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Cloning and expression of a trehalose synthase from Pseudomonas stutzeri CJ38 in Escherichia coli for the production of trehalose

  • Applied Genetics and Molecular Biotechnology
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Abstract

A novel strain was isolated, Pseudomonas stutzeri CJ38, that enabled direct transformation of maltose to trehalose. In comparison with others reported to date, CJ38 provided a novel trehalose synthase (TSase) without any byproduct, including glucose. Activity analysis, using either maltose or trehalose as a substrate, showed a reversible reaction. There was also no detectable activity of related enzymes with liquid starch and maltooligosaccharides as substrates. Using a malPQ-negative host and MacConkey medium, the TSase gene was cloned in Escherichia coli from CJ38. The resulting sequence contained an open reading frame consisted of 689 amino acids with a calculated molecular mass of 76 kDa. A search for related sequences in various gene and protein data banks revealed a novel family of enzymes that was predicted putatively as a glycosidase or TSase family, with no biochemical evidence. The recombinant enzyme exhibited a high activity toward the substrate maltose, about 50-fold higher than the parent strain and resulted in a high conversion yield (72%) at a relatively high substrate concentration (20%). These results provided the possibility that the strain was effectively used as a potential biocatalyst for the production of trehalose from maltose in a one-step reaction.

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Acknowledgements

We thank E.K. Song and Y.J. Jeon for many helpful comments. This work was supported by the Development Program for Industrial Platform Technology of the Ministry of Commerce, Industry and Energy of Korea

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Authors and Affiliations

  1. R&D Center for Bioproducts, CJ Corporation, 522-1 Dokpyong-ri, Majand-myon, Ichon-si, Kyonggi-do, 467-812, Korea

    Jin-Ho Lee, Kwang-Ho Lee, Chang-Gyeom Kim, Young-Hoon Park & Sung-Oh Chung

  2. Macrogen Incorporated, Seoul, Korea

    Se-Young Lee

  3. Institute of Industrial Biotechnology, Inha University, Incheon, Korea

    Geun-Joong Kim

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  1. Jin-Ho Lee
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  2. Kwang-Ho Lee
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  3. Chang-Gyeom Kim
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  5. Geun-Joong Kim
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Correspondence to Sung-Oh Chung.

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Lee, JH., Lee, KH., Kim, CG. et al. Cloning and expression of a trehalose synthase from Pseudomonas stutzeri CJ38 in Escherichia coli for the production of trehalose. Appl Microbiol Biotechnol 68, 213–219 (2005). https://doi.org/10.1007/s00253-004-1862-5

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  • DOI: https://doi.org/10.1007/s00253-004-1862-5

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