Applied Microbiology and Biotechnology

, Volume 68, Issue 2, pp 213–219

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

  • Jin-Ho Lee
  • Kwang-Ho Lee
  • Chang-Gyeom Kim
  • Se-Young Lee
  • Geun-Joong Kim
  • Young-Hoon Park
  • Sung-Oh Chung
Applied Genetics and Molecular Biotechnology

DOI: 10.1007/s00253-004-1862-5

Cite this article as:
Lee, J., Lee, K., Kim, C. et al. Appl Microbiol Biotechnol (2005) 68: 213. doi:10.1007/s00253-004-1862-5

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.

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Jin-Ho Lee
    • 1
  • Kwang-Ho Lee
    • 1
  • Chang-Gyeom Kim
    • 1
  • Se-Young Lee
    • 2
  • Geun-Joong Kim
    • 3
  • Young-Hoon Park
    • 1
  • Sung-Oh Chung
    • 1
  1. 1.R&D Center for BioproductsCJ CorporationIchon-siKorea
  2. 2.Macrogen IncorporatedSeoulKorea
  3. 3.Institute of Industrial BiotechnologyInha UniversityIncheonKorea