ORIGINAL PAPERA new strain of Bjerkandera sp. production, purificationand characterization of versatile peroxidaseRoberto Taboada-Puig • Thelmo Lu´-Chau •Mar? ´a Teresa Moreira • Gumersindo Feijoo •Mar? ´a Jesu´s Mart? ´nez • Juan Manuel LemaReceived: 25 November 2009/Accepted: 29 April 2010/Published online: 14 May 2010? Springer Science+Business Media B.V. 2010Abstract The lignin modifying enzymes (LMEs) secretedby a new white rot fungus isolated from Chile were studiedin this work. This fungus has been identified as a new ana-morph of Bjerkandera sp. based on the sequences of theribosomal DNA and morphological analysis at lightmicroscopy showing hyaline hyphae without clamp con-nection,cylindricalconidiaandlackofsexualforms,similarto those reported in other Bjerkandera anamorphs. ThecharacterizationoftheculturemediumforthehighestLMEsproduction was performed in flask cultures, with a formu-lation of the culture medium containing high levels of glu-cose and peptone. The highest Mn-oxidizing peroxidaseactivity (1,400 U/L) was achieved on day 6 in Erlenmeyerflasks. Four peroxidases (named R1B1, R1B2, R1B3 andR1B4), have been purified by using ion-exchange andexclusion molar chromatographies. All of them showedtypical activity on Mn 2? and exhibited Mn-independentactivity against 2,6-dimethoxyphenol. R1B4 showed alsoactivity on veratryl alcohol (pH 3) indicating that thisenzyme belongs to the versatile peroxidase family. The highVP production capacities of this strain, as well asthe enzymatic characteristics of the LMEs suggest that itmay be successfully used in the degradation of recalcitrantcompounds.Keywords Bjerkandera sp. ? Anamorph ? ITS analysis ?Versatile peroxidaseIntroductionThe degradation of compounds introduced by mankind inthe environment means an important ecological challengeas these compounds have complex structure and low bio-availability. Conventional physicochemical or biologicaltreatment plants are only able of a partial degradation whileadvanced oxidation processes such as UV exposure orozonation render variable yields of degradation. A bio-logical alternative may be based on the use of certainfungal strains, known as white rot fungi. It has beenreported that these microorganisms degrade a wide rangeof organic compounds such as dyes (Wesenberg et al.2003), polycyclic aromatic compounds (Valent? ´n et al.2007), etc. This ability is attributed to lignin modifyingenzymes (LMEs) synthesized during secondary metabo-lism in response to nutrient limitation. Two ligninolyticperoxidases, lignin peroxidase (LiP) (Tien and Kirk 1988)and manganese peroxidase (MnP) (Kuwahara et al. 1984),were described in Phanerochaete chrysosporium andreported in other white-rot fungi from the group of basid-iomycetes (Hatakka 1994). In addition, some fungi containanother lignin-degrading enzyme, versatile peroxidase(VP), also known as hybrid Mn-peroxidase, whichcombines properties of LiP and MnP (Mart? ´nez 2002;Ruiz-Duenas et al. 2009), thus, they are able to oxidizeMn 2? to Mn 3? , as well as non-phenolic aromatic com-pounds. However, its catalytic cycle in presence of Mn 2? isR. Taboada-Puig ? M. T. Moreira (&) ? G. Feijoo ? J. M. LemaDepartment of Chemical Engineering, School of Engineering,University of Santiagode Compostela, 15782 Santiago de Compostela, Spaine-mail: maite.moreira@usc.esT. Lu´-ChauDepartment of Chemical Engineering, Institute of Technology,15706 Santiago de Compostela, SpainM. J. Mart? ´nezCentro de Investigaciones Biolo´gicas, Consejo Superior deInvestigaciones Cient? ´ficas, Ramiro de Maeztu 28040 Madrid,Spain123World J Microbiol Biotechnol (2011) 27:115–122DOI 10.1007/s11274-010-0435-2
