Applied Microbiology and Biotechnology

, Volume 102, Issue 21, pp 9207–9220 | Cite as

Levansucrase from Halomonas smyrnensis AAD6T: first halophilic GH-J clan enzyme recombinantly expressed, purified, and characterized

  • Onur Kirtel
  • Carmen Menéndez
  • Maxime Versluys
  • Wim Van den Ende
  • Lázaro Hernández
  • Ebru Toksoy ÖnerEmail author
Biotechnologically relevant enzymes and proteins


Fructans, homopolymers of fructose produced by fructosyltransferases (FTs), are emerging as intriguing components in halophiles since they are thought to be associated with osmotic stress tolerance and overall fitness of microorganisms and plants under high-salinity conditions. Here, we report on the full characterization of the first halophilic FT, a levansucrase from Halomonas smyrnensis AAD6T (HsLsc; EC The encoding gene (lsc) was cloned into a vector with a 6xHis Tag at its C-terminus, then expressed in Escherichia coli. The purified recombinant enzyme (47.3 kDa) produces levan and a wide variety of fructooligosaccharides from sucrose, but only in the presence of high salt concentrations (> 1.5 M NaCl). HsLsc showed Hill kinetics and pH and temperature optima of 5.9 and 37 °C, respectively. Interestingly, HsLsc was still very active at salt concentrations close to saturation (4.5 M NaCl) and was selectively inhibited by divalent cations. The enzyme showed high potential in producing novel saccharides derived from raffinose as both fructosyl donor and acceptor and cellobiose, lactose, galactose, and ʟ-arabinose as fructosyl acceptors. With its unique biochemical characteristics, HsLsc is an important enzyme for future research and potential industrial applications in a world faced with drought and diminishing freshwater supplies.


Levansucrase Fructosyltransferase Halophilic enzyme Halomonas smyrnensis Levan Fructan 


Funding information

WVdE and MV are supported by funds from FWO Vlaanderen. The financial support of The Scientific and Technological Research Council of Turkey (TUBITAK) (grant number: 115O495) and Marmara University Scientific Research Fund (grant number: FEN-A-130515-0178) are gratefully acknowledged.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2018_9311_MOESM1_ESM.pdf (493 kb)
ESM 1 (PDF 492 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.IBSB—Industrial Biotechnology and Systems Biology Research Group, Department of BioengineeringMarmara UniversityIstanbulTurkey
  2. 2.Enzyme Technology Group, Center for Genetic Engineering and Biotechnology (CIGB)HavanaCuba
  3. 3.Laboratory of Molecular Plant BiologyKU LeuvenLeuvenBelgium

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