Gene Sequence, Bioinformatics and Enzymatic Characterization of α-Amylase from Saccharomycopsis fibuligera KZ
A fragment coding for a putative extracellular α-amylase, from the genomic library of the yeast Saccharomycopsis fibuligera KZ, has been subcloned into yeast expression vector pVT100L and sequenced. The nucleotide sequence revealed an ORF of 1,485 bp coding for a 494 amino acid residues long protein...
Gespeichert in:
| Veröffentlicht in: | The Protein Journal 2010-07, Vol.29 (5), p.355-364 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 364 |
|---|---|
| container_issue | 5 |
| container_start_page | 355 |
| container_title | The Protein Journal |
| container_volume | 29 |
| creator | Hostinová, Eva Janeček, Štefan Gašperík, Juraj |
| description | A fragment coding for a putative extracellular α-amylase, from the genomic library of the yeast
Saccharomycopsis fibuligera
KZ, has been subcloned into yeast expression vector pVT100L and sequenced. The nucleotide sequence revealed an ORF of 1,485 bp coding for a 494 amino acid residues long protein with 99% identity to the α-amylase Sfamy from
S. fibuligera
HUT 7212. The
S. fibuligera
KZ α-amylase (Sfamy KZ) belongs to typical extracellular fungal α-amylases classified in the glycoside hydrolase family 13, subfamily 1, as supported also by clustering observed in the evolutionary tree. Sfamy KZ, in addition to the essential GH13 α-amylase three-domain arrangement (catalytic TIM barrel plus domains B and C), does not contain any distinct starch-binding domain. Sfamy KZ was expressed as a recombinant protein in
Saccharomyces cerevisiae
and purified to electrophoretic homogeneity. The enzyme had a molecular mass 53 kDa and contained about 2.5% of carbohydrate. The enzyme exhibited pH and temperature optima in the range of 5–6 and 40–50 °C, respectively. Stable adsorption of the enzyme to starch granules was not detected but a low degradation of raw starch in a concentration-dependent manner was observed. |
| doi_str_mv | 10.1007/s10930-010-9260-6 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_733479451</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>733479451</sourcerecordid><originalsourceid>FETCH-LOGICAL-c376t-636e37d31f768e012501bf624d14abbbeaa38a9bf4b2cf188ca54cef2442ab363</originalsourceid><addsrcrecordid>eNp9kL1OHDEURq2IKMAmD5AGuaPBxH8zninJagMIJIolTRrL9l4Hoxl7sXeK5a3yInmmeFlISWVf3XM_6TsIfWX0nFGqvhVGe0EJZZT0vKWk_YCOWKMEkb0UBy9_TkTXqUN0XMojpbzrFf-EDjltGl4vjlC8hAh4CU8TRAdn-HtIIfqUR7MJrmATV3gRn7cvI54_mGzcBnJ4rnOKOHn89w-5GLeDKYB9TiNeGucqlsatS-sSCvbBTkP4Ddngm1-f0UdvhgJfXt8Z-vljcT-_Ird3l9fzi1vihGo3pBUtCLUSzKu2A8p4Q5n1LZcrJo21FowRnemtl5Y7z7rOmUY68FxKbqxoxQyd7nPXOdVqZaPHUBwMg4mQpqKVEFL1smGVZHvS5VRKBq_XOYwmbzWjemdZ7y3ralnvLOtd-slr-mRHWP2_eNNaAb4HSl3F2l0_pinH2vid1H_I9IoF</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>733479451</pqid></control><display><type>article</type><title>Gene Sequence, Bioinformatics and Enzymatic Characterization of α-Amylase from Saccharomycopsis fibuligera KZ</title><source>MEDLINE</source><source>SpringerLink Journals - AutoHoldings</source><creator>Hostinová, Eva ; Janeček, Štefan ; Gašperík, Juraj</creator><creatorcontrib>Hostinová, Eva ; Janeček, Štefan ; Gašperík, Juraj</creatorcontrib><description>A fragment coding for a putative extracellular α-amylase, from the genomic library of the yeast
Saccharomycopsis fibuligera
KZ, has been subcloned into yeast expression vector pVT100L and sequenced. The nucleotide sequence revealed an ORF of 1,485 bp coding for a 494 amino acid residues long protein with 99% identity to the α-amylase Sfamy from
S. fibuligera
HUT 7212. The
S. fibuligera
