Redesigning Aldolase Stereoselectivity by Homologous Grafting
The 2-deoxy-d-ribose-5-phosphate aldolase (DERA) offers access to highly desirable building blocks for organic synthesis by catalyzing a stereoselective C-C bond formation between acetaldehyde and certain electrophilic aldehydes. DERA´s potential is particularly highlighted by the ability to catalyz...
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| creator | Bisterfeld, Carolin Classen, Thomas Küberl, Irene Henßen, Birgit Metz, Alexander Gohlke, Holger Pietruszka, Jörg |
| description | The 2-deoxy-d-ribose-5-phosphate aldolase (DERA) offers access to highly desirable building blocks for organic synthesis by catalyzing a stereoselective C-C bond formation between acetaldehyde and certain electrophilic aldehydes. DERA´s potential is particularly highlighted by the ability to catalyze sequential, highly enantioselective aldol reactions. However, its synthetic use is limited by the absence of an enantiocomplementary enzyme. Here, we introduce the concept of homologous grafting to identify stereoselectivity-determining amino acid positions in DERA. We identified such positions by structural analysis of the homologous aldolases 2-keto-3-deoxy-6-phosphogluconate aldolase (KDPG) and the enantiocomplementary enzyme 2-keto-3-deoxy-6-phosphogalactonate aldolase (KDPGal). Mutation of these positions led to a slightly inversed enantiopreference of both aldolases to the same extent. By transferring these sequence motifs onto DERA we achieved the intended change in enantioselectivity. |
| doi_str_mv | 10.1371/journal.pone.0156525 |
| format | Article |
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DERA´s potential is particularly highlighted by the ability to catalyze sequential, highly enantioselective aldol reactions. However, its synthetic use is limited by the absence of an enantiocomplementary enzyme. Here, we introduce the concept of homologous grafting to identify stereoselectivity-determining amino acid positions in DERA. We identified such positions by structural analysis of the homologous aldolases 2-keto-3-deoxy-6-phosphogluconate aldolase (KDPG) and the enantiocomplementary enzyme 2-keto-3-deoxy-6-phosphogalactonate aldolase (KDPGal). Mutation of these positions led to a slightly inversed enantiopreference of both aldolases to the same extent. By transferring these sequence motifs onto DERA we achieved the intended change in enantioselectivity.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0156525</identifier><identifier>PMID: 27327271</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Acetaldehyde ; Aldehyde-Lyases - chemistry ; Aldehyde-Lyases - metabolism ; Aldehydes ; Aldolase ; Aldolases ; Amino Acid Sequence ; Amino acids ; Amino Acids - metabolism ; Analysis ; Biocatalysis ; Biocatalysts ; Biochemistry ; Biology and Life Sciences ; Carbon-carbon composites ; Chemical reactions ; Chemical synthesis ; Chromatography ; Crystal structure ; D-Ribose ; Enantiomers ; Enzymes ; Escherichia coli - enzymology ; Fructose-Bisphosphate Aldolase - chemistry ; Fructose-Bisphosphate Aldolase - metabolism ; Grafting ; Homology ; Influence ; Kinetics ; Models, Molecular ; Monosaccharides ; Mutation ; Pharmaceutical sciences ; Phylogeny ; Physical Sciences ; Protein Engineering ; Protein Structure, Secondary ; Proteins ; Pyruvates - metabolism ; Research and Analysis Methods ; Ribose ; Stereoisomerism ; Stereoselectivity ; Structural analysis ; Substrate Specificity</subject><ispartof>PloS one, 2016-06, Vol.11 (6), p.e0156525-e0156525</ispartof><rights>COPYRIGHT 2016 Public Library of Science</rights><rights>2016 Bisterfeld et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2016 Bisterfeld et al 2016 Bisterfeld et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c725t-5c46709dcc87e351acac84f98cc3ef80c525ff2af2720427a3b27526b97a99d93</citedby><cites>FETCH-LOGICAL-c725t-5c46709dcc87e351acac84f98cc3ef80c525ff2af2720427a3b27526b97a99d93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4915726/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4915726/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27327271$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Parker, Emily J.