performed to identify functionally relevant mutations that enhance pH performance...

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Main Authors: Khairul Anuar, Nurul Fatin Syamimi, Abdul Wahab, Roswanira, Huyop, Fahrul, Abd. Halim, Khairul Bariyyah, Abdul Hamid, Azzmer Azzar
Format: Article
Published: Taylor and Francis Ltd. 2020
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QD Chemistry
Online Access:http://eprints.utm.my/id/eprint/91099/
http://dx.doi.org/10.1080/07391102.2019.1683074
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spelling my.utm.910992021-05-31T13:45:13Z http://eprints.utm.my/id/eprint/91099/ In silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability Khairul Anuar, Nurul Fatin Syamimi Abdul Wahab, Roswanira Huyop, Fahrul Abd. Halim, Khairul Bariyyah Abdul Hamid, Azzmer Azzar QD Chemistry Alkaline-stable lipases are highly valuable biocatalysts that catalyze reactions under highly basic conditions. Herein, computational predictions of lipase from Acinetobacter haemolyticus and its mutant, Mut-LipKV1 was performed to identify functionally relevant mutations that enhance pH performance under increasing basicity. Mut-LipKV1 was constructed by in silico site directed mutagenesis of several outer loop acidic residues, aspartic acid (Asp) into basic ones, lysine (Lys) at positions 51, 122 and 247, followed by simulation under extreme pH conditions (pH 8.0–pH 12.0). The energy minimized Mut-LipKV1 model exhibited good quality as shown by PROCHECK, ERRAT and Verify3D data that corresponded to 79.2, 88.82 and 89.42% in comparison to 75.2, 86.15, and 95.19% in the wild-type. Electrostatic surface potentials and charge distributions of the Mut-LipKV1 model was more stable and better adapted to conditions of elevated pHs (pH 8.0−10.0). Mut-LipKV1 exhibited a mixture of neutral and positive surface charge distribution compared to the predominantly negative charge in the wild-type lipase at pH 8.0. Data of molecular dynamics simulations also supported the increased alkaline-stability of Mut-LipKV1, wherein the lipase was more stable at a higher pH 9.0 (RMSD = ∼0.3nm, RMSF = ∼0.05–0.2nm), over the optimal pH 8.0 of the wild-type lipase (RMSD = 0.3nm, RMSF = 0.05–0.20nm). Thus, the adaptive strategy of replacing surface aspartic acid to lysine in lipase was successful in yielding a more alkaline-stable Mut-LipKV1 under elevated basic conditions. Communicated by Ramaswamy H. Sarma. Taylor and Francis Ltd. 2020-10-12 Article PeerReviewed Khairul Anuar, Nurul Fatin Syamimi and Abdul Wahab, Roswanira and Huyop, Fahrul and Abd. Halim, Khairul Bariyyah and Abdul Hamid, Azzmer Azzar (2020) In silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability. Journal of Biomolecular Structure and Dynamics, 38 (15). pp. 4493-4507. ISSN 0739-1102 http://dx.doi.org/10.1080/07391102.2019.1683074 DOI:10.1080/07391102.2019.1683074
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
topic QD Chemistry
spellingShingle QD Chemistry
Khairul Anuar, Nurul Fatin Syamimi
Abdul Wahab, Roswanira
Huyop, Fahrul
Abd. Halim, Khairul Bariyyah
Abdul Hamid, Azzmer Azzar
In silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability
description Alkaline-stable lipases are highly valuable biocatalysts that catalyze reactions under highly basic conditions. Herein, computational predictions of lipase from Acinetobacter haemolyticus and its mutant, Mut-LipKV1 was performed to identify functionally relevant mutations that enhance pH performance under increasing basicity. Mut-LipKV1 was constructed by in silico site directed mutagenesis of several outer loop acidic residues, aspartic acid (Asp) into basic ones, lysine (Lys) at positions 51, 122 and 247, followed by simulation under extreme pH conditions (pH 8.0–pH 12.0). The energy minimized Mut-LipKV1 model exhibited good quality as shown by PROCHECK, ERRAT and Verify3D data that corresponded to 79.2, 88.82 and 89.42% in comparison to 75.2, 86.15, and 95.19% in the wild-type. Electrostatic surface potentials and charge distributions of the Mut-LipKV1 model was more stable and better adapted to conditions of elevated pHs (pH 8.0−10.0). Mut-LipKV1 exhibited a mixture of neutral and positive surface charge distribution compared to the predominantly negative charge in the wild-type lipase at pH 8.0. Data of molecular dynamics simulations also supported the increased alkaline-stability of Mut-LipKV1, wherein the lipase was more stable at a higher pH 9.0 (RMSD = ∼0.3nm, RMSF = ∼0.05–0.2nm), over the optimal pH 8.0 of the wild-type lipase (RMSD = 0.3nm, RMSF = 0.05–0.20nm). Thus, the adaptive strategy of replacing surface aspartic acid to lysine in lipase was successful in yielding a more alkaline-stable Mut-LipKV1 under elevated basic conditions. Communicated by Ramaswamy H. Sarma.
format Article
author Khairul Anuar, Nurul Fatin Syamimi
Abdul Wahab, Roswanira
Huyop, Fahrul
Abd. Halim, Khairul Bariyyah
Abdul Hamid, Azzmer Azzar
author_facet Khairul Anuar, Nurul Fatin Syamimi
Abdul Wahab, Roswanira
Huyop, Fahrul
Abd. Halim, Khairul Bariyyah
Abdul Hamid, Azzmer Azzar
author_sort Khairul Anuar, Nurul Fatin Syamimi
title In silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability
title_short In silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability
title_full In silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability
title_fullStr In silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability
title_full_unstemmed In silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability
title_sort in silico mutation on a mutant lipase from acinetobacter haemolyticus towards enhancing alkaline stability
publisher Taylor and Francis Ltd.
publishDate 2020
url http://eprints.utm.my/id/eprint/91099/
http://dx.doi.org/10.1080/07391102.2019.1683074
_version_ 1702169645167935488
score 13.211869

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