Semi-rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide.
Identifieur interne : 000013 ( Main/Exploration ); précédent : 000012; suivant : 000014Semi-rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide.
Auteurs : Ana Marija Balaž [Serbie] ; Jelena Stevanovi [Serbie] ; Raluca Ostafe [États-Unis] ; Marija Blazi [Serbie] ; Karla Ili Ur I [Serbie] ; Rainer Fischer [Allemagne, États-Unis] ; Radivoje Prodanovi [Serbie]Source :
- Molecular diversity [ 1573-501X ] ; 2020.
Abstract
Cellobiose dehydrogenase (CDH, EC 1.1.99.18) from white rot fungi Phanerochaete chrysosporium can be used for constructing biosensors and biofuel cells, for bleaching cotton in textile industry, and recently, the enzyme has found an important application in biomedicine as an antimicrobial and antibiofilm agent. Stability and activity of the wild-type (wt) CDH and mutants at methionine residues in the presence of hydrogen peroxide were investigated. Saturation mutagenesis libraries were made at the only methionine in heme domain M65 and two methionines M685 and M738 in the flavin domain that were closest to the active site. After screening the libraries, three mutants with increased activity and stability in the presence of peroxide were found, M65F with 70% of residual activity after 6 h of incubation in 0.3 M hydrogen peroxide, M738S with 80% of residual activity and M685Y with over 90% of residual activity compared to wild-type CDH that retained 40% of original activity. Combined mutants showed no activity. The most stable mutant M685Y with 5.8 times increased half-life in the presence of peroxide showed also 2.5 times increased kcat for lactose compared to wtCDH and could be good candidate for applications in biofuel cells and biocatalysis for lactobionic acid production.
DOI: 10.1007/s11030-019-09965-0
PubMed: 31154590
Affiliations:
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16, 11000, Belgrade, Serbia. rprodano@chem.bg.ac.rs.</nlm:affiliation><country xml:lang="fr">Serbie</country><wicri:regionArea>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade</wicri:regionArea><wicri:noRegion>Belgrade</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2020">2020</date><idno type="RBID">pubmed:31154590</idno><idno type="pmid">31154590</idno><idno type="doi">10.1007/s11030-019-09965-0</idno><idno type="wicri:Area/Main/Corpus">000066</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000066</idno><idno type="wicri:Area/Main/Curation">000066</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000066</idno><idno type="wicri:Area/Main/Exploration">000066</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Semi-rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide.</title><author><name sortKey="Balaz, Ana Marija" sort="Balaz, Ana Marija" uniqKey="Balaz A" first="Ana Marija" last="Balaž">Ana Marija Balaž</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Njegoseva 12, 11000, Belgrade, Serbia.</nlm:affiliation><country xml:lang="fr">Serbie</country><wicri:regionArea>Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Njegoseva 12, 11000, Belgrade</wicri:regionArea><wicri:noRegion>Belgrade</wicri:noRegion></affiliation></author><author><name sortKey="Stevanovi, Jelena" sort="Stevanovi, Jelena" uniqKey="Stevanovi J" first="Jelena" last="Stevanovi">Jelena Stevanovi</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade, Serbia.</nlm:affiliation><country xml:lang="fr">Serbie</country><wicri:regionArea>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade</wicri:regionArea><wicri:noRegion>Belgrade</wicri:noRegion></affiliation></author><author><name sortKey="Ostafe, Raluca" sort="Ostafe, Raluca" uniqKey="Ostafe R" first="Raluca" last="Ostafe">Raluca Ostafe</name><affiliation wicri:level="1"><nlm:affiliation>Hall for Discovery and Learning Research, Purdue University, 207 S. Martin Jischke Dr., West Lafayette, IN, 47907, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Hall for Discovery and Learning Research, Purdue University, 207 S. Martin Jischke Dr., West Lafayette, IN, 47907</wicri:regionArea><wicri:noRegion>47907</wicri:noRegion></affiliation></author><author><name sortKey="Blazi, Marija" sort="Blazi, Marija" uniqKey="Blazi M" first="Marija" last="Blazi">Marija Blazi</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Njegoseva 12, 11000, Belgrade, Serbia.</nlm:affiliation><country xml:lang="fr">Serbie</country><wicri:regionArea>Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Njegoseva 12, 11000, Belgrade</wicri:regionArea><wicri:noRegion>Belgrade</wicri:noRegion></affiliation></author><author><name sortKey="Ili Ur I, Karla" sort="Ili Ur I, Karla" uniqKey="Ili Ur I K" first="Karla" last="Ili Ur I">Karla Ili Ur I</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade, Serbia.