{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T07:57:54Z","timestamp":1791187074986,"version":"4.1.0"},"reference-count":47,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Biotechnol"],"published-print":{"date-parts":[[2013,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>Fungal laccases are multicopper oxidases with huge applicability in different sectors. Here, we describe the development of a set of high-throughput colorimetric assays for screening laccase libraries in directed evolution studies.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>Firstly, we designed three colorimetric assays based on the oxidation of sinapic acid, acetosyringone and syringaldehyde with \u03bb<jats:sub>max<\/jats:sub> of 512, 520 and 370\u00a0nm, respectively. These syringyl-type phenolic compounds are released during the degradation of lignocellulose and can act as laccase redox mediators. The oxidation of the three compounds by low and high-redox potential laccases evolved in <jats:italic>Saccharomyces cerevisiae<\/jats:italic> produced quantifiable and linear responses, with detection limits around 1\u00a0mU\/mL and CV values below 16%. The phenolic substrates were also suitable for pre-screening mutant libraries on solid phase format. Intense colored-halos were developed around the yeast colonies secreting laccase. Furthermore, the oxidation of violuric acid to its iminoxyl radical (\u03bb<jats:sub>max<\/jats:sub> of 515\u00a0nm and CV below 15%) was devised as reporter assay for laccase redox potential during the screening of mutant libraries from high-redox potential laccases. Finally, we developed three dye-decolorizing assays based on the enzymatic oxidation of Methyl Orange (470\u00a0nm), Evans Blue (605\u00a0nm) and Remazol Brilliant Blue (640\u00a0nm) giving up to 40% decolorization yields and CV values below 18%. The assays were reliable for direct measurement of laccase activity or to indirectly explore the oxidation of mediators that do not render colored products (but promote dye decolorization). Every single assay reported in this work was tested by exploring mutant libraries created by error prone PCR of fungal laccases secreted by yeast.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusions<\/jats:title>\n            <jats:p>The high-throughput screening methods reported in this work could be useful for engineering laccases for different purposes. The assays based on the oxidation of syringyl-compounds might be valuable tools for tailoring laccases precisely enhanced to aid biomass conversion processes. The violuric assay might be useful to preserve the redox potential of laccase whilst evolving towards new functions. The dye-decolorizing assays are useful for engineering <jats:italic>ad hoc<\/jats:italic> laccases for detoxification of textile wastewaters, or as indirect assays to explore laccase activity on other natural mediators.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1472-6750-13-90","type":"journal-article","created":{"date-parts":[[2013,10,26]],"date-time":"2013-10-26T09:03:32Z","timestamp":1382778212000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":51,"title":["New colorimetric screening assays for the directed evolution of fungal laccases to improve the conversion of plant