Document Detail

Purification and characterization of thermostable endo-1,5-alpha-L-arabinase from a strain of Bacillus thermodenitrificans.
MedLine Citation:
PMID:  11916679     Owner:  NLM     Status:  MEDLINE    
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 and two arabinofuranosidases (EC extracellularly when grown at 60 degrees 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 degrees C. The enzyme had optimal activity at 70 degrees C and pH 6.0. The enzyme had apparent K(m) values of 8.5 and 45 mg/ml and apparent V(max) 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.
Makoto Takao; Kana Akiyama; Takuo Sakai
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  68     ISSN:  0099-2240     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2002 Apr 
Date Detail:
Created Date:  2002-03-27     Completed Date:  2002-07-12     Revised Date:  2013-04-18    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1639-46     Citation Subset:  IM    
Department of Food and Nutrition, Faculty of Agriculture, Kinki University, 3327-204 Naka-machi, Nara 631-8505, Japan.
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MeSH Terms
Bacillus / enzymology*,  growth & development
Enzyme Stability
Genes, rRNA
Glycoside Hydrolases* / chemistry,  genetics,  isolation & purification,  metabolism
Hot Temperature
Molecular Sequence Data
Polysaccharides / metabolism
RNA, Ribosomal, 16S / genetics
Sequence Analysis, DNA
Substrate Specificity
Reg. No./Substance:
0/Polysaccharides; 0/RNA, Ribosomal, 16S; 11078-27-6/araban; EC 3.2.1.-/Glycoside Hydrolases; EC endo-1,5-alpha-L-arabinosidase

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