Invertase secretion in Hansenula polymorpha under the AOX1 promoter from Pichia pastoris.
A DNA fragment containing a transcription regulating region of the alcohol oxidase (AOX1) gene from the methylotrophic yeast Pichia pastoris was used in the construction of a vector for the expression of heterologous proteins in the methylotrophic yeast Hansenula polymorpha. We used this vector to clone the SUC2 gene from Saccharomyces cerevisiae into H. polymorpha yeast. The culture conditions for invertase production using a fed-batch culture were studied. More than 1.5 x 10(3) U/ml of biologically active invertase (1 g/l) were secreted to the cellular periplasmic space. The fermentative process was scaled up to 50 l. Invertase produced from H. polymorpha was glycosylated, but it contained significantly less carbohydrate than protein produced by S. cerevisiae. Using the Western-blot technique, it was observed that invertase secreted from H. polymorpha and invertase secreted from S. cerevisiae showed common antigenic determinants.
Rodriguez
L
L
Bioindustry Division, Center for Genetic Engineering & Biotechnology, La Habana, Cuba.
Narciandi
R E
RE
Roca
H
H
Cremata
J
J
Montesinos
R
R
Rodriguez
E
E
Grillo
J M
JM
Muzio
V
V
Herrera
L S
LS
Delgado
J M
JM
D000429:Alcohol Oxidoreductases / Q000235:genetics; D002451:Cell Compartmentation; D003470:Culture Media; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005822:Genetic Vectors; D005990:Glycerol / Q000494:pharmacology; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D000432:Methanol / Q000378:metabolism; D010843:Pichia / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D043324:beta-Fructofuranosidase
D003470:Culture Media; D011993:Recombinant Fusion Proteins; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D005990:Glycerol; D000432:Methanol