Role of surface tryptophan for peroxidase oxidation of nonphenolic lignin
13 p.-4 fig.-2 tab.
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Other Authors: | |
| Format: | artículo biblioteca |
| Language: | English |
| Published: |
BioMed Central
2016-09-17
|
| Subjects: | Ligninolytic peroxidases, Single-electron transfer, Catalytic tryptophan, Directed mutagenesis, Transient-state kinetics, Methylation, Acetylation, Nonphenolic lignin, Enzymatic delignification, NMR spectroscopy, |
| Online Access: | http://hdl.handle.net/10261/137338 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003329 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| id |
dig-irnas-es-10261-137338 |
|---|---|
| record_format |
koha |
| institution |
IRNAS ES |
| collection |
DSpace |
| country |
España |
| countrycode |
ES |
| component |
Bibliográfico |
| access |
En linea |
| databasecode |
dig-irnas-es |
| tag |
biblioteca |
| region |
Europa del Sur |
| libraryname |
Biblioteca del IRNAS España |
| language |
English |
| topic |
Ligninolytic peroxidases Single-electron transfer Catalytic tryptophan Directed mutagenesis Transient-state kinetics Methylation Acetylation Nonphenolic lignin Enzymatic delignification NMR spectroscopy Ligninolytic peroxidases Single-electron transfer Catalytic tryptophan Directed mutagenesis Transient-state kinetics Methylation Acetylation Nonphenolic lignin Enzymatic delignification NMR spectroscopy |
| spellingShingle |
Ligninolytic peroxidases Single-electron transfer Catalytic tryptophan Directed mutagenesis Transient-state kinetics Methylation Acetylation Nonphenolic lignin Enzymatic delignification NMR spectroscopy Ligninolytic peroxidases Single-electron transfer Catalytic tryptophan Directed mutagenesis Transient-state kinetics Methylation Acetylation Nonphenolic lignin Enzymatic delignification NMR spectroscopy Sáez-Jiménez, Verónica Rencoret, Jorge Rodríguez-Carvajal, M.A. Gutiérrez Suárez, Ana Ruiz-Dueñas, F. J. Martínez, Ángel T. Role of surface tryptophan for peroxidase oxidation of nonphenolic lignin |
| description |
13 p.-4 fig.-2 tab. |
| author2 |
European Commission |
| author_facet |
European Commission Sáez-Jiménez, Verónica Rencoret, Jorge Rodríguez-Carvajal, M.A. Gutiérrez Suárez, Ana Ruiz-Dueñas, F. J. Martínez, Ángel T. |
| format |
artículo |
| topic_facet |
Ligninolytic peroxidases Single-electron transfer Catalytic tryptophan Directed mutagenesis Transient-state kinetics Methylation Acetylation Nonphenolic lignin Enzymatic delignification NMR spectroscopy |
| author |
Sáez-Jiménez, Verónica Rencoret, Jorge Rodríguez-Carvajal, M.A. Gutiérrez Suárez, Ana Ruiz-Dueñas, F. J. Martínez, Ángel T. |
| author_sort |
Sáez-Jiménez, Verónica |
| title |
Role of surface tryptophan for peroxidase oxidation of nonphenolic lignin |
| title_short |
Role of surface tryptophan for peroxidase oxidation of nonphenolic lignin |
| title_full |
Role of surface tryptophan for peroxidase oxidation of nonphenolic lignin |
| title_fullStr |
Role of surface tryptophan for peroxidase oxidation of nonphenolic lignin |
| title_full_unstemmed |
Role of surface tryptophan for peroxidase oxidation of nonphenolic lignin |
| title_sort |
role of surface tryptophan for peroxidase oxidation of nonphenolic lignin |
| publisher |
BioMed Central |
| publishDate |
2016-09-17 |
| url |
http://hdl.handle.net/10261/137338 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003329 |
| work_keys_str_mv |
AT saezjimenezveronica roleofsurfacetryptophanforperoxidaseoxidationofnonphenoliclignin AT rencoretjorge roleofsurfacetryptophanforperoxidaseoxidationofnonphenoliclignin AT rodriguezcarvajalma roleofsurfacetryptophanforperoxidaseoxidationofnonphenoliclignin AT gutierrezsuarezana roleofsurfacetryptophanforperoxidaseoxidationofnonphenoliclignin AT ruizduenasfj roleofsurfacetryptophanforperoxidaseoxidationofnonphenoliclignin AT martinezangelt roleofsurfacetryptophanforperoxidaseoxidationofnonphenoliclignin |
| _version_ |
1777664884059668480 |
| spelling |
