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Studies on the Pycnoporus sanguineus CCT-4518 laccase purified by hydrophobic interaction chromatography

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Abstract

A laccase from Pycnoporus sanguineus was purified by two steps using phenyl-Sepharose columm. A typical procedure provided 54.1-fold purification, with a yield of 8.37%, using syringaldazine as substrate. The molecular weight of the purified laccase was 69 and 68 kDa as estimated by 12% (w/v) SDS-PAGE gel and by gel filtration, respectively. The K m values for the substrates ABTS, syringaldazine, and guaiacol were 58, 8.3, and 370 μM, respectively. The enzyme’s pH optimum for syringaldazine was 4.2 and optimal activity was 50°C. The enzyme showed to be thermostable because when kept at 50°C for 24 and 48 h it retained 93 and 76% activity. This laccase was inhibited by l-cysteine, β-mercaptoethanol, NaN3, NaF, and HgCl2.

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Acknowledgments

This work was supported by Grants from CNPq, FUNAPE/UFG, and International Foundation for Science, Stockholm, Sweden, through a grant to Dr MFS (IFS-W/3433-1). TAG was supported by CAPES/Brazil.

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Authors and Affiliations

  1. Faculdade de Farmácia, Universidade Federal de Goiás, 74001-970, Goiânia, GO, Brazil

    Telma Alves Garcia & Mariângela Fontes Santiago

  2. Departamento de Bioquímica -ICB, Universidade Federal de Goiás, 74001-970, Goiânia, GO, Brazil

    Cirano José Ulhoa

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  1. Telma Alves Garcia
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  2. Mariângela Fontes Santiago
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  3. Cirano José Ulhoa
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Correspondence to Cirano José Ulhoa.

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Garcia, T.A., Santiago, M.F. & Ulhoa, C.J. Studies on the Pycnoporus sanguineus CCT-4518 laccase purified by hydrophobic interaction chromatography. Appl Microbiol Biotechnol 75, 311–318 (2007). https://doi.org/10.1007/s00253-006-0817-4

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  • DOI: https://doi.org/10.1007/s00253-006-0817-4

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