biomass"],"prefix":"10.1186","volume":"13","author":[{"given":"Isabel","family":"Pardo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiomara","family":"Chanag\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Isabel","family":"Vicente","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miguel","family":"Alcalde","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susana","family":"Camarero","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2013,10,26]]},"reference":[{"key":"990_CR1","doi-asserted-by":"publisher","first-page":"32","DOI":"10.1186\/1475-2859-7-32","volume":"7","author":"A Kunamneni","year":"2008","unstructured":"Kunamneni A, Camarero S, Garcia-Burgos C, Plou FJ, Ballesteros A, Alcalde M: Engineering and applications of fungal laccases for organic synthesis. Microb Cell Fact. 2008, 7: 32-10.1186\/1475-2859-7-32.","journal-title":"Microb Cell Fact"},{"key":"990_CR2","doi-asserted-by":"publisher","first-page":"500","DOI":"10.1016\/j.biotechadv.2006.04.003","volume":"24","author":"S Rodr\u00edguez-Couto","year":"2006","unstructured":"Rodr\u00edguez-Couto S, Toca JL: Industrial and biotechnological applications of laccases: a review. Biotechnol Adv. 2006, 24: 500-513. 10.1016\/j.biotechadv.2006.04.003.","journal-title":"Biotechnol Adv"},{"key":"990_CR3","doi-asserted-by":"publisher","first-page":"835","DOI":"10.1016\/j.enzmictec.2006.01.009","volume":"39","author":"S Shleev","year":"2006","unstructured":"Shleev S, Persson P, Shumakovich G, Mazhugo Y, Yaropolov A, Ruzgas T, Gorton L: Laccase-based biosensors for monitoring lignin. Enzyme Microb Technol. 2006, 39: 835-840. 10.1016\/j.enzmictec.2006.01.009.","journal-title":"Enzyme Microb Technol"},{"key":"990_CR4","doi-asserted-by":"publisher","first-page":"12372","DOI":"10.1074\/jbc.274.18.12372","volume":"274","author":"F Xu","year":"1999","unstructured":"Xu F, Palmer AE, Yaver DS, Berka RM, Gambetta GA, Brown SH, Solomon EI: Targeted mutations in a Trametes villosa laccase - Axial perturbations of the T1 copper. J Biol Chem. 1999, 274: 12372-12375. 10.1074\/jbc.274.18.12372.","journal-title":"J Biol Chem"},{"key":"990_CR5","doi-asserted-by":"crossref","first-page":"4509","DOI":"10.1128\/aem.60.12.4509-4516.1994","volume":"60","author":"S Camarero","year":"1994","unstructured":"Camarero S, Galletti GC, Mart\u00ednez AT: Preferential degradation of phenolic lignin-units by two white-rot fungi. Appl Environ Microbiol. 1994, 60: 4509-4516.","journal-title":"Appl Environ Microbiol"},{"key":"990_CR6","doi-asserted-by":"publisher","first-page":"2964","DOI":"10.1021\/es062328j","volume":"41","author":"A Ca\u00f1as","year":"2007","unstructured":"Ca\u00f1as A, Alcalde M, Plou FJ, Mart\u00ednez MJ, Mart\u00ednez AT, Camarero S: Transformation of polycyclic aromatic hydrocarbons by laccase is strongly enhanced by phenolic compounds present in soil. Environ Sci Technol. 2007, 41: 2964-2971. 10.1021\/es062328j.","journal-title":"Environ Sci Technol"},{"key":"990_CR7","doi-asserted-by":"publisher","first-page":"1775","DOI":"10.1128\/AEM.71.4.1775-1784.2005","volume":"71","author":"S Camarero","year":"2005","unstructured":"Camarero S, Ibarra D, Mart\u00ednez MJ, Mart\u00ednez AT: Lignin-derived compounds as efficient laccase mediators for decolorization of different types of recalcitrant dyes. Appl Environ Microbiol. 2005, 71: 1775-1784. 10.1128\/AEM.71.4.1775-1784.2005.","journal-title":"Appl Environ Microbiol"},{"key":"990_CR8","doi-asserted-by":"publisher","first-page":"699","DOI":"10.1515\/HF.2009.066","volume":"63","author":"P Nousiainen","year":"2009","unstructured":"Nousiainen P, Maijala P, Hatakka A, Mart\u00ednez AT, Sipila J: Syringyl-type simple plant phenolics as mediating oxidants in laccase catalyzed degradation of lignocellulosic materials: model compound studies. Holzforschung. 