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Applied and Environmental Microbiology, April 2003, p. 1953-1958, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.1953-1958.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Kraft Pulp Biobleaching and Mediated Oxidation of a Nonphenolic Substrate by Laccase from Streptomyces cyaneus CECT 3335

M. Enriqueta Arias,1* María Arenas,1 Juana Rodríguez,1 Juan Soliveri,1 Andrew S. Ball,2 and Manuel Hernández1

Departamento de Microbiología y Parasitología, Universidad de Alcalá, 28871 Alcalá de Henares, Madrid, Spain,1 Department of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, United Kingdom2

Received 5 August 2002/ Accepted 6 January 2003

A new laccase (EC 1.10.3.2) produced by Streptomyces cyaneus CECT 3335 in liquid media containing soya flour (20 g per liter) was purified to homogeneity. The physicochemical, catalytic, and spectral characteristics of this enzyme, as well as its suitability for biobleaching of eucalyptus kraft pulps, were assessed. The purified laccase had a molecular mass of 75 kDa and an isoelectric point of 5.6, and its optimal pH and temperature were 4.5 and 70°C, respectively. The activity was strongly enhanced in the presence of Cu2+, Mn2+, and Mg2+ and was completely inhibited by EDTA and sodium azide. The purified laccase exhibited high levels of activity against 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) and 2,6-dimethoxyphenol and no activity against tyrosine. The UV-visible spectrum of the purified laccase was the typical spectrum of the blue laccases, with an absorption peak at 600 nm and a shoulder around 330 to 340 nm. The ability of the purified laccase to oxidize a nonphenolic compound, such as veratryl alcohol, in the presence of ABTS opens up new possibilities for the use of bacterial laccases in the pulp and paper industry. We demonstrated that application of the laccase from S. cyaneus in the presence of ABTS to biobleaching of eucalyptus kraft pulps resulted in a significant decrease in the kappa number (2.3 U) and an important increase in the brightness (2.2%, as determined by the International Standard Organization test) of pulps, showing the suitability of laccases produced by streptomycetes for industrial purposes.

* Corresponding author. Mailing address: Departamento de Microbiología y Parasitología, Universidad de Alcalá, 28871 Alcalá de Henares, Madrid, Spain. Phone: 34 91 8854633. Fax: 34 91 8854623. E-mail: enriqueta.arias{at}uah.es.

Applied and Environmental Microbiology, April 2003, p. 1953-1958, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.1953-1958.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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