KZ α-amylase (Sfamy KZ) belongs to typical extracellular fungal α-amylases classified in the glycoside hydrolase family 13, subfamily 1, as supported also by clustering observed in the evolutionary tree. Sfamy KZ, in addition to the essential GH13 α-amylase three-domain arrangement (catalytic TIM barrel plus domains B and C), does not contain any distinct starch-binding domain. Sfamy KZ was expressed as a recombinant protein in
Saccharomyces cerevisiae
and purified to electrophoretic homogeneity. The enzyme had a molecular mass 53 kDa and contained about 2.5% of carbohydrate. The enzyme exhibited pH and temperature optima in the range of 5–6 and 40–50 °C, respectively. Stable adsorption of the enzyme to starch granules was not detected but a low degradation of raw starch in a concentration-dependent manner was observed.</description><identifier>ISSN: 1572-3887</identifier><identifier>EISSN: 1573-4943</identifier><identifier>EISSN: 1875-8355</identifier><identifier>DOI: 10.1007/s10930-010-9260-6</identifier><identifier>PMID: 20552260</identifier><language>eng</language><publisher>Boston: Springer US</publisher><subject>alpha-Amylases - chemistry ; alpha-Amylases - genetics ; alpha-Amylases - metabolism ; Amino Acid Sequence ; Animal Anatomy ; Base Sequence ; Biochemistry ; Bioorganic Chemistry ; Chemistry ; Chemistry and Materials Science ; Cloning, Molecular ; Computational Biology ; Fungal Proteins - chemistry ; Fungal Proteins - genetics ; Fungal Proteins - metabolism ; Histology ; Hydrogen-Ion Concentration ; Hydrolysis ; Models, Molecular ; Molecular Sequence Data ; Morphology ; Organic Chemistry ; Phylogeny ; Recombinant Proteins - chemistry ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Saccharomyces cerevisiae - genetics ; Saccharomycopsis - enzymology ; Saccharomycopsis - genetics ; Starch - metabolism ; Temperature ; Zea mays</subject><ispartof>The Protein Journal, 2010-07, Vol.29 (5), p.355-364</ispartof><rights>Springer Science+Business Media, LLC 2010</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c376t-636e37d31f768e012501bf624d14abbbeaa38a9bf4b2cf188ca54cef2442ab363</citedby><cites>FETCH-LOGICAL-c376t-636e37d31f768e012501bf624d14abbbeaa38a9bf4b2cf188ca54cef2442ab363</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10930-010-9260-6$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10930-010-9260-6$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27923,27924,41487,42556,51318</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20552260$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hostinová, Eva</creatorcontrib><creatorcontrib>Janeček, Štefan</creatorcontrib><creatorcontrib>Gašperík, Juraj</creatorcontrib><title>Gene Sequence, Bioinformatics and Enzymatic Characterization of α-Amylase from Saccharomycopsis fibuligera KZ</title><title>The Protein Journal</title><addtitle>Protein J</addtitle><addtitle>Protein J</addtitle><description>A fragment coding for a putative extracellular α-amylase, from the genomic library of the yeast
Saccharomycopsis fibuligera
KZ, has been subcloned into yeast expression vector pVT100L and sequenced. The nucleotide sequence revealed an ORF of 1,485 bp coding for a 494 amino acid residues long protein with 99% identity to the α-amylase Sfamy from
S. fibuligera
HUT 7212. The
S. fibuligera
KZ α-amylase (Sfamy KZ) belongs to typical extracellular fungal α-amylases classified in the glycoside hydrolase family 13, subfamily 1, as supported also by clustering observed in the evolutionary tree. Sfamy KZ, in addition to the essential GH13 α-amylase three-domain arrangement (catalytic TIM barrel plus domains B and C), does not contain any distinct starch-binding domain. Sfamy KZ was expressed as a recombinant protein in
Saccharomyces cerevisiae