2009, 63: 699-704.","journal-title":"Holzforschung"},{"key":"990_CR9","doi-asserted-by":"publisher","first-page":"687","DOI":"10.1016\/j.chemosphere.2009.07.039","volume":"77","author":"C Torres-Duarte","year":"2009","unstructured":"Torres-Duarte C, Roman R, Tinoco R, V\u00e1zquez-Duhalt R: Halogenated pesticide transformation by a laccase-mediator system. Chemosphere. 2009, 77: 687-692. 10.1016\/j.chemosphere.2009.07.039.","journal-title":"Chemosphere"},{"key":"990_CR10","doi-asserted-by":"publisher","first-page":"694","DOI":"10.1016\/j.biotechadv.2010.05.002","volume":"28","author":"AI Ca\u00f1as","year":"2010","unstructured":"Ca\u00f1as AI, Camarero S: Laccases and their natural mediators: biotechnological tools for sustainable eco-friendly processes. Biotechnol Adv. 2010, 28: 694-705. 10.1016\/j.biotechadv.2010.05.002.","journal-title":"Biotechnol Adv"},{"key":"990_CR11","doi-asserted-by":"publisher","first-page":"2161","DOI":"10.1021\/jf011348w","volume":"50","author":"H Kikuzaki","year":"2002","unstructured":"Kikuzaki H, Hisamoto M, Hirose K, Akiyama K, Taniguchi H: Antioxidant properties of ferulic acid and its related compounds. J Agric Food Chem. 2002, 50: 2161-2168. 10.1021\/jf011348w.","journal-title":"J Agric Food Chem"},{"key":"990_CR12","doi-asserted-by":"publisher","first-page":"130","DOI":"10.1016\/j.bbrc.2004.12.153","volume":"328","author":"CJ Baker","year":"2005","unstructured":"Baker CJ, Mock NM, Whitaker BD, Roberts DP, Rice CP, Deahl KL, Aver'yanov AA: Involvement of acetosyringone in plant-pathogen recognition. Biochem Biophys Res Commun. 2005, 328: 130-136. 10.1016\/j.bbrc.2004.12.153.","journal-title":"Biochem Biophys Res Commun"},{"key":"990_CR13","doi-asserted-by":"publisher","first-page":"8211","DOI":"10.1016\/j.biortech.2010.05.080","volume":"101","author":"E Aracri","year":"2010","unstructured":"Aracri E, Fillat A, Colom JF, Guti\u00e9rrez A, del R\u00edo JC, Mart\u00ednez AT, Vidal T: Enzymatic grafting of simple phenols on flax and sisal pulp fibres using laccases. Bioresour Technol. 2010, 101: 8211-8216. 10.1016\/j.biortech.2010.05.080.","journal-title":"Bioresour Technol"},{"key":"990_CR14","doi-asserted-by":"publisher","first-page":"718","DOI":"10.1021\/ie00027a034","volume":"33","author":"GX Wu","year":"1994","unstructured":"Wu GX, Heitz M, Chornet E: Improved alkaline oxidation process for the production of aldehydes (vanillin and syringaldehyde) from steam-explosion hardwood lignin. Ind Eng Chem Res. 1994, 33: 718-723. 10.1021\/ie00027a034.","journal-title":"Ind Eng Chem Res"},{"key":"990_CR15","doi-asserted-by":"publisher","first-page":"741","DOI":"10.1021\/ie102132a","volume":"50","author":"PCR Pinto","year":"2011","unstructured":"Pinto PCR, da Silva EAB, Rodrigues AE: Insights into oxidative conversion of lignin to high-added-value phenolic aldehydes. Ind Eng Chem Res. 2011, 50: 741-748. 10.1021\/ie102132a.","journal-title":"Ind Eng Chem Res"},{"key":"990_CR16","doi-asserted-by":"publisher","first-page":"1264","DOI":"10.1016\/j.enzmictec.2006.09.016","volume":"40","author":"S Camarero","year":"2007","unstructured":"Camarero S, Ibarra D, Mart\u00ednez AT, Romero J, Guti\u00e9rrez A, del R\u00edo JC: Paper pulp delignification using laccase and natural mediators. Enzyme Microb Technol. 