MnP Production By Phanerochaete Chrysosporium And Agrobacterium Tumefaciens Mediated Transformation Of Vgb Gene

Phanerochaete chrysosporium is frequently investigated as a model organism for the production of ligninolytic enzymes, such as manganese peroxidase (MnP), which can degrade lignin and other environment pollutants, like aromatic compounds. Trichoderma reesei is also an important strain to produce cellulase.In this study, the effects of inoculation amount, Tween80, carbon and nitrogen sources and shear force on MnP production were investigated. The results demonstrated that the optimal inoculation was105spores/mL. The addition of1g/LTween80benefited for protection of MnP against the damage from the shear force and stimulated the secretion of MnP. Increasing the glucose concentration could maintain the activity of MnP. The high shear force delayed the secretion of MnP and reduced its production by influencing the morphology of pellets. Based on the plasmid pCAMBIA1300, four binary plasmids were constructed, which contained different promoters, CaMV35S, pki, gpd and gpdA. respectively. The vgb gene was also introduced into these vectors. P. chrysosporium and T. reesei were transfected by Agrobacterium tumefaciens mediated transformation. However, the exogenous vgb gene and resistance gene hph were only integrated into the genome of T. reesei.21transformants of T. reesei were obtained. The results of PCR and qRT-PCR indicated the successful expression of vgb gene. Under limited oxygen condition, there were significant difference of biomass and FPA between the wild type strain and transfomants.