Purification and biochemical characteristics of a novel fructosyltransferase with a high FOS transfructosylation activity from Aspergillus oryzae S719

Protein Expr Purif. 2020 Mar;167:105549. doi: 10.1016/j.pep.2019.105549. Epub 2019 Dec 2.


Fructooligosaccharides (FOS) have widely used for the manufacture of low-calorie and functional foods, because they can inhibit intestinal pathogenic microorganism growth and increase the absorption of Ca2+ and Mg2+. In this study, the novel fructosyltransferase (FTase) from Aspergillus oryzae strain S719 was successfully purified and characterized. The specific activity of the final purified material was 4200 mg-1 with purification ratio of 66 times and yield of 26%. The molecular weight of FTase of A. oryzae S719 was around 95 kDa by SDS-PAGE, which was identified as a type of FTase by Mass Spectrometry (MS). The purified FTase had optimum temperature and pH of 55 °C and 6.0, respectively. The FTase showed to be stable with more than 80% of its original activity at room temperature after 12 h and maintaining activity above 90% at pH 4.0-11.0. The Km and kcat values of the FTase were 310 mmol L-1 and 2.0 × 103 min-1, respectively. The FTase was activated by 5 mmol L-1 Mg2+ and 10 mmol L-1 Na+ (relative activity of 116 and 114%, respectively), indicating that the enzyme was Mg2+ and Na+ dependent. About 64% of FOS was obtained by the purified FTase under 500 g L-1 sucrose within 4 h of reaction time, which was the shortest reaction time to be reported regarding the purified enzyme production of FOS. Together, these results indicated that the FTase of A. oryzae S719 is an excellent candidate for the industrial production of FOS.

Keywords: Aspergillus oryzae; Enzyme purification; Fructooligosaccharides production; Fructosyltransferase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspergillus oryzae / enzymology*
  • Food Industry
  • Hexosyltransferases* / biosynthesis
  • Hexosyltransferases* / chemistry
  • Hexosyltransferases* / isolation & purification
  • Oligosaccharides / metabolism*


  • Oligosaccharides
  • fructooligosaccharide
  • Hexosyltransferases
  • inulosucrase