Open AccessIdentification of Pantoea ananatis gene encoding membrane pyrroloquinoline quinone (PQQ)‐dependent glucose dehydrogenase and pqqABCDEF operon essential for PQQ biosynthesis
Author(s) -
Andreeva Irina G.,
Golubeva Lyubov I.,
Kuvaeva Tatiana M.,
Gak Evgueni R.,
Katashkina Joanna I.,
Mashko Sergey V.
Publication year - 2011
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2011.02240.x
Subject(s) - pyrroloquinoline quinone , operon , biology , escherichia coli , biochemistry , phosphoenolpyruvate carboxykinase , gene , microbiology and biotechnology , enzyme , cofactor
Pantoea ananatis accumulates gluconate during aerobic growth in the presence of glucose. Computer analysis of the P. ananatis SC17(0) sequenced genome revealed an ORF encoding a homologue (named gcd ) of the mGDH (EC 1.1.99.17) apoenzyme from Escherichia coli and a putative pyrroloquinoline quinone (PQQ) biosynthetic operon homologous to pqqABCDEF from Klebsiella pneumoniae . Construction of Δ gcd and Δ pqq mutants of P. ananatis confirmed the proposed functions of these genetic elements. The P. ananatis pqqABCDEF was cloned in vivo and integrated into the chromosomes of P. ananatis and E. coli according to the Dual In/Out strategy. Introduction of a second copy of pqqABCDEF to P. ananatis SC17(0) doubled the accumulation of PQQ. Integration of the operon into E. coli MG1655Δ ptsG Δ manXY restored the growth of bacteria on glucose. The obtained data show the essential role of pqqABCDEF in PQQ biosynthesis in P. ananatis and E. coli . We propose that the cloned operon could be useful for an efficient phosphoenolpyruvate‐independent glucose consumption pathway due to glucose oxidation and construction of E. coli strains with the advantage of phosphoenolpyruvate‐derived metabolite production.