</nlm:affiliation><country xml:lang="fr">Serbie</country><wicri:regionArea>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade</wicri:regionArea><wicri:noRegion>Belgrade</wicri:noRegion></affiliation></author><author><name sortKey="Fischer, Rainer" sort="Fischer, Rainer" uniqKey="Fischer R" first="Rainer" last="Fischer">Rainer Fischer</name><affiliation wicri:level="3"><nlm:affiliation>Institute of Molecular Biotechnology, RWTH Aachen University, Worringerweg 1, 52074, Aachen, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute of Molecular Biotechnology, RWTH Aachen University, Worringerweg 1, 52074, Aachen</wicri:regionArea><placeName><region type="land" nuts="1">Rhénanie-du-Nord-Westphalie</region><region type="district" nuts="2">District de Cologne</region><settlement type="city">Aix-la-Chapelle</settlement></placeName></affiliation><affiliation wicri:level="1"><nlm:affiliation>Single Cell Analytics Center, Indiana Bioscience Research Institute, 1345 W. 16th St. Suite 300, Indianapolis, IN, 46202, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Single Cell Analytics Center, Indiana Bioscience Research Institute, 1345 W. 16th St. Suite 300, Indianapolis, IN, 46202</wicri:regionArea><wicri:noRegion>46202</wicri:noRegion></affiliation></author><author><name sortKey="Prodanovi, Radivoje" sort="Prodanovi, Radivoje" uniqKey="Prodanovi R" first="Radivoje" last="Prodanovi">Radivoje Prodanovi</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade, Serbia. rprodano@chem.bg.ac.rs.</nlm:affiliation><country xml:lang="fr">Serbie</country><wicri:regionArea>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade</wicri:regionArea><wicri:noRegion>Belgrade</wicri:noRegion></affiliation></author></analytic><series><title level="j">Molecular diversity</title><idno type="eISSN">1573-501X</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Cellobiose dehydrogenase (CDH, EC 1.1.99.18) from white rot fungi Phanerochaete chrysosporium can be used for constructing biosensors and biofuel cells, for bleaching cotton in textile industry, and recently, the enzyme has found an important application in biomedicine as an antimicrobial and antibiofilm agent. Stability and activity of the wild-type (wt) CDH and mutants at methionine residues in the presence of hydrogen peroxide were investigated. Saturation mutagenesis libraries were made at the only methionine in heme domain M65 and two methionines M685 and M738 in the flavin domain that were closest to the active site. After screening the libraries, three mutants with increased activity and stability in the presence of peroxide were found, M65F with 70% of residual activity after 6 h of incubation in 0.3 M hydrogen peroxide, M738S with 80% of residual activity and M685Y with over 90% of residual activity compared to wild-type CDH that retained 40% of original activity. Combined mutants showed no activity. The most stable mutant M685Y with 5.8 times increased half-life in the presence of peroxide showed also 2.5 times increased k<sub>cat</sub> for lactose compared to wtCDH and could be good candidate for applications in biofuel cells and biocatalysis for lactobionic acid production.</div></front></TEI><pubmed><MedlineCitation Status="In-Process" Owner="NLM"><PMID Version="1">31154590</PMID><DateRevised><Year>2020</Year><Month>07</Month><Day>14</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1573-501X</ISSN><JournalIssue CitedMedium="Internet"><Volume>24</Volume><Issue>3</Issue><PubDate><Year>2020</Year><Month>Aug</Month></PubDate></JournalIssue><Title>Molecular diversity</Title><ISOAbbreviation>Mol Divers</ISOAbbreviation></Journal><ArticleTitle>Semi-rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide.</ArticleTitle><Pagination><MedlinePgn>593-601</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s11030-019-09965-0</ELocationID><Abstract><AbstractText>Cellobiose dehydrogenase (CDH, EC 1.1.99.18) from white rot fungi Phanerochaete chrysosporium can be used for constructing biosensors and biofuel cells, for bleaching cotton in textile industry, and recently, the enzyme has found an important application in biomedicine as an antimicrobial and antibiofilm agent. Stability and activity of the wild-type (wt) CDH and mutants at methionine residues in the presence of hydrogen peroxide were investigated. Saturation mutagenesis libraries were made at the only methionine in heme domain M65 and two methionines M685 and M738 in the flavin domain that were closest to the active site. After screening the libraries, three mutants with increased activity and stability in the presence of peroxide were found, M65F with 70% of residual activity after 6 h of incubation in 0.3 M hydrogen peroxide, M738S with 80% of residual activity and M685Y with over 90% of residual activity compared to wild-type CDH that retained 40% of original activity. Combined mutants showed no activity. The most stable mutant M685Y with 5.8 times increased half-life in the presence of peroxide showed also 2.5 times increased k<sub>cat</sub> for lactose compared to wtCDH and could be good candidate for applications in biofuel cells and biocatalysis for lactobionic acid production.