Expression of human serum - albumins in pichia pastoris

摘要

A novel expression cassette, vectors and process for the secretionof HSA in Pichia pastoris cells. In accordance with the present inventionthere has been discovered an improved expression cassette for the productionof HSA in Pichia pastoris comprisinga) a Pichia pastoris 5' regulatory region having a 5' end and a 3'end selected from the group consisting of the Pichia pastoris AOX1 5'regulatory region and the Pichia pastoris DAS1 5' regulatory region whereinthe 3' end of the 5' regulatory region is operably linked to;b) an HSA structural gene encoding a signal sequence and matureprotein having a 5' end and a 3' end wherein the HSA structural gene has anATG start codon within about 11 deoxyribonucleotides of the 5' end of saidHSA structural gene; and operably linked toc) a 3' termination sequence.In accordance with the present invention there has also discoveredPichia pastoris cells transformed with an improved expression cassette forthe production of HSA in Pichia pastoris comprisinga) a Pichia pastoris 5' regulatory region having a 5' end and a 3'end selected from the group consisting of the Pichia pastoris AOX1 5'regulatory region and the Pichia pastoris DAS1 5' regulatory region whereinthe 3' end of the 5' regulatory region is operably linked to;b) an HSA structural gene encoding a signal sequence and matureprotein having a 5' end and a 3' end wherein the HSA structural gene has anATG start codon within about 11 deoxyribonucleotides of the 5' end of saidHSA structural gene; and operably linked toc) a 3' termination sequence.In a further embodiment of the present invention, there has alsodiscovered a process for the secretion of HSA from transformed Pichiapastoris cells comprisinga) transforming a Pichia pastoris cell with at least one vectorhaving at least one expression cassette comprisingi) a Pichia pastoris 5' regulatory region having a 5' end anda 3' end selected from the group consisting of the Pichia pastoris AOX1 5'regulatory region and the Pichia pastoris DAS1 5' regulatory region whereinthe 3' end of the 5' regulatory region is operably toii) an HSA structural gene encoding a signal sequence andmature protein having a 5' end and a 3' end wherein the HSA structural genesaid HSA structural gene; and operably linked toiii) a 3' termination sequence; and thenb) culturing the resulting transformed Pichia pastoris suitableconditions to obtain the secretion of HSA.