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Cloning and characterization of an exoinulinase from Bacillus polymyxa

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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 K m and V max 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.

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Authors and Affiliations

  1. Mitsubishi Kagaku Institute of Life Sciences, Tokyo, 194-8511, Japan

    Hyun-Ju Kwon

  2. The Special Division for Human Life Technology, National Institute of Advanced Industrial Science and Technology (Kansai), Osaka, 563-8577, Japan

    Sung-Jong Jeon

  3. Department of Microbiology, Graduate School, Dongeui University, Pusan, 614-714, Korea

    Dong-Ju You, Kwang-Hyeon Kim, Yong-Kee Jeong, Young-Hee Kim & Byung-Woo Kim

  4. Department of Food Science and Nutrition, Graduate School, Dongeui University, Pusan, 614-714, Korea

    Young-Man Kim

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  1. Hyun-Ju Kwon
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  2. Sung-Jong Jeon
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  3. Dong-Ju You
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  4. Kwang-Hyeon Kim
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  5. Yong-Kee Jeong
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  6. Young-Hee Kim
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  7. Young-Man Kim
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  8. Byung-Woo Kim
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Correspondence to Byung-Woo Kim.

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Kwon, HJ., Jeon, SJ., You, DJ. et al. Cloning and characterization of an exoinulinase from Bacillus polymyxa . Biotechnology Letters 25, 155–159 (2003). https://doi.org/10.1023/A:1021987923630

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  • DOI: https://doi.org/10.1023/A:1021987923630

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