Abstract
Fructo-oligosaccharides (FOS) are linear fructans comprising 2–5 fructose units linked to a terminal glucose residue. They are widely used as food and feed additives due to their sweetness, low calorific value, and prebiotic properties. Here we describe the synthesis of FOS catalyzed by a cell-free crude enzyme solution containing recombinant fructosyltransferase (1-FFT) produced in the yeast Kluyveromyces lactis. During the enzyme catalysis, glucose accumulates as a by-product and eventually inhibits FOS production. We therefore used an enzyme membrane reactor (EMR) to achieve the continuous removal of glucose and the simultaneous replenishment of sucrose. We observed a loss of flux during the reaction with the characteristics of complete pore blocking, probably caused by a combination of proteins (enzyme molecules) and polysaccharides (FOS). Such complex fouling mechanisms must be overcome to achieve the efficient production of FOS using EMR systems.
Highlights
Fructo-oligosaccharides (FOS) are linear fructans containing 2–5 β(2→1)-linked D-fructose units joined to a terminal α(1→2)-linked glucose residue
We investigated investigatedthe the relationship between fluxfouling and fouling in the enzyme membrane reactor (EMR) toindetermine order to the conditions the highest flux after a filtration lasting 8 run h
Our preliminary experiments in an EMR containing the enzyme 1-FFT revealed an unavoidable decline in flux, so we applied a D-optimal design using the permeate flux after 8 h as a parameter and statistically optimized the model with respect to the enzyme activity
Summary
Fructo-oligosaccharides (FOS) are linear fructans containing 2–5 β(2→1)-linked D-fructose units joined to a terminal α(1→2)-linked glucose residue. FOS are water-soluble molecules that have a low calorific value but 30–50% of the sweetness of sucrose, depending on their chemical structure and degree of polymerization [1,2]. Short-chain FOS in particular have similar organoleptic properties to sucrose and are widely used as low-calorie sweeteners [3]. FOS occur naturally in many plants, including onions, bananas, artichokes, tomatoes, and asparagus. They can be extracted from these sources or produced by the limited enzymatic hydrolysis of inulin, which is abundant in chicory and Jerusalem artichoke [6,7]. FOS can be produced de novo by the enzymatic biotransformation of sucrose. The transferase activity of 1-FFT, which has been isolated from Aspergillus niger, Aspergillus terreus, and Aureobasidium pullulans, among other sources, is stronger
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