Biochemical Characterization of Soluble Acid and Alkaline Invertases from Shoots of Etiolated Pea Seedlings
Soluble invertase was purified from pea （Pisum sativum L.） by sequential procedures entailing ammonium sulfate precipitation, DEAE-Sepharose column, Con-A- and Green 19-Sepharose affinity columns, hydroxyapatite column, ultra-filtration, and Sephacryl 300 gel filtration. The purified soluble acid （S...
Gespeichert in:
| Veröffentlicht in: | Journal of integrative plant biology 2010-06, Vol.52 (6), p.536-548 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 548 |
|---|---|
| container_issue | 6 |
| container_start_page | 536 |
| container_title | Journal of integrative plant biology |
| container_volume | 52 |
| creator | Kim, Donggiun Park, So Yun Chung, Youngjae Park, Jongbum Lee, Sukchan Lee, Taek-Kyun |
| description | Soluble invertase was purified from pea （Pisum sativum L.） by sequential procedures entailing ammonium sulfate precipitation, DEAE-Sepharose column, Con-A- and Green 19-Sepharose affinity columns, hydroxyapatite column, ultra-filtration, and Sephacryl 300 gel filtration. The purified soluble acid （SAC） and alkaline （SALK） invertases had a pH optimum of 5.3 and 7.3, respectively. The temperature optimum of two invertases was 37 ℃. The effects of various concentrations of Tris-HCI, HgCI2, and CuSO4 on the activities of the two purified enzymes were examined. Tris-HCI and HgCI2 did not affect SAC activity, whereas 10 mM Tris-HCI and 0.05 mM HgCI2 inhibited SALK activity by about 50%. SAC and SALK were inhibited by 4.8 mM and 0.6 mM CuSO4 by 50%, respectively. The enzymes display typical hyperbolic saturation kinetics for sucrose hydrolysis. The Kms of SAC and SALK were determined to be 1.8 and 38.6 mM, respectively. The molecular masses of SAC shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting were 22 kDa and 45 kDa. The molecular mass of SALK was 30 kDa. Iso-electric points of the SAC and SALK were estimated to be about pH 7.0 and pH 5.7, respectively. |
| doi_str_mv | 10.1111/j.1744-7909.2010.00937.x |
| format | Article |
| fullrecord | <record><control><sourceid>wanfang_jour_proqu</sourceid><recordid>TN_cdi_wanfang_journals_zwxb201006003</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><cqvip_id>34284572</cqvip_id><wanfj_id>zwxb201006003</wanfj_id><sourcerecordid>zwxb201006003</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4877-ed85d7cc34937d4aeecd5e44c44210f55f0c73b1f9eb0382d5ebb8dee15036473</originalsourceid><addsrcrecordid>eNqNkV1v0zAUhiMEYmPwF8BCmrhKcWwnTi-46KptFA2YKBOXR45z0rpN481OWbdfzwktu8Y3tnye93y8J0lYxkcZnY-rUaaVSvWYj0eC0y_nY6lHu2fJ8VPgOb0LLdIx1-IoeRXjinNZ8kK8TI4Ez4ko1XGyPnPeLnHjrGnZdGmCsT0G92h65zvmGzb37bZqkU2sq5npajZp16Z1HbJZ9xtDbyJG1gS_YfOl930cNOckbk2PNbtGw-aINQkW8XXyojFtxDeH-yS5uTj_Of2cXn2_nE0nV6lVpdYp1mVea2uloplqZRBtnaNSVimR8SbPG261rLJmjBVNJChYVWWNmOVcFkrLk-R0n_fedI3pFrDy29BRRXi831WDX7wgL4j7sOdug7_bYuxh46LFtjUd-m0ELWVelLkoiCz3pA0-xoAN3Aa3MeEBMg7DRmAFg_EwGA9DBfi7EdiR9O2hyLbaYP0k_LcCAj4dunUtPvx3Yvgyuz6jF-nTvd7FHndPehPWUGipc_j17RLyi_nXH5mYgiD-3Z5vjAezCC7CzZwyS56VZJ7iRLw_TLv03eKOVgeVseuG-gOpRKlyLeQf-ei9mA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>733568526</pqid></control><display><type>article</type><title>Biochemical Characterization of Soluble Acid and Alkaline Invertases