2007, 40: 1264-1271. 10.1016\/j.enzmictec.2006.09.016.","journal-title":"Enzyme Microb Technol"},{"key":"990_CR17","doi-asserted-by":"publisher","first-page":"20","DOI":"10.3109\/07388551003757816","volume":"31","author":"W Parawira","year":"2011","unstructured":"Parawira W, Tekere M: Biotechnological strategies to overcome inhibitors in lignocellulose hydrolysates for ethanol production: review. Crit Rev Biotechnol. 2011, 31: 20-31. 10.3109\/07388551003757816.","journal-title":"Crit Rev Biotechnol"},{"key":"990_CR18","doi-asserted-by":"publisher","first-page":"6378","DOI":"10.1016\/j.biortech.2009.07.049","volume":"100","author":"M Jurado","year":"2009","unstructured":"Jurado M, Prieto A, Mart\u00ednez-Alcal\u00e1 MA, Mart\u00ednez AT, Mart\u00ednez MJ: Laccase detoxification of steam-exploded wheat straw for second generation bioethanol. Bioresour Technol. 2009, 100: 6378-6384. 10.1016\/j.biortech.2009.07.049.","journal-title":"Bioresour Technol"},{"key":"990_CR19","doi-asserted-by":"publisher","first-page":"74","DOI":"10.1002\/btpr.1666","volume":"29","author":"P Alvira","year":"2013","unstructured":"Alvira P, Moreno AD, Ibarra D, S\u00e1ez F, Ballesteros M: Improving the fermentation performance of saccharomyces cerevisiae by laccase during ethanol production from steam-exploded wheat straw at high-substrate loadings. Biotechnol Prog. 2013, 29: 74-82. 10.1002\/btpr.1666.","journal-title":"Biotechnol Prog"},{"key":"990_CR20","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1016\/S1367-5931(99)80010-6","volume":"3","author":"FH Arnold","year":"1999","unstructured":"Arnold FH, Volkov AA: Directed evolution of biocatalysts. Curr Opin Chem Biol. 1999, 3: 54-59. 10.1016\/S1367-5931(99)80010-6.","journal-title":"Curr Opin Chem Biol"},{"key":"990_CR21","doi-asserted-by":"publisher","first-page":"1370","DOI":"10.1128\/AEM.07530-11","volume":"78","author":"S Camarero","year":"2012","unstructured":"Camarero S, Pardo I, Ca\u00f1as AI, Molina P, Record E, Mart\u00ednez AT, Mart\u00ednez MJ, Alcalde M: Engineering platforms for directed evolution of laccase from Pycnoporus cinnabarinus. Appl Environ Microbiol. 2012, 78: 1370-1384. 10.1128\/AEM.07530-11.","journal-title":"Appl Environ Microbiol"},{"key":"990_CR22","doi-asserted-by":"publisher","first-page":"1030","DOI":"10.1016\/j.chembiol.2010.07.010","volume":"17","author":"D Mat\u00e9","year":"2010","unstructured":"Mat\u00e9 D, Garc\u00eda-Burgos C, Garc\u00eda-Ruiz E, Ballesteros A, Camarero S, Alcalde M: Laboratory evolution of high redox potential laccases. Chem Biol. 2010, 17: 1030-1041. 10.1016\/j.chembiol.2010.07.010.","journal-title":"Chem Biol"},{"key":"990_CR23","doi-asserted-by":"publisher","first-page":"2978","DOI":"10.1002\/bit.24588","volume":"109","author":"I Pardo","year":"2012","unstructured":"Pardo I, Vicente AI, Mate DM, Alcalde M, Camarero S: Development of chimeric laccases by directed evolution. Biotechnol Bioeng. 2012, 109: 2978-2986. 10.1002\/bit.24588.","journal-title":"Biotechnol Bioeng"},{"key":"990_CR24","doi-asserted-by":"publisher","first-page":"441","DOI":"10.1093\/jxb\/ern261","volume":"60","author":"FJ Ruiz-Due\u00f1as","year":"2009","unstructured":"Ruiz-Due\u00f1as FJ, Morales M, Garc\u00eda E, Miki Y, Mart\u00ednez MJ, Mart\u00ednez AT: Substrate oxidation sites in versatile peroxidase and other basidiomycete peroxidases. J Exp Bot. 2009, 60: 441-452. 