Efficient expression and secretion of Aspergillus niger RH5344 polygalacturonase in Saccharomyces cerevisiae
An Aspergillus niger endopolygalacturonase (EC 3.2.1.15) cDNA was expressed in the yeast Saccharomyces cerevisiae. Secretion of the protein into the growth medium was efficiently directed by the fungal leader sequence, and processing occurred at the same site as in Aspergillus. The expression level...
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| Veröffentlicht in: | Applied microbiology and biotechnology 1995-12, Vol.44 (1/2), p.147-156 |
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| container_title | Applied microbiology and biotechnology |
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| creator | Lang, C Looman, A.C |
| description | An Aspergillus niger endopolygalacturonase (EC 3.2.1.15) cDNA was expressed in the yeast Saccharomyces cerevisiae. Secretion of the protein into the growth medium was efficiently directed by the fungal leader sequence, and processing occurred at the same site as in Aspergillus. The expression level was significantly enhanced by using a "short" version of the yeast ADHI promoter. An additional increase in the yield of heterologous protein was due to a higher plasmid stability and a rise in plasmid copy number. This was achieved by deleting most of the bacterial sequences from the expression vector. The yeast-derived enzyme showed the same enzymatic and biochemical properties as the fungal polygalacturonase, such as substrate specificity, pH and temperature optima and pI value. The yeast-derived enzyme, however, showed a higher degree of glycosylation and exhibited a more pronounced temperature stability than the fungal enzyme. |
| doi_str_mv | 10.1007/bf00164494 |
| format | Article |
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Secretion of the protein into the growth medium was efficiently directed by the fungal leader sequence, and processing occurred at the same site as in Aspergillus. The expression level was significantly enhanced by using a "short" version of the yeast ADHI promoter. An additional increase in the yield of heterologous protein was due to a higher plasmid stability and a rise in plasmid copy number. This was achieved by deleting most of the bacterial sequences from the expression vector. The yeast-derived enzyme showed the same enzymatic and biochemical properties as the fungal polygalacturonase, such as substrate specificity, pH and temperature optima and pI value. The yeast-derived enzyme, however, showed a higher degree of glycosylation and exhibited a more pronounced temperature stability than the fungal enzyme.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/bf00164494</identifier><identifier>PMID: 8579828</identifier><identifier>CODEN: AMBIDG</identifier><language>eng</language><publisher>Berlin: Springer</publisher><subject>Alcohol Dehydrogenase - genetics ; Aspergillus niger - enzymology ; Base Sequence ; Biological and medical sciences ; Biotechnology ; Fermentation ; food microbiology ; food processing ; Fundamental and applied biological sciences. Psychology ; Genetic engineering ; Genetic technics ; Glycosylation ; horticultural crops ; Methods. Procedures. Technologies ; Modification of gene expression level ; Molecular Sequence Data ; Plasmids ; Polygalacturonase - biosynthesis ; Promoter Regions, Genetic ; Recombinant Proteins - biosynthesis ; Saccharomyces cerevisiae - genetics</subject><ispartof>Applied microbiology and biotechnology, 1995-12, Vol.44 (1/2), p.147-156</ispartof><rights>1996 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c442t-d8fa8fb076175c740b64c66b8b3de8afc9c1b2d1b183282ffd0ddfe9a3abb813</citedby><cites>FETCH-LOGICAL-c442t-d8fa8fb076175c740b64c66b8b3de8afc9c1b2d1b183282ffd0ddfe9a3abb813</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2935918$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8579828$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lang, C</creatorcontrib><creatorcontrib>Looman, A.C</creatorcontrib><title>Efficient expression and secretion of Aspergillus niger RH5344 polygalacturonase in Saccharomyces cerevisiae</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><description>An Aspergillus niger endopolygalacturonase (EC 3.2.1.15) cDNA was expressed in the yeast Saccharomyces cerevisiae. Secretion of the protein into the growth medium was efficiently directed by the fungal leader sequence, and processing occurred at the same site as in Aspergillus. The expression level was significantly enhanced by using a "short" version of the yeast ADHI promoter. An additional increase in the yield of heterologous protein was due to a higher plasmid stability and a rise in plasmid copy number. This was achieved by deleting most of the bacterial sequences from the expression vector. The yeast-derived enzyme showed the same enzymatic and biochemical properties as the fungal polygalacturonase, such as substrate specificity, pH and temperature optima and pI value. The yeast-derived enzyme, however, showed a higher degree of glycosylation and exhibited a more pronounced temperature stability than the fungal enzyme.