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Expression and Secretion of Bifidobacterium  adolescentis Amylase by Bifidobacterium  Longum

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Abstract

Bifidobacterium  adolescentis Int-57 (INT57), isolated from human feces, secretes an amylase. We have shot-gun cloned, sequence analyzed and expressed the gene encoding this amylase in B. longum. The sequenced 2477 bp fragment was homologous to other extracellular amylases. The encoded protein was predicted to be composed of 595 amino acids with a molecular weight of 64 kDa, and was designated AmyB. Highly conserved amylase domains were found in AmyB. The signal sequence and cleavage site was predicted by sequence analysis. AmyB was subcloned into pBES2, a novel E. coli–Bifidobacterium shuttle vector, to construct pYBamy59. Subsequently, B. longum, with no apparent amylase activity, was transformed with pYBamy59. More than 90% of the amylase activity was detected in the culture broth. This approach may open the way for the development of more efficient expression and secretion systems for Bifidobacterium.

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Authors and Affiliations

  1. Research Center, BIFIDO Co., Ltd., Seoul, 151-818, Korea

    Myeong Soo Park & Geun Eog Ji

  2. Department of Genetic Engineering, Hallym University, Kangwon-Do, Chuncheon, 200-702, Korea

    Seong Lyul Rhim

  3. Department of Food Science and Nutrition, Seoul National University, Seoul, 151-742, Korea

    Geun Eog Ji

  4. Department of Nutrition & Hotel Culinary Art, Anyang Technical College, Gyeonggi, 430-749, Korea

    Myeong Soo Park

Authors
  1. Seong Lyul Rhim
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  2. Myeong Soo Park
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  3. Geun Eog Ji
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Correspondence to Geun Eog Ji.

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Both authors contributed equally

Received 17 June 2005; Revisions requested 13 July 2005 and 26 September 2005; Revisions received 12 September 2005 and 8 November 2005; Accepted 11 November 2005

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Rhim, S.L., Park, M.S. & Ji, G.E. Expression and Secretion of Bifidobacterium  adolescentis Amylase by Bifidobacterium  Longum. Biotechnol Lett 28, 163–168 (2006). https://doi.org/10.1007/s10529-005-5330-9

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  • DOI: https://doi.org/10.1007/s10529-005-5330-9

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