Enhanced bioaccumulation of heavy metals by bacterial cells displaying synthetic phytochelatins
A novel strategy using synthetic phytochelatins is described for the purpose of developing microbial agents for enhanced bioaccumulation of toxic metals. Synthetic genes encoding for several metal‐chelating phytochelatin analogs (Glu‐Cys)nGly (EC8 (n = 8), EC11 (n = 11), and EC20 (n = 20)) were synt...
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| Veröffentlicht in: | Biotechnology and bioengineering 2000-12, Vol.70 (5), p.518-524 |
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| creator | Bae, Weon Chen, Wilfred Mulchandani, Ashok Mehra, Rajesh K. |
| description | A novel strategy using synthetic phytochelatins is described for the purpose of developing microbial agents for enhanced bioaccumulation of toxic metals. Synthetic genes encoding for several metal‐chelating phytochelatin analogs (Glu‐Cys)nGly (EC8 (n = 8), EC11 (n = 11), and EC20 (n = 20)) were synthesized, linked to a lpp‐ompA fusion gene, and displayed on the surface of E. coli. For comparison, EC20 was also expressed periplasmically as a fusion with the maltose‐binding protein (MBP‐EC20). Purified MBP‐EC20 was shown to accumulate more Cd2+ per peptide than typical mammalian metallothioneins with a stoichiometry of 10 Cd2+/peptide. Cells displaying synthetic phytochelatins exhibited chain‐length dependent increase in metal accumulation. For example, 18 nmoles of Cd2+/mg dry cells were accumulated by cells displaying EC8, whereas cells exhibiting EC20 accumulated a maximum of 60 nmoles of Cd2+/mg dry cells. Moreover, cells with surface‐expressed EC20 accumulated twice the amount of Cd2+ as cells expressing EC20 periplasmically. The ability to genetically engineer ECs with precisely defined chain length could provide an attractive strategy for developing high‐affinity bioadsorbents suitable for heavy metal removal. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 70: 518–524, 2000. |
| doi_str_mv | 10.1002/1097-0290(20001205)70:5<518::AID-BIT6>3.0.CO;2-5 |
| format | Article |
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Synthetic genes encoding for several metal‐chelating phytochelatin analogs (Glu‐Cys)nGly (EC8 (n = 8), EC11 (n = 11), and EC20 (n = 20)) were synthesized, linked to a lpp‐ompA fusion gene, and displayed on the surface of E. coli. For comparison, EC20 was also expressed periplasmically as a fusion with the maltose‐binding protein (MBP‐EC20). Purified MBP‐EC20 was shown to accumulate more Cd2+ per peptide than typical mammalian metallothioneins with a stoichiometry of 10 Cd2+/peptide. Cells displaying synthetic phytochelatins exhibited chain‐length dependent increase in metal accumulation. For example, 18 nmoles of Cd2+/mg dry cells were accumulated by cells displaying EC8, whereas cells exhibiting EC20 accumulated a maximum of 60 nmoles of Cd2+/mg dry cells. Moreover, cells with surface‐expressed EC20 accumulated twice the amount of Cd2+ as cells expressing EC20 periplasmically. The ability to genetically engineer ECs with precisely defined chain length could provide an attractive strategy for developing high‐affinity bioadsorbents suitable for heavy metal removal. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 70: 518–524, 2000.</description><identifier>ISSN: 0006-3592</identifier><identifier>EISSN: 1097-0290</identifier><identifier>DOI: 10.1002/1097-0290(20001205)70:5<518::AID-BIT6>3.0.CO;2-5</identifier><identifier>PMID: 11042548</identifier><identifier>CODEN: BIBIAU</identifier><language>eng</language><publisher>New York: John Wiley & Sons, Inc</publisher><subject>Absorption ; Bacteria ; Bacterial Outer Membrane Proteins - genetics ; Bacterial Outer Membrane Proteins - metabolism ; Base Sequence ; bioadsorbents ; Biological and medical sciences ; Biotechnology ; Cadmium - metabolism ; Cell Membrane - genetics ; Cell Membrane - metabolism ; Cells ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Fundamental and applied biological sciences. Psychology ; Genes ; Genetic engineering ; Genetic Engineering - methods ; Genetic technics ; Glutathione ; Heavy metals ; Lipoproteins - genetics ; Lipoproteins - metabolism ; Lpp-OmpA ; Metalloproteins - genetics ; Metalloproteins - metabolism ; Metals, Heavy - metabolism ; Methods. Procedures. Technologies ; Modification of gene expression level ; Molecular Sequence Data ; phytochelatin analogs ; Phytochelatins ; Plant Proteins - genetics ; Plant Proteins - metabolism ; Proteins ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - metabolism ; Stoichiometry ; Synthesis (chemical)</subject><ispartof>Biotechnology and bioengineering, 2000-12, Vol.70 (5), p.518-524</ispartof><rights>Copyright © 2000 John Wiley & Sons, Inc.