dig-irnas-es-10261-1373382021-12-27T15:37:32Z Role of surface tryptophan for peroxidase oxidation of nonphenolic lignin Sáez-Jiménez, Verónica Rencoret, Jorge Rodríguez-Carvajal, M.A. Gutiérrez Suárez, Ana Ruiz-Dueñas, F. J. Martínez, Ángel T. European Commission Ministerio de Economía y Competitividad (España) CSIC - Unidad de Recursos de Información Científica para la Investigación (URICI) Ligninolytic peroxidases Single-electron transfer Catalytic tryptophan Directed mutagenesis Transient-state kinetics Methylation Acetylation Nonphenolic lignin Enzymatic delignification NMR spectroscopy 13 p.-4 fig.-2 tab. [Background]: Despite claims as key enzymes in enzymatic delignification, very scarce information on the reaction rates between the ligninolytic versatile peroxidase (VP) and lignin peroxidase (LiP) and the lignin polymer is available, due to methodological difficulties related to lignin heterogeneity and low solubility [Results]: Two water-soluble sulfonated lignins (from Picea abies and Eucalyptus grandis) were chemically characterized and used to estimate single electron-transfer rates to the H2O2-activated Pleurotus eryngii VP (native enzyme and mutated variant) transient states (compounds I and II bearing two- and one-electron deficiencies, respectively). When the rate-limiting reduction of compound II was quantified by stopped-flow rapid spectrophotometry, from fourfold (softwood lignin) to over 100-fold (hardwood lignin) lower electron-transfer efficiencies (k 3app values) were observed for the W164S variant at surface Trp164, compared with the native VP. These lignosulfonates have ~20–30 % phenolic units, which could be responsible for the observed residual activity. Therefore, methylated (and acetylated) samples were used in new stopped-flow experiments, where negligible electron transfer to the W164S compound II was found. This revealed that the residual reduction of W164S compound II by native lignin was due to its phenolic moiety. Since both native lignins have a relatively similar phenolic moiety, the higher W164S activity on the softwood lignin could be due to easier access of its mono-methoxylated units for direct oxidation at the heme channel in the absence of the catalytic tryptophan. Moreover, the lower electron transfer rates from the derivatized lignosulfonates to native VP suggest that peroxidase attack starts at the phenolic lignin moiety. In agreement with the transient-state kinetic data, very low structural modification of lignin, as revealed by size-exclusion chromatography and two-dimensional nuclear magnetic resonance, was obtained during steady-state treatment (up to 24 h) of native lignosulfonates with the W164S variant compared with native VP and, more importantly, this activity disappeared when nonphenolic lignosulfonates were used. [Conclusions]: We demonstrate for the first time that the surface tryptophan conserved in most LiPs and VPs (Trp164 of P. eryngii VPL) is strictly required for oxidation of the nonphenolic moiety, which represents the major and more recalcitrant part of the lignin polymer. This work was supported by the INDOX (KBBE-2013-613549 to ATM) and EnzOx2 (H2020-BBI-PPP-2015-RIA-720297 to ATM) EU projects, and the NOESIS (BIO2014-56388-R to FJR-D), BIORENZYMERY (AGL2014-53730-R to AG) and LIGNIN (CTQ2014-60764-JIN to JR) projects of the Spanish Ministry of Economy and Competitiveness (MINECO) co-financed by FEDER funds. We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). Peer reviewed 2016-09-26T11:50:54Z 2016-09-26T11:50:54Z 2016-09-17 artículo http://purl.org/coar/resource_type/c_6501 Biotechnology for Biofuels 9: 198 (2016) 1754-6834 http://hdl.handle.net/10261/137338 10.1186/s13068-016-0615-x http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003329 28616078 en #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/EC/H2020/720297 info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2014-56388-R info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/AGL2014-53730-R info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/CTQ2014-60764-JIN Publisher's version http://dx.doi.org/ 10.1186/s13068-016-0615-x Sí open BioMed Central |