ArticleThe Journal of biological chemistry2014
Enhanced enzyme kinetic stability by increasing rigidity within the active site.
Article in The Journal of biological chemistry, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 78 papers, 2 of them syntheses that pooled it.
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Who cites it
78 citing papers in PubMed, 2 syntheses or guidelines pooled it.
- The diversity of PET degrading enzymes: A systematic review of sequence, structure, and function.Protein science : a publication of the Protein Society · 2025Pooled it
- A meta-analysis of the activity, stability, and mutational characteristics of temperature-adapted enzymes.Bioscience reports · 2021Pooled it
- Semi-Rational Engineering of Formate Dehydrogenase Reveals Loop Dynamics as a Key Determinant of Thermostability.Chembiochem : a European journal of chemical biology · 2026Article
- Rational structural design for the dual enhancement of catalytic activity and thermal stability of hyaluronidase from Citrobacter portucalensis.Biotechnology for biofuels and bioproducts · 2026Article
- Modulation of Fructose Transfer Process for Promoting Apparent Isomerization Activity of Amylosucrase fromJournal of agricultural and food chemistry · 2026Article
- Investigation of regioselectivity and thermostability of free and immobilized Pleurotus citrinopileatus lipase.Applied microbiology and biotechnology · 2026Article
- Temperature-Dependent Active-Site Rearrangements of PETaseSM14: Insights from Molecular Dynamics Simulations.International journal of molecular sciences · 2026Article
- Computational Design of a Thermo-Acidostable Endo-Polygalacturonase for Efficient Juice Extraction.Foods (Basel, Switzerland) · 2026Article
- Green bioconversion of insoluble chitin: chitinase development pathways via multi-strategy synergy.Bioresources and bioprocessing · 2026Review
- A CALB-like Cold-Active Lipolytic Enzyme fromMarine drugs · 2025Article
- Engineering a novel carbaryl-degrading esterase from Bacillus velezensis for enhanced degradability via semi-rational design and whole-cell biocatalysis.World journal of microbiology & biotechnology · 2025Article
- Catalytic His-loop flexibility drives high activity in hyperthermophilic esterase EstE1 while preserving structural stability.Microbiology spectrum · 2025Article
- pH Controlled Activation and Stabilization of Enzymes Using Responsive Polymer-Bioconjugates.Biomacromolecules · 2025Article
- Exploring sequence- and structure-based fitness landscapes to enhance thermal resistance and activity of endoglucanase II with minimal experimental effort.RSC chemical biology · 2025Article
- Analysis of epitopes and structural responses in egg allergen Gal d 1 using bioinformatic tools and molecular dynamics simulation.Current research in food science · 2025Article
- Structural Insights into Cold-Active Lipase from Glaciozyma antarctica PI12: Alphafold2 Prediction and Molecular Dynamics Simulation.Journal of molecular evolution · 2024Article
- Regiocontrol of the Bulk Polymerization of Lysine Ethyl Ester by the Selection of Suitable Immobilized Enzyme Catalysts.Biomacromolecules · 2024Article
- Improvement of thermostability by increasing rigidity in the finger regions and flexibility in the catalytic pocket area of Pseudoalteromonas porphyrae κ-carrageenase.World journal of microbiology & biotechnology · 2024Article
- In silico design of multipoint mutants for enhanced performance of Thermomyces lanuginosus lipase for efficient biodiesel production.Biotechnology for biofuels and bioproducts · 2024Article
- Revealing the Role of the Arg and Lys in Shifting Paradigm from BTK Selective Inhibition to the BTK/HCK Dual Inhibition - Delving into the Inhibitory Activity of KIN-8194 against BTK, and HCK in the Treatment of MutatedAnti-cancer agents in medicinal chemistry · 2024Article
18 more citing papers are in PubMed but not listed here.
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Enzyme stability is an important issue for protein engineers. Understanding how rigidity in the active site affects protein kinetic stability will provide new insight into enzyme stabilization. In this study, we demonstrated enhanced kinetic stability of Candida antarctica lipase B (CalB) by mutating the structurally flexible residues within the active site. Six residues within 10 Å of the catalytic Ser(105) residue with a high B factor were selected for iterative saturation mutagenesis. After screening 2200 colonies, we obtained the D223G/L278M mutant, which exhibited a 13-fold increase in half-life at 48 °C and a 12 °C higher T50(15), the temperature at which enzyme activity is reduced to 50% after a 15-min heat treatment. Further characterization showed that global unfolding resistance against both thermal and chemical denaturation also improved. Analysis of the crystal structures of wild-type CalB and the D223G/L278M mutant revealed that the latter formed an extra main chain hydrogen bond network with seven structurally coupled residues within the flexible α10 helix that are primarily involved in forming the active site. Further investigation of the relative B factor profile and molecular dynamics simulation confirmed that the enhanced rigidity decreased fluctuation of the active site residues at high temperature. These results indicate that enhancing the rigidity of the flexible segment within the active site may provide an efficient method for improving enzyme kinetic stability.
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