Heterologous production of Rhizopus oryzae lipase in Pichia pastoris using the alcohol oxidase and formaldehyde dehydrogenase promoters in batch and fed-batch cultures

摘要

A Rhizopus oryzae lipase(ROL)has been expressed in Pichia pastoris as a reporter gene using two different regulated promoters.Both phenotypes,Mut~s and Mut~+,have been used for the expression under the control of alcohol oxidase 1 promoter(PAOX1).Moreover,the new formaldehyde dehydrogenase 1 promoter(PFLD1)has been tested.PFLD1 allows the design of methanol-free culture strategies,being methylamine a less volatile and flammable inducer.The comparison in batch and fed-batch cultures of the two promoters has shown that Mut~s(PAOX1)is somewhat more efficient than PFLD1 and Mut~+(PAOX1)systems,with a 1.1-and 1.4-fold higher specific productivity(U lipase/g biomass h),respectively.However,the productivity in all three systems was very similar(2500U lipase/lh).These results demonstrate that the expression of heterologous proteins under the transcriptional control of PFLD1 is a promising alternative to the well-known PAOX1.