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

Nguyen Duc Huy; Saravanakumar Thiyagarajan; Yoon E. Choi; Dae Hyuk Kim; Seung Moon Park

doi:10.1007/s00449-013-0891-9

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

Nguyen Duc Huy, Saravanakumar Thiyagarajan, Yoon E. Choi, Dae Hyuk Kim, Seung Moon Park

Research output: Contribution to journal › Article › peer-review

13 Citations (Scopus)

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.

Original language	English
Pages (from-to)	677-685
Number of pages	9
Journal	Bioprocess and Biosystems Engineering
Volume	36
Issue number	6
DOIs	https://doi.org/10.1007/s00449-013-0891-9
Publication status	Published - 2013 Jun
Externally published	Yes

Bibliographical note

Funding Information:
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.

Keywords

Arabinanase
Barley straw
Hemicellulose
Phanerochaete chrysosporium
Pichia pastoris

ASJC Scopus subject areas

Biotechnology
Bioengineering

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10.1007/s00449-013-0891-9

Cite this

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

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.",

keywords = "Arabinanase, Barley straw, Hemicellulose, Phanerochaete chrysosporium, Pichia pastoris",

author = "Huy, {Nguyen Duc} and Saravanakumar Thiyagarajan and Choi, {Yoon E.} and Kim, {Dae Hyuk} and Park, {Seung Moon}",

note = "Funding Information: 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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AU - Huy, Nguyen Duc

AU - Thiyagarajan, Saravanakumar

AU - Choi, Yoon E.

AU - Kim, Dae Hyuk

AU - Park, Seung Moon

N1 - Funding Information: 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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N2 - 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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Cloning and characterization of a thermostable endo-arabinanase from Phanerochaete chrysosporium and its synergistic action with endo-xylanase

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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