</rights><rights>2001 INIST-CNRS</rights><rights>Copyright 2000 John Wiley & Sons, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c5866-c6f25d07751a098b7cd9030ce7e656d0566026e4284d6f42519b41c16b23278f3</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%2F1097-0290%2820001205%2970%3A5%3C518%3A%3AAID-BIT6%3E3.0.CO%3B2-5$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2F1097-0290%2820001205%2970%3A5%3C518%3A%3AAID-BIT6%3E3.0.CO%3B2-5$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=823747$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11042548$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bae, Weon</creatorcontrib><creatorcontrib>Chen, Wilfred</creatorcontrib><creatorcontrib>Mulchandani, Ashok</creatorcontrib><creatorcontrib>Mehra, Rajesh K.</creatorcontrib><title>Enhanced bioaccumulation of heavy metals by bacterial cells displaying synthetic phytochelatins</title><title>Biotechnology and bioengineering</title><addtitle>Biotechnol. Bioeng</addtitle><description>A novel strategy using synthetic phytochelatins is described for the purpose of developing microbial agents for enhanced bioaccumulation of toxic metals. Synthetic genes encoding for several metal‐chelating phytochelatin analogs (Glu‐Cys)nGly (EC8 (n = 8), EC11 (n = 11), and EC20 (n = 20)) were synthesized, linked to a lpp‐ompA fusion gene, and displayed on the surface of E. coli. For comparison, EC20 was also expressed periplasmically as a fusion with the maltose‐binding protein (MBP‐EC20). Purified MBP‐EC20 was shown to accumulate more Cd2+ per peptide than typical mammalian metallothioneins with a stoichiometry of 10 Cd2+/peptide. Cells displaying synthetic phytochelatins exhibited chain‐length dependent increase in metal accumulation. For example, 18 nmoles of Cd2+/mg dry cells were accumulated by cells displaying EC8, whereas cells exhibiting EC20 accumulated a maximum of 60 nmoles of Cd2+/mg dry cells. Moreover, cells with surface‐expressed EC20 accumulated twice the amount of Cd2+ as cells expressing EC20 periplasmically. The ability to genetically engineer ECs with precisely defined chain length could provide an attractive strategy for developing high‐affinity bioadsorbents suitable for heavy metal removal. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 70: 518–524, 2000.</description><subject>Absorption</subject><subject>Bacteria</subject><subject>Bacterial Outer Membrane Proteins - genetics</subject><subject>Bacterial Outer Membrane Proteins - metabolism</subject><subject>Base Sequence</subject><subject>bioadsorbents</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cadmium - metabolism</subject><subject>Cell Membrane - genetics</subject><subject>Cell Membrane - metabolism</subject><subject>Cells</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes</subject><subject>Genetic engineering</subject><subject>Genetic Engineering - methods</subject><subject>Genetic technics</subject><subject>Glutathione</subject><subject>Heavy metals</subject><subject>Lipoproteins - genetics</subject><subject>Lipoproteins - metabolism</subject><subject>Lpp-OmpA</subject><subject>Metalloproteins - genetics</subject><subject>Metalloproteins - metabolism</subject><subject>Metals, Heavy - metabolism</subject><subject>Methods. Procedures. Technologies</subject><subject>Modification of gene expression level</subject><subject>Molecular Sequence Data</subject><subject>phytochelatin analogs</subject><subject>Phytochelatins</subject><subject>Plant Proteins - genetics</subject><subject>Plant Proteins - metabolism</subject><subject>Proteins</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Stoichiometry</subject><subject>Synthesis (chemical)</subject><issn>0006-3592</issn><issn>1097-0290</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkV1v0zAUhiMEYt3gL6BISIhdpBzbsZ0UhDTC2IoqOrQhdnfkOA4x5KPEKZB_j0NLuUFCXFm2nvP41XmDICEwJwD0GYFURkBTeEoBgFDgpxIW_AUnyWJxtnwdvVreiJdsDvNs_ZxG_E4wO4zcDWZ-RkSMp_QoOHbus7_KRIj7wREhEFMeJ7MAz9tKtdoUYW47pfW22dZqsF0bdmVYGfVtDBszqNqF-RjmSg-mt6oOtan9U2HdplajbT-FbmyHygxWh5tqHDpdmUnTugfBvdJPm4f78yT48Ob8JruMVuuLZXa2ijT3kSItSsoLkJITBWmSS12kwEAbaQQXBXAhgAoT0yQuROmzkzSPiSYip4zKpGQnwZOdd9N3X7fGDdhYN6VUrem2DimJYyJS-k-QSMkY5eDBqx2o-8653pS46W2j-hEJ4NQOTqvGadX4ux2UgBx9O4i-HZzaQYaA2Ropcq98tP97mzem-CPc1-GBx3tAOa3qsvfVWHfgEspkLD31fkd9t7UZ_yvWX1L9untntHNaN5gfB6fqv6CQTHL8-O4CL99er26zq2u8ZT8BcoLCrw</recordid><startdate>20001205</startdate><enddate>20001205</enddate><creator>Bae, Weon</creator><creator>Chen, Wilfred</creator><creator>Mulchandani, Ashok</creator><creator>Mehra, Rajesh K.</creator><general>John Wiley & Sons, Inc</general><general>Wiley</general><scope>BSCLL</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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20001205</creationdate><title>Enhanced bioaccumulation of heavy metals by bacterial cells displaying synthetic phytochelatins</title><author>Bae, Weon ; Chen, Wilfred ; Mulchandani, Ashok ; Mehra, Rajesh K.