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Applied and Environmental Microbiology, April 2002, p. 1639-1646, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1639-1646.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Purification and Characterization of Thermostable Endo-1,5-{alpha}-L-Arabinase from a Strain of Bacillus thermodenitrificans

Makoto Takao, Kana Akiyama, and Takuo Sakai*

Department of Food and Nutrition, Faculty of Agriculture, Kinki University, 3327-204 Naka-machi, Nara 631-8505, Japan

Received 27 August 2001/ Accepted 8 January 2002

A strain of a thermophilic bacterium, tentatively designated Bacillus thermodenitrificans TS-3, with arabinan-degrading activity was isolated. It produced an endo-arabinase (ABN) (EC 3.2.1.99) and two arabinofuranosidases (EC 3.2.1.55) extracellularly when grown at 60°C on a medium containing sugar beet arabinan. The ABN (tentatively called an ABN-TS) was purified 7,417-fold by anion-exchange, hydrophobic, size exclusion, and hydroxyapatite chromatographies. The molecular mass of ABN-TS was 35 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the isoelectric point was pH 4.5. The enzyme was observed to be more thermostable than known ABNs; it had a half-life of 4 h at 75°C. The enzyme had optimal activity at 70°C and pH 6.0. The enzyme had apparent Km values of 8.5 and 45 mg/ml and apparent Vmax values of 1.6 and 1.1 mmol/min/mg of protein against debranched arabinan ({alpha}-1,5-arabinan) and arabinan, respectively. The enzyme had no pectin-releasing activity (protopectinase activity) from sugar beet protopectin, differing from an ABN (protopectinase-C) from mesophilic Bacillus subtilis IFO 3134. The pattern of degradation of debranched arabinan by ABN-TS indicated that the enzyme was an endo-acting enzyme and the main end products were arabinobiose and arabinose. The results of preliminary experiments indicated that the culture filtrate of strain TS-3 is suitable for L-arabinose production from sugar beet pulp at high temperature.

* Corresponding author. Mailing address: Department of Food and Nutrition, Faculty of Agriculture, Kinki University, 3327-204 Naka-machi, Nara 631-8505, Japan. Phone: 81-742-43-1511. Fax: 81-742-43-2252. E-mail: tsakaiku{at}nara.kindai.ac.jp.

Applied and Environmental Microbiology, April 2002, p. 1639-1646, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1639-1646.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Inacio, J. M., de Sa-Nogueira, I. (2008). Characterization of abn2 (yxiA), Encoding a Bacillus subtilis GH43 Arabinanase, Abn2, and Its Role in Arabino-Polysaccharide Degradation. J. Bacteriol. 190: 4272-4280 [Abstract] [Full Text]  
  • Yamaguchi, A., Tada, T., Wada, K., Nakaniwa, T., Kitatani, T., Sogabe, Y., Takao, M., Sakai, T., Nishimura, K. (2005). Structural Basis for Thermostability of Endo-1,5-{alpha}-l-Arabinanase from Bacillus thermodenitrificans TS-3. J Biochem 137: 587-592 [Abstract] [Full Text]  
  • Margolles, A., de los Reyes-Gavilan, C. G. (2003). Purification and Functional Characterization of a Novel {alpha}-L-Arabinofuranosidase from Bifidobacterium longum B667. Appl. Environ. Microbiol. 69: 5096-5103 [Abstract] [Full Text]  


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