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Balaž</LastName><ForeName>Ana Marija</ForeName><Initials>AM</Initials><AffiliationInfo><Affiliation>Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Njegoseva 12, 11000, Belgrade, Serbia.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Stevanović</LastName><ForeName>Jelena</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade, Serbia.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ostafe</LastName><ForeName>Raluca</ForeName><Initials>R</Initials><AffiliationInfo><Affiliation>Hall for Discovery and Learning Research, Purdue University, 207 S. Martin Jischke Dr., West Lafayette, IN, 47907, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Blazić</LastName><ForeName>Marija</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Njegoseva 12, 11000, Belgrade, Serbia.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ilić Đurđić</LastName><ForeName>Karla</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade, Serbia.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Fischer</LastName><ForeName>Rainer</ForeName><Initials>R</Initials><AffiliationInfo><Affiliation>Institute of Molecular Biotechnology, RWTH Aachen University, Worringerweg 1, 52074, Aachen, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Single Cell Analytics Center, Indiana Bioscience Research Institute, 1345 W. 16th St. Suite 300, Indianapolis, IN, 46202, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Prodanović</LastName><ForeName>Radivoje</ForeName><Initials>R</Initials><Identifier Source="ORCID">http://orcid.org/0000-0003-4662-1825</Identifier><AffiliationInfo><Affiliation>Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12 - 16, 11000, Belgrade, Serbia. rprodano@chem.bg.ac.rs.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>III46010</GrantID><Agency>Ministarstvo Prosvete, Nauke i Tehnološkog Razvoja</Agency><Country></Country></Grant><Grant><GrantID>ON172049</GrantID><Agency>Ministarstvo Prosvete, Nauke i Tehnološkog Razvoja</Agency><Country></Country></Grant><Grant><GrantID>ON173017</GrantID><Agency>Ministarstvo Prosvete, Nauke i Tehnološkog Razvoja</Agency><Country></Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>06</Month><Day>01</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>Mol Divers</MedlineTA><NlmUniqueID>9516534</NlmUniqueID><ISSNLinking>1381-1991</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Hydrogen peroxide</Keyword><Keyword MajorTopicYN="N">Library</Keyword><Keyword MajorTopicYN="N">Methionine</Keyword><Keyword MajorTopicYN="N">Protein engineering</Keyword><Keyword MajorTopicYN="N">Yeast</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>03</Month><Day>04</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2019</Year><Month>05</Month><Day>25</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2019</Year><Month>6</Month><Day>4</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2019</Year><Month>6</Month><Day>4</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2019</Year><Month>6</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">31154590</ArticleId><ArticleId IdType="doi">10.1007/s11030-019-09965-0</ArticleId><ArticleId IdType="pii">10.1007/s11030-019-09965-0</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Allemagne</li><li>Serbie</li><li>États-Unis</li></country><region><li>District de Cologne</li><li>Rhénanie-du-Nord-Westphalie</li></region><settlement><li>Aix-la-Chapelle</li></settlement></list><tree><country name="Serbie"><noRegion><name sortKey="Balaz, Ana Marija" sort="Balaz, Ana Marija" uniqKey="Balaz A" first="Ana Marija" last="Balaž">Ana Marija Balaž</name></noRegion><name sortKey="Blazi, Marija" sort="Blazi, Marija" uniqKey="Blazi M" first="Marija" last="Blazi">Marija Blazi</name><name sortKey="Ili Ur I, Karla" sort="Ili Ur I, Karla" uniqKey="Ili Ur I K" first="Karla" last="Ili Ur I">Karla Ili Ur I</name><name sortKey="Prodanovi, Radivoje" sort="Prodanovi, Radivoje" uniqKey="Prodanovi R" first="Radivoje" last="Prodanovi">Radivoje Prodanovi</name><name sortKey="Stevanovi, Jelena" sort="Stevanovi, Jelena" uniqKey="Stevanovi J" first="Jelena" last="Stevanovi">Jelena Stevanovi</name></country><country name="États-Unis"><noRegion><name sortKey="Ostafe, Raluca" sort="Ostafe, Raluca" uniqKey="Ostafe R" first="Raluca" last="Ostafe">Raluca Ostafe</name></noRegion><name sortKey="Fischer, Rainer" sort="Fischer, Rainer" uniqKey="Fischer R" first="Rainer" last="Fischer">Rainer Fischer</name></country><country name="Allemagne"><region name="Rhénanie-du-Nord-Westphalie"><name sortKey="Fischer, Rainer" sort="Fischer, Rainer" uniqKey="Fischer R" first="Rainer" last="Fischer">Rainer Fischer</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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