10.1093\/jxb\/ern261.","journal-title":"J Exp Bot"},{"key":"990_CR25","first-page":"51","volume":"72","author":"J Bidlack","year":"1992","unstructured":"Bidlack J, Malone M, Benson R: Molecular structure and component integration of secondary cell walls in plants. Proc Okla Acad Sci. 1992, 72: 51-56.","journal-title":"Proc Okla Acad Sci"},{"key":"990_CR26","doi-asserted-by":"publisher","first-page":"6703","DOI":"10.1021\/es8008979","volume":"42","author":"S Camarero","year":"2008","unstructured":"Camarero S, Ca\u00f1as AI, Nousiainen P, Record E, Lomascolo A, Mart\u00ednez MJ, Mart\u00ednez AT: p-Hydroxycinnamic acids as natural mediators for laccase oxidation of recalcitrant compounds. Environ Sci Technol. 2008, 42: 6703-6709. 10.1021\/es8008979.","journal-title":"Environ Sci Technol"},{"key":"990_CR27","doi-asserted-by":"publisher","first-page":"4124","DOI":"10.1021\/es062723+","volume":"41","author":"A Guti\u00e9rrez","year":"2007","unstructured":"Guti\u00e9rrez A, Rencoret J, Ibarra D, Molina S, Camarero S, Romero J, del R\u00edo JC, Mart\u00ednez AT: Removal of lipophilic extractives from paper pulp by laccase and lignin-derived phenols as natural mediators. Environ Sci Technol. 2007, 41: 4124-4129. 10.1021\/es062723+.","journal-title":"Environ Sci Technol"},{"key":"990_CR28","doi-asserted-by":"publisher","first-page":"694","DOI":"10.1002\/(SICI)1097-0290(19980320)57:6<694::AID-BIT7>3.0.CO;2-G","volume":"57","author":"K Lacki","year":"1998","unstructured":"Lacki K, Duvnjak Z: Transformation of 3,5-dimethoxy,4-hydroxy cinnamic acid by polyphenol oxidase from the fungus Trametes versicolor: product elucidation studies. Biotechnol Bioeng. 1998, 57: 694-703. 10.1002\/(SICI)1097-0290(19980320)57:6<694::AID-BIT7>3.0.CO;2-G.","journal-title":"Biotechnol Bioeng"},{"key":"990_CR29","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1007\/BF02268570","volume":"36","author":"K-S Shin","year":"1995","unstructured":"Shin K-S: Oxidation of syringic acid by extracellular peroxidase of white-rot fungus, Pleurotus ostreatus. Mycoscience. 1995, 36: 31-35. 10.1007\/BF02268570.","journal-title":"Mycoscience"},{"key":"990_CR30","doi-asserted-by":"publisher","first-page":"371","DOI":"10.1016\/j.biortech.2012.08.023","volume":"124","author":"T Rosado","year":"2012","unstructured":"Rosado T, Bernardo P, Koci K, Coelho AV, Robalo MP, Martins LO: Methyl syringate: an efficient phenolic mediator for bacterial and fungal laccases. Bioresour Technol. 2012, 124: 371-378.","journal-title":"Bioresour Technol"},{"key":"990_CR31","doi-asserted-by":"publisher","first-page":"1052","DOI":"10.1016\/j.chembiol.2007.08.010","volume":"14","author":"M Zum\u00e1rraga","year":"2007","unstructured":"Zum\u00e1rraga M, Bulter T, Shleev S, Polaina J, Martinez-Arias A, Plou FJ, Ballesteros A, Alcalde M: In vitro evolution of a fungal laccase in high concentrations of organic cosolvents. Chem Biol. 2007, 14: 1052-1064. 10.1016\/j.chembiol.2007.08.010.","journal-title":"Chem Biol"},{"key":"990_CR32","doi-asserted-by":"crossref","first-page":"2654","DOI":"10.1128\/AEM.65.6.2654-2660.1999","volume":"65","author":"KC Li","year":"1999","unstructured":"Li KC, Xu F, Eriksson KEL: Comparison of fungal laccases and redox mediators in oxidation of a nonphenolic lignin model compound. Appl Environ Microbiol. 