</description><subject>Alcohol Dehydrogenase - genetics</subject><subject>Aspergillus niger - enzymology</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Fermentation</subject><subject>food microbiology</subject><subject>food processing</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Glycosylation</subject><subject>horticultural crops</subject><subject>Methods. Procedures. Technologies</subject><subject>Modification of gene expression level</subject><subject>Molecular Sequence Data</subject><subject>Plasmids</subject><subject>Polygalacturonase - biosynthesis</subject><subject>Promoter Regions, Genetic</subject><subject>Recombinant Proteins - biosynthesis</subject><subject>Saccharomyces cerevisiae - genetics</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo90M9rFDEUB_AgSt1WL97FHKQHYTS_Jskca2mtUBBsPQ8vmZc1kp2MyYy4_72z7NrT4_E-fHl8CXnD2UfOmPnkAmNcK9WpZ2TDlRQN01w9JxvGTduYtrMvyXmtv1YlrNZn5My2prPCbki6CSH6iONM8e9UsNaYRwrjQCv6gvNhy4Fe1QnLNqa0VDrGLRb6_a6VStEpp_0WEvh5KXmEijSO9AG8_wkl7_YeK_VY8E-sEfAVeREgVXx9mhfk8fbm8fquuf_25ev11X3jlRJzM9gANjhm9Pq-N4o5rbzWzjo5oIXgO8-dGLjjVgorQhjYMATsQIJzlssLcnmMnUr-vWCd-12sHlOCEfNSe2NMZ4y1K_xwhL7kWguGfipxB2Xfc9Yfmu0_3_5vdsVvT6mL2-HwRE9Vrvf3pztUDykUGH2sT0x0su34gb07sgC5h21ZyY8HwbhkvBVGGyP_AZR6i3s</recordid><startdate>19951201</startdate><enddate>19951201</enddate><creator>Lang, C</creator><creator>Looman, A.C</creator><general>Springer</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19951201</creationdate><title>Efficient expression and secretion of Aspergillus niger RH5344 polygalacturonase in Saccharomyces cerevisiae</title><author>Lang, C ; Looman, A.C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c442t-d8fa8fb076175c740b64c66b8b3de8afc9c1b2d1b183282ffd0ddfe9a3abb813</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>Alcohol Dehydrogenase - genetics</topic><topic>Aspergillus niger - enzymology</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Fermentation</topic><topic>food microbiology</topic><topic>food processing</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Glycosylation</topic><topic>horticultural crops</topic><topic>Methods. Procedures. Technologies</topic><topic>Modification of gene expression level</topic><topic>Molecular Sequence Data</topic><topic>Plasmids</topic><topic>Polygalacturonase - biosynthesis</topic><topic>Promoter Regions, Genetic</topic><topic>Recombinant Proteins - biosynthesis</topic><topic>Saccharomyces cerevisiae - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lang, C</creatorcontrib><creatorcontrib>Looman, A.C</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lang, C</au><au>Looman, A.C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Efficient expression and secretion of Aspergillus niger RH5344 polygalacturonase in Saccharomyces cerevisiae</atitle><jtitle>Applied microbiology and biotechnology</jtitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>1995-12-01</date><risdate>1995</risdate><volume>44</volume><issue>1/2</issue><spage>147</spage><epage>156</epage><pages>147-156</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><coden>AMBIDG</coden><abstract>An Aspergillus niger endopolygalacturonase (EC 3.2.1.15) cDNA was expressed in the yeast Saccharomyces cerevisiae. Secretion of the protein into the growth medium was efficiently directed by the fungal leader sequence, and processing occurred at the same site as in Aspergillus. The expression level was significantly enhanced by using a "short" version of the yeast ADHI promoter. An additional increase in the yield of heterologous protein was due to a higher plasmid stability and a rise in plasmid copy number. This was achieved by deleting most of the bacterial sequences from the expression vector. The yeast-derived enzyme showed the same enzymatic and biochemical properties as the fungal polygalacturonase, such as substrate specificity, pH and temperature optima and pI value. The yeast-derived enzyme, however, showed a higher degree of glycosylation and exhibited a more pronounced temperature stability than the fungal enzyme.</abstract><cop>Berlin</cop><pub>Springer</pub><pmid>8579828</pmid><doi>10.1007/bf00164494</doi><tpages>10</tpages></addata></record> |
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| identifier | ISSN: 0175-7598 |
| ispartof | Applied microbiology and biotechnology, 1995-12, Vol.44 (1/2), p.147-156 |
| issn | 0175-7598 1432-0614 |
| language | eng |
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| source | MEDLINE; SpringerNature Journals |
| subjects | Alcohol Dehydrogenase - genetics Aspergillus niger - enzymology Base Sequence Biological and medical sciences Biotechnology Fermentation food microbiology food processing Fundamental and applied biological sciences. Psychology Genetic engineering Genetic technics Glycosylation horticultural crops Methods. Procedures. Technologies Modification of gene expression level Molecular Sequence Data Plasmids Polygalacturonase - biosynthesis Promoter Regions, Genetic Recombinant Proteins - biosynthesis Saccharomyces cerevisiae - genetics |
| title | Efficient expression and secretion of Aspergillus niger RH5344 polygalacturonase in Saccharomyces cerevisiae |
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