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J. Bacteriol., Sep 1995, 5088-5098, Vol 177, No. 17
JS Velterop, E Sellink, JJ Meulenberg, S David, I Bulder and PW Postma
In Klebsiella pneumoniae, six genes, constituting the pqqABCDEF operon,
which are required for the synthesis of the cofactor pyrroloquinoline
quinone (PQQ) have been identified. The role of each of these K. pneumoniae
Pqq proteins was examined by expression of the cloned pqq genes in
Escherichia coli, which cannot synthesize PQQ. All six pqq genes were
required for PQQ biosynthesis and excretion into the medium in sufficient
amounts to allow growth of E. coli on glucose via the PQQ- dependent
glucose dehydrogenase. Mutants lacking the PqqB or PqqF protein synthesized
small amounts of PQQ, however. PQQ synthesis was also studied in cell
extracts. Extracts made from cells containing all Pqq proteins contained
PQQ. Lack of each of the Pqq proteins except PqqB resulted in the absence
of PQQ. Extracts lacking PqqB synthesized PQQ slowly. Complementation
studies with extracts containing different Pqq proteins showed that an
extract lacking PqqC synthesized an intermediate which was also detected in
the culture medium of pqqC mutants. It is proposed that PqqC catalyzes the
last step in PQQ biosynthesis. Studies with cells lacking PqqB suggest that
the same intermediate might be accumulated in these mutants. By using
pqq-lacZ protein fusions, it was shown that the expression of the putative
precursor of PQQ, the small PqqA polypeptide, was much higher than that of
the other Pqq proteins. Synthesis of PQQ most likely requires molecular
oxygen, since PQQ was not synthesized under anaerobic conditions, although
the pqq genes were expressed.
Copyright © 1995, American Society for Microbiology
Synthesis of pyrroloquinoline quinone in vivo and in vitro and detection of an intermediate in the biosynthetic pathway
E. C. Slater Institute, BioCentrum Amsterdam, University of Amsterdam, The Netherlands.
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