Bioprocess and Biosystems Engineering

, Volume 38, Issue 12, pp 2417–2426 | Cite as

Kinetics and model development for enzymatic synthesis of fructo-oligosaccharides using fructosyltransferase

  • Rahul Kashyap
  • Tapas Palai
  • Prashant K. Bhattacharya
Original Paper

Abstract

Experimental investigations were made to synthesize fructo-oligosaccharides (FOS) from sucrose using fructosyltransferase. The influence of various parameters such as temperature (45–55 °C), pH (4–5), initial sucrose concentration (ISC: 300–500 g/L) and enzyme concentration (4–32 U/mL) were varied. A maximum FOS yield of 60 % was observed at ISC 500 g/L, pH 4.5 with enzyme activity 32 U/mL and at 55 °C. It was confirmed that 1-kestose (tri-) was the major product of FOS as compared to nystose (tetra-) and fructosylnystose (penta-saccharides). Further, the reaction rate increases with increase in temperature. From separate sets of experiments, it was observed that FOS formation was affected by glucose inhibition. Apart from the increase in the rate of FOS formation with increasing enzyme activity, the final values of FOS yield increase though till 16 U/mL and thereafter attain plateau. A kinetic model was also developed, based on Michaelis–Menten kinetics, and a five-step ten-parameter model, including glucose inhibition, was obtained. Model was solved using COPASI® (version 4.8) solver for kinetic parameter estimations followed by time course simulations.

Keywords

Sucrose Fructosyl-transferase Enzymatic reaction Fructo-oligosaccharide Michaelis–Menten kinetics Kinetic model 

Abbreviations

E

Enzyme

FTase

Fructosyltransferase

f(k)

Objective function of rate constants

Glu

Glucose

Fru

Fructose

k

Rate constants for step reactions

Suc

Sucrose

y and y

Experimental and simulated data

U

Unit of enzyme

BCA

Bicinchoninic acid

FOS

Fructo-oligosaccharides

HPLC

High performance liquid chromatography

IGC

Initial glucose concentration

ISC

Initial sucrose concentration

PAHBAH

p-Hydroxybenzhydrazide

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Rahul Kashyap
    • 1
  • Tapas Palai
    • 2
  • Prashant K. Bhattacharya
    • 1
  1. 1.Department of Chemical EngineeringIndian Institute of Technology KanpurKanpurIndia
  2. 2.Department of Chemical Engineering and TechnologyBirla Institute of Technology MesraRanchiIndia

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