Abstract

A new industrial monosaccharide, anhydro-fructose (AF), will be produced from starch by a new biocatalytic process using glucan lyase. Screening, in vitro evolution and rational enzyme design will be used to produce glucan lyase with high performance in the industrial bioconversion of starch to AF. In a concerted action the market potential of AF compound will be exploited by systematic enzymatic and chemical modification of this compound to produce a line of high-value added products, and to study their benefits on human health. One can envisage the production of AF both as bulk product and as a fine chemical dependent on its application in chemical synthesis, for therapy and as a food ingredient. The commercial development of AF and derivatives is envisaged by traditional and start-up bioindustries that participate in this transdisciplinary project.

Objectives

• To develop an environment-friendly biocatalytic process for an economic and sustainable production of the new industrial monosaccharide anhydro-fructose (AF).
• To develop a process-robust glucan lyase using in vitro evolution and gene shuffling involving structurally related starch-active enzymes.
• To exploit the market potential of AF by systematic enzymatic and chemical modification of this compound to create a new line of high value-added products for applications in the food, cosmetics and pharmaceutical industries.

Description of the work

The project work will begin with the production of the new sugar anhydro-fructose (AF) by treatment of starch with the enzyme glucan lyase. This will be made available to the partners and the project will use established methods to produce new and high value-added products derived from AF economically, including the medical and nutritional evaluation of such products. It will finish with an evaluation of the achievements of the project in terms of its beneficial socio-economic effects. A very broad array of diverse methodologies will be employed to achieve the objectives of the proposal, which is inherent to the highly integrated transdisciplinary approach pursued by the academic and industrial participants.

It is essential to improve the catalytic properties of glucan lyase for the financially feasible industrial production of AF. The screening of natural resources as well as advanced biotechnological methodologies assisted by bioinformatics strategy will be used to obtain enzymes with improved properties. In addition, the starch degradation process will be developed using biochemical engineering to provide the project partners with sufficient amounts of AF.

Concurrently, AF will be used as the starting material for extensive biocatalytic and chemical modification to produce a line of high value-added products. In particular the synthesis of an array of monomeric, oligomeric and polymeric compounds from AF are envisaged which may include new sweeteners, antioxidants, emulsifiers, nutritional oligosaccharides, and new polymeric biomaterials. While AF as a monosugar is not sweet in itself, it has proved to be an antioxidant, a humectant and a calorie-free sweetener filler in in vitro tests. This will be further investigated using food models for its shelf-life and its organoleptic effect. The expected market potentials of these compounds are in the food, cosmetics and pharmaceutical industries.

Deliverables

• Providing glucan lyase variants with improved characteristics for industrial applications.
• Providing new insights into the atomic resolution of the mechanism of action of glucan lyase and members of the same structural family involved in a large number of biological processes.
• Biocatalytic and chemical synthesis of an array of new monomeric and oligomeric compounds from AF with expected applications in the food, cosmetics and pharmaceutical industries.
• The production of copolymers from AF with favourable hydrogel properties will create new biomaterials for cosmetics and pharmaceutical applications.
• A possible beneficial effect of AF on Type 2 diabetes will be evaluated using animal models, and promising derivatives are included in these studies.

Contacts

Coordinator

• Friedrich GIFFHORN / University Saarland Department of Applied Microbiology / GERMANY

Participant

• Carlsberg Laboratory Deptment of Chemistry / DENMARK
• Shukun YU / Danisco Biotechnology DENMARK
• INSA Bioprocess Laboratory / FRANCE
• Prof Dr Klaus Buchholz / Institut für Technologie der Kohlenhydrate Technical University Braunschweig / GERMANY
• Bo AHRÉN / Lund University Hospital Department of Medicine / SWEDEN
• Inge LUNDT / Technical University of Denmark Department of Chemistry / DENMARK
• Sine LARSEN / University of Copenhagen Centre for Crystallographic Studies / DENMARK