1999, 65: 2654-2660.","journal-title":"Appl Environ Microbiol"},{"key":"990_CR33","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/j.bej.2012.05.010","volume":"67","author":"U Fillat","year":"2012","unstructured":"Fillat U, Prieto A, Camarero S, Mart\u00ednez AT, Mart\u00ednez MJ: Biodeinking of flexographic inks by fungal laccases using synthetic and natural mediators. Biochem Eng J. 2012, 67: 97-103.","journal-title":"Biochem Eng J"},{"key":"990_CR34","doi-asserted-by":"publisher","first-page":"6717","DOI":"10.1016\/j.biortech.2011.03.100","volume":"102","author":"ED Babot","year":"2011","unstructured":"Babot ED, Rico A, Rencoret J, Kalum L, Lund H, Romero J, del R\u00edo JC, Mart\u00ednez AT, Guti\u00e9rrez A: Towards industrially feasible delignification and pitch removal by treating paper pulp with Myceliophthora thermophila laccase and a phenolic mediator. Bioresour Technol. 2011, 102: 6717-6722. 10.1016\/j.biortech.2011.03.100.","journal-title":"Bioresour Technol"},{"key":"990_CR35","doi-asserted-by":"publisher","first-page":"823","DOI":"10.1007\/s11274-006-9305-3","volume":"23","author":"P Sharma","year":"2007","unstructured":"Sharma P, Goel R, Capalash N: Bacterial laccases. World J Microbiol Biotechnol. 2007, 23: 823-832. 10.1007\/s11274-006-9305-3.","journal-title":"World J Microbiol Biotechnol"},{"key":"990_CR36","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1080\/07391102.2012.674275","volume":"30","author":"MT Cambria","year":"2012","unstructured":"Cambria MT, Gullotto D, Garavaglia S, Cambria A: In silico study of structural determinants modulating the redox potential of Rigidoporus lignosus and other fungal laccases. J Biomol Struct Dyn. 2012, 30: 89-101. 10.1080\/07391102.2012.674275.","journal-title":"J Biomol Struct Dyn"},{"key":"990_CR37","doi-asserted-by":"publisher","first-page":"410","DOI":"10.1016\/S0009-2614(03)00305-1","volume":"371","author":"H Kim","year":"2003","unstructured":"Kim H, Mickel M, Hampp N: Molecular origin of the stability of violuric acid radicals at high pH-values. Chem Phys Lett. 2003, 371: 410-416. 10.1016\/S0009-2614(03)00305-1.","journal-title":"Chem Phys Lett"},{"key":"990_CR38","doi-asserted-by":"publisher","first-page":"5663","DOI":"10.1021\/ja01554a021","volume":"80","author":"PA Leermakers","year":"1958","unstructured":"Leermakers PA, Hoffman WA: Chelates of violuric acid. J Am Chem Soc. 1958, 80: 5663-5667. 10.1021\/ja01554a021.","journal-title":"J Am Chem Soc"},{"key":"990_CR39","unstructured":"Drumond FM, Junqueira D, Palma De Oliveira D: Azo Dyes and Their Metabolites: Does the Discharge of the Azo Dyeinto Water Bodies Represent Humanand Ecological Risks?. Advances in Treating Textile Effluent. Edited by: Hause P. doi:10.5772\/19872http:\/\/www.intechopen.com\/books\/advances-in-treating-textile-effluent\/azo-dyes-and-their-metabolites-does-the-discharge-of-the-azo-dye-into-water-bodies-represent-human-aISBN 978-953-307-704-8"},{"key":"990_CR40","doi-asserted-by":"publisher","first-page":"1588","DOI":"10.1021\/bp049963i","volume":"20","author":"A Zille","year":"2004","unstructured":"Zille A, Ramalho P, Tzanov T, Millward R, Aires V, Cardoso MH, Ramalho MT, G\u00fcbitz GM, Cavaco-Paulo A: Predicting dye biodegradation from redox potentials. Biotechnol Prog. 2004, 20: 1588-1592. 10.1021\/bp049963i.","journal-title":"Biotechnol Prog"},{"key":"990_CR41","doi-asserted-by":"crossref","first-page":"4374","DOI":"10.1128\/aem.61.12.4374-4377.1995","volume":"61","author":"M Chivukula","year":"1995","unstructured":"Chivukula M, Renganathan V: Phenolic azo dye oxidation by laccase from Pyricularia oryzae. Appl Environ Microbiol. 