Rational Engineering of a Thermostable Dipeptidase from Thermobrachium celere for Efficient Biosynthesis of l -Carnosine
Abstract
(TcPepD) was identified and engineered for efficient l-carnosine synthesis. Wild-type TcPepD exhibited an optimal temperature of 70 °C and high thermal stability. Structure-guided mutagenesis based on AlphaFold modeling and molecular docking led to a double mutant, M2 (N117D/I169L), with a 2.7-fold increase in synthetic activity compared to the wild-type. Kinetic analysis showed that M2 displayed enhanced substrate affinity, particularly toward l-histidine, and improved catalytic efficiency. Molecular dynamics simulations revealed synergistic effects of the two mutations, resulting in improved substrate coordination and optimized active-site flexibility. Under high-substrate conditions at 65-70 °C, M2 produced up to 84.3 mM l-carnosine, demonstrating strong potential for high-temperature and high-efficiency biocatalytic production.
Fields of Study & Subjects
Biochemical effects in animals
Aldose Reductase and Taurine
Healthcare and Venom Research
Publication details
- Journal / source
- Journal of Agricultural and Food Chemistry
- Publisher
- American Chemical Society (ACS)
- Volume
- 74
- Issue
- 9
- Pages
- 7743–7751
- Publication date
- 26 February 2026
- Language
- EN