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Analyzing and engineering the product selectivity of a 2-methylenebornane synthase
Publikationstyp
Journal Article
Date Issued
2020-05-04
Sprache
English
TORE-URI
Journal
Volume
9
Issue
5
Start Page
981
End Page
986
Citation
ACS Synthetic Biology 5 (9): 981 - 986 (2020)
Publisher DOI
Publisher
ACS
Copyright © 2020 American Chemical Society. Terpenes constitute the largest class of natural products with more than 70 »000 compounds. Many different terpenes find applications in the flavor and fragrance industry or can be used as fine chemicals or drugs. In some bacteria, noncanonical terpenes with 11 carbon atoms are synthesized via a GPP-C2-methyltransferase and the subsequent conversion of 2-methyl-GPP by certain terpene synthases into mainly 2-methylisoborneol and 2-methylenebornane. Many other C11-terpenes were reported as side products, but they are synthesized only in minor amounts by the bacterial C11-terpene biosynthesis pathway. To enable biotechnological synthesis of these largely unexplored natural products, we changed the product selectivity of the 2-methylenebornane synthase from Pseudomonas fluorescens by a semirational protein engineering approach. Active site amino acids with impact on the product selectivity were identified and variants with completely altered product spectra could be identified and characterized. The gathered data provide new insights into the structure-function relationship for C11-terpene synthases and demonstrate the production of formerly inaccessible noncanonical terpenes.
Subjects
2-methyl-GPP
2-methylenebornane
C11-terpenes
noncanonical terpenoids
Pseudomonas fluorescens
terpene synthase
DDC Class
540: Chemie
570: Biowissenschaften, Biologie