1995, 61: 4374-4377.","journal-title":"Appl Environ Microbiol"},{"key":"990_CR42","doi-asserted-by":"publisher","first-page":"2230","DOI":"10.1016\/j.biortech.2009.11.066","volume":"101","author":"PP Champagne","year":"2010","unstructured":"Champagne PP, Ramsay JA: Dye decolorization and detoxification by laccase immobilized on porous glass beads. Bioresour Technol. 2010, 101: 2230-2235. 10.1016\/j.biortech.2009.11.066.","journal-title":"Bioresour Technol"},{"key":"990_CR43","doi-asserted-by":"publisher","first-page":"558","DOI":"10.1002\/biot.200800248","volume":"4","author":"V Brissos","year":"2009","unstructured":"Brissos V, Pereira L, Munteanu F-D, Cavaco-Paulo A, Martins LO: Expression system of CotA-laccase for directed evolution and high-throughput screenings for the oxidation of high-redox potential dyes. Biotechnol J. 2009, 4: 558-563. 10.1002\/biot.200800248.","journal-title":"Biotechnol J"},{"key":"990_CR44","doi-asserted-by":"publisher","first-page":"425","DOI":"10.1016\/S0969-806X(97)00318-6","volume":"53","author":"WZ Lin","year":"1998","unstructured":"Lin WZ, Navaratnam S, Yao SD, Lin NY: Antioxidative properties of hydroxycinnamic acid derivatives and a phenylpropanoid glycoside. A pulse radiolysis study. Radiat Phys Chem. 1998, 53: 425-430. 10.1016\/S0969-806X(97)00318-6.","journal-title":"Radiat Phys Chem"},{"key":"990_CR45","first-page":"33","volume":"37","author":"B Moodley","year":"2011","unstructured":"Moodley B, Mulholland DA, Brookes HC: The electro-oxidation of lignin in Sappi Saiccor dissolving pulp mill effluent. Water Sa. 2011, 37: 33-40.","journal-title":"Water Sa"},{"key":"990_CR46","doi-asserted-by":"publisher","first-page":"4595","DOI":"10.1021\/jf0352879","volume":"52","author":"K Aaby","year":"2004","unstructured":"Aaby K, Hvattum E, Skrede G: Analysis of flavonoids and other phenolic compounds using high-performance liquid chromatography with coulometric array detection: relationship to antioxidant activity. J Agric Food Chem. 2004, 52: 4595-4603. 10.1021\/jf0352879.","journal-title":"J Agric Food Chem"},{"key":"990_CR47","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1042\/bj3340063","volume":"334","author":"F Xu","year":"1998","unstructured":"Xu F, Berka RM, Wahleithner JA, Nelson BA, Shuster JR, Brown SH, Palmer AE, Solomon EI: Site-directed mutations in fungal laccase: effect on redox potential, activity and pH profile. Biochem J. 1998, 334: 63-70.","journal-title":"Biochem J"}],"container-title":["BMC Biotechnology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1472-6750-13-90.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,2]],"date-time":"2021-09-02T20:06:39Z","timestamp":1630613199000},"score":1,"resource":{"primary":{"URL":"https:\/\/bmcbiotechnol.biomedcentral.com\/articles\/10.1186\/1472-6750-13-90"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,10,26]]},"references-count":47,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2013,12]]}},"alternative-id":["990"],"URL":"https:\/\/doi.org\/10.1186\/1472-6750-13-90","relation":{},"ISSN":["1472-6750"],"issn-type":[{"value":"1472-6750","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,10,26]]},"assertion":[{"value":"19 May 2013","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 October 2013","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 October 2013","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"90"}}