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5866-c6f25d07751a098b7cd9030ce7e656d0566026e4284d6f42519b41c16b23278f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Absorption</topic><topic>Bacteria</topic><topic>Bacterial Outer Membrane Proteins - genetics</topic><topic>Bacterial Outer Membrane Proteins - metabolism</topic><topic>Base Sequence</topic><topic>bioadsorbents</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cadmium - metabolism</topic><topic>Cell Membrane - genetics</topic><topic>Cell Membrane - metabolism</topic><topic>Cells</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes</topic><topic>Genetic engineering</topic><topic>Genetic Engineering - methods</topic><topic>Genetic technics</topic><topic>Glutathione</topic><topic>Heavy metals</topic><topic>Lipoproteins - genetics</topic><topic>Lipoproteins - metabolism</topic><topic>Lpp-OmpA</topic><topic>Metalloproteins - genetics</topic><topic>Metalloproteins - metabolism</topic><topic>Metals, Heavy - metabolism</topic><topic>Methods. Procedures. Technologies</topic><topic>Modification of gene expression level</topic><topic>Molecular Sequence Data</topic><topic>phytochelatin analogs</topic><topic>Phytochelatins</topic><topic>Plant Proteins - genetics</topic><topic>Plant Proteins - metabolism</topic><topic>Proteins</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Stoichiometry</topic><topic>Synthesis (chemical)</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bae, Weon</creatorcontrib><creatorcontrib>Chen, Wilfred</creatorcontrib><creatorcontrib>Mulchandani, Ashok</creatorcontrib><creatorcontrib>Mehra, Rajesh K.</creatorcontrib><collection>Istex</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>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Biotechnology and bioengineering</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bae, Weon</au><au>Chen, Wilfred</au><au>Mulchandani, Ashok</au><au>Mehra, Rajesh K.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enhanced bioaccumulation of heavy metals by bacterial cells displaying synthetic phytochelatins</atitle><jtitle>Biotechnology and bioengineering</jtitle><addtitle>Biotechnol. Bioeng</addtitle><date>2000-12-05</date><risdate>2000</risdate><volume>70</volume><issue>5</issue><spage>518</spage><epage>524</epage><pages>518-524</pages><issn>0006-3592</issn><eissn>1097-0290</eissn><coden>BIBIAU</coden><abstract>A novel strategy using synthetic phytochelatins is described for the purpose of developing microbial agents for enhanced bioaccumulation of toxic metals. Synthetic genes encoding for several metal‐chelating phytochelatin analogs (Glu‐Cys)nGly (EC8 (n = 8), EC11 (n = 11), and EC20 (n = 20)) were synthesized, linked to a lpp‐ompA fusion gene, and displayed on the surface of E. coli. For comparison, EC20 was also expressed periplasmically as a fusion with the maltose‐binding protein (MBP‐EC20). Purified MBP‐EC20 was shown to accumulate more Cd2+ per peptide than typical mammalian metallothioneins with a stoichiometry of 10 Cd2+/peptide. Cells displaying synthetic phytochelatins exhibited chain‐length dependent increase in metal accumulation. For example, 18 nmoles of Cd2+/mg dry cells were accumulated by cells displaying EC8, whereas cells exhibiting EC20 accumulated a maximum of 60 nmoles of Cd2+/mg dry cells. Moreover, cells with surface‐expressed EC20 accumulated twice the amount of Cd2+ as cells expressing EC20 periplasmically. The ability to genetically engineer ECs with precisely defined chain length could provide an attractive strategy for developing high‐affinity bioadsorbents suitable for heavy metal removal. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 70: 518–524, 2000.</abstract><cop>New York</cop><pub>John Wiley & Sons, Inc</pub><pmid>11042548</pmid><doi>10.1002/1097-0290(20001205)70:5<518::AID-BIT6>3.0.CO;2-5</doi><tpages>7</tpages></addata></record> |
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| subjects | Absorption Bacteria Bacterial Outer Membrane Proteins - genetics Bacterial Outer Membrane Proteins - metabolism Base Sequence bioadsorbents Biological and medical sciences Biotechnology Cadmium - metabolism Cell Membrane - genetics Cell Membrane - metabolism Cells Escherichia coli - genetics Escherichia coli - metabolism Fundamental and applied biological sciences. Psychology Genes Genetic engineering Genetic Engineering - methods Genetic technics Glutathione Heavy metals Lipoproteins - genetics Lipoproteins - metabolism Lpp-OmpA Metalloproteins - genetics Metalloproteins - metabolism Metals, Heavy - metabolism Methods. Procedures. Technologies Modification of gene expression level Molecular Sequence Data phytochelatin analogs Phytochelatins Plant Proteins - genetics Plant Proteins - metabolism Proteins Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Stoichiometry Synthesis (chemical) |
| title | Enhanced bioaccumulation of heavy metals by bacterial cells displaying synthetic phytochelatins |
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