from Shoots of Etiolated Pea Seedlings</title><source>MEDLINE</source><source>Wiley Online Library</source><source>Alma/SFX Local Collection</source><creator>Kim, Donggiun ; Park, So Yun ; Chung, Youngjae ; Park, Jongbum ; Lee, Sukchan ; Lee, Taek-Kyun</creator><creatorcontrib>Kim, Donggiun ; Park, So Yun ; Chung, Youngjae ; Park, Jongbum ; Lee, Sukchan ; Lee, Taek-Kyun</creatorcontrib><description>Soluble invertase was purified from pea （Pisum sativum L.） by sequential procedures entailing ammonium sulfate precipitation, DEAE-Sepharose column, Con-A- and Green 19-Sepharose affinity columns, hydroxyapatite column, ultra-filtration, and Sephacryl 300 gel filtration. The purified soluble acid （SAC） and alkaline （SALK） invertases had a pH optimum of 5.3 and 7.3, respectively. The temperature optimum of two invertases was 37 ℃. The effects of various concentrations of Tris-HCI, HgCI2, and CuSO4 on the activities of the two purified enzymes were examined. Tris-HCI and HgCI2 did not affect SAC activity, whereas 10 mM Tris-HCI and 0.05 mM HgCI2 inhibited SALK activity by about 50%. SAC and SALK were inhibited by 4.8 mM and 0.6 mM CuSO4 by 50%, respectively. The enzymes display typical hyperbolic saturation kinetics for sucrose hydrolysis. The Kms of SAC and SALK were determined to be 1.8 and 38.6 mM, respectively. The molecular masses of SAC shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting were 22 kDa and 45 kDa. The molecular mass of SALK was 30 kDa. Iso-electric points of the SAC and SALK were estimated to be about pH 7.0 and pH 5.7, respectively.</description><identifier>ISSN: 1672-9072</identifier><identifier>EISSN: 1744-7909</identifier><identifier>DOI: 10.1111/j.1744-7909.2010.00937.x</identifier><identifier>PMID: 20590984</identifier><language>eng</language><publisher>Melbourne, Australia: Melbourne, Australia : Blackwell Publishing Asia</publisher><subject>beta-Fructofuranosidase - metabolism ; Electrophoresis, Polyacrylamide Gel ; Hydrogen-Ion Concentration ; Isoelectric Point ; Molecular Weight ; Pisum sativum - enzymology ; Plant Proteins - antagonists & inhibitors ; Plant Proteins - chemistry ; Plant Proteins - genetics ; Plant Proteins - metabolism ; Plant Shoots - enzymology ; Seedlings - enzymology ; Sepharose ; Substrate Specificity ; Sucrose - metabolism ; 十二烷基硫酸钠 ; 可溶性 ; 生化特性 ; 聚丙烯酰胺凝胶电泳 ; 豌豆苗 ; 转化酶</subject><ispartof>Journal of integrative plant biology, 2010-06, Vol.52 (6), p.536-548</ispartof><rights>2010 Institute of Botany, Chinese Academy of Sciences</rights><rights>Copyright © Wanfang Data Co. Ltd. All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4877-ed85d7cc34937d4aeecd5e44c44210f55f0c73b1f9eb0382d5ebb8dee15036473</citedby><cites>FETCH-LOGICAL-c4877-ed85d7cc34937d4aeecd5e44c44210f55f0c73b1f9eb0382d5ebb8dee15036473</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://image.cqvip.com/vip1000/qk/94176A/94176A.jpg</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1744-7909.2010.00937.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1744-7909.2010.00937.