Enzymatic versatility and thermostability of a new aryl-alcohol oxidase from Thermothelomyces thermophilus M77
| العنوان: | Enzymatic versatility and thermostability of a new aryl-alcohol oxidase from Thermothelomyces thermophilus M77 |
|---|---|
| المؤلفون: | Andre S. Godoy, Marco Antonio Seiki Kadowaki, Mariana Ortiz de Godoy, Paula Higasi, Rolf A. Prade, Igor Polikarpov, Evandro Ares de Araújo |
| المصدر: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Models, Molecular, Glycosylation, Stereochemistry, Protein Conformation, Biophysics, Crystallography, X-Ray, Biochemistry, Cofactor, Substrate Specificity, 03 medical and health sciences, chemistry.chemical_compound, Ascomycota, Oxidoreductase, Enzyme Stability, Aryl-alcohol oxidase, Molecular Biology, 030304 developmental biology, Thermostability, chemistry.chemical_classification, Flavin adenine dinucleotide, 0303 health sciences, Binding Sites, biology, 030306 microbiology, Temperature, Active site, Hydrogen Peroxide, biology.organism_classification, BIOTECNOLOGIA, Alcohol Oxidoreductases, chemistry, biology.protein, Calcium, Myceliophthora thermophila |
| الوصف: | Background Fungal aryl-alcohol oxidases (AAOx) are extracellular flavoenzymes that belong to glucose-methanol-choline oxidoreductase family and are responsible for the selective conversion of primary aromatic alcohols into aldehydes and aromatic aldehydes to their corresponding acids, with concomitant production of hydrogen peroxide (H2O2) as by-product. The H2O2 can be provided to lignin degradation pathway, a biotechnological property explored in biofuel production. In the thermophilic fungus Thermothelomyces thermophilus (formerly Myceliophthora thermophila), just one AAOx was identified in the exo-proteome. Methods The glycosylated and non-refolded crystal structure of an AAOx from T. thermophilus at 2.6 A resolution was elucidated by X-ray crystallography combined with small-angle X-ray scattering (SAXS) studies. Moreover, biochemical analyses were carried out to shed light on enzyme substrate specificity and thermostability. Results This flavoenzyme harbors a flavin adenine dinucleotide as a cofactor and is able to oxidize aromatic substrates and 5-HMF. Our results also show that the enzyme has similar oxidation rates for bulky or simple aromatic substrates such as cinnamyl and veratryl alcohols. Moreover, the crystal structure of MtAAOx reveals an open active site, which might explain observed specificity of the enzyme. Conclusions MtAAOx shows previously undescribed structural differences such as a fully accessible catalytic tunnel, heavy glycosylation and Ca2+ binding site providing evidences for thermostability and activity of the enzymes from AA3_2 subfamily. General significance Structural and biochemical analyses of MtAAOx could be important for comprehension of aryl-alcohol oxidases structure-function relationships and provide additional molecular tools to be used in future biotechnological applications. |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::49dc87b4b00ffe2414042450696adab0 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....49dc87b4b00ffe2414042450696adab0 |
| قاعدة البيانات: | OpenAIRE |
| ResultId |
1 |
|---|---|
| Header |
edsair OpenAIRE edsair.doi.dedup.....49dc87b4b00ffe2414042450696adab0 832 3 unknown 832.017883300781 |
| PLink |
https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&scope=site&db=edsair&AN=edsair.doi.dedup.....49dc87b4b00ffe2414042450696adab0&custid=s6537998&authtype=sso |
| FullText |
Array
(
[Availability] => 0
)
