Miguel Alcalde *

Autor invitado por SIIC

El trabajo contribuye al diseño de plataformas evolutivas para el diseño de lacasas con alto potencial redox

DISEÑO DE PLATAFORMAS PAR LA EVOLUCION DIRIGIDA DE LA LACASA DE PYCNOPORUS CINNABARINUS

Se ha abordado el diseño de lacasas de alto potencial redox mediante ciclos iterativos de evolución dirigida con el fin de conseguir enzimas sumamente estables, solubles y activas. Los mutantes diseñados son una plataforma ideal con la que afrontar nuevos retos en biología sintética.

*Miguel Alcalde
describe para SIIC los aspectos relevantes de su trabajo
ENGINEERING PLATFORMS FOR DIRECTED EVOLUTION OF LACCASE FROM PYCNOPORUS CINNABARINUS
Institución principal de la investigación
*Instituto de Catálisis, Consejo Superior de Investigaciones Cientificas (CSIC), Madrid, España
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Otros artículos escogidos

Referencias bibliográficas
1. González-Pérez D, García-Ruiz E, Alcalde M. Saccharomyces cerevisiae in directed evolution: an efficient tool to improve enzymes. Bioengineering Bugs 3:1-6, 2012.
2. Camarero S, Pardo I, Cañas AI, Molina P, Record E, Martínez AT, Martínez MJ, Alcalde M. Engineering platforms for directed evolution of laccase from Pycnoporus cinnabarinus. Applied & Environmental Microbiology 78:1370-1384, 2012.
3. García-Ruiz E, González-Pérez D, Ruiz-Dueñas FJ, Martínez AT, Alcalde M. Directed evolution of a temperature, peroxide and alkaline pH tolerant versatile peroxidase. Biochemical Journal 441:487-498, 2012.
4. Alcalde M. Fundamentos de evolución molecular dirigida de enzimas. In: Monografías de la Real Academia Nacional de Farmacia. Sánchez-Montero JM ed. En prensa.
5. De Abreu MA, Alvaro-Benito M, Plou FJ, Fernández-Lobato M, Alcalde M. Screening -fructofuranosidases mutant libraries to enhance the transglycosylation rates of β-(26) fructooligosaccharides. Combinatorial Chemistry & High-Throughput Screening 14:730-738, 2011.
6. Mate D, García-Ruiz E, Camarero S, Alcalde M. Directed evolution of fungal laccases. Current Genomics 12:113-122, 2011.
7. Mate D, García-Burgos C, García E, Ballesteros A, Camarero S, Alcalde M. Laboratory evolution of high redox potential laccases. Chemistry & Biology 17:1030-1041, 2010. Full paper seleccionado para preview en Chemistry & Biology, Ref: Sayut DJ, Sun L. Creating designer laccases. Chem Biol 27:918-920, 2010.
8. García E, Maté D, Ballesteros A, Martínez AT, Alcalde M. Evolving thermostability in mutant libraries of ligninolytic oxidoreductases expressed in yeast. Microbial Cell Factories 9:17, 2010.
9. Alcalde M. Mutagenesis protocols in Saccharomyces cerevisiae by in vivo overlap extension. In: In vitro mutagenesis protocols, 3rd ed. Methods in Molecular Biology 634. (Bramman J. ed) Totowa, New Jersey: Springer-Humana Press. 3-15. ISBN: 978-1-60761-651-1.
10. Zumárraga M, Vaz Domínguez C, Camarero S, Shleev S, Polaina J, Martínez-Arias A, Ferrer M, De Lacey A, Fernández V, Ballesteros A, Plou FJ, Alcalde M. Combinatorial saturation mutagenesis of the Myceliophthora thermophila laccase T2 mutant: the connection between the C-terminal plug and the conserved 509VSG511 tripeptide. Combinatorial Chemistry & High-Throughput Screening 11:807-816, 2008.
11. Zumárraga M, Camarero S, Martínez-Arias A, Ballesteros A, Plou FJ, Alcalde M. Altering the laccase functionality by in vivo assembly of mutant libraries with different mutational spectra. Proteins: Structure, Function and Bioinformatics 71:250-260, 2010.
12. Kunamneni A, Camarero S, García C, Plou FJ, Ballesteros A, Alcalde M. Engineering and applications of fungal laccases for organic sinthesis. Microbial Cell Factories 7(32):1-17, 2008.
13. Kunamneni A, Plou FJ, Ballesteros A, Alcalde M. Laccase and their applications: a patent review. Recent Patents on Biotechnology 2:10-24, 2008.
14. Ferrer M, Beloqui A, Zumárraga M, Alcalde M, Golyshin PN. Microbes and enzymes: recent trends and new directions to expand protein space. In: Protein engineering handbook. Eds.: Lutz S, Bornscheuer UT, Wiley-VCH Verlag GMBH & Co; 1 pp. 233-264, 2008.
15. Zumárraga M, Bulter T, Shleev S, Polaina J, Plou FJ, Ballesteros A, Alcalde M. In vitro evolution of a fungal laccase in high concentrations of organic cosolvents. Chemistry & Biology 14:1052-1064, 2007.
16. Cañas AI, Alcalde M, Plou FJ, Martínez MJ, Martínez AT, Camarero S. Transformation of polycyclic aromatic hydrocarbons by laccase is strongly enhanced by phenolic compounds present in soil Environmental Science and Technology 47:2964-2971, 2007.
