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Phospholipase C (PLC) enzymes are key virulence factors in several pathogenic bacteria.  Burkholderia pseudomallei , the causative agent of melioidosis, possesses at least three  plc  genes ( plc1 ,  plc2  and  plc3 ). We found that in culture medium  plc1  gene expression increased with increasing pH, whilst expression of the  plc3  gene was pH (4.5 to 9.0) independent. Expression of the  plc2  gene was not detected in culture medium. All three  plc  genes were expressed during macrophage infection by  B. pseudomallei  K96243. Comparing  B. pseudomallei  wild-type with  plc  mutants revealed that  plc2, plc12  or  plc123  mutants showed reduced intracellular survival in macrophages and reduced plaque formation in HeLa cells. However,  plc1  or  plc3  mutants showed no significant differences in plaque formation compared to wild-type bacteria. These findings suggest that Plc2, but not Plc1 or Plc3 are required for infection of host cells. In  Galleria mellonella ,  plc1, plc2  or  plc3  mutants were not attenuated compared to the wild-type strain, but multiple  plc  mutants showed reduced virulence. These findings indicate functional redundancy of the  B. pseudomallei  phospholipases in virulence.

Functional redundancy of Burkholderia pseudomallei phospholipase C enzymes and their role in virulence