Corner Engineering: Tailoring Enzymes for Enhanced Resistance and Thermostability in Deep Eutectic Solvents
Angewandte Chemie, vol. 136
Abstract
Abstract Deep eutectic solvents (DESs), heralded for their synthesis simplicity, economic viability, and reduced volatility and flammability, have found increasing application in biocatalysis. However, challenges persist due to a frequent diminution in enzyme activity and stability. Herein, we developed a general protein engineering strategy, termed corner engineering, to acquire DES‐resistant and thermostable enzymes via precise tailoring of the transition region in enzyme structure. Employing Bacillus subtilis lipase A (BSLA) as a model, we delineated the engineering process, yielding five multi‐DESs resistant variants with highly improved thermostability, such as K88E/N89 K exhibited up to a 10.0‐fold catalytic efficiency (kcat/KM) increase in 30 % (v/v) choline chloride (ChCl): acetamide and 4.1‐fold in 95 % (v/v) ChCl: ethylene glycol accompanying 6.7‐fold thermal resistance improvement than wild type at ≈50 °C. The generality of the optimized approach was validated by two extra industrial enzymes, endo‐β‐1,4‐glucanase PvCel5A (used for biofuel production) and esterase Bs2Est (used for plastics degradation). The molecular investigations revealed that increased water molecules at substrate binding cleft and finetuned helix formation at the corner region are two dominant determinants governing elevated resistance and thermostability. This study, coupling corner engineering with obtained molecular insights, illuminates enzyme‐DES interaction patterns and fosters the rational design of more DES‐resistant and thermostable enzymes in biocatalysis and biotransformation.
Authors 6
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Affiliation as printed
School of Food Science and Pharmaceutical Engineering Nanjing Normal University No. 2 Xuelin Road Nanjing 210097 China
Nanjing Normal University, School of Food and Pharmaceutical Engineering, No.2 Xuelin Road, Nanjing, 210023, China, 210097 Nanjing, CHINA
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Affiliation as printed
School of Food Science and Pharmaceutical Engineering Nanjing Normal University No. 2 Xuelin Road Nanjing 210097 China
Nanjing Normal University, School of Food and Pharmaceutical Engineering, CHINA
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Haiyang Cui Aachen
University of Illinois Urbana-Champaign · Carl R. Woese Institute for Genomic Biology · RWTH Aachen University
Affiliation as printed
Current address: Carl R. Woese Institute for Genomic Biology University of Illinois at Urbana-Champaign 1206 West Gregory Drive Urbana IL 61801 USA
RWTH Aachen University Templergraben 55 52062 Aachen Germany
University of Illinois Urbana-Champaign, Carl R. Woese Institute for Genomic Biology, UNITED STATES
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Affiliation as printed
School of Food Science and Pharmaceutical Engineering Nanjing Normal University No. 2 Xuelin Road Nanjing 210097 China
Nanjing Normal University, School of Food and Pharmaceutical Engineering, CHINA
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Xiujuan Li corresponding
Affiliation as printed
School of Food Science and Pharmaceutical Engineering Nanjing Normal University No. 2 Xuelin Road Nanjing 210097 China
Nanjing Normal University, School of Food Science and Pharmaceutical Engineering, No. 2 Xuelin Road, 210023 Nanjing, CHINA
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He Huang corresponding
Affiliation as printed
School of Food Science and Pharmaceutical Engineering Nanjing Normal University No. 2 Xuelin Road Nanjing 210097 China
Nanjing Normal University, School of Food and Pharmaceutical Engineering, CHINA
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