Array ( [0] => Array ( [Url] => https://explore.openaire.eu/search/publication?articleId=doi_dedup___::49dc87b4b00ffe2414042450696adab0# [Name] => EDS - OpenAIRE [Category] => fullText [Text] => View record in OpenAIRE [MouseOverText] => View record in OpenAIRE ) ) |
| Items |
Array
(
[Name] => Title
[Label] => Title
[Group] => Ti
[Data] => Enzymatic versatility and thermostability of a new aryl-alcohol oxidase from Thermothelomyces thermophilus M77
)
Array ( [Name] => Author [Label] => Authors [Group] => Au [Data] => <searchLink fieldCode="AR" term="%22Andre+S%2E+Godoy%22">Andre S. Godoy</searchLink><br /><searchLink fieldCode="AR" term="%22Marco+Antonio+Seiki+Kadowaki%22">Marco Antonio Seiki Kadowaki</searchLink><br /><searchLink fieldCode="AR" term="%22Mariana+Ortiz+de+Godoy%22">Mariana Ortiz de Godoy</searchLink><br /><searchLink fieldCode="AR" term="%22Paula+Higasi%22">Paula Higasi</searchLink><br /><searchLink fieldCode="AR" term="%22Rolf+A%2E+Prade%22">Rolf A. Prade</searchLink><br /><searchLink fieldCode="AR" term="%22Igor+Polikarpov%22">Igor Polikarpov</searchLink><br /><searchLink fieldCode="AR" term="%22Evandro+Ares+de+Araújo%22">Evandro Ares de Araújo</searchLink> ) Array ( [Name] => TitleSource [Label] => Source [Group] => Src [Data] => Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual)<br />Universidade de São Paulo (USP)<br />instacron:USP ) Array ( [Name] => DatePubCY [Label] => Publication Year [Group] => Date [Data] => 2020 ) Array ( [Name] => Subject [Label] => Subject Terms [Group] => Su [Data] => <searchLink fieldCode="DE" term="%22Models%2C+Molecular%22">Models, Molecular</searchLink><br /><searchLink fieldCode="DE" term="%22Glycosylation%22">Glycosylation</searchLink><br /><searchLink fieldCode="DE" term="%22Stereochemistry%22">Stereochemistry</searchLink><br /><searchLink fieldCode="DE" term="%22Protein+Conformation%22">Protein Conformation</searchLink><br /><searchLink fieldCode="DE" term="%22Biophysics%22">Biophysics</searchLink><br /><searchLink fieldCode="DE" term="%22Crystallography%2C+X-Ray%22">Crystallography, X-Ray</searchLink><br /><searchLink fieldCode="DE" term="%22Biochemistry%22">Biochemistry</searchLink><br /><searchLink fieldCode="DE" term="%22Cofactor%22">Cofactor</searchLink><br /><searchLink fieldCode="DE" term="%22Substrate+Specificity%22">Substrate Specificity</searchLink><br /><searchLink fieldCode="DE" term="%2203+medical+and+health+sciences%22">03 medical and health sciences</searchLink><br /><searchLink fieldCode="DE" term="%22chemistry%2Echemical%5Fcompound%22">chemistry.chemical_compound</searchLink><br /><searchLink fieldCode="DE" term="%22Ascomycota%22">Ascomycota</searchLink><br /><searchLink fieldCode="DE" term="%22Oxidoreductase%22">Oxidoreductase</searchLink><br /><searchLink fieldCode="DE" term="%22Enzyme+Stability%22">Enzyme Stability</searchLink><br /><searchLink fieldCode="DE" term="%22Aryl-alcohol+oxidase%22">Aryl-alcohol oxidase</searchLink><br /><searchLink fieldCode="DE" term="%22Molecular+Biology%22">Molecular Biology</searchLink><br /><searchLink fieldCode="DE" term="%22030304+developmental+biology%22">030304 developmental biology</searchLink><br /><searchLink fieldCode="DE" term="%22Thermostability%22">Thermostability</searchLink><br /><searchLink fieldCode="DE" term="%22chemistry%2Echemical%5Fclassification%22">chemistry.chemical_classification</searchLink><br /><searchLink fieldCode="DE" term="%22Flavin+adenine+dinucleotide%22">Flavin adenine dinucleotide</searchLink><br /><searchLink fieldCode="DE" term="%220303+health+sciences%22">0303 health sciences</searchLink><br /><searchLink fieldCode="DE" term="%22Binding+Sites%22">Binding Sites</searchLink><br /><searchLink