Cloning and mutational analysis of pyrroquinoline quinone(PQQ) genes from a phosphate - solubilizing biocontrol bacterium Enterobacter intermedium.

E. intermedium 60-2G possessing a strong ability to solubilize insoluble phosphate, has plant growth-promoting activity, induced systemic resistance activity against scab pathogen in cucumber, and antifungal activity against various phytopathogenic fungi. The phosphate solubilizing activity of 60-2G may be mainly accomplished by production of gluconic acid through a direct extracellular oxidation of glucose by glucose dehydrogenase that required a PQQ cofactor for its activation. A pqq gene cluster conferred Phosphate-solubilizing activity in E. coli DH5${\alpha}$ was cloned and sequenced. The 6,783 bP pqq sequence had six open reading frames (from A to F) and showed 50-95％ homology to pqq genes from other bacteria. The E. coli strain expressing the pqq genes solubilized phosphate from hydroxyapatite after a pH drop to 4.0, which paralleled in time the secretion of gluconic acid. To study the role of PQQ in biocontrol traits of E. intermedium, PQQ mutants of 60-2G were constructed by marker exchangee mutagenesis. The PQQ mutants of E. intermedium were lost activities of solubilizing phosphate, growth inhibition of phytopathogenic fungi, and plant growth promotion. These findings suggest that PQQ plays an important role, possibly activation of certain enzymes, in several beneficial bacterial traits of E. intermedium by as yet an unknown mechanism.