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@PHDTHESIS{Haase:1047604,
author = {Haase, Mona},
title = {{A}pplication of {C}3-methyltransferases for natural
product synthesis},
volume = {52},
school = {Düsseldorf},
type = {Dissertation},
address = {Jülich},
publisher = {Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag},
reportid = {FZJ-2025-04406},
isbn = {978-3-95806-862-9},
series = {Bioorganische Chemie an der Heinrich-Heine-Universität im
Forschungszentrum Jülich},
pages = {432},
year = {2025},
note = {Dissertation, Düsseldorf, 2025},
abstract = {Natural products play a vital role in drug development,
either directly as pharmaceuticals or as templates for
designing target-specific therapeutics. The chemical
synthesis of these compounds often presents significant
challenges, requiring multiple reaction steps and hazardous
reagents. One prominent structural motif found in numerous
bioactive natural products is the hexahydropyrrolo[2,3-
b]indole (HPI) framework. Its unique tricyclic structure and
diverse biological activities have driven significant
synthetic interest, yet traditional methods struggle to
achieve stereoselectivity efficiently. This thesis aims to
develop a biocatalytic route for synthesising the HPI motif
using methyltransferases (MTases), which catalyses
C3-methylation of tryptophan-based cyclic dipeptides (DKPs).
The MTase StspM1 from Streptomyces sp. HPH0547 was selected
as the starting point for these studies: Optimised reaction
conditions, enzyme immobilisation, and S-adenosyl methionine
(SAM) cofactor recycling were investigated to create a
scalable, efficient method. To broaden the understanding of
their catalytic functions, homologs of StspM1 were explored,
focusing on their structure and mechanism. This
investigation was supported by two crystal structures
obtained during the study. Computational docking and
mutagenesis confirmed key residues critical for activity.
Among these homologs, SgMT from Streptomyces griseoviridis
was successfully integrated into the total synthesis of the
natural product lansai B, demonstrating its synthetic
utility. The gene clusters linked to these C3-MTases involve
additional enzymes contributing to the biosynthesis of
lansai B, including a second MTase, a cyclodipeptide
synthase (CDPS) and a prenyltransferase (PTase). The second
MTase was confirmed to function as an N-MTase. The PTase was
found to prenylate the C5-position, representing the final
step in the biosynthesis of lansai B This work advances
biocatalytic strategies for synthesising complex natural
product frameworks, offering new methodologies for
pharmaceutical development.},
cin = {IBOC},
cid = {I:(DE-Juel1)IBOC-20090406},
pnm = {2172 - Utilization of renewable carbon and energy sources
and engineering of ecosystem functions (POF4-217)},
pid = {G:(DE-HGF)POF4-2172},
typ = {PUB:(DE-HGF)3 / PUB:(DE-HGF)11},
url = {https://juser.fz-juelich.de/record/1047604},
}