ArticleApplied and environmental microbiology2014
Thermostability improvement of a streptomyces xylanase by introducing proline and glutamic acid residues.
Article in Applied and environmental microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 121 citations in OpenAlex.
- Influence of a Thermostable Fungal Xylanase Combined With Multi-Enzyme Complexes on the Structure, Physicochemical, and Functional Properties of Dietary Fiber From Hawthorn Pomace.Journal of food science · 2026Article
- Thermostable Proteases fromMicroorganisms · 2025Review
- Network Dynamics as Fingerprints of Thermostability in an In Silico-Engineered DyP-Type Peroxidase.ACS catalysis · 2025Article
- Discovery andACS omega · 2025Article
- Generation and characterization of two acid-resistant macrocinComputational and structural biotechnology journal · 2024Article
- Thermostability modification of β-mannanase fromFrontiers in microbiology · 2023Article
- Engineering the Thermostability of the Mono- and Diacylglycerol Lipase SMG1 for the Synthesis of Diacylglycerols.Foods (Basel, Switzerland) · 2022Article
- Thermostability engineering of industrial enzymes through structure modification.Applied microbiology and biotechnology · 2022Review
- High-temperature behavior of hyperthermostable Thermotoga maritima xylanase XYN10B after designed and evolved mutations.Applied microbiology and biotechnology · 2022Article
- Improvement of XYL10C_∆N catalytic performance through loop engineering for lignocellulosic biomass utilization in feed and fuel industries.Biotechnology for biofuels · 2021Article
- Significantly improving the thermostability of a hyperthermophilic GH10 family xylanase XynAF1 by semi-rational design.Applied microbiology and biotechnology · 2021Article
- A Thermally Stable Protein EPP1 of Corn Borer Ostrinia furnacalis Regulates Hemocytic Encapsulation.Journal of innate immunity · 2021Article
- Novel Proteome and N-Glycoproteome of the Thermophilic FungusFrontiers in microbiology · 2021Article
- Improving the thermostability of a GH97 dextran glucosidase by rational design.Biotechnology letters · 2020Article
- Improving the catalytic thermostability of3 Biotech · 2020Article
- Isolation and Characterization of a Novel Cold-Active, Halotolerant Endoxylanase fromMarine drugs · 2020Article
- Site-directed mutagenesis of coenzyme-independent carotenoid oxygenase CSO2 to enhance the enzymatic synthesis of vanillin.Applied microbiology and biotechnology · 2020Article
- Genetically Engineered Proteins to Improve Biomass Conversion: New Advances and Challenges for Tailoring Biocatalysts.Molecules (Basel, Switzerland) · 2019Review
- Biochemical Characteristics of Microbial Enzymes and Their Significance from Industrial Perspectives.Molecular biotechnology · 2019Review
- Discovery of a Thermostable GH10 Xylanase with Broad Substrate Specificity from the Arctic Mid-Ocean Ridge Vent System.Applied and environmental microbiology · 2019Article
Corrections and comments
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Authors and funding
10 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Protein engineering is commonly used to improve the robustness of enzymes for activity and stability at high temperatures. In this study, we identified four residues expected to affect the thermostability of Streptomyces sp. strain S9 xylanase XynAS9 through multiple-sequence analysis (MSA) and molecular dynamic simulations (MDS). Site-directed mutagenesis was employed to construct five mutants by replacing these residues with proline or glutamic acid (V81P, G82E, V81P/G82E, D185P/S186E, and V81P/G82E/D185P/S186E), and the mutant and wild-type enzymes were expressed in Pichia pastoris. Compared to the wild-type XynAS9, all five mutant enzymes showed improved thermal properties. The activity and stability assays, including circular dichroism and differential scanning calorimetry, showed that the mutations at positions 81 and 82 increased the thermal performance more than the mutations at positions 185 and 186. The mutants with combined substitutions (V81P/G82E and V81P/G82E/D185P/S186E) showed the most pronounced shifts in temperature optima, about 17°C upward, and their half-lives for thermal inactivation at 70°C and melting temperatures were increased by >9 times and approximately 7.0°C, respectively. The mutation combination of V81P and G82E in adjacent positions more than doubled the effect of single mutations. Both mutation regions were at the end of long secondary-structure elements and probably rigidified the local structure. MDS indicated that a long loop region after positions 81 and 82 located at the end of the inner β-barrel was prone to unfold. The rigidified main chain and filling of a groove by the mutations on the bottom of the active site canyon may stabilize the mutants and thus improve their thermostability.
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