Affiliation:
1. Centro de Neurociências e Biologia Celular, Departamento de Zoologia, Universidade de Coimbra, 3004-517 Coimbra, Portugal
2. Departamento de Bioquímica, Universidade de Coimbra, 3001-401 Coimbra, Portugal
Abstract
ABSTRACT
The pathway for the synthesis of glucosylglycerate (GG) in the thermophilic bacterium
Persephonella marina
is proposed based on the activities of recombinant glucosyl-3-phosphoglycerate (GPG) synthase (GpgS) and glucosyl-3-phosphoglycerate phosphatase (GpgP). The sequences of
gpgS
and
gpgP
from the cold-adapted bacterium
Methanococcoides burtonii
were used to identify the homologues in the genome of
P. marina
, which were separately cloned and overexpressed as His-tagged proteins in
Escherichia coli
. The recombinant GpgS protein of
P. marina
, unlike the homologue from
M. burtonii
, which was specific for GDP-glucose, catalyzed the synthesis of GPG from UDP-glucose, GDP-glucose, ADP-glucose, and TDP-glucose (in order of decreasing efficiency) and from
d
-3-phosphoglycerate, with maximal activity at 90°C. The recombinant GpgP protein, like the
M. burtonii
homologue, dephosphorylated GPG and mannosyl-3-phosphoglycerate (MPG) to GG and mannosylglycerate, respectively, yet at high temperatures the hydrolysis of GPG was more efficient than that of MPG. Gel filtration indicates that GpgS is a dimeric protein, while GpgP is monomeric. This is the first characterization of genes and enzymes for the synthesis of GG in a thermophile.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
Cited by
30 articles.
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