Abstract
Native 3-hydroxybutyrate dehydrogenase from Alcaligenes faecalis can catalyze the reversible reduction of acetoacetate, a four carbon chain oxo acid. This enzyme has been engineered to enable the reduction of levulinic acid, with one carbon longer than acetoacetate. In this study, the native and engineered enzymes were subjected to the catalysis of oxo acids with a carbon chain length of 3 to 8, in order to examine the capability of the enzyme to work on various platform chemicals. The engineered enzyme could reduce the C7 and C8 oxo acids whereas the wild-type had no activity on these substrates. Docking simulation has indicated Tyr155 and Ser142 are key residues for the catalysis. In addition, stable hydrogen bond formation between Gln196 and the substrates affects the turnover rate. Mutation sites in the engineered enzyme were focused on creating larger active site volume for substrates with extended chain lengths. Both qualitative and quantitative structural basis for the enzyme substrate specificity on alpha, beta, gamma and omega hydroxy acids could be elucidated.
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 callDisclaimer: 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.
