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Purification and characterization of a β-1,4-glucosidase from a newly isolated strain of Fomitopsis pinicola

  • Biotechnologically Relevant Enzymes and Proteins
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Abstract

An efficient ß-1,4-glucosidase (BGL) producing strain, Fomitopsis pinicola KMJ812, was isolated and identified based on morphological features and sequence analysis of internal transcribed spacer rDNA. An extracellular BGL was purified to homogeneity by sequential chromatography of F. pinicola culture supernatants on a DEAE-sepharose column, a gel filtration column, and then on a Mono Q column with fast protein liquid chromatography. The relative molecular weight of F. pinicola BGL was determined to be 105 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis, or 110 kDa by size exclusion chromatography, indicating that the enzyme is a monomer. The hydrolytic activity of the BGL had a pH optimum of 4.5 and a temperature optimum of 50°C. The enzyme showed high substrate specificity and high catalytic efficiency (k cat = 2,990 s−1, K m = 1.76 mM, k cat/K m = 1,700 mM−1 s−1) for p-nitrophenyl-β-d-glucopyranoside. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase family 3, indicating that the F. pinicola BGL is a member of glycoside hydrolase family 3. Although BGLs have been purified and characterized from several other sources, F. pinicola BGL is distinguished from other BGLs by its high catalytic efficiency and strict substrate specificity.

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Acknowledgment

This study was supported by a grant (S210707L010120) from Korea Forest Service.

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Authors and Affiliations

  1. Department of Chemical Engineering, Konkuk University, Seoul, 143-701, Korea

    Ah-Reum Joo, Marimuthu Jeya, Kyoung-Mi Lee, Won-Il Sim, Jung-Soo Kim, In-Won Kim & Jung-Kul Lee

  2. Institute of Biomedical Science and Technology, Konkuk University, Seoul, 143-701, Korea

    Kyoung-Mi Lee & Jung-Kul Lee

  3. Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, Korea

    Ah-Reum Joo & Deok-Kun Oh

  4. Department of Forest Products, College of Forest Science, Kookmin University, Seoul, 136-702, Korea

    Yeong-Suk Kim

  5. Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, Madurai, 625021, India

    Paramasamy Gunasekaran

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  1. Ah-Reum Joo
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  2. Marimuthu Jeya
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Correspondence to Jung-Kul Lee.

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Joo, AR., Jeya, M., Lee, KM. et al. Purification and characterization of a β-1,4-glucosidase from a newly isolated strain of Fomitopsis pinicola . Appl Microbiol Biotechnol 83, 285–294 (2009). https://doi.org/10.1007/s00253-009-1861-7

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  • DOI: https://doi.org/10.1007/s00253-009-1861-7

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