Synergy Between an α-L-Arabinofuranosidase from Aspergillus oryzae and an Endo-Arabinanase from Streptomyces coelicolor for Degradation of Arabinan

An α-L-arabinofuranosidase gene of Aspergillus oryzae was expressed in Pichia pastoris. The re-combinant enzyme released L-arabinose from arabinose-containing polysaccharides such as lupin pectic galactan, corn hull arabinoxylan, sugar beet arabinan, and potato pectic galactan. The enzyme displayed an optimum activity at 45°C and pH 4.0. The enzyme was slowly inactivated above pH 6.0 and below pH 3.0, and was stable at temperatures up to 40°C. On the other hand, a putative endo-arabinanase gene of Strep-tomyces coelicolor was cloned and expressed in Escherichia coli. The recombinant enzyme hydrolyzed linear arabinans and produced α-1,5-arabinooligosaccharides. The enzyme displayed an optimum activity at 45°C and pH 6.0. The enzyme was slowly inactivated above pH 10.0 and below pH 4.0, and it was stable at temperatures up to 35°C. Synergisms between the α-L-arabinofuranosidase and the endo-arabinanase for the degradation of arabinan and debranched arabinan were observed. The hydrolysis was most efficient when α-L-arabinofuranosidase and endo-arabinanase were in a ratio of 95 : 5.

[1]  A. Kuno,et al.  An Exo-β-1,3-galactanase Having a Novel β-1,3-Galactan-binding Module from Phanerochaete chrysosporium* , 2005, Journal of Biological Chemistry.

[2]  H. Matsuzawa,et al.  Crystal Structure of a Family 54 α-l-Arabinofuranosidase Reveals a Novel Carbohydrate-binding Module That Can Bind Arabinose* , 2004, Journal of Biological Chemistry.

[3]  K. Yura,et al.  Structure and Function of a Family 10 β-Xylanase Chimera of Streptomyces olivaceoviridis E-86 FXYN and Cellulomonas fimi Cex* , 2004, Journal of Biological Chemistry.

[4]  A. Kuno,et al.  Purification, characterization and gene cloning of two alpha-L-arabinofuranosidases from streptomyces chartreusis GS901. , 2000, The Biochemical journal.

[5]  A. Kuno,et al.  Substrate Specificity of the α-L-Arabinofuranosidase from Trichoderma reesei , 1998 .

[6]  I. Kusakabe,et al.  Purification and Substrate Specificities of Two α-l-Arabinofuranosidases from Aspergillus awamori IFO 4033 , 1998, Applied and Environmental Microbiology.

[7]  S. Kaneko,et al.  Substrate Specificities of α-L-Arabinofuranosidases Produced by Two Species of Aspergillus niger , 1998 .

[8]  S. Kaneko,et al.  Substrate specificity of α-L-arabinofuranosidase from Streptomyces diastatochromogenes 065 toward arabinose-containing oligosaccharides , 1998 .

[9]  Kazuko Sanai,et al.  Inhibition of sucrose digestion and absorption by L-arabinose in rats , 1997 .

[10]  K Kawakubo,et al.  L-arabinose selectively inhibits intestinal sucrase in an uncompetitive manner and suppresses glycemic response after sucrose ingestion in animals. , 1996, Metabolism: clinical and experimental.

[11]  S. Kaneko,et al.  Substrate specificity of alpha-L-arabinofuranosidase from Bacillus subtilis 3-6 toward arabinofurano-oligosaccharides. , 1995, Bioscience, biotechnology, and biochemistry.

[12]  S. Kaneko,et al.  The core trisaccharide of alpha-L-arabinofuranan: synthesis of methyl 3,5-di-O-alpha-L-arabinofuranosyl-alpha-L-arabinofuranoside. , 1995, Carbohydrate research.

[13]  S. Kaneko,et al.  Synthesis of regioisomeric methyl α-l-arabinofuranobiosides , 1995 .

[14]  M. Sano,et al.  Purification and some properties of alpha-L-arabinofuranosidase from Bacillus subtilis 3-6 , 1994, Applied and environmental microbiology.

[15]  S. Kaneko,et al.  Detection of α-L-Arabinofuranosidase Activity in Isoelectric Focused Gels using 6-Bromo-2-naphthyl-α-L-arabinofuranoside , 1994 .

[16]  A Bairoch,et al.  New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. , 1993, The Biochemical journal.

[17]  S. Kaneko,et al.  Purification and some properties of intracellular alpha-L-arabinofuranosidase from Aspergillus niger 5-16. , 1993, Bioscience, biotechnology, and biochemistry.

[18]  M. Tanaka,et al.  alpha-l-Arabinofuranosidase from Radish (Raphanus sativus L.) Seeds. , 1992, Plant physiology.

[19]  Antony Bacic,et al.  8 – Structure and Function of Plant Cell Walls , 1988 .

[20]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[21]  Norton Nelson,et al.  A PHOTOMETRIC ADAPTATION OF THE SOMOGYI METHOD FOR THE DETERMINATION OF GLUCOSE , 1944 .

[22]  K. Hata,et al.  a-L-Arabinofuranosidase from Radish (Raphanus sativus 1.) Seeds , 2022 .