17. Zumárraga M, Plou FJ, García H, Ballesteros A, Alcalde M. Biorremediation of polycyclic aromatic hydrocarbons by a fungal laccases engineered by directed evolution. Biocatalysis and Biotransformation 25:219-228, 2007.
18. Alcalde M, Ferrer M, Plou FJ. Environmental biocatalysis: from remediation with enzymes to novel green processes. Biocatalysis and Biotransformation 25:113, 2007.
19. Kunamneni A, Ballesteros A, Plou FJ, Alcalde M. Fungal laccase: a versatile enzyme for biotechnological applications. In: Communicating current research and educational topics and trends in applied microbiology. Ed: Méndez-Vilas. Microbiology Book Series, Formatex Publishers, Spain: 233-245, 2007.
20. Alcalde M. Laccase: biological functions, molecular structure and industrial applications. In: Industrial enzymes: structure, function and applications. Eds: Polaina J, MacCabe AP. Springer, New York, pp. 459-474, 2007.
21. Alcalde M, Zumárraga M, Polaina J, Ballesteros A, Plou FJ. Combinatorial saturation mutagenesis by in vivo overlap extension for the engineering of fungal laccases. Combinatorial Chemistry and High-Throughput Screening 9:719-727, 2006.
22. Alcalde M, Ferrer M, Plou FJ, Ballesteros A. Environmental biocatalysis: from remediation with enzymes to novel green processes. Trends in Biotechnology 24:281-287, 2006.
23. Beloqui A, Pita M, Polaina J, Martínez A, Golyshina O, Zumárraga M, Yakimov M, García H, Alcalde M, Fernández V, Elborough K, Andreu J, Ballesteros A, Plou FJ, Timmis K, Ferrer M, Golyshin P. Novel polyphenol oxidase mined from a metagenome expression library of bovine rumen. The Journal of Biological Chemistry 281:22933-22942, 2006.
24. Mencía M, Martínez A, Alcalde M, De Lorenzo V. Identification of a -hexachlorocyclohexane dehydrochlorinase (LinA) variant with improved expression and solubility properties. Biocatalysis and Biotransformation 24:223-230, 2006.
25. Reyes-Duarte D, Polaina J, López-Cortés N, Alcalde M, Plou FJ, Elborough K, Ballesteros A, Timmis KN, Golyshin PN, Ferrer M. Conversion of a carboxylesterase into a triacylglycerol lipase by random mutation. Angewandte Chemie-International Edition 44:7553-7557, 2005.
26. Alcalde M, Bulter T, Zumárraga M, García H, Mencía M, Plou FJ, Ballesteros A. Screening mutant libraries of fungal laccases in the presence of organic solvents. Journal of Biomolecular Screening 10:624-631, 2005.
27. García-Arellano H, Alcalde M, Ballesteros A. Use and improvement of microbial redox enzymes for environmental purposes. Microbial Cell Factories 3:10-13, 2004.
28. Alcalde M, Farinas ET, Arnold FH. Colorimetric high-throughput assay for alkene epoxidation catalyzed by cytochrome P450 BM-3 variant 139-3. Journal of Biomolecular Screening 9:141-146, 2004.
29. Farinas ET, Alcalde M, Arnold FH. Alkene epoxidation catalysed by cytochrome P450 BM-3 mutant 139-3. Tetrahedron 60:525-528, 2004.
30. Bulter T, Alcalde M, Sieber V, Meinhold P, Schlachtbauer C, Arnold FH. Functional expression of a fungal laccase in Saccharomyces cerevisiae by directed evolution. Applied Environmental Microbiology 69:987-995, 2003.
31. Bulter T, Alcalde M. Preparing libraries in Saccharomyces cerevisiae. In: Directed evolution library creation. Methods and protocols. Methods in molecular biology Vol. 231. Eds: Arnold FH, Gergiou G. Humana Press, Totowa, New Jersey: 17-22, 2003.
32. Bulter T, Sieber V, Alcalde M. Screening mutant libraries in Saccharomyces cerevisiae. In: Directed enzyme evolution. Screening and selection methods. Methods in molecular biology Vol. 230. Eds: Arnold FH, Gergiou G. Humana Press, Totowa, New Jersey, pp. 99-108, 2003.
33. Alcalde M, Bulter T. Colorimetric assays for screening laccases. In: Directed enzyme evolution. Screening and selection methods. Methods in molecular biology Vol. 230. Eds: Arnold FH, Gergiou G. Humana Press, Totowa, New Jersey, pp. 193-202, 2003.
34. Alcalde M. Evolución molecular dirigida. Investigación y Ciencia. (Versión española de Scientific American) 326:69-77, 2003.
35. Alcalde M, Bulter T, Arnold FH. Colorimetric assays for biodegradation of polycyclic aromatic hydrocarbons by fungal laccases. Journal of Biomolecular Screening 7:537-543, 2002.
36. Sun L, Bulter T, Alcalde M, Petronia I, Arnold FH. Modification of galactose oxidase to introduce glucose 6-oxidase activity. ChemBioChem 8:781-783, 2002.

