| High-level secretion of hirudin by Hansenula polymorpha--authentic processing of three different preprohirudins. | |
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MedLine Citation:
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PMID: 8597538 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A DNA sequence coding for a subtype of the hirudin variant HV1 was expressed in the methylotrophic yeast Hansenula polymorpha from a strongly inducible promoter element derived from a gene of the methanol metabolism pathway. For secretion, the coding sequence was fused to the KEX2 recognition site of three different prepro segments engineered from the MF alpha 1 gene of Saccharomyces cerevisiae, the glucoamylase (GAM1) gene of Schwanniomyces occidentalis and the gene for a crustacean hyperglycemic hormone from the shore crab Carcinus maenas. In all three cases, correct processing of the precursor molecule and efficient secretion of the mature protein were observed. In fermentations on a 10-1 scale of a transformant strain harbouring a MF alpha 1/hirudin-gene fusion yields in the range of grams per litre could be obtained. The majority of the secreted product was identified as the full-length 65-amino-acid hirudin. Only small amounts of a truncated 63-amino- acid product, frequently observed in S. cerevisiae-based expression systems, could be detected. |
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Authors:
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U Weydemann; P Keup; M Piontek; A W Strasser; J Schweden; G Gellissen; Z A Janowicz |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Applied microbiology and biotechnology Volume: 44 ISSN: 0175-7598 ISO Abbreviation: Appl. Microbiol. Biotechnol. Publication Date: 1995 Dec |
Date Detail:
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Created Date: 1996-04-22 Completed Date: 1996-04-22 Revised Date: 2008-11-21 |
Medline Journal Info:
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Nlm Unique ID: 8406612 Medline TA: Appl Microbiol Biotechnol Country: GERMANY |
Other Details:
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Languages: eng Pagination: 377-85 Citation Subset: B |
Affiliation:
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Rhein Biotech GinbH, Düsseldorf, Germany. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Animals Brachyura / genetics Fermentation Fungal Proteins / biosynthesis, genetics Glucan 1,4-alpha-Glucosidase / biosynthesis, genetics Hirudins / biosynthesis, genetics, secretion* Invertebrate Hormones / biosynthesis, genetics Molecular Sequence Data Nerve Tissue Proteins / biosynthesis, genetics Peptide Biosynthesis Peptides / genetics Pichia / metabolism* Promoter Regions, Genetic Protein Precursors / metabolism* Protein Processing, Post-Translational* Recombinant Fusion Proteins / biosynthesis, genetics, secretion* Saccharomyces cerevisiae / genetics |
| Chemical | |
Reg. No./Substance:
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0/Fungal Proteins; 0/Hirudins; 0/Invertebrate Hormones; 0/Nerve Tissue Proteins; 0/Peptides; 0/Protein Precursors; 0/Recombinant Fusion Proteins; 0/hyperglycemic hormone, crustacean; 61194-02-3/mating factor; EC 3.2.1.3/Glucan 1,4-alpha-Glucosidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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