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20590984$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kim, Donggiun</creatorcontrib><creatorcontrib>Park, So Yun</creatorcontrib><creatorcontrib>Chung, Youngjae</creatorcontrib><creatorcontrib>Park, Jongbum</creatorcontrib><creatorcontrib>Lee, Sukchan</creatorcontrib><creatorcontrib>Lee, Taek-Kyun</creatorcontrib><title>Biochemical Characterization of Soluble Acid and Alkaline Invertases from Shoots of Etiolated Pea Seedlings</title><title>Journal of integrative plant biology</title><addtitle>Journal of Integrative Plant Biology</addtitle><description>Soluble invertase was purified from pea （Pisum sativum L.） by sequential procedures entailing ammonium sulfate precipitation, DEAE-Sepharose column, Con-A- and Green 19-Sepharose affinity columns, hydroxyapatite column, ultra-filtration, and Sephacryl 300 gel filtration. The purified soluble acid （SAC） and alkaline （SALK） invertases had a pH optimum of 5.3 and 7.3, respectively. The temperature optimum of two invertases was 37 ℃. The effects of various concentrations of Tris-HCI, HgCI2, and CuSO4 on the activities of the two purified enzymes were examined. Tris-HCI and HgCI2 did not affect SAC activity, whereas 10 mM Tris-HCI and 0.05 mM HgCI2 inhibited SALK activity by about 50%. SAC and SALK were inhibited by 4.8 mM and 0.6 mM CuSO4 by 50%, respectively. The enzymes display typical hyperbolic saturation kinetics for sucrose hydrolysis. The Kms of SAC and SALK were determined to be 1.8 and 38.6 mM, respectively. The molecular masses of SAC shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting were 22 kDa and 45 kDa. The molecular mass of SALK was 30 kDa. Iso-electric points of the SAC and SALK were estimated to be about pH 7.0 and pH 5.7, respectively.</description><subject>beta-Fructofuranosidase - metabolism</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Hydrogen-Ion Concentration</subject><subject>Isoelectric Point</subject><subject>Molecular Weight</subject><subject>Pisum sativum - enzymology</subject><subject>Plant Proteins - antagonists & inhibitors</subject><subject>Plant Proteins - chemistry</subject><subject>Plant Proteins - genetics</subject><subject>Plant Proteins - metabolism</subject><subject>Plant Shoots - enzymology</subject><subject>Seedlings - enzymology</subject><subject>Sepharose</subject><subject>Substrate Specificity</subject><subject>Sucrose - metabolism</subject><subject>十二烷基硫酸钠</subject><subject>可溶性</subject><subject>生化特性</subject><subject>聚丙烯酰胺凝胶电泳</subject><subject>豌豆苗</subject><subject>转化酶</subject><issn>1672-9072</issn><issn>1744-7909</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkV1v0zAUhiMEYmPwF8BCmrhKcWwnTi-46KptFA2YKBOXR45z0rpN481OWbdfzwktu8Y3tnye93y8J0lYxkcZnY-rUaaVSvWYj0eC0y_nY6lHu2fJ8VPgOb0LLdIx1-IoeRXjinNZ8kK8TI4Ez4ko1XGyPnPeLnHjrGnZdGmCsT0G92h65zvmGzb37bZqkU2sq5npajZp16Z1HbJZ9xtDbyJG1gS_YfOl930cNOckbk2PNbtGw-aINQkW8XXyojFtxDeH-yS5uTj_Of2cXn2_nE0nV6lVpdYp1mVea2uloplqZRBtnaNSVimR8SbPG261rLJmjBVNJChYVWWNmOVcFkrLk-R0n_fedI3pFrDy29BRRXi831WDX7wgL4j7sOdug7_bYuxh46LFtjUd-m0ELWVelLkoiCz3pA0-xoAN3Aa3MeEBMg7DRmAFg_EwGA9DBfi7EdiR9O2hyLbaYP0k_LcCAj4dunUtPvx3Yvgyuz6jF-nTvd7FHndPehPWUGipc_j17RLyi_nXH5mYgiD-3Z5vjAezCC7CzZwyS56VZJ7iRLw_TLv03eKOVgeVseuG-gOpRKlyLeQf-ei9mA</recordid><startdate>201006</startdate><enddate>201006</enddate><creator>Kim, Donggiun</creator><creator>Park, So Yun</creator><creator>Chung, Youngjae</creator><creator>Park, Jongbum</creator><creator>Lee, Sukchan</creator><creator>Lee, Taek-Kyun</creator><general>Melbourne, Australia : Blackwell Publishing Asia</general><general>Blackwell Publishing Asia</general><general>Department of Biological Science, Silla University, Busan, 617-736, Korea%South Sea Environment Research Department, Korea Ocean Research and Development Institute, Geoje, 656-830, Korea%Department of Life Science and Biotechnology, Shin Gyeong University, Hwaseong 445-741, Korea%Department of Genetic Engineering, Sungkyunkwan University, Suwon, 440-746, Korea</general><scope>2RA</scope><scope>92L</scope><scope>CQIGP</scope><scope>W94</scope><scope>WU4</scope><scope>~WA</scope><scope>FBQ</scope><scope>BSCLL</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><scope>2B.</scope><scope>4A8</scope><scope>92I</scope><scope>93N</scope><scope>PSX</scope><scope>TCJ</scope></search><sort><creationdate>201006</creationdate><title>Biochemical Characterization of Soluble Acid and Alkaline Invertases from Shoots of Etiolated Pea Seedlings</title><author>Kim, Donggiun ; Park, So Yun ; Chung, Youngjae ; Park, Jongbum ; Lee, Sukchan ; Lee, Taek-Kyun</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4877-ed85d7cc34937d4aeecd5e44c44210f55f0c73b1f9eb0382d5ebb8dee15036473</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>beta-Fructofuranosidase - metabolism</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Hydrogen-Ion Concentration</topic><topic>Isoelectric Point</topic><topic>Molecular Weight</topic><topic>Pisum sativum - enzymology</topic><topic>Plant Proteins - antagonists & inhibitors</topic><topic>Plant Proteins - chemistry</topic><topic>Plant Proteins - genetics</topic><topic>Plant Proteins - metabolism</topic><topic>Plant Shoots - enzymology</topic><topic>Seedlings - enzymology</topic><topic>Sepharose</topic><topic>Substrate Specificity</topic><topic>Sucrose - metabolism</topic><topic>十二烷基硫酸钠</topic><topic>可溶性</topic><topic>生化特性</topic><topic>聚丙烯酰胺凝胶电泳</topic><topic>豌豆苗</topic><topic>转化酶</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kim, Donggiun</creatorcontrib><creatorcontrib>Park, So Yun</creatorcontrib><creatorcontrib>Chung, Youngjae</creatorcontrib><creatorcontrib>Park, Jongbum</creatorcontrib><creatorcontrib>Lee, Sukchan</creatorcontrib><creatorcontrib>Lee, Taek-Kyun</creatorcontrib><collection>维普_期刊</collection><collection>中文科技期刊数据库-CALIS站点</collection><collection>维普中文期刊数据库</collection><collection>中文科技期刊数据库-自然科学</collection><collection>中文科技期刊数据库-自然科学-生物科学</collection><collection>中文科技期刊数据库- 镜像站点</collection><collection>AGRIS</collection><collection>Istex</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><collection>Wanfang Data Journals - Hong Kong</collection><collection>WANFANG Data Centre</collection><collection>Wanfang Data Journals</collection><collection>万方数据期刊 - 香港版</collection><collection>China Online Journals (COJ)</collection><collection>China Online Journals (COJ)</collection><jtitle>Journal of integrative plant biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kim, Donggiun</au><au>Park, So Yun</au><au>Chung, Youngjae</au><au>Park, Jongbum</au><au>Lee, Sukchan</au><au>Lee, Taek-Kyun</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Biochemical Characterization of Soluble Acid and Alkaline Invertases from Shoots of Etiolated Pea Seedlings</atitle><jtitle>Journal of integrative plant