Unleashing the Power of Evolution in Xylanase Engineering: Investigating the Role of Distal Mutation Regulation

• DOI: 10.1021/acs.jafc.4c03245
• Publication Date: 2024-07-31T17:30:26Z
• Authors: Ya Wu; Yu Yang; Gen Lu; Wan-Lu Xiang; Tian-Yu Sun; Ke-Wei Chen; Xiang Lv; Yi-Fan Gui; Rui-Qi Zeng; Yi-Kai Du; Chun-Hua Fu; Jian-Wen Huang; Chun-Chi Chen; Rey-Ting Guo; Long-Jiang Yu
• Keywords: Thermal Stability; Protein Engineering; Molecular mechanics

The drive to enhance enzyme performance in industrial applications frequently clashes with the practical limitations of exhaustive experimental screening, underscoring urgency for more refined and strategic methodologies engineering. In this study, xylanase Xyl-1 was used as model, coupling evolutionary insights energy functions obtain theoretical potential mutants, which were subsequently validated experimentally. We observed that mutations nonloop region primarily aimed at enhancing stability also encountered selective pressure activity. Notably, simultaneously boosted activity, achieving a 65% success rate. Using greedy strategy, mutant M4 developed, 12 °C higher melting temperature doubled By integration spectroscopy, crystallography, quantum mechanics/molecular mechanics molecular dynamics, mechanism behind enhanced thermal elucidated. It determined activity differences between wild type driven by dynamic factors influenced distal mutations. conclusion, study emphasizes pivotal role evolution-based approaches augmenting enzymes. sheds light on unique adaptive mechanisms employed various structural regions proteins expands our understanding intricate relationship distant dynamics.