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- Title
Cloning and characterization of an exoinulinase from Bacillus polymyxa.
- Authors
Hyun-Ju Kwon; Sung-Jong Jeon; Dong-Ju You; Kwang-Hyeon Kim; Yong-Kee Jeong; Young-Hee Kim; Young-Man Kim; Byung-Woo Kim
- Abstract
A gene encoding an exoinulinase (inu) from Bacillus polymyxa MGL21 was cloned and sequenced. It is composed of 1455 nucleotides, encoding a protein (485 amino acids) with a molecular mass of 55 522 Da. Inu was expressed in Escherichia coli and the His-tagged exoinulinase was purified. The purified enzyme hydrolyzed sucrose, levan and raffinose, in addition to inulin, with a sucrose/inulin ratio of 2. Inulinase activity was optimal at 35 °C and pH 7, was completely inactivated by 1 mM Ag+ or Hg2+. The Km and Vmax values for inulin hydrolysis were 0.7 mM and 2500 μM min-1 mg-1 protein. The enzyme acted on inulin via an exo-attack to produce fructose mainly.
- Subjects
BACILLUS (Bacteria); ENTEROBACTERIACEAE; HYDROGEN-ion concentration; INULIN; ESCHERICHIA coli; GENETIC engineering
- Publication
Biotechnology Letters, 2003, Vol 25, Issue 2, p155
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1023/A:1021987923630