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Overproduction of BiP negatively affects the secretion of Aspergillus niger glucose oxidase by the yeast Hansenula polymorpha

van der Heide, M., Hollenberg, CP., van der Klei, IJ. & Veenhuis, M., Mar-2002, In : Applied Microbiology and Biotechnology. 58, 4, p. 487-494 8 p.

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DOI

We have cloned the Hansenula polymorpha BIP gene from genomic DNA using a PCR-based strategy. H. polymorpha BIP encodes a protein of 665 amino acids, which shows very high homology to Saccharomyces cerevisiae KAR2p. KAR2p belongs to the Hsp70 family of molecular chaperones and resides in the endoplasmic reticulum (ER)-lumen. H. polymorpha BiP contains a putative N-terminal signal sequence of 30 amino acids together with the conserved -HDEL sequence, the typical ER retention signal, at the extreme C-terminus. We have analysed the effect of BIP overexpression, placing the gene under control of the strong alcohol oxidase promoter (P-MOX), on the secretion of artificially produced Aspergillus niger glucose oxidase (GOX) by H. polymorpha. BiP overproduction did not lead to any growth defects of the cells; at the subcellular level, proliferation of ER-like vesicles was observed. However, artificially enhanced BiP levels strongly affected GOX secretion and led to accumulation of this protein in the ER-like vesicles. This was not simply due to the high BiP overproduction, because it was also observed under conditions of low P-MOX induction during growth of cells on glycerol. Vacuolar carboxypeptidase Y was properly sorted to its target organelle in the BiP overproducing strains.

Original languageEnglish
Pages (from-to)487-494
Number of pages8
JournalApplied Microbiology and Biotechnology
Volume58
Issue number4
Publication statusPublished - Mar-2002

    Keywords

  • CHAIN BINDING-PROTEIN, ENDOPLASMIC-RETICULUM, SACCHAROMYCES-CEREVISIAE, GENE, TRANSLOCATION, OVEREXPRESSION, DEGRADATION, EXPRESSION, MEMBRANE, CUTINASE

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