Crystal Structure and Biophysical Analysis of Furfural-Detoxifying Aldehyde Reductase from Clostridium beijerinckii
Many aldehydes, such as furfural, are present in high quantities in lignocellulose lysates and are fermentation inhibitors, which makes biofuel production from this abundant carbon source extremely challenging. Cbei_3974 has recently been identified as an aldo-keto reductase responsible for partial...
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description | Many aldehydes, such as furfural, are present in high quantities in lignocellulose lysates and are fermentation inhibitors, which makes biofuel production from this abundant carbon source extremely challenging. Cbei_3974 has recently been identified as an aldo-keto reductase responsible for partial furfural resistance in
Rational engineering of this enzyme could enhance the furfural tolerance of this organism, thereby improving biofuel yields. We report an extensive characterization of Cbei_3974 and a single-crystal X-ray structure of Cbei_3974 in complex with NADPH at a resolution of 1.75 Å. Docking studies identified residues involved in substrate binding, and an activity screen revealed the substrate tolerance of the enzyme. Hydride transfer, which is partially rate limiting under physiological conditions, occurs from the pro-
hydrogen of NADPH. Enzyme isotope labeling revealed a temperature-independent enzyme isotope effect of unity, indicating that the enzyme does not use dynamic coupling for catalysis and suggesting that the active site of the enzyme is optimally configured for catalysis with the substrate tested.
Here we report the crystal structure and biophysical properties of an aldehyde reductase that can detoxify furfural, a common inhibitor of biofuel fermentation found in lignocellulose lysates. The data contained here will serve as a guide for protein engineers to develop improved enzyme variants that would impart furfural resistance to the microorganisms used in biofuel production and thus lead to enhanced biofuel yields from this sustainable resource. |
doi_str_mv | 10.1128/AEM.00978-19 |
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Rational engineering of this enzyme could enhance the furfural tolerance of this organism, thereby improving biofuel yields. We report an extensive characterization of Cbei_3974 and a single-crystal X-ray structure of Cbei_3974 in complex with NADPH at a resolution of 1.75 Å. Docking studies identified residues involved in substrate binding, and an activity screen revealed the substrate tolerance of the enzyme. Hydride transfer, which is partially rate limiting under physiological conditions, occurs from the pro-
hydrogen of NADPH. Enzyme isotope labeling revealed a temperature-independent enzyme isotope effect of unity, indicating that the enzyme does not use dynamic coupling for catalysis and suggesting that the active site of the enzyme is optimally configured for catalysis with the substrate tested.
Here we report the crystal structure and biophysical properties of an aldehyde reductase that can detoxify furfural, a common inhibitor of biofuel fermentation found in lignocellulose lysates. The data contained here will serve as a guide for protein engineers to develop improved enzyme variants that would impart furfural resistance to the microorganisms used in biofuel production and thus lead to enhanced biofuel yields from this sustainable resource.</description><identifier>ISSN: 0099-2240</identifier><identifier>EISSN: 1098-5336</identifier><identifier>DOI: 10.1128/AEM.00978-19</identifier><identifier>PMID: 31101612</identifier><language>eng</language><publisher>United States: American Society for Microbiology</publisher><subject>Aldehyde reductase ; Aldehydes ; Aldo-keto reductase ; Bacteria ; Biodiesel fuels ; Biofuels ; Biomass ; Carbon sources ; Catalysis ; Clostridium beijerinckii ; Crystal structure ; Docking ; Enzymes ; Enzymology and Protein Engineering ; Fermentation ; Fuels ; Furfural ; Isotope effect ; Lignocellulose ; Lysates ; NADP ; Reductases ; Single crystals ; Substrates</subject><ispartof>Applied and environmental microbiology, 2019-08, Vol.85 (15), p.1</ispartof><rights>Copyright © 2019 American Society for Microbiology.</rights><rights>Copyright American Society for Microbiology Aug 2019</rights><rights>Copyright © 2019 American Society for Microbiology. 