fieldCode="DE" term="%22biology%22">biology</searchLink><br /><searchLink fieldCode="DE" term="%22030306+microbiology%22">030306 microbiology</searchLink><br /><searchLink fieldCode="DE" term="%22Temperature%22">Temperature</searchLink><br /><searchLink fieldCode="DE" term="%22Active+site%22">Active site</searchLink><br /><searchLink fieldCode="DE" term="%22Hydrogen+Peroxide%22">Hydrogen Peroxide</searchLink><br /><searchLink fieldCode="DE" term="%22biology%2Eorganism%5Fclassification%22">biology.organism_classification</searchLink><br /><searchLink fieldCode="DE" term="%22BIOTECNOLOGIA%22">BIOTECNOLOGIA</searchLink><br /><searchLink fieldCode="DE" term="%22Alcohol+Oxidoreductases%22">Alcohol Oxidoreductases</searchLink><br /><searchLink fieldCode="DE" term="%22chemistry%22">chemistry</searchLink><br /><searchLink fieldCode="DE" term="%22biology%2Eprotein%22">biology.protein</searchLink><br /><searchLink fieldCode="DE" term="%22Calcium%22">Calcium</searchLink><br /><searchLink fieldCode="DE" term="%22Myceliophthora+thermophila%22">Myceliophthora thermophila</searchLink> ) Array ( [Name] => Abstract [Label] => Description [Group] => Ab [Data] => Background Fungal aryl-alcohol oxidases (AAOx) are extracellular flavoenzymes that belong to glucose-methanol-choline oxidoreductase family and are responsible for the selective conversion of primary aromatic alcohols into aldehydes and aromatic aldehydes to their corresponding acids, with concomitant production of hydrogen peroxide (H2O2) as by-product. The H2O2 can be provided to lignin degradation pathway, a biotechnological property explored in biofuel production. In the thermophilic fungus Thermothelomyces thermophilus (formerly Myceliophthora thermophila), just one AAOx was identified in the exo-proteome. Methods The glycosylated and non-refolded crystal structure of an AAOx from T. thermophilus at 2.6 A resolution was elucidated by X-ray crystallography combined with small-angle X-ray scattering (SAXS) studies. Moreover, biochemical analyses were carried out to shed light on enzyme substrate specificity and thermostability. Results This flavoenzyme harbors a flavin adenine dinucleotide as a cofactor and is able to oxidize aromatic substrates and 5-HMF. Our results also show that the enzyme has similar oxidation rates for bulky or simple aromatic substrates such as cinnamyl and veratryl alcohols. Moreover, the crystal structure of MtAAOx reveals an open active site, which might explain observed specificity of the enzyme. Conclusions MtAAOx shows previously undescribed structural differences such as a fully accessible catalytic tunnel, heavy glycosylation and Ca2+ binding site providing evidences for thermostability and activity of the enzymes from AA3_2 subfamily. General significance Structural and biochemical analyses of MtAAOx could be important for comprehension of aryl-alcohol oxidases structure-function relationships and provide additional molecular tools to be used in future biotechnological applications. ) Array ( [Name] => URL [Label] => Access URL [Group] => URL [Data] => <link linkTarget="URL" linkTerm="https://explore.openaire.eu/search/publication?articleId=doi_dedup___::49dc87b4b00ffe2414042450696adab0" linkWindow="_blank">https://explore.openaire.eu/search/publication?articleId=doi_dedup___::49dc87b4b00ffe2414042450696adab0</link> ) Array ( [Name] => Copyright [Label] => Rights [Group] => Cpyrght [Data] => OPEN ) Array ( [Name] => AN [Label] => Accession Number [Group] => ID [Data] => edsair.doi.dedup.....49dc87b4b00ffe2414042450696adab0 ) |
| RecordInfo |
Array
(
[BibEntity] => Array
(
[Languages] => Array
(
[0] => Array
