AIR1-CT92-0001 Valorisation of Sugar Beet Pulp by Solid State Fermentation - Proteins and Probiotics

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AIR Cluster III - Bioconversion : Agricultural Residues : Biological Conversion : Biotechnology : Composts/Fertilisers : Fine Chemicals : Process Engineering : Protein/Amino Acid : Sugar

	Contract No	AIR1-CT92-0001
	Total Cost	2 118 477
	EC Contribution	878 500
	Start Date	01/01/1993
	Duration	28 months

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SUMMARY

This project is a continuation of the previous project under the ECLAIR programme - AGRE-0038, valorization of sugar beet pulp by solid state fermentation, conducted from 1st November 1990 to 31st December 1991. The above mentioned programme led to promising potential for the production and commercial utilization of proteins and probiotics. The first topic, protein enrichment of sugar beet pulp by solid state fermentation with filamentous fungi was treated by Südzucker. The second topic, development of a microbial feed additive with probiotic characteristics was treated by the subcontractor Enzymatix Ltd, Cambridge, UK. The two parallel studies conducted during the ECLAIR contract up to the end of 1991 will be carried on. It has examined the use of solid state fermentation with sugar beet pulp as the main substrate, as well as evaluating maize grits and wheat bran. The objective is the production and assessment of fungal enzymes. The work includes enzyme extraction and purification, as well as parallel studies covering applications in baking and animal nutrition. A novel solid state fermentation system has been developed, from which improved xylanases have been obtained. Note: during 1995, studies were carried out up to the end of June by Generale Sucriere associated with Lyven and Ebro Agricolas associated with IRTA. No study was carried out by Solvay during this period.

OBJECTIVE

Protein enrichment : The scale up of fermentation from laboratory to pilot will be carried out in order to:

• Test and optimize the fermentation process which has been developed at laboratory scale.

• Produce sufficient quantities of fermented pulp in order to conduct nutritional tests on animals (mainly pigs and poultry).

• Evaluate the economy of the process. It will be necessary, starting from the pilot fermenter operating conditions, to evaluate the potential industrial costs (investment, raw materials and nutrients, energy, manpower, etc.). The production costs will be compared to the potential valorization for animal feed uses.

Probiotics : The six previously selected strains will be tested for their resistance to gastrointestinal conditions (resistance to antibiotics and bile acids), then through animal feed trials. A first trial with piglets will be carried out at the University of Stuttgart-Hohenheim. A comparison will be made between a control diet (with no additive), a diet containing a classical antibiotic (tylosin) and a diet containing a mixture of the 6 selected probiotics. Further trials will be carried out at IRTA Reus. The potential industrial production of the selected strains will be evaluated. SSF can be used only for 2 of the 6 selected microorganisms.

Production of hemicellulases : The basic purpose of the proposed project is to apply SSF, using sugar beet pulp as the main substrate, for the manufacture of a range of hemicellulases. The hemicellulolytic activities selected presently are endo-xylanase, endo-beta-glucanase, arabanase and arabunofuranosidase. Other hemicellulolytic activities could be also studied if necessary. The improvement of feed digestibility (fodder cereals, cellulose containing feeds) will be the main subject of application studies. Other applications will be considered such as fodder silage and composing.

Production of phytase : The objectives will focus on three major activities:

• Genetic engineering:

  This improvement of the recombinant clones will be done through the construction of new cloning vectors in which the phytase gene is under the control of different constitutive promoters. Two host strains will be used for comparison purposes, namely Asp. ficuum NRRL3135 and an industrial Aspergillus strain of the niger group. The host strains will be transformed with the new vectors; transformants will be assessed under SSF and liquid fermentation.

• Fermentation:

  The first objective will be to assess, both in SSF and in liquid, the recombinant clones obtained. The second will be to optimize the fermentation conditions with the most productive clone of each strain in order to compare the performances of both processes.

• Nutrition test:

  The product of the previous points, i.e. phytase samples, will be assessed in vitro and in vivo.

Contacts

Coordinator

• Michel BRUDER / Generale Sucriere FRANCE

EC Scientific Officer

• Ciarán MANGAN / European Commission - DG Research Quality of Life Programme - Unit B1.2 / BELGIUM

Participant

• Mr Teodoro NOGUERAS / Compañia de Industrias Agricolas S A EBRO-AGRICOLAS / SPAIN
• MR. P D'OULTREMONT / SOLVAY S.A. RESEARCH AND TECHNOLOGY BRUSSELS CENTE / BELGIUM