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Download fileCryptic Oxidative Transamination of Hydroxynaphthoquinone in Natural Product Biosynthesis
journal contribution
posted on 2022-03-16, 13:39 authored by Tomohiro Noguchi, Shota Isogai, Tohru Terada, Makoto Nishiyama, Tomohisa KuzuyamaPyridoxal
5′-phosphate (PLP)-dependent enzymes are a group
of versatile enzymes that catalyze various reactions, but only a small
number of them react with O2. Here, we report an unprecedented
PLP-dependent enzyme, NphE, that catalyzes both transamination and
two-electron oxidation using O2 as an oxidant. Our intensive
analysis reveals that NphE transfers the l-glutamate-derived
amine to 1,3,6,8-tetrahydroxynaphthalene-derived mompain to form 8-amino-flaviolin
(8-AF) via a highly conjugated quinonoid intermediate that is reactive
with O2. During the NphE reaction, O2 is reduced
to yield H2O2. An integrated technique involving
NphE structure prediction by AlphaFold v2.0 and molecular dynamics
simulation suggested the O2-accessible cavity. Our in vivo
results demonstrated that 8-AF is a genuine biosynthetic intermediate
for the 1,3,6,8-tetrahydroxynaphthalene-derived meroterpenoid naphterpin
without an amino group, which was supported by site-directed mutagenesis.
This study clearly establishes the NphE reaction product 8-AF as a
common intermediate with a cryptic amino group for the biosynthesis
of terpenoid–polyketide hybrid natural products.
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vivo results demonstratedstudy clearly establishesintensive analysis revealselectron oxidation usingcatalyze various reactionsgenuine biosynthetic intermediatecryptic oxidative transamination2 </ subcryptic amino groupcommon intermediateyield hversatile enzymesunprecedented plpsmall numberdirected mutagenesisderived mompainderived aminedependent enzymeamino groupalphafold v2