Abstract

New high quality and thereby added-value end-products are produced from cruciferous oilseeds, principally of rapeseed, in Pilot Plant Scale by use of a developed ”Green Chemistry” Biorefining Process technique. Separation of the seed constituents occur in an aqueous emulsion followed by decanting, sieving, centrifugations, and FC-techniques at gentle conditions without use of organic solvents. The gentle and special isolation techniques allow isolation of glucosinolates and other bioactive compounds, production of special oil-lipids and protein products with a high content of natural antioxidants, as well as various other types of speciality products. Thereby, the biorefining techniques give an attractive supplement to traditional processing techniques as revealed from market research. Combined results of utilization of this biorefining techniques and biorefined natural products give the needed basis for:

• Increased value of agricultural crops

• Optimal utilization of renewable resources

• Benefits to producers, end-users, and the nature

INTRODUCTION

Double low oilseed rape and other cruciferous crops are well suited for agricultural production of high quality vegetable oil and proteins (Bjergegaard and Sørensen, 1996). In addition, the seeds of these plants contain a great number of other compounds, which have a potential high value as food and for production of non-food products. However, some of these constituents in cruciferous seeds, especially glucosinolates and dietary fibres, create problems and limitations with respect to production of high quality oils and proteins by use of traditional oil mill processing (Bjergegaard et al., 1999a and refs. cited therein). The aim of the project now ongoing is therefore not only elimination or reduction of these problems and limitations, but the glucosinolates and dietary fibres are also the target for development of added-value products. The project is at present running as a EU-supported programme (FAIR CT 95-0260), and it is based on work directed at development of aqueous processing of oil and protein crops at gentle conditions (Jensen et al., 1990; Bagger et al., 1998).

The technical approach is based on development of advanced but relatively simple aqueous and “Green Chemistry” processing techniques. This biorefining process allows separation and isolation of the various constituents accumulated in cruