Production of two aprotinin variants in Hansenula polymorpha

DNA-sequences coding for two DesPro(2) aprotinin variants were expressed in the methylotrophic yeast Hansenula polymorpha from a strong inducible promoter element derived from the MOX gene, a key gene of methanol metabolism. For secretion the coding sequences were fused to the KEX2 recognition site...

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Bibliographic Details
Published in:Process biochemistry (1991) 1996-09, Vol.31 (7), p.679-689
Main Authors: Zurek, Christian, Kubis, Edward, Keup, Peter, Hörlein, Dietrich, Beunink, Jürgen, Thömmes, Jörg, Kula, Maria-Regina, Hollenberg, Cornelis P., Gellissen, Gerd
Format: Article
Language:English
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Summary:DNA-sequences coding for two DesPro(2) aprotinin variants were expressed in the methylotrophic yeast Hansenula polymorpha from a strong inducible promoter element derived from the MOX gene, a key gene of methanol metabolism. For secretion the coding sequences were fused to the KEX2 recognition site of the S. cerevisiae-derived MFα1 preproleader sequence. Correct processing of the precursor molecules and efficient secretion of the mature proteins was observed. A pH/pO 2-controlled C-source feeding mode was applied for fermentations on a 10 litre scale. In cultures of a transformant strain harbouring 20 copies of the aprotinin expression cassette a yield of 350 mg/litre could be obtained. The secreted recombinant products were purified in a simple two-step isolation procedure employing an expanded bed adsorption as an initial purification step.
ISSN:1359-5113
1873-3298
DOI:10.1016/S0032-9592(96)00018-0