Cleaning protocols for crystallization robots: preventing protease contamination
The protease in the commonly used commercial low‐foam enzyme cleaner Zymit cannot be completely blocked by EDTA, a widely used inhibitor of metalloproteases, at concentrations of up to 5 mM. Severe protein degradation was observed in crystallization drops after EDTA‐containing wash steps unless resi...
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| Veröffentlicht in: | Acta crystallographica. Section F, Structural biology communications Structural biology communications, 2015-01, Vol.71 (1), p.100-102 |
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| container_title | Acta crystallographica. Section F, Structural biology communications |
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| creator | Naschberger, Andreas Fürnrohr, Barbara G. Dunzendorfer-Matt, Theresia Bonagura, Christopher A. Wright, David Scheffzek, Klaus Rupp, Bernhard |
| description | The protease in the commonly used commercial low‐foam enzyme cleaner Zymit cannot be completely blocked by EDTA, a widely used inhibitor of metalloproteases, at concentrations of up to 5 mM. Severe protein degradation was observed in crystallization drops after EDTA‐containing wash steps unless residual Zymit protease was removed with NaOH at a concentration of at least 0.1 M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1 M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity. |
| doi_str_mv | 10.1107/S2053230X14026053 |
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Severe protein degradation was observed in crystallization drops after EDTA‐containing wash steps unless residual Zymit protease was removed with NaOH at a concentration of at least 0.1 M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1 M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity.</description><identifier>ISSN: 2053-230X</identifier><identifier>EISSN: 2053-230X</identifier><identifier>DOI: 10.1107/S2053230X14026053</identifier><identifier>PMID: 25615978</identifier><language>eng</language><publisher>5 Abbey Square, Chester, Cheshire CH1 2HU, England: International Union of Crystallography</publisher><subject>Automation, Laboratory ; cleaning protocol ; Crystallization ; crystallization robotics ; Crystallography, X-Ray ; Equipment Contamination ; Peptide Hydrolases - chemistry ; protease ; protease inhibitor ; Protease Inhibitors - chemistry ; Research Communications ; Robotics ; Zymit</subject><ispartof>Acta crystallographica. 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Protocols including wash steps with at least 0.1 M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity.</description><subject>Automation, Laboratory</subject><subject>cleaning protocol</subject><subject>Crystallization</subject><subject>crystallization robotics</subject><subject>Crystallography, X-Ray</subject><subject>Equipment Contamination</subject><subject>Peptide Hydrolases - chemistry</subject><subject>protease</subject><subject>protease inhibitor</subject><subject>Protease Inhibitors - chemistry</subject><subject>Research Communications</subject><subject>Robotics</subject><subject>Zymit</subject><issn>2053-230X</issn><issn>2053-230X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUFP3DAQha2KChDwA7hUkbhwSRnbcZxwqIRWZUFCdCVAQC-W4zjU1GsvdpZ2-fV1urCi5cDJo5nvPc3zILSL4TPGwA8uCDBKKNzgAkiZ6g9oc2jlQ2_tVb2BdmK8B4BBhnm9jjYIKzGrebWJJiOrpTPuLpsF33vlbcw6HzIVFrGX1pon2RvvsuAb38fDROlH7foXgZZRZ8q7Xk6N-0tuo4-dtFHvPL9b6Or46-XoJD_7Nj4dHZ3limHG864rm7LR0Fa85nULZc0ZVLJluGxxW7FCs0I1isgCQEIjZVt1jcId4ZyylINuoS9L39m8mepWpaWCtGIWzFSGhfDSiH8nzvwQd_5RFBQKzgaD_WeD4B_mOvZiaqLS1kqn_TwKXDJSUEzqKqF7_6H3fh5cipeoouK0SssnCi8pFXyMQXerZTCI4e_Fm5MlzafXKVaKlwMloF4Cv4zVi_cdxdHtMbm4ZsCHhPlSa2Kvf6-0MvwUJaecievzsYDxhIy_04kA-gclZLJn</recordid><startdate>20150101</startdate><enddate>20150101</enddate><creator>Naschberger, Andreas</creator><creator>Fürnrohr, Barbara G.</creator><creator>Dunzendorfer-Matt, Theresia</creator><creator>Bonagura, Christopher A.</creator><creator>Wright, David</creator><creator>Scheffzek, Klaus</creator><creator>Rupp, Bernhard</creator><general>International Union of Crystallography</general><general>Wiley Subscription Services, Inc</general><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>7QL</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20150101</creationdate><title>Cleaning protocols for crystallization robots: preventing protease contamination</title><author>Naschberger, Andreas ; Fürnrohr, Barbara G. ; Dunzendorfer-Matt, Theresia ; Bonagura, Christopher A. ; Wright, David ; Scheffzek, Klaus ; Rupp, Bernhard</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5157-ff6b6be0d87979d0697508ad516d1d854e54cbc2a400a0baad8fbc1f277351073</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Automation, Laboratory</topic><topic>cleaning protocol</topic><topic>Crystallization</topic><topic>crystallization robotics</topic><topic>Crystallography, X-Ray</topic><topic>Equipment Contamination</topic><topic>Peptide Hydrolases - chemistry</topic><topic>protease</topic><topic>protease inhibitor</topic><topic>Protease Inhibitors - chemistry</topic><topic>Research Communications</topic><topic>Robotics</topic><topic>Zymit</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Naschberger, Andreas</creatorcontrib><creatorcontrib>Fürnrohr, Barbara G.</creatorcontrib><creatorcontrib>Dunzendorfer-Matt, Theresia</creatorcontrib><creatorcontrib>Bonagura, Christopher A.</creatorcontrib><creatorcontrib>Wright, David</creatorcontrib><creatorcontrib>Scheffzek, Klaus</creatorcontrib><creatorcontrib>Rupp, Bernhard</creatorcontrib><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>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Acta crystallographica. 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Section F, Structural biology communications</jtitle><addtitle>Acta Crystallographica Section F</addtitle><date>2015-01-01</date><risdate>2015</risdate><volume>71</volume><issue>1</issue><spage>100</spage><epage>102</epage><pages>100-102</pages><issn>2053-230X</issn><eissn>2053-230X</eissn><abstract>The protease in the commonly used commercial low‐foam enzyme cleaner Zymit cannot be completely blocked by EDTA, a widely used inhibitor of metalloproteases, at concentrations of up to 5 mM. Severe protein degradation was observed in crystallization drops after EDTA‐containing wash steps unless residual Zymit protease was removed with NaOH at a concentration of at least 0.1 M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1 M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity.</abstract><cop>5 Abbey Square, Chester, Cheshire CH1 2HU, England</cop><pub>International Union of Crystallography</pub><pmid>25615978</pmid><doi>10.1107/S2053230X14026053</doi><tpages>3</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Automation, Laboratory cleaning protocol Crystallization crystallization robotics Crystallography, X-Ray Equipment Contamination Peptide Hydrolases - chemistry protease protease inhibitor Protease Inhibitors - chemistry Research Communications Robotics Zymit |
| title | Cleaning protocols for crystallization robots: preventing protease contamination |
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