biology</jtitle><addtitle>Journal of Integrative Plant Biology</addtitle><date>2010-06</date><risdate>2010</risdate><volume>52</volume><issue>6</issue><spage>536</spage><epage>548</epage><pages>536-548</pages><issn>1672-9072</issn><eissn>1744-7909</eissn><abstract>Soluble invertase was purified from pea （Pisum sativum L.） by sequential procedures entailing ammonium sulfate precipitation, DEAE-Sepharose column, Con-A- and Green 19-Sepharose affinity columns, hydroxyapatite column, ultra-filtration, and Sephacryl 300 gel filtration. The purified soluble acid （SAC） and alkaline （SALK） invertases had a pH optimum of 5.3 and 7.3, respectively. The temperature optimum of two invertases was 37 ℃. The effects of various concentrations of Tris-HCI, HgCI2, and CuSO4 on the activities of the two purified enzymes were examined. Tris-HCI and HgCI2 did not affect SAC activity, whereas 10 mM Tris-HCI and 0.05 mM HgCI2 inhibited SALK activity by about 50%. SAC and SALK were inhibited by 4.8 mM and 0.6 mM CuSO4 by 50%, respectively. The enzymes display typical hyperbolic saturation kinetics for sucrose hydrolysis. The Kms of SAC and SALK were determined to be 1.8 and 38.6 mM, respectively. The molecular masses of SAC shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting were 22 kDa and 45 kDa. The molecular mass of SALK was 30 kDa. Iso-electric points of the SAC and SALK were estimated to be about pH 7.0 and pH 5.7, respectively.</abstract><cop>Melbourne, Australia</cop><pub>Melbourne, Australia : Blackwell Publishing Asia</pub><pmid>20590984</pmid><doi>10.1111/j.1744-7909.2010.00937.x</doi><tpages>13</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1672-9072 |
| ispartof | Journal of integrative plant biology, 2010-06, Vol.52 (6), p.536-548 |
| issn | 1672-9072 1744-7909 |
| language | eng |
| recordid | cdi_wanfang_journals_zwxb201006003 |
| source | MEDLINE; Wiley Online Library; Alma/SFX Local Collection |
| subjects | beta-Fructofuranosidase - metabolism Electrophoresis, Polyacrylamide Gel Hydrogen-Ion Concentration Isoelectric Point Molecular Weight Pisum sativum - enzymology Plant Proteins - antagonists & inhibitors Plant Proteins - chemistry Plant Proteins - genetics Plant Proteins - metabolism Plant Shoots - enzymology Seedlings - enzymology Sepharose Substrate Specificity Sucrose - metabolism 十二烷基硫酸钠 可溶性 生化特性 聚丙烯酰胺凝胶电泳 豌豆苗 转化酶 |
| title | Biochemical Characterization of Soluble Acid and Alkaline Invertases from Shoots of Etiolated Pea Seedlings |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-14T13%3A03%3A28IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-wanfang_jour_proqu&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Biochemical%20Characterization%20of%20Soluble%20Acid%20and%20Alkaline%20Invertases%20from%20Shoots%20of%20Etiolated%20Pea%20Seedlings&rft.jtitle=Journal%20of%20integrative%20plant%20biology&rft.au=Kim,%20Donggiun&rft.date=2010-06&rft.volume=52&rft.issue=6&rft.spage=536&rft.epage=548&rft.pages=536-548&rft.issn=1672-9072&rft.eissn=1744-7909&rft_id=info:doi/10.1111/j.1744-7909.2010.00937.x&rft_dat=%3Cwanfang_jour_proqu%3Ezwxb201006003%3C/wanfang_jour_proqu%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=733568526&rft_id=info:pmid/20590984&rft_cqvip_id=34284572&rft_wanfj_id=zwxb201006003&rfr_iscdi=true |