So, you want to know about

siderophore synthesis
Presented by:
Steven Backues
Brooks Maki
and
Donnie Berkholz 'The Invisible Man
ydroxamic Acid Groups
N-Alkylation oI O-substituted ydroxamic
acid.
Formation oI an oxime Irom an aldehyde and
a hydroxylamine. Followed by reduction and
acylation
These derivatives allowed synthesis oI
several siderophores and their analogues
Alternative Methods
Oxidation oI Lysine and OrnithineWith
DMD (dimethyldioxarine)
Conversion oI primary amines to imines,
with oxidation oI imines to oxaziridines.
ydrolysis leads to hydroxylamines.
D-Ferrichrome synthesized by this method
Danoxamine
Composed oI a linear series oI three
hydroxamic acids.
Composed oI two major groups linked by
succinic acid.
Biosynthesis oI Siderophores
ow it`s really done.
ydroxamate Siderophores
Step 1: Ornithine N
5
-
Oxygenase
The Iormation oI the N-O bond is the Iirst
committed step in hydroxamate synthesis
Ornithine, an amino acid used in the urea
cycle, is reacted with O
2
and NADP to give
an N-O bond at the end oI its side chain.
Step 2: N
5-
Transacylase
The nitrogen modiIied in this way is
additionally attached to an acyl group carried
by coenzyme A
This completes the hydroxamate prosthetic
group
The ydroxamate prosthetic group:
N
NH
3
+
H
COO-
HO
O
H
3
C
Step 3: Non-ribosomal Peptide
Synthetase
This synthetase is a large complex with many
subdomains, including an adenylation
domain, a thiolation domain, and a
condensation domain.
Adenylation
First, the hydroxamate is is activated by
addition oI an adenylate group at its C
terminus
The source oI the adenylate group is ATP,
and the reaction occurs with production oI
pyrophosphate
Thiolation
The hydroxamate group is then transIerred to
the enzyme through the Iormation oI a
thioester linkage with displacement oI the
adenylate group.
Condensation
Finally, the hydroxamate group is attached to
another molecule (perhaps another
hydroxamate group, or else a growing chain)
by the nucleophilic attack oI an O or N
Irom the chain on the S-linked carbonyl,
displacing the sulIur.
Cathechols: Vibriobactin
Vibriobactin is a siderophore used by Jibrio
cholerae
Its synthesis also involves a large, non-
ribosomal peptide synthetase, and Iollows
many oI the same pathways as the synthesis
oI hydroxamate siderophores outlined above.
Vibriobactin
OH
OH
O
N
NH
Me
O
N
HO
HO
O
N
Me
O
NH
O OH
OH
The Cathechol Prosthetic
Group
The cathechol prosthetic group is 2,3-
dihydroxybenzoic acid, which is Iormed
Irom chorismic acid
Nonribosomal Peptide
Synthetase
2,3-dihydrobenzoic acid then acts as a
substrate Ior Vibirobactin Syntetase
It is Iirst activated by adenylation, then
transIerred to the enzyme with Iormation oI a
thioester
TransIer to Norspermidine
This thioester complex then undergoes
nucleophilic attack by a primary amine on
norspermidine.
The norspermidine/cathechol complex goes
on to react with two more cathechol
prosthetic groups (these, however, attached
by threonine derived linkages) to Iorm the
Iinal siderophore
Norspermidinecathecol
OH
OH
O
N
NH
Me
O
H
N NH
2
ersiniabactin
OH
S
N
S
N
H
Me
Me
S
N
Me
COOH
OH
Synthesis
Although it has neither hydroxamate nor
cathechol groups, ersiniabactin Iollows
some oI the same synthesis pathways, using
a nonribosomal peptide sythetase that has
clear homologies with, Ior example,
vibriobactin sythetase
Beginnings
Synthesis begins with salicylic acid (2-
hydroxy-benzoic acid)
This is activated by the attachment oI an
adenylate group, then loaded onto the
enzyme by the Iormation oI a thioester, as
beIore
longation
At the same time, two cystines are also activated
then loaded onto the same enzyme, also via a
thioester linkage
Then, in the condensation/cyclization domain, the
salicyate group is transIerred onto one oI the
cystines, which is then cyclized.
This cyclization is an unusual property oI this
particular synthetaes
Completion
A second cystine is added, and also cyclized,
and the resulting molecule undergoes the
addition oI a malonyl group, S-
adenosylmethionine, and an additional
cystine to complete the synthesis
Overview:
The use oI a large, multidomain nonribosomal
peptide synthetase was a common element oI all oI
these syntheses.
All oI these processes included the activation oI a
substrate by adenylation and the transIer to a
thioester linkage with the enzyme, Iollowed by
condensation to Iorm a longer chain. This is similar
to the process Iollowed in biosynthesis oI Iatty
acids.
#eIerences
#oosenberg, J.M. and Miller, M.J. Total Synthesis oI the Siderophore
Danoxamine. J. Org. Chem. 2000 Vol. 65 No. 16. 4833 4838.
Lin, . and Miller, M.J. Synthesis oI Siderophore Components by and
Indirect Oxidation Method. J. Org. Chem. 1999 Vol. 64 No. 20.
7451 7458.
Gaspar, M., Grazina, #., Bodor, A., Farkas, ., and Santos, M.A.
Macrocyclic tetraamine tris(hydroxamate) ligand. J. Chem Soc.
1999 799 806.
Duhme, A.K. Synthesis oI two dioxomolybdenum complexes oI a
siderophore analogue. J. Chem. Soc. 1997 773 778.
Atkinson, A. Bacterial Iron Transport. Biochemistry. 1998. 15965 -
15973