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 journalArticle

6 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 languageEnglish
Pages (from-to)677-685
Number of pages9
JournalBioprocess and Biosystems Engineering
Volume36
Issue number6
DOIs
Publication statusPublished - 2013 Jun 1
Externally publishedYes

Fingerprint

Phanerochaete
Cloning
Amino acids
Organism Cloning
Amino Acids
Complementary DNA
Thin layer chromatography
Pichia
Glycoside Hydrolases
Polypeptides
Straw
Molecular mass
Propellers
Hordeum
Thin Layer Chromatography
Nucleotides
Protein Sorting Signals
Sugars
Polyacrylamide Gel Electrophoresis
Hydrolysis

Keywords

  • Arabinanase
  • Barley straw
  • Hemicellulose
  • Phanerochaete chrysosporium
  • Pichia pastoris

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering

Cite this

Cloning and characterization of a thermostable endo-arabinanase from Phanerochaete chrysosporium and its synergistic action with endo-xylanase. / Huy, Nguyen Duc; Thiyagarajan, Saravanakumar; Choi, Yoon-E; Kim, Dae Hyuk; Park, Seung Moon.

In: Bioprocess and Biosystems Engineering, Vol. 36, No. 6, 01.06.2013, p. 677-685.

Research output: Contribution to journalArticle

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