(
[Text] => Undetermined
)
)
[Subjects] => Array
(
[0] => Array
(
[SubjectFull] => Models, Molecular
[Type] => general
)
[1] => Array
(
[SubjectFull] => Glycosylation
[Type] => general
)
[2] => Array
(
[SubjectFull] => Stereochemistry
[Type] => general
)
[3] => Array
(
[SubjectFull] => Protein Conformation
[Type] => general
)
[4] => Array
(
[SubjectFull] => Biophysics
[Type] => general
)
[5] => Array
(
[SubjectFull] => Crystallography, X-Ray
[Type] => general
)
[6] => Array
(
[SubjectFull] => Biochemistry
[Type] => general
)
[7] => Array
(
[SubjectFull] => Cofactor
[Type] => general
)
[8] => Array
(
[SubjectFull] => Substrate Specificity
[Type] => general
)
[9] => Array
(
[SubjectFull] => 03 medical and health sciences
[Type] => general
)
[10] => Array
(
[SubjectFull] => chemistry.chemical_compound
[Type] => general
)
[11] => Array
(
[SubjectFull] => Ascomycota
[Type] => general
)
[12] => Array
(
[SubjectFull] => Oxidoreductase
[Type] => general
)
[13] => Array
(
[SubjectFull] => Enzyme Stability
[Type] => general
)
[14] => Array
(
[SubjectFull] => Aryl-alcohol oxidase
[Type] => general
)
[15] => Array
(
[SubjectFull] => Molecular Biology
[Type] => general
)
[16] => Array
(
[SubjectFull] => 030304 developmental biology
[Type] => general
)
[17] => Array
(
[SubjectFull] => Thermostability
[Type] => general
)
[18] => Array
(
[SubjectFull] => chemistry.chemical_classification
[Type] => general
)
[19] => Array
(
[SubjectFull] => Flavin adenine dinucleotide
[Type] => general
)
[20] => Array
(
[SubjectFull] => 0303 health sciences
[Type] => general
)
[21] => Array
(
[SubjectFull] => Binding Sites
[Type] => general
)
[22] => Array
(
[SubjectFull] => biology
[Type] => general
)
[23] => Array
(
[SubjectFull] => 030306 microbiology
[Type] => general
)
[24] => Array
(
[SubjectFull] => Temperature
[Type] => general
)
[25] => Array
(
[SubjectFull] => Active site
[Type] => general
)
[26] => Array
(
[SubjectFull] => Hydrogen Peroxide
[Type] => general
)
[27] => Array
(
[SubjectFull] => biology.organism_classification
[Type] => general
)
[28] => Array
(
[SubjectFull] => BIOTECNOLOGIA
[Type] => general
)
[29] => Array
(
[SubjectFull] => Alcohol Oxidoreductases
[Type] => general
)
[30] => Array
(
[SubjectFull] => chemistry
[Type] => general
)
[31] => Array
(
[SubjectFull] => biology.protein
[Type] => general
)
[32] => Array
(
[SubjectFull] => Calcium
[Type] => general
)
[33] => Array
(
[SubjectFull] => Myceliophthora thermophila
[Type] => general
)
)
[Titles] => Array
(
[0] => Array
(
[TitleFull] => Enzymatic versatility and thermostability of a new aryl-alcohol oxidase from Thermothelomyces thermophilus M77
[Type] => main
)
)
)
[BibRelationships] => Array
(
[HasContributorRelationships] => Array
(
[0] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Andre S. Godoy
)
)
)
[1] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Marco Antonio Seiki Kadowaki
)
)
)
[2] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Mariana Ortiz de Godoy
)
)
)
[3] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Paula Higasi
)
)
)
[4] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Rolf A. Prade
)
)
)
[5] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Igor Polikarpov
)
)
)
[6] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Evandro Ares de Araújo
)
)
)
)
[IsPartOfRelationships] => Array
(
[0] => Array
(
[BibEntity] => Array
(
[Dates] => Array
(
[0] => Array
(
[D] => 01
[M] => 01
[Type] => published
[Y] => 2020
)
)
[Identifiers] => Array
(
[0] => Array
(
[Type] => issn-locals
[Value] => edsair
)
[1] => Array
(
[Type] => issn-locals
[Value] => edsairFT
)
)
)
)
)
)
)
|
| IllustrationInfo |