Publication

Redirection of peroxisomal alcohol oxidase of Hansenula polymorpha to the secretory pathway

van der Heide, M., Leão, A. N., van der Klei, I. J. & Veenhuis, M., Oct-2007, In : Fems Yeast Research. 7, 7, p. 1093-1102 10 p.

Research output: Contribution to journalArticleAcademicpeer-review

Copy link to clipboard

Documents

DOI

We report on the rerouting of peroxisomal alcohol oxidase (AO) to the secretory pathway of Hansenula polymorpha. Using the leader sequence of the Saccharomyces cerevisiae mating factor alpha (MF alpha) as sorting signal, AO was correctly sorted to the endoplasmic reticulum (ER), which strongly proliferated in these cells. The MF alpha presequence, but not the prosequence, was cleaved from the protein. AO protein was present in the ER as monomers that lacked FAD, and hence was enzymatically inactive. Furthermore, the recombinant AO protein was subject to gradual degradation, possibly because the protein did not fold properly. However, when the S. cerevisiae invertase signal sequence (ISS) was used, secretion of AO protein was observed in conjunction with bulk of the protein being localized to the ER. The amount of secreted AO protein increased with increasing copy numbers of the AO expression cassette integrated into the genome. The secreted AO protein was correctly processed and displayed enzyme activity.

Original languageEnglish
Pages (from-to)1093-1102
Number of pages10
JournalFems Yeast Research
Volume7
Issue number7
Publication statusPublished - Oct-2007
Event4th Hansenula Polymorpha Worldwide Network Conference (HPWN) - , Netherlands
Duration: 3-Sep-20065-Sep-2006

Event

4th Hansenula Polymorpha Worldwide Network Conference (HPWN)

03/09/200605/09/2006

Netherlands

Event: Other

    Keywords

  • protein secretion, yeast, Hansenula polymorpha, alcohol oxidase, heterologous gene expression, FLAVIN ADENINE-DINUCLEOTIDE, METHYLOTROPHIC YEAST, ENDOPLASMIC-RETICULUM, METHANOL METABOLISM, PROTEIN, GENE, DEGRADATION, BIOGENESIS, PEXOPHAGY, ENCODES

Download statistics

No data available

ID: 1897718