Computationally Informed Redesign of Levansucrase from Erwinia tasmaniensis to Enhance Its Thermostability for Levan Biosynthesis
Issued Date
2025-12-10
Resource Type
ISSN
00218561
eISSN
15205118
Scopus ID
2-s2.0-105024477917
Pubmed ID
41288511
Journal Title
Journal of Agricultural and Food Chemistry
Volume
73
Issue
49
Start Page
31523
End Page
31532
Rights Holder(s)
SCOPUS
Bibliographic Citation
Journal of Agricultural and Food Chemistry Vol.73 No.49 (2025) , 31523-31532
Suggested Citation
Charoenwongpaiboon T., Srichompoo Y., Wangpaiboon K., Benini S., Field R.A., Lorthongpanich C., Pongsawasdi P., Pichyangkura R. Computationally Informed Redesign of Levansucrase from Erwinia tasmaniensis to Enhance Its Thermostability for Levan Biosynthesis. Journal of Agricultural and Food Chemistry Vol.73 No.49 (2025) , 31523-31532. 31532. doi:10.1021/acs.jafc.5c11841 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/113571
Title
Computationally Informed Redesign of Levansucrase from Erwinia tasmaniensis to Enhance Its Thermostability for Levan Biosynthesis
Corresponding Author(s)
Other Contributor(s)
Abstract
Levan is a versatile biomaterial because of its unique physicochemical properties and bioactivities. To synthesize levan efficiently, it is important to improve the thermostability of levansucrase. This study presents the first engineering study on Erwinia tasmaniensis levansucrase (EtLsc) employing a rational protein design approach. Molecular dynamics (MD) simulations were used to identify thermally sensitive regions of EtLsc, and thermostable variants were designed by using FireProt folding energy calculations. Among the designed candidates, the A197P and S239P mutants had largely higher melting temperatures (T<inf>m</inf>) and half-life (t<inf>1/2</inf>) compared to the wild type. The double variant A197P/S239P exhibited a 7.9 °C increase in T<inf>m</inf>and a 48-fold extension of t<inf>1/2</inf>at 50 °C, which represents a more significant enhancement than previous studies. Kinetic and product analyses using HPSEC, HPAEC-PAD, and<sup>1</sup>H NMR demonstrated that these mutations did not alter the catalytic efficiency or levan structure. The results demonstrate the potential of MD-aided energy-based engineering for thermostable EtLsc designs.