2019 American Society for Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c412t-fb5f30826aab46980618698ce9fe126a43d9120b069d3b93d388de52d0af28073</citedby><cites>FETCH-LOGICAL-c412t-fb5f30826aab46980618698ce9fe126a43d9120b069d3b93d388de52d0af28073</cites><orcidid>0000-0002-5008-8958</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6643236/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6643236/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,3188,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31101612$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Cann, Isaac</contributor><creatorcontrib>Scott, Alan F</creatorcontrib><creatorcontrib>Cresser-Brown, Joel</creatorcontrib><creatorcontrib>Williams, Thomas L</creatorcontrib><creatorcontrib>Rizkallah, Pierre J</creatorcontrib><creatorcontrib>Jin, Yi</creatorcontrib><creatorcontrib>Luk, Louis Y-P</creatorcontrib><creatorcontrib>Allemann, Rudolf K</creatorcontrib><title>Crystal Structure and Biophysical Analysis of Furfural-Detoxifying Aldehyde Reductase from Clostridium beijerinckii</title><title>Applied and environmental microbiology</title><addtitle>Appl Environ Microbiol</addtitle><description>Many aldehydes, such as furfural, are present in high quantities in lignocellulose lysates and are fermentation inhibitors, which makes biofuel production from this abundant carbon source extremely challenging. Cbei_3974 has recently been identified as an aldo-keto reductase responsible for partial furfural resistance in
Rational engineering of this enzyme could enhance the furfural tolerance of this organism, thereby improving biofuel yields. We report an extensive characterization of Cbei_3974 and a single-crystal X-ray structure of Cbei_3974 in complex with NADPH at a resolution of 1.75 Å. Docking studies identified residues involved in substrate binding, and an activity screen revealed the substrate tolerance of the enzyme. Hydride transfer, which is partially rate limiting under physiological conditions, occurs from the pro-
hydrogen of NADPH. Enzyme isotope labeling revealed a temperature-independent enzyme isotope effect of unity, indicating that the enzyme does not use dynamic coupling for catalysis and suggesting that the active site of the enzyme is optimally configured for catalysis with the substrate tested.
Here we report the crystal structure and biophysical properties of an aldehyde reductase that can detoxify furfural, a common inhibitor of biofuel fermentation found in lignocellulose lysates. The data contained here will serve as a guide for protein engineers to develop improved enzyme variants that would impart furfural resistance to the microorganisms used in biofuel production and thus lead to enhanced biofuel yields from this sustainable resource.</description><subject>Aldehyde reductase</subject><subject>Aldehydes</subject><subject>Aldo-keto reductase</subject><subject>Bacteria</subject><subject>Biodiesel fuels</subject><subject>Biofuels</subject><subject>Biomass</subject><subject>Carbon sources</subject><subject>Catalysis</subject><subject>Clostridium beijerinckii</subject><subject>Crystal structure</subject><subject>Docking</subject><subject>Enzymes</subject><subject>Enzymology and Protein Engineering</subject><subject>Fermentation</subject><subject>Fuels</subject><subject>Furfural</subject><subject>Isotope effect</subject><subject>Lignocellulose</subject><subject>Lysates</subject><subject>NADP</subject><subject>Reductases</subject><subject>Single crystals</subject><subject>Substrates</subject><issn>0099-2240</issn><issn>1098-5336</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><recordid>eNpdkc1v1DAQxS1ERZfCjTOyxIUDKWM767UvSMvS0kpFSHycLSced70k8dZOEPnv8dJSQU8zmvnpad48Ql4wOGWMq7frs0-nAHqlKqYfkQUDraqlEPIxWZSxrjiv4Zg8zXkHADVI9YQcC8aAScYXJG_SnEfb0a9jmtpxSkjt4Oj7EPfbOYe2bNaD7UqbafT0fEp-SrarPuAYfwU_h-GarjuH29kh_YKuaNiM1KfY000X85iCC1NPGww7TGFof4TwjBx522V8fldPyPfzs2-bi-rq88fLzfqqamvGx8o3Sy9AcWltU0utQDJVSovaIyvTWjjNODQgtRONFk4o5XDJHVjPFazECXl3q7ufmh5di8NYTjf7FHqbZhNtMP9vhrA11_GnkbIWXMgi8PpOIMWbCfNo-pBb7Do7YJyy4VxwWNaqhoK-eoDu4pTK5w7UivPiA3Sh3txSbYo5J_T3xzAwhzRNSdP8SdOwA_7yXwP38N_4xG9dApwz</recordid><startdate>20190801</startdate><enddate>20190801</enddate><creator>Scott, Alan F</creator><creator>Cresser-Brown, Joel</creator><creator>Williams, Thomas L</creator><creator>Rizkallah, Pierre J</creator><creator>Jin, Yi</creator><creator>Luk, Louis Y-P</creator><creator>Allemann, Rudolf K</creator><general>American Society for Microbiology</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7SN</scope><scope>7SS</scope><scope>7ST</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>RC3</scope><scope>SOI</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-5008-8958</orcidid></search><sort><creationdate>20190801</creationdate><title>Crystal Structure and Biophysical Analysis of Furfural-Detoxifying Aldehyde Reductase