Inducible expression of a heterologous protein in Hansenula polymorpha using the alcohol oxidase 1 promoter of Pichia pastoris.
Pichia pastoris (Pp) and Hansenula polymorpha (Hp) are methylotrophic yeasts commonly used for industrial purposes. Growth of either of these yeasts in the presence of methanol as the carbon source results in high-level induction of alcohol oxidase expression. The respective alcohol oxidase genes, AOX1 in Pp and MOX in Hp, have similar regulatory characteristics. Our studies show that the Pp AOX1 promoter (AOX1p) can be used for methanol-induced expression of a heterologous gene in Hp. Furthermore, the size of an AOX1p-heterologous gene-AOX1 terminator cassette transcript synthesized in Hp is indistinguishable from that synthesized in Pp suggesting that transcription both initiates and terminates at the same sites in both yeast species. Induction of AOX1p in Hp demonstrates that the methanol-inducible regulatory mechanism in Hp is able to recognize and activate the Pp promoter in spite of extensive sequence variations between AOX1p and MOXp.
Raschke
W C
WC
Salk Institute Biotechnology/Industrial Associates, Inc. (SIBIA), La Jolla, CA 92037, USA.
Neiditch
B R
BR
Hendricks
M
M
Cregg
J M
JM
D000429:Alcohol Oxidoreductases / Q000235:genetics*; D007611:Aprotinin / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D000432:Methanol / Q000378:metabolism; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics; D013045:Species Specificity; D014158:Transcription, Genetic
D004271:DNA, Fungal; D011994:Recombinant Proteins; C098973:recombinant Kunitz protease inhibitor; D007611:Aprotinin; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D000432:Methanol