and purified to electrophoretic homogeneity. The enzyme had a molecular mass 53 kDa and contained about 2.5% of carbohydrate. The enzyme exhibited pH and temperature optima in the range of 5–6 and 40–50 °C, respectively. Stable adsorption of the enzyme to starch granules was not detected but a low degradation of raw starch in a concentration-dependent manner was observed.</description><subject>alpha-Amylases - chemistry</subject><subject>alpha-Amylases - genetics</subject><subject>alpha-Amylases - metabolism</subject><subject>Amino Acid Sequence</subject><subject>Animal Anatomy</subject><subject>Base Sequence</subject><subject>Biochemistry</subject><subject>Bioorganic Chemistry</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Cloning, Molecular</subject><subject>Computational Biology</subject><subject>Fungal Proteins - chemistry</subject><subject>Fungal Proteins - genetics</subject><subject>Fungal Proteins - metabolism</subject><subject>Histology</subject><subject>Hydrogen-Ion Concentration</subject><subject>Hydrolysis</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Morphology</subject><subject>Organic Chemistry</subject><subject>Phylogeny</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomycopsis - enzymology</subject><subject>Saccharomycopsis - genetics</subject><subject>Starch - metabolism</subject><subject>Temperature</subject><subject>Zea mays</subject><issn>1572-3887</issn><issn>1573-4943</issn><issn>1875-8355</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kL1OHDEURq2IKMAmD5AGuaPBxH8zninJagMIJIolTRrL9l4Hoxl7sXeK5a3yInmmeFlISWVf3XM_6TsIfWX0nFGqvhVGe0EJZZT0vKWk_YCOWKMEkb0UBy9_TkTXqUN0XMojpbzrFf-EDjltGl4vjlC8hAh4CU8TRAdn-HtIIfqUR7MJrmATV3gRn7cvI54_mGzcBnJ4rnOKOHn89w-5GLeDKYB9TiNeGucqlsatS-sSCvbBTkP4Ddngm1-f0UdvhgJfXt8Z-vljcT-_Ird3l9fzi1vihGo3pBUtCLUSzKu2A8p4Q5n1LZcrJo21FowRnemtl5Y7z7rOmUY68FxKbqxoxQyd7nPXOdVqZaPHUBwMg4mQpqKVEFL1smGVZHvS5VRKBq_XOYwmbzWjemdZ7y3ralnvLOtd-slr-mRHWP2_eNNaAb4HSl3F2l0_pinH2vid1H_I9IoF</recordid><startdate>20100701</startdate><enddate>20100701</enddate><creator>Hostinová, Eva</creator><creator>Janeček, Štefan</creator><creator>Gašperík, Juraj</creator><general>Springer US</general><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>20100701</creationdate><title>Gene Sequence, Bioinformatics and Enzymatic Characterization of α-Amylase from Saccharomycopsis fibuligera KZ</title><author>Hostinová, Eva ; Janeček, Štefan ; Gašperík, Juraj</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c376t-636e37d31f768e012501bf624d14abbbeaa38a9bf4b2cf188ca54cef2442ab363</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>alpha-Amylases - chemistry</topic><topic>alpha-Amylases - genetics</topic><topic>alpha-Amylases - metabolism</topic><topic>Amino Acid Sequence</topic><topic>Animal Anatomy</topic><topic>Base Sequence</topic><topic>Biochemistry</topic><topic>Bioorganic Chemistry</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Cloning, Molecular</topic><topic>Computational Biology</topic><topic>Fungal Proteins - chemistry</topic><topic>Fungal Proteins - genetics</topic><topic>Fungal Proteins - metabolism</topic><topic>Histology</topic><topic>Hydrogen-Ion Concentration</topic><topic>Hydrolysis</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Morphology</topic><topic>Organic Chemistry</topic><topic>Phylogeny</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><topic>Saccharomyces cerevisiae - genetics</topic><topic>Saccharomycopsis - enzymology</topic><topic>Saccharomycopsis - genetics</topic><topic>Starch - metabolism</topic><topic>Temperature</topic><topic>Zea mays</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hostinová, Eva</creatorcontrib><creatorcontrib>Janeček, Štefan</creatorcontrib><creatorcontrib>Gašperík, Juraj</creatorcontrib><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>The