Abstract
One of the most desirable properties for industrial enzymes is high thermotolerance, which can reduce the amount of biocatalyst used and lower the production cost. Aiming to improve the thermotolerance of trehalose synthase (TreS, EC 5.4.99.16) from Thermomonospora curvata, four mutants (G78D, V289L, G322A, I323L) and four cyclized TreS variants fused using different Tag/Catcher pairs (SpyTag-TreS-SpyCatcher, SpyTag-TreS-KTag, SnoopTag-TreS-SnoopCatcher, SnoopTagJR-TreS-DogTag) were constructed. The results showed that cyclization led to a much larger increase of thermostability than that achieved via site-directed mutagenesis. The t1/2 of all four cyclized TreS variants at 55 °C increased 2- to 3- fold, while the analysis of kinetic and thermodynamic stability indicated that the T50 of the different cyclized TreS variants increased by between 7.5 °C and 15.5 °C. Molecular dynamics simulations showed that the Rg values of cyclized TreS decreased significantly, indicating that the protein maintained a tight tertiary structure at high temperatures, avoiding exposure of the hydrophobic core to the solvent. Cyclization using a Tag/Catcher pair is a simple and effective method for improving the thermotolerance of enzymes.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: International Journal of Biological Macromolecules
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
