Abstract
A recombinant putative glycoside hydrolase from Caldicellulosiruptor saccharolyticus was purified with a specific activity of 12 U mg−1 by heat treatment and His-Trap affinity chromatography, and identified as a single 56 kDa band upon SDS-PAGE. The native enzyme is a dimer with a molecular mass of 112 kDa as determined by gel filtration. The enzyme exhibited its highest activity when debranched arabinan (1,5-α-l-arabinan) was used as the substrate, demonstrating that the enzyme was an endo-1,5-α-l-arabinanase. The K m, k cat, and k cat/K m values were 18 mg ml−1, 50 s−1, and a 2.8 mg ml−1 s−1, respectively. Maximum enzyme activity was at pH 6.5 and 75°C. The half-lives of the enzyme at 65, 70 and 75°C were 2440, 254 and 93 h, respectively, indicating that it is the most thermostable of the known endo-1,5-α-l-arabinanases.
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Acknowledgments
This study was carried out with the support of ‘Forest Science and Technology Projects (Project No. S210707L010120)’ provided by Korea Forest Service and by a grant from Agricultural R&D Promotion Center, Korea Rural Economic Institute, Republic of Korea.
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Hong, MR., Park, CS. & Oh, DK. Characterization of a thermostable endo-1,5-α-l-arabinanase from Caldicellulorsiruptor saccharolyticus . Biotechnol Lett 31, 1439–1443 (2009). https://doi.org/10.1007/s10529-009-0019-0
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DOI: https://doi.org/10.1007/s10529-009-0019-0