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Cloning and characterization of a thermostable endo-arabinanase from Phanerochaete chrysosporium and its synergistic action with endo-xylanase

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Abstract

Putative arabinanase (PcARA) was cloned from cDNA of Phanerochaete chrysosporium. The gene sequencing indicated that PcARA consisted of 939 nucleotides that encodes for 312 amino acid arabinanase-polypeptide chain, including a signal peptide of 19 amino acids. Three-dimensional homology indicated that this enzyme is a five-bladed β-propeller, belonging to glycosidase family 43 and its secondary structure is consisted of 24 β-sheets. The PcARA-cDNA was expressed in Pichia pastoris using pPICZαC. SDS-PAGE of purified arabinanase showed a single band of 33 kDa that is very close to theoretical molecular mass of 33.9 kDa calculated by its amino acid content. Recombinant arabinanase (rPcARA) exhibited maximum activity at pH and temperature of 5.0 and 60 °C, respectively. End-product analysis of debranched arabinan hydrolysis by thin-layer chromatography indicated that rPcARA acted as endo-type. The synergistic action of rPcARA with recombinant xylanase resulted in 72 and 9.3 % release of total soluble sugar of arabinoxylan and NaOH-pretreated barley straw, respectively.

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Acknowledgments

This research was supported by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea. In partly, this research was also supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2010-0015666). We thank the Research Institute of Bioindustry at Chonbuk National University for kindly providing the facilities for this research.

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Authors and Affiliations

  1. Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk, 570-752, Korea

    Nguyen Duc Huy, Saravanakumar Thiyagarajan & Seung-Moon Park

  2. Institute of Resources, Environment and Biotechnology, Hue University, Hue, Vietnam

    Nguyen Duc Huy

  3. LED-Agribio Fusion Technology Research Center, Chonbuk National University, Iksan, Jeonbuk, 570-752, Korea

    Yoon-E Choi & Seung-Moon Park

  4. Department of Molecular Biology, College of Natural Sciences, Chonbuk National University, Jeonju, Jeonbuk, 561-756, Korea

    Dae-Hyuk Kim

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  1. Nguyen Duc Huy
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  2. Saravanakumar Thiyagarajan
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Correspondence to Seung-Moon Park.

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Huy, N.D., Thiyagarajan, S., Choi, YE. et al. Cloning and characterization of a thermostable endo-arabinanase from Phanerochaete chrysosporium and its synergistic action with endo-xylanase. Bioprocess Biosyst Eng 36, 677–685 (2013). https://doi.org/10.1007/s00449-013-0891-9

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  • DOI: https://doi.org/10.1007/s00449-013-0891-9

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