Premium
Mechanistic Characterisation of Two Sesquiterpene Cyclases from the Plant Pathogenic Fungus Fusarium fujikuroi
Author(s) -
Burkhardt Immo,
Siemon Thomas,
Henrot Matthias,
Studt Lena,
Rösler Sarah,
Tudzynski Bettina,
Christmann Mathias,
Dickschat Jeroen S.
Publication year - 2016
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201603782
Subject(s) - sesquiterpene , stereochemistry , escherichia coli , mutagenesis , terpene , enzyme , chemistry , biochemistry , biology , mutation , gene
Two sesquiterpene cyclases from Fusarium fujikuroi were expressed in Escherichia coli and purified. The first enzyme was inactive because of a critical mutation, but activity was restored by sequence correction through site‐directed mutagenesis. The mutated enzyme and two naturally functional homologues from other fusaria converted farnesyl diphosphate into guaia‐6,10(14)‐diene. The second enzyme produced eremophilene. The absolute configuration of guaia‐6,10(14)‐diene was elucidated by enantioselective synthesis, while that of eremophilene was evident from the sign of its optical rotation and is opposite to that in plants but the same as in Sorangium cellulosum. The mechanisms of both terpene cyclases were studied with various 13 C‐ and 2 H‐labelled FPP isotopomers.