Effect Of Animal Endogenous Polypeptide CGA-N46on Cell Walls And Mitochondria Of Candidas

CGA-N46is an N terminal derived peptide of human Chromogranin A (CGA).Researching the effect of CGA-N46to Candida proliferation and the impact onCandida cell walls and mitochondria could clarify the inhibitory mechanism of CGA-N46, and provide theoretical basis for its functional targets research.In this paper, CGA-N46sample was fermented and purificated by engineeredbacteria Bacillus subtilis DB1342(p-3N46), which was used as the experimentalstrains. It was confirmed in the CGA-N46inhibitory properties of experiments that,CGA-N46for Candida krusei had a good inhibition of cell proliferation, whichmainly inhibit krusei Candida logarithmic growth phase, and for Candida krusei cellinhibition rate in a concentration-dependent increase; observed by transmissionelectron microscopy CGA-N46can Candida krusei mitochondria vacuolization ofmedullary cytoplasm becomes uneven, resulting in white, the film dissolves part ofthe nucleus, not continuous; through detection of β-1,3-glucanase found CGA-N46does not affect the integrity of the cell wall of Candida krusei; through the fluorescentprobe DCFH-DA and DHE detection CGA-N46for Candida krusei reactive oxygenspecies (ROS) effects found with increasing concentrations of CGA-N46, Candidakrusei intracellular reactive oxygen species (H2O2and02-) levels were significantlyreduced; Rh-123fluorescent probe to detect mitochondrial membrane potentialexperiments showed that the mitochondrial membrane potential with CGA-N46concentrations increase gradually reduced; research CGA-N46for Candida kruseiCa2+-Mg2+-ATPase affected, indicating that CGA-N46allows increased Ca2+-Mg2+-ATPase enzyme activity; SDS-PAGE protein electrophoresis detection ofmitochondrial proteins results showed that the CGA-N46role Candida kruseimolecular size of13KDa,43KDa,84KDa protein upregulation; Caspase3activityassay experiments confirmed, CGA-N463has no effect on Candida krusei Caspase.In summary, the animal endogenous polypeptide CGA-N46can make Candidakrusei ultrastructural change, affect the function of Candida krusei mitochondrial, butdoes not affect the integrity of their cell walls. Therefore, CGA-N46may inhibitCandida proliferation through mitochondrial pathway.