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Production and Characterization of Cellobiohydrolase from a Novel Strain of Penicillium purpurogenum KJS506

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Abstract

A high cellobiohydrolase (CBH)-producing strain was isolated and identified as Penicillium purpurogenum KJS506 according to the morphology and comparison of internal transcribed spacer rDNA gene sequence. When rice straw and corn steep powder were used as carbon and nitrogen sources, respectively, a maximum CBH activity of 2.6 U mg-protein−1, one of the highest among CBH-producing microorganisms, was obtained. The optimum temperature and pH for CBH production were 30 °C and 4.0, respectively. The increased production of CBH in P. purpurogenum culture at 30 °C was confirmed by two-dimensional electrophoresis followed by MS/MS sequencing of the partial peptide. The internal amino acid sequences of P. purpurogenum CBH showed a significant homology with hydrolases from glycoside hydrolase family 7. The extracellular CBH was purified to homogeneity by sequential chromatography of P. purpurogenum culture supernatants on a DEAE-sepharose column, a gel filtration column, and then on a Mono Q column with fast-protein liquid chromatography. The purified CBH was a monomeric protein with a molecular weight of 60 kDa and showed broad substrate specificity with maximum activity towards p-nitrophenyl β-d-cellobiopyranoside. P. purpurogenum CBH showed t 1/2 value of 4 h at 60 °C and V max value of 11.9 μmol min−1 mg-protein−1 for p-nitrophenyl-d-cellobiopyranoside. Although CBHs have been reported, the high specific activity distinguishes P. purpurogenum CBH.

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Acknowledgments

This study was supported by a grant (code 2008A0080126) from ARPC. It was also supported by a grant (Code 20070301034024) from BioGreen 21 Program, Rural Development Administration, Republic of Korea.

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Authors and Affiliations

  1. Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, 143-701, South Korea

    Kyoung-Mi Lee, Marimuthu Jeya, Kyoung-Min Lee & Jung-Kul Lee

  2. Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, 143-701, South Korea

    Ah-Reum Joo & Hee-Jung Moon

  3. Institute of SK-KU Biomaterials, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, 143-701, South Korea

    Jung-Kul Lee

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  1. Kyoung-Mi Lee
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Lee, KM., Joo, AR., Jeya, M. et al. Production and Characterization of Cellobiohydrolase from a Novel Strain of Penicillium purpurogenum KJS506. Appl Biochem Biotechnol 163, 25–39 (2011). https://doi.org/10.1007/s12010-010-9013-1

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