Protein Journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hostinová, Eva</au><au>Janeček, Štefan</au><au>Gašperík, Juraj</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Gene Sequence, Bioinformatics and Enzymatic Characterization of α-Amylase from Saccharomycopsis fibuligera KZ</atitle><jtitle>The Protein Journal</jtitle><stitle>Protein J</stitle><addtitle>Protein J</addtitle><date>2010-07-01</date><risdate>2010</risdate><volume>29</volume><issue>5</issue><spage>355</spage><epage>364</epage><pages>355-364</pages><issn>1572-3887</issn><eissn>1573-4943</eissn><eissn>1875-8355</eissn><abstract>A fragment coding for a putative extracellular α-amylase, from the genomic library of the yeast
Saccharomycopsis fibuligera
KZ, has been subcloned into yeast expression vector pVT100L and sequenced. The nucleotide sequence revealed an ORF of 1,485 bp coding for a 494 amino acid residues long protein with 99% identity to the α-amylase Sfamy from
S. fibuligera
HUT 7212. The
S. fibuligera
KZ α-amylase (Sfamy KZ) belongs to typical extracellular fungal α-amylases classified in the glycoside hydrolase family 13, subfamily 1, as supported also by clustering observed in the evolutionary tree. Sfamy KZ, in addition to the essential GH13 α-amylase three-domain arrangement (catalytic TIM barrel plus domains B and C), does not contain any distinct starch-binding domain. Sfamy KZ was expressed as a recombinant protein in
Saccharomyces cerevisiae
and purified to electrophoretic homogeneity. The enzyme had a molecular mass 53 kDa and contained about 2.5% of carbohydrate. The enzyme exhibited pH and temperature optima in the range of 5–6 and 40–50 °C, respectively. Stable adsorption of the enzyme to starch granules was not detected but a low degradation of raw starch in a concentration-dependent manner was observed.</abstract><cop>Boston</cop><pub>Springer US</pub><pmid>20552260</pmid><doi>10.1007/s10930-010-9260-6</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1572-3887 |
| ispartof | The Protein Journal, 2010-07, Vol.29 (5), p.355-364 |
| issn | 1572-3887 1573-4943 1875-8355 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_733479451 |
| source | MEDLINE; SpringerLink Journals - AutoHoldings |
| subjects | alpha-Amylases - chemistry alpha-Amylases - genetics alpha-Amylases - metabolism Amino Acid Sequence Animal Anatomy Base Sequence Biochemistry Bioorganic Chemistry Chemistry Chemistry and Materials Science Cloning, Molecular Computational Biology Fungal Proteins - chemistry Fungal Proteins - genetics Fungal Proteins - metabolism Histology Hydrogen-Ion Concentration Hydrolysis Models, Molecular Molecular Sequence Data Morphology Organic Chemistry Phylogeny Recombinant Proteins - chemistry Recombinant Proteins - genetics Recombinant Proteins - metabolism Saccharomyces cerevisiae - genetics Saccharomycopsis - enzymology Saccharomycopsis - genetics Starch - metabolism Temperature Zea mays |
| title | Gene Sequence, Bioinformatics and Enzymatic Characterization of α-Amylase from Saccharomycopsis fibuligera KZ |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-11T00%3A54%3A04IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Gene%20Sequence,%20Bioinformatics%20and%20Enzymatic%20Characterization%20of%20%CE%B1-Amylase%20from%20Saccharomycopsis%20fibuligera%20KZ&rft.jtitle=The%20Protein%20Journal&rft.au=Hostinov%C3%A1,%20Eva&rft.date=2010-07-01&rft.volume=29&rft.issue=5&rft.spage=355&rft.epage=364&rft.pages=355-364&rft.issn=1572-3887&rft.eissn=1573-4943&rft_id=info:doi/10.1007/s10930-010-9260-6&rft_dat=%3Cproquest_cross%3E733479451%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=733479451&rft_id=info:pmid/20552260&rfr_iscdi=true |