Di-glycosyl-stevioside production via Leuconostoc citreum sk24.002 alternansucrase enzymatic reaction and structural characterization

  • Abubakr MusaEmail author
  • Bo JiangEmail author
  • Haile Ma
  • Mohammed A. A. Gasmalla
  • Mandour H. Abdalhai
  • Ammar Al-alfarga
Original Paper


In this work, alternansucrase enzyme from Leuconostoc citreum SK24.002 was used to produce di-glycosyl-stevioside through acceptor reaction. This was detected by high-performance liquid chromatography (HPLC) and was isolated using AB-8 macroporous resin and semi-preparative HPLC. With the help of mass spectroscopy, 1D NMR (H1 and C13) and 2D NMR (COSY, HMQC, HMBC) the di-glycosyl-stevioside structure was identified to be 13-{[α-D-glucopyranosyl-(1→3)-α-D-glucopyranosyl-(1→6)-β-D glucopyranosyl-(1→2)-β-D-glucopyranosyl]oxy} kaur-16-en-19-oic acid β-D glucopyranosyl eater. In order to increase the modified stevioside (di-glycosyl-stevioside) product and reduce production time, the impact of velocity of shaking on the di-glycosyl-stevioside production was also evaluated, using 75–150 rpm for 24 h at 25 °C. The results showed that the di-glycosyl-stevioside production increased significantly (p < 0.05) from 0.35 ± 0.0.01 mg/mL for 75 rpm to 1.0667 ± 0.0152 mg/mL for 150 rpm at 6 h during the glycosylation reaction.


Stevioside Acceptor reaction Shaking velocity Di-glycosyl-stevioside Structure characterization 



This research was supported financially by the National Natural Science Foundation of China (Grant Nos. 31000764, 20976073, 31230057), the National High Technology Research and Development Program of China (Grant No. 2013AA102102) and the Science & Technology Pillar Program of Jiangsu Province (Grant Nos. BE2013647, BE2012613, BY2012049).


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Authors and Affiliations

  • Abubakr Musa
    • 1
    • 3
  • Bo Jiang
    • 2
  • Haile Ma
    • 1
  • Mohammed A. A. Gasmalla
    • 1
    • 4
  • Mandour H. Abdalhai
    • 1
  • Ammar Al-alfarga
    • 5
  1. 1.School of Food and Biological EngineeringJiangnan UniversityZhenjiangChina
  2. 2.State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxiChina
  3. 3.Sugar InstituteUniversity of GeziraWad MadaniSudan
  4. 4.Nutrition & Food Technology, Faculty of Science and TechnologyOmdurman Islamic UniversityKhartoumSudan
  5. 5.Department of Chemistry, Faculty of ScienceUniversity of JeddahJeddahSaudi Arabia

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