Purification and biochemical characterization of an extracellular fructosyltransferase enzyme from Aspergillus niger sp.
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ORIGINAL ARTICLE
Purification and biochemical characterization of an extracellular fructosyltransferase enzyme from Aspergillus niger sp. XOBP48: implication in fructooligosaccharide production Jeff Ojwach1 · Ajit Kumar1 · Samson Mukaratirwa1,3 · Taurai Mutanda2 Received: 13 March 2020 / Accepted: 15 September 2020 © King Abdulaziz City for Science and Technology 2020
Abstract An extracellular fructosyltransferase (Ftase) enzyme with a molar mass of ≈70 kDa from a newly isolated indigenous coprophilous fungus Aspergillus niger sp. XOBP48 is purified to homogeneity and characterized in this study. The enzyme was purified to 4.66-fold with a total yield of 15.53% and specific activity of 1219.17 U mg−1 of protein after a three-step procedure involving (NH4)2SO4 fractionation, dialysis and anion exchange chromatography. Ftase showed optimum activity at pH 6.0 and temperature 50 °C. Ftase exhibited over 80% residual activity at pH range of 4.0–10.0 and ≈90% residual activity at temperature range of 40–60 °C for 6 h. Metal ion inhibitors H g2+ and A g+ significantly inhibited Ftase activity at 1 mmol concentration. Ftase showed Km, vmax and kcat values of 79.51 mmol, 45.04 µmol min−1 and 31.5 min−1, respectively, with a catalytic efficiency (kcat/Km) of 396 µmol−1 min−1 for the substrate sucrose. HPLC-RI experiments identified the end products of fructosyltransferase activity as monomeric glucose, 1-kestose ( GF2), and 1,1-kestotetraose ( GF3). This study evaluates the feasibility of using this purified extracellular Ftase for the enzymatic synthesis of biofunctional fructooligosaccharides. Keywords Fructooligosaccharides · Fructosyltransferase · Aspergillus niger · 1-Kestose · 1,1-Kestotetraose · Zymography
Introduction The biofunctional food products which are oligosaccharide in nature such as fructooligosaccharides (FOS), inulooligosaccharides (IOS), galactooligosaccharides (GOS), Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13205-020-02440-w) contains supplementary material, which is available to authorized users. * Jeff Ojwach [email protected] 1
Discipline of Microbiology, School of Life Sciences, College of Agriculture, Engineering and Science, University of KwaZulu-Natal (Westville Campus), Private Bag X54001, Durban 4000, South Africa
2
Department of Nature Conservation, Faculty of Natural Sciences, Centre for Algal Biotechnology, Mangosuthu University of Technology, P.O. Box 12363, Jacobs 4026, Durban, South Africa
3
Present Address: One Health Center for Zoonoses and Tropical Veterinary Medicine, Ross University School of Veterinary Medicine, Basseterre, West Indies, Saint Kitts and Nevis
xylooligosaccharides (XOS), isomaltooligosaccharides (IMOS) and mannan oligosaccharides (MOS) confer both nutritional and health benefits have received greater attention over the last decade (Pearson 2018). These foods with nutraceutical health properties are termed prebiotics and novel to study (do Prado et al. 2018). Among these prebiotics, F
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