Study on expression of fusion hydrophobin in Pichia pastoris and its properties
XU Danyun
WANG Wei
MA Aimin
Abstract：Two hydrophobin genes, Po. hyd1 of Pleurotus ostreatus and Le. hyd of Lentinula edodes were ligated to construct the expression vector pPIC9k-Po. hyd1-Le. hyd, and then it was transferred into the Pichia pastoris GS115 competent cells by electroporation. After PCR validation and geneticin G418 resistance screening, three highly expressed Pichia pastoris transformants were obtained. After separation and purification of fusion hydrophobin Po. hyd1-Le. hyd from fermentation supernatant by the trifluoroacetic acid method, it showed that its monomer was about 26 kD. The fermentation conditions of P. pastoris were optimized by single factor and response surface analysis and the best fermentation conditions were showed as followed: the mass fraction of carbon source( methanol) was 0. 62 ％, the mass concentration of nitrogen source( m( yeast extract) ∶m( peptone) = 1∶2) was 8. 39 g/L, the initial pH value of the fermentation medium was 6. 05, the mass concentration of YNB was 100 mg/L, the fluid amount was 50 m L and the initial OD600 value was 1. 2. Under the above conditions, the maximum yield of the fusion hydrophobin Po. hyd1-Le. hyd could be up to 30 mg/L.The foaming and emulsifying properties of the fusion hydrophobin Po. hyd1-Le. hyd were investigated and they were all better than that of fusion hydrophobin extracted from the Pleurotus ostreatus mycelium.
Keywords：fusion hydrophobinPichia pastoristransformantresponse surface optimizationemulsibilityfoamability
Publication Date：2018-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：11( 71-81 )