Purification, characterization and crystallization of an extracellular alkaline protease from Aspergillus nidulans HA-10
Aspergillus nidulans is a highly potent fungus used in the production of alkaline protease. Extracellular alkaline protease was purified from A. nidulans in a two-step procedure involving ammonium sulphate precipitation and Sephadex G-100 column chromatography. The molecular mass of the enzyme was d...
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| Veröffentlicht in: | Journal of basic microbiology 2008-10, Vol.48 (5), p.347-352 |
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| container_title | Journal of basic microbiology |
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| creator | Charles, P Devanathan, V Anbu, Periasamy Ponnuswamy, M.N Kalaichelvan, P.T Hur, Byung-Ki |
| description | Aspergillus nidulans is a highly potent fungus used in the production of alkaline protease. Extracellular alkaline protease was purified from A. nidulans in a two-step procedure involving ammonium sulphate precipitation and Sephadex G-100 column chromatography. The molecular mass of the enzyme was determined to be 42 kDa by SDS-PAGE. The enzyme activity was also analyzed by zymogram with gelatin. The enzyme was more stable over a wide range of pH (6-10) and the temperatures up to 50 °C. It showed optimum enzyme activity at pH 8.0 and a temperature of 35 °C. The protease enzyme was completely inhibited by the serine protease inhibitor of phenylmethylsulfonyl fluoride (PMSF). The crystallization of the purified enzyme was performed by hanging drop vapour diffusion method using PEG 6000 as the precipitant. The micro crystals occurred in 40% of PEG 6000. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) |
| doi_str_mv | 10.1002/jobm.200800043 |
| format | Article |
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Extracellular alkaline protease was purified from A. nidulans in a two-step procedure involving ammonium sulphate precipitation and Sephadex G-100 column chromatography. The molecular mass of the enzyme was determined to be 42 kDa by SDS-PAGE. The enzyme activity was also analyzed by zymogram with gelatin. The enzyme was more stable over a wide range of pH (6-10) and the temperatures up to 50 °C. It showed optimum enzyme activity at pH 8.0 and a temperature of 35 °C. The protease enzyme was completely inhibited by the serine protease inhibitor of phenylmethylsulfonyl fluoride (PMSF). The crystallization of the purified enzyme was performed by hanging drop vapour diffusion method using PEG 6000 as the precipitant. The micro crystals occurred in 40% of PEG 6000. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)</description><identifier>ISSN: 0233-111X</identifier><identifier>EISSN: 1521-4028</identifier><identifier>DOI: 10.1002/jobm.200800043</identifier><identifier>PMID: 18702069</identifier><language>eng</language><publisher>Weinheim: Wiley-VCH Verlag</publisher><subject>Alkaline protease ; Aspergillus nidulans ; Aspergillus nidulans - enzymology ; Crystallization ; Fungal Proteins - isolation & purification ; Fungal Proteins - metabolism ; Hydrogen-Ion Concentration ; Molecular Weight ; Phenylmethylsulfonyl Fluoride - pharmacology ; Purification ; Serine Endopeptidases - drug effects ; Serine Endopeptidases - isolation & purification ; Serine Endopeptidases - metabolism ; Serine Proteinase Inhibitors - pharmacology ; Temperature</subject><ispartof>Journal of basic microbiology, 2008-10, Vol.48 (5), p.347-352</ispartof><rights>Copyright © 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><rights>(c) 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4713-2138778249ef00ecaee83927bd243f44c6a02eb6dd71e2f14de55507e6d8edfe3</citedby><cites>FETCH-LOGICAL-c4713-2138778249ef00ecaee83927bd243f44c6a02eb6dd71e2f14de55507e6d8edfe3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fjobm.200800043$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fjobm.200800043$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18702069$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Charles, P</creatorcontrib><creatorcontrib>Devanathan, V</creatorcontrib><creatorcontrib>Anbu, Periasamy</creatorcontrib><creatorcontrib>Ponnuswamy, M.N</creatorcontrib><creatorcontrib>Kalaichelvan, P.T</creatorcontrib><creatorcontrib>Hur, Byung-Ki</creatorcontrib><title>Purification, characterization and crystallization of an extracellular alkaline protease from Aspergillus nidulans HA-10</title><title>Journal of basic microbiology</title><addtitle>J. Basic Microbiol</addtitle><description>Aspergillus nidulans is a highly potent fungus used in the production of alkaline protease. Extracellular alkaline protease was purified from A. nidulans in a two-step procedure involving ammonium sulphate precipitation and Sephadex G-100 column chromatography. The molecular mass of the enzyme was determined to be 42 kDa by SDS-PAGE. The enzyme activity was also analyzed by zymogram with gelatin. The enzyme was more stable over a wide range of pH (6-10) and the temperatures up to 50 °C. It showed optimum enzyme activity at pH 8.0 and a temperature of 35 °C. The protease enzyme was completely inhibited by the serine protease inhibitor of phenylmethylsulfonyl fluoride (PMSF). The crystallization of the purified enzyme was performed by hanging drop vapour diffusion method using PEG 6000 as the precipitant. The micro crystals occurred in 40% of PEG 6000. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)</description><subject>Alkaline protease</subject><subject>Aspergillus nidulans</subject><subject>Aspergillus nidulans - enzymology</subject><subject>Crystallization</subject><subject>Fungal Proteins - isolation & purification</subject><subject>Fungal Proteins - metabolism</subject><subject>Hydrogen-Ion Concentration</subject><subject>Molecular Weight</subject><subject>Phenylmethylsulfonyl Fluoride - pharmacology</subject><subject>Purification</subject><subject>Serine Endopeptidases - drug effects</subject><subject>Serine Endopeptidases - isolation & purification</subject><subject>Serine Endopeptidases - metabolism</subject><subject>Serine Proteinase Inhibitors - pharmacology</subject><subject>Temperature</subject><issn>0233-111X</issn><issn>1521-4028</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0E1v1DAQBmALgehSuHIEnziRZfyRODluK9iCCi0qLb1Z3nhc3OZjsROxy6_HS5bCjZPl0TOvRi8hzxnMGQB_c9uv2jkHKAFAigdkxnLOMgm8fEhmwIXIGGPXB-RJjLeJVBWvHpMDVirgUFQzsjkfg3e-NoPvu9e0_maCqQcM_ufvCTWdpXXYxsE0zZ9Z79KY4mZIFJtmbEygprkzje-QrkM_oIlIXehbuohrDDc-oUg7bxPtIj1ZZAyekkfONBGf7d9Dcvnu7Zfjk-z0bPn-eHGa1VIxkXEmSqVKLit0AFgbxFJUXK0sl8JJWRcGOK4KaxVD7pi0mOc5KCxsidahOCSvptx02PcR46BbH3dnmw77MeqiKkTOpExwPsE69DEGdHodfGvCVjPQu671rmt933VaeLFPHlct2r98X24C1QR--Aa3_4nTH86OPv4bnk27Pg64ud814U4XSqhcf_201FdXy8_nF9fpk_zLyTvTa3MTfNSXFxyYAJbLsioL8QuEKqaK</recordid><startdate>200810</startdate><enddate>200810</enddate><creator>Charles, P</creator><creator>Devanathan, V</creator><creator>Anbu, Periasamy</creator><creator>Ponnuswamy, M.N</creator><creator>Kalaichelvan, P.T</creator><creator>Hur, Byung-Ki</creator><general>Wiley-VCH Verlag</general><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</general><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></search><sort><creationdate>200810</creationdate><title>Purification, characterization and crystallization of an extracellular alkaline protease from Aspergillus nidulans HA-10</title><author>Charles, P ; Devanathan, V ; Anbu, Periasamy ; Ponnuswamy, M.N ; Kalaichelvan, P.T ; Hur, Byung-Ki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4713-2138778249ef00ecaee83927bd243f44c6a02eb6dd71e2f14de55507e6d8edfe3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Alkaline protease</topic><topic>Aspergillus nidulans</topic><topic>Aspergillus nidulans - enzymology</topic><topic>Crystallization</topic><topic>Fungal Proteins - isolation & purification</topic><topic>Fungal Proteins - metabolism</topic><topic>Hydrogen-Ion Concentration</topic><topic>Molecular Weight</topic><topic>Phenylmethylsulfonyl Fluoride - pharmacology</topic><topic>Purification</topic><topic>Serine Endopeptidases - drug effects</topic><topic>Serine Endopeptidases - isolation & purification</topic><topic>Serine Endopeptidases - metabolism</topic><topic>Serine Proteinase Inhibitors - pharmacology</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Charles, P</creatorcontrib><creatorcontrib>Devanathan, V</creatorcontrib><creatorcontrib>Anbu, Periasamy</creatorcontrib><creatorcontrib>Ponnuswamy, M.N</creatorcontrib><creatorcontrib>Kalaichelvan, P.T</creatorcontrib><creatorcontrib>Hur, Byung-Ki</creatorcontrib><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><jtitle>Journal of basic microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Charles, P</au><au>Devanathan, V</au><au>Anbu, Periasamy</au><au>Ponnuswamy, M.N</au><au>Kalaichelvan, P.T</au><au>Hur, Byung-Ki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification, characterization and crystallization of an extracellular alkaline protease from Aspergillus nidulans HA-10</atitle><jtitle>Journal of basic microbiology</jtitle><addtitle>J. Basic Microbiol</addtitle><date>2008-10</date><risdate>2008</risdate><volume>48</volume><issue>5</issue><spage>347</spage><epage>352</epage><pages>347-352</pages><issn>0233-111X</issn><eissn>1521-4028</eissn><abstract>Aspergillus nidulans is a highly potent fungus used in the production of alkaline protease. Extracellular alkaline protease was purified from A. nidulans in a two-step procedure involving ammonium sulphate precipitation and Sephadex G-100 column chromatography. The molecular mass of the enzyme was determined to be 42 kDa by SDS-PAGE. The enzyme activity was also analyzed by zymogram with gelatin. The enzyme was more stable over a wide range of pH (6-10) and the temperatures up to 50 °C. It showed optimum enzyme activity at pH 8.0 and a temperature of 35 °C. The protease enzyme was completely inhibited by the serine protease inhibitor of phenylmethylsulfonyl fluoride (PMSF). The crystallization of the purified enzyme was performed by hanging drop vapour diffusion method using PEG 6000 as the precipitant. The micro crystals occurred in 40% of PEG 6000. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)</abstract><cop>Weinheim</cop><pub>Wiley-VCH Verlag</pub><pmid>18702069</pmid><doi>10.1002/jobm.200800043</doi><tpages>6</tpages></addata></record> |
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| subjects | Alkaline protease Aspergillus nidulans Aspergillus nidulans - enzymology Crystallization Fungal Proteins - isolation & purification Fungal Proteins - metabolism Hydrogen-Ion Concentration Molecular Weight Phenylmethylsulfonyl Fluoride - pharmacology Purification Serine Endopeptidases - drug effects Serine Endopeptidases - isolation & purification Serine Endopeptidases - metabolism Serine Proteinase Inhibitors - pharmacology Temperature |
| title | Purification, characterization and crystallization of an extracellular alkaline protease from Aspergillus nidulans HA-10 |
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