Substitution of Two Active-Site Residues Alters C9-Hydroxylation in a Class II Diterpene Synthase

Diterpenes form a vast and diverse class of natural products of both ecological and economic importance. Class II diterpene synthase (diTPS) enzymes control the committed biosynthetic reactions underlying diterpene chemical diversity. Homology modelling with site‐directed mutagenesis identified two...

Full description

Saved in:
Bibliographic Details
Published in:Chembiochem : a European journal of chemical biology 2016-12, Vol.17 (24), p.2304-2307
Main Authors: Mafu, Sibongile, Fischer, Emil, Addison, J. Bennett, Riberio Barbosana, Isabel, Zerbe, Philipp
Format: Article
Language:English
Subjects:
Alkyl and Aryl Transferases - genetics
Alkyl and Aryl Transferases - metabolism
Amino Acid Sequence
Biocatalysis
Catalytic Domain
diterpene synthase
Diterpenes - chemistry
enzyme catalysis
Gas Chromatography-Mass Spectrometry
Hydroxylation
Magnetic Resonance Spectroscopy
Marrubium - enzymology
Molecular Sequence Data
Mutagenesis, Site-Directed
natural products
peregrinol diphosphate
Plant Proteins - genetics
Plant Proteins - metabolism
Sequence Alignment
Stereoisomerism
terpenes
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Diterpenes form a vast and diverse class of natural products of both ecological and economic importance. Class II diterpene synthase (diTPS) enzymes control the committed biosynthetic reactions underlying diterpene chemical diversity. Homology modelling with site‐directed mutagenesis identified two active‐site residues in the horehound (Marrubium vulgare) class II diTPS peregrinol diphosphate synthase (MvCPS1); residue substitutions abolished the unique MvCPS1‐catalysed water‐capture reaction at C9 and redirected enzyme activity toward formation of an alternative product, halima‐5(10),13‐dienyl diphosphate. These findings contributed new insight into the steric interactions that govern diTPS‐catalysed regiospecific oxygenation reactions and highlight the feasibility of diTPS engineering to provide a broader spectrum of bioactive diterpene natural products. Diterpene synthase catalytic plasticity: Structure‐function analysis of the Marrubium vulgare class II diterpene synthase peregrinol diphosphate synthase (MvCPS1) identified two active‐site residues that contribute to the enzyme's product specificity by redirecting catalysis from peregrinol diphosphate to the formation of an alternate diterpene scaffold, halima‐5(10),13‐dienyl diphosphate.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.201600419