from Clostridium beijerinckii</title><author>Scott, Alan F ; Cresser-Brown, Joel ; Williams, Thomas L ; Rizkallah, Pierre J ; Jin, Yi ; Luk, Louis Y-P ; Allemann, Rudolf K</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c412t-fb5f30826aab46980618698ce9fe126a43d9120b069d3b93d388de52d0af28073</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Aldehyde reductase</topic><topic>Aldehydes</topic><topic>Aldo-keto reductase</topic><topic>Bacteria</topic><topic>Biodiesel fuels</topic><topic>Biofuels</topic><topic>Biomass</topic><topic>Carbon sources</topic><topic>Catalysis</topic><topic>Clostridium beijerinckii</topic><topic>Crystal structure</topic><topic>Docking</topic><topic>Enzymes</topic><topic>Enzymology and Protein Engineering</topic><topic>Fermentation</topic><topic>Fuels</topic><topic>Furfural</topic><topic>Isotope effect</topic><topic>Lignocellulose</topic><topic>Lysates</topic><topic>NADP</topic><topic>Reductases</topic><topic>Single crystals</topic><topic>Substrates</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Scott, Alan F</creatorcontrib><creatorcontrib>Cresser-Brown, Joel</creatorcontrib><creatorcontrib>Williams, Thomas L</creatorcontrib><creatorcontrib>Rizkallah, Pierre J</creatorcontrib><creatorcontrib>Jin, Yi</creatorcontrib><creatorcontrib>Luk, Louis Y-P</creatorcontrib><creatorcontrib>Allemann, Rudolf K</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Environment Abstracts</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>Genetics Abstracts</collection><collection>Environment Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Applied and environmental microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Scott, Alan F</au><au>Cresser-Brown, Joel</au><au>Williams, Thomas L</au><au>Rizkallah, Pierre J</au><au>Jin, Yi</au><au>Luk, Louis Y-P</au><au>Allemann, Rudolf K</au><au>Cann, Isaac</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Crystal Structure and Biophysical Analysis of Furfural-Detoxifying Aldehyde Reductase from Clostridium beijerinckii</atitle><jtitle>Applied and environmental microbiology</jtitle><addtitle>Appl Environ Microbiol</addtitle><date>2019-08-01</date><risdate>2019</risdate><volume>85</volume><issue>15</issue><spage>1</spage><pages>1-</pages><issn>0099-2240</issn><eissn>1098-5336</eissn><abstract>Many aldehydes, such as furfural, are present in high quantities in lignocellulose lysates and are fermentation inhibitors, which makes biofuel production from this abundant carbon source extremely challenging. Cbei_3974 has recently been identified as an aldo-keto reductase responsible for partial furfural resistance in
Rational engineering of this enzyme could enhance the furfural tolerance of this organism, thereby improving biofuel yields. We report an extensive characterization of Cbei_3974 and a single-crystal X-ray structure of Cbei_3974 in complex with NADPH at a resolution of 1.75 Å. Docking studies identified residues involved in substrate binding, and an activity screen revealed the substrate tolerance of the enzyme. Hydride transfer, which is partially rate limiting under physiological conditions, occurs from the pro-
hydrogen of NADPH. Enzyme isotope labeling revealed a temperature-independent enzyme isotope effect of unity, indicating that the enzyme does not use dynamic coupling for catalysis and suggesting that the active site of the enzyme is optimally configured for catalysis with the substrate tested.
Here we report the crystal structure and biophysical properties of an aldehyde reductase that can detoxify furfural, a common inhibitor of biofuel fermentation found in lignocellulose lysates. The data contained here will serve as a guide for protein engineers to develop improved enzyme variants that would impart furfural resistance to the microorganisms used in biofuel production and thus lead to enhanced biofuel yields from this sustainable resource.</abstract><cop>United States</cop><pub>American Society for Microbiology</pub><pmid>31101612</pmid><doi>10.1128/AEM.00978-19</doi><orcidid>https://orcid.org/0000-0002-5008-8958</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | Aldehyde reductase Aldehydes Aldo-keto reductase Bacteria Biodiesel fuels Biofuels Biomass Carbon sources Catalysis Clostridium beijerinckii Crystal structure Docking Enzymes Enzymology and Protein Engineering Fermentation Fuels Furfural Isotope effect Lignocellulose Lysates NADP Reductases Single crystals Substrates |
title | Crystal Structure and Biophysical Analysis of Furfural-Detoxifying Aldehyde Reductase from Clostridium beijerinckii |
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