Otros artículos de Miguel Alcalde

González-Pérez D, García-Ruiz E, Alcalde M. Saccharomyces cerevisiae in directed evolution: an efficient tool to improve enzymes. Bioengineering Bugs 3:1-6, 2012.
Mate D, García-Burgos C, García E, Ballesteros A, Camarero S, Alcalde M. Laboratory evolution of high redox potential laccases. Chemistry & Biology 17:1030-1041, 2010.
Sayut DJ, Sun L. Creating designer laccases. Chem Biol 27:918-920, 2010.
Camarero S, Pardo I, Cañas AI, Molina P, Record E, Martínez AT, Martínez MJ, Alcalde M. Engineering platforms for directed evolution of laccase from Pycnoporus cinnabarinus. Applied & Environmental Microbiology 78:1370-1384, 2012.
García-Ruiz E, González-Pérez D, Ruiz-Dueñas FJ, Martínez AT, Alcalde M. Directed evolution of a temperature, peroxide and alkaline pH tolerant versatile peroxidase. Biochemical Journal 441:487-498, 2012.

Mate D, García-Ruiz E, Camarero S, Alcalde M. Directed evolution of fungal laccases. Current Genomics 12:113-122, 2012.
García E, Maté D, Ballesteros A, Martínez AT, Alcalde M. Evolving thermostability in mutant libraries of ligninolytic oxidoreductases expressed in yeast. Microbial Cell Factories 9:17, 2010.
Zumárraga M, Vaz Domínguez C, Camarero S, Shleev S, Polaina J, Martínez-Arias A, Ferrer M, De Lacey A, Fernández V, Ballesteros A, Plou FJ, Alcalde M. Combinatorial saturation mutagenesis of the Myceliophthora thermophila laccase T2 mutant: the connection between the C-terminal plug and the conserved 509VSG511 tripeptide. Combinatorial Chemistry & High-Throughput Screening 11:807-816, 2008.
Zumárraga M, Camarero S, Martínez-Arias A, Ballesteros A, Plou FJ, Alcalde M. Altering the laccase functionality by in vivo assembly of mutant libraries with different mutational spectra. Proteins: Structure, Function and Bioinformatics 71:250-260, 2008.
Zumárraga M, Bulter T, Shleev S, Polaina J, Plou FJ, Ballesteros A, Alcalde M. In vitro evolution of a fungal laccase in high concentrations of organic cosolvents. Chemistry & Biology 14:1052-1064, 2007.
Zumárraga M, Plou FJ, García H, Ballesteros A, Alcalde M. Biorremediation of polycyclic aromatic hydrocarbons by a fungal laccases engineered by directed evolution. Biocatalysis and Biotransformation 25:219-228, 2007.
Alcalde M, Zumárraga M, Polaina J, Ballesteros A, Plou FJ. Combinatorial saturation mutagenesis by in vivo overlap extension for the engineering of fungal laccases. Combinatorial Chemistry and High-Throughput Screening 9:719-727, 2006.
Alcalde M, Ferrer M, Plou FJ, Ballesteros A. Environmental biocatalysis: from remediation with enzymes to novel green processes. Trends in Biotechnology 24:281-287, 2006.
Bulter T, Alcalde M, Sieber V, Meinhold P, Schlachtbauer C, Arnold FH. Functional expression of a fungal laccase in Saccharomyces cerevisiae by directed evolution. Applied Environmental Microbiology 69:987-995, 2003.

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DISEÑO DE PLATAFORMAS PAR LA EVOLUCION DIRIGIDA DE LA LACASA DE PYCNOPORUS CINNABARINUS

Título original en castellano
DISEÑO DE PLATAFORMAS PAR LA EVOLUCION DIRIGIDA DE LA LACASA DE PYCNOPORUS CINNABARINUS

Autor
Miguel Alcalde¹
1 Científico en Plantilla Csic, Instituto de Catálisis, Consejo Superior de Investigaciones Cientificas (CSIC), Madrid, España, Cientifico Titular

Acceso a la fuente original
Applied and Environmental Microbiology
http://aem.asm.org/

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