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

• Main Authors: Khairul Anuar, Nurul Fatin Syamimi, Abdul Wahab, Roswanira, Huyop, Fahrul Zaman, Abd Halim, Khairul Bariyyah, Abdul Hamid, Azzmer Azzar
• Format: Article
• Language: English; English; English
• Published: Taylor and Francis Ltd. 2020
• Subjects: QD Chemistry
• Online Access: http://irep.iium.edu.my/88006/1/88006_In%20silico%20mutation%20on%20a%20mutant%20lipase.pdf; http://irep.iium.edu.my/88006/2/88006_In%20silico%20mutation%20on%20a%20mutant%20lipase_SCOPUS.pdf; http://irep.iium.edu.my/88006/3/88006_In%20silico%20mutation%20on%20a%20mutant%20lipase_WoS.pdf; http://irep.iium.edu.my/88006/; https://www.tandfonline.com/doi/full/10.1080/07391102.2019.1683074

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 = ∼0.3 nm, RMSF = ∼0.05–0.2 nm), over the optimal pH 8.0 of the wild-type lipase (RMSD = 0.3 nm, RMSF = 0.05–0.20 nm). 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