Identification of D۷۵C, E۲۹۶L as the most promising thermostable mutant of the acetylesterase enzyme from Trichoderma harzianum IOC-۳۸۴۴ by an in-silico approach

سال انتشار: 1402
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 109

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شناسه ملی سند علمی:

IBIS12_210

تاریخ نمایه سازی: 5 دی 1403

چکیده مقاله:

Introduction: Trichoderma harzianum IOC-۳۸۴۴ has good characteristics for biomass degradation Its acetylesterase (EC ۳.۱.۱.۶) [۲] hydrolyzes glucuronoxylan acetyl side groups and plays a role in the biodegradation of xylan [۳]. It’s a safe alternative to harmful chemicals like chlorine as a bleaching agent in the pulp and paper industries [۴]. This study aims to improve the enzyme's thermal stability using an in-silico approach, making it suitable for high-temperature industrial applications. Methods: The study used Augustus gene prediction to identify the enzyme's coding gene and created a ۳D model using the Alphafold server. FoldX۵ was used for stability measurement, and key residues were found through the NCBI CDD database. Alanine scanning identified the best enzyme-stabilizing mutations, creating single, double, triple, and quadruple mutants while keeping key residues intact. and molecular docking and MM-GBSA scoring were performed on the wild-type enzyme and over thirty mutants. Four top candidates were chosen for molecular dynamics simulation to assess mutations' effect on enzyme thermal stability over time. Results: The enzyme model's stability was found to be -۳۶.۱۷ kcal/mol. However, all the mutants picked for docking showed better stability scores. This enzyme belongs to the SGNH hydrolase family and has four key residues: Ser۲۱, Gly۵۴, Asn۱۲۱, and His۲۹۳. The wild-type enzyme had an average docking and MM-GBSA score of -۲۱.۵۴ kcal/mol, with all four top mutants performing better. After running molecular dynamics and comparing geometrical parameters, including RMSD, RMSF, H-bonds, SASA, and Rg, for the wild-type enzyme and all four mutants at temperatures of ۳۷ and ۷۰ °C, we identified a double mutant (D۷۵C, E۲۹۶L) as the most promising variant for temperature stability. Conclusion: The study identifies D۷۵C, E۲۹۶L as the most thermostable and effective mutant of the acetylesterase enzyme, making it a safer alternative to hazardous chemicals and a viable solution for industrial applications.

نویسندگان

Ali Abolhasanzadeh Parizi

Department of Biology, Faculty of Science, University of Sistan and Baluchestan, Zahedan, Iran

Milad Lagzian

Department of Biology, Faculty of Science, University of Sistan and Baluchestan, Zahedan, Iran