</contributor><creatorcontrib>Bisterfeld, Carolin</creatorcontrib><creatorcontrib>Classen, Thomas</creatorcontrib><creatorcontrib>Küberl, Irene</creatorcontrib><creatorcontrib>Henßen, Birgit</creatorcontrib><creatorcontrib>Metz, Alexander</creatorcontrib><creatorcontrib>Gohlke, Holger</creatorcontrib><creatorcontrib>Pietruszka, Jörg</creatorcontrib><title>Redesigning Aldolase Stereoselectivity by Homologous Grafting</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>The 2-deoxy-d-ribose-5-phosphate aldolase (DERA) offers access to highly desirable building blocks for organic synthesis by catalyzing a stereoselective C-C bond formation between acetaldehyde and certain electrophilic aldehydes. DERA´s potential is particularly highlighted by the ability to catalyze sequential, highly enantioselective aldol reactions. However, its synthetic use is limited by the absence of an enantiocomplementary enzyme. Here, we introduce the concept of homologous grafting to identify stereoselectivity-determining amino acid positions in DERA. We identified such positions by structural analysis of the homologous aldolases 2-keto-3-deoxy-6-phosphogluconate aldolase (KDPG) and the enantiocomplementary enzyme 2-keto-3-deoxy-6-phosphogalactonate aldolase (KDPGal). Mutation of these positions led to a slightly inversed enantiopreference of both aldolases to the same extent. By transferring these sequence motifs onto DERA we achieved the intended change in enantioselectivity.</description><subject>Acetaldehyde</subject><subject>Aldehyde-Lyases - chemistry</subject><subject>Aldehyde-Lyases - metabolism</subject><subject>Aldehydes</subject><subject>Aldolase</subject><subject>Aldolases</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Amino Acids - metabolism</subject><subject>Analysis</subject><subject>Biocatalysis</subject><subject>Biocatalysts</subject><subject>Biochemistry</subject><subject>Biology and Life Sciences</subject><subject>Carbon-carbon composites</subject><subject>Chemical reactions</subject><subject>Chemical synthesis</subject><subject>Chromatography</subject><subject>Crystal structure</subject><subject>D-Ribose</subject><subject>Enantiomers</subject><subject>Enzymes</subject><subject>Escherichia coli - enzymology</subject><subject>Fructose-Bisphosphate Aldolase - chemistry</subject><subject>Fructose-Bisphosphate Aldolase - metabolism</subject><subject>Grafting</subject><subject>Homology</subject><subject>Influence</subject><subject>Kinetics</subject><subject>Models, Molecular</subject><subject>Monosaccharides</subject><subject>Mutation</subject><subject>Pharmaceutical sciences</subject><subject>Phylogeny</subject><subject>Physical Sciences</subject><subject>Protein Engineering</subject><subject>Protein Structure, Secondary</subject><subject>Proteins</subject><subject>Pyruvates - metabolism</subject><subject>Research and Analysis Methods</subject><subject>Ribose</subject><subject>Stereoisomerism</subject><subject>Stereoselectivity</subject><subject>Structural analysis</subject><subject>Substrate Specificity</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNk11rFDEUhgdR7If-A9EFoejFrvnO5EJhKdouFAqtehsymcxsluxkm2SK--_NutOyIwUlkITked-cnOQUxRsIZhBz-Gnl-9ApN9v4zswApIwi-qw4hgKjKUMAPz-YHxUnMa4AoLhk7GVxhDhGHHF4XHy-MbWJtu1s107mrvZORTO5TSYYH40zOtl7m7aTaju59GvvfOv7OLkIqklZ8ap40SgXzethPC1-fPv6_fxyenV9sTifX001RzRNqSaMA1FrXXKDKVRa6ZI0otQam6YEOofeNEg1OShAEFe4QpwiVgmuhKgFPi3e7X03zkc53DxKyEXJOROAZ2KxJ2qvVnIT7FqFrfTKyj8LPrRShWS1M7KCmjKADUGCEJ57lYPTqlK0JIJhkr2-DKf11drU2nQpKDcyHe90dilbfy-JgJQjlg0-DAbB3_UmJrm2URvnVGdy-iQsQckIpUT8G-VCIEg4hBl9_xf6dCIGqlX5rrZrfA5R70zlnFBGMBagzNTsCSq32qytzj-qsXl9JPg4EmQmmV-pVX2McnF78__s9c8xe3bALo1yaRm965P1XRyDZA_q4GMMpnl8DwjkriAesiF3BSGHgsiyt4dv-Sh6qAD8G3LrA8c</recordid><startdate>20160621</startdate><enddate>20160621</enddate><creator>Bisterfeld, Carolin</creator><creator>Classen, Thomas</creator><creator>Küberl, Irene</creator><creator>Henßen, Birgit</creator><creator>Metz, Alexander</creator><creator>Gohlke, Holger</creator><creator>Pietruszka, Jörg</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20160621</creationdate><title>Redesigning Aldolase Stereoselectivity by Homologous Grafting</title><author>Bisterfeld, Carolin ; Classen, Thomas ; Küberl, Irene ; Henßen, Birgit ; Metz, Alexander ; Gohlke, Holger ; Pietruszka, Jörg</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c725t-5c46709dcc87e351acac84f98cc3ef80c525ff2af2720427a3b27526b97a99d93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Acetaldehyde</topic><topic>Aldehyde-Lyases - chemistry</topic><topic>Aldehyde-Lyases - metabolism</topic><topic>Aldehydes</topic><topic>Aldolase</topic><topic>Aldolases</topic><topic>Amino Acid Sequence</topic><topic>Amino acids</topic><topic>Amino Acids - metabolism</topic><topic>Analysis</topic><topic>Biocatalysis</topic><topic>Biocatalysts</topic><topic>Biochemistry</topic><topic>Biology and Life Sciences</topic><topic>Carbon-carbon composites</topic><topic>Chemical reactions</topic><topic>Chemical synthesis</topic><topic>Chromatography</topic><topic>Crystal structure</topic><topic>D-Ribose</topic><topic>Enantiomers</topic><topic>Enzymes</topic><topic>Escherichia coli - enzymology</topic><topic>Fructose-Bisphosphate Aldolase - chemistry</topic><topic>Fructose-Bisphosphate Aldolase - metabolism</topic><topic>Grafting</topic><topic>Homology</topic><topic>Influence</topic><topic>Kinetics</topic><topic>Models, Molecular</topic><topic>Monosaccharides</topic><topic>Mutation</topic><topic>Pharmaceutical sciences</topic><topic>Phylogeny</topic><topic>Physical Sciences</topic><topic>Protein Engineering</topic><topic>Protein Structure, Secondary</topic><topic>Proteins</topic><topic>Pyruvates - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bisterfeld, Carolin</au><au>Classen, Thomas</au><au>Küberl, Irene</au><au>Henßen, Birgit</au><au>Metz, Alexander</au><au>Gohlke, Holger</au><au>Pietruszka, Jörg</au><au>Parker, Emily J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Redesigning Aldolase Stereoselectivity by Homologous Grafting</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2016-06-21</date><risdate>2016</risdate><volume>11</volume><issue>6</issue><spage>e0156525</spage><epage>e0156525</epage><pages>e0156525-e0156525</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>The 2-deoxy-d-ribose-5-phosphate aldolase (DERA) offers access to highly desirable building blocks for organic synthesis by catalyzing a stereoselective C-C bond formation between acetaldehyde and certain electrophilic aldehydes. DERA´s potential is particularly highlighted by the ability to catalyze sequential, highly enantioselective aldol reactions. However, its synthetic use is limited by the absence of an enantiocomplementary enzyme. Here, we introduce the concept of homologous grafting to identify stereoselectivity-determining amino acid positions in DERA. We identified such positions by structural analysis of the homologous aldolases 2-keto-3-deoxy-6-phosphogluconate aldolase (KDPG) and the enantiocomplementary enzyme 2-keto-3-deoxy-6-phosphogalactonate aldolase (KDPGal). Mutation of these positions led to a slightly inversed enantiopreference of both aldolases to the same extent. By transferring these sequence motifs onto DERA we achieved the intended change in enantioselectivity.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>27327271</pmid><doi>10.1371/journal.pone.0156525</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Acetaldehyde Aldehyde-Lyases - chemistry Aldehyde-Lyases - metabolism Aldehydes Aldolase Aldolases Amino Acid Sequence Amino acids Amino Acids - metabolism Analysis Biocatalysis Biocatalysts Biochemistry Biology and Life Sciences Carbon-carbon composites Chemical reactions Chemical synthesis Chromatography Crystal structure D-Ribose Enantiomers Enzymes Escherichia coli - enzymology Fructose-Bisphosphate Aldolase - chemistry Fructose-Bisphosphate Aldolase - metabolism Grafting Homology Influence Kinetics Models, Molecular Monosaccharides Mutation Pharmaceutical sciences Phylogeny Physical Sciences Protein Engineering Protein Structure, Secondary Proteins Pyruvates - metabolism Research and Analysis Methods Ribose Stereoisomerism Stereoselectivity Structural analysis Substrate Specificity |
| title | Redesigning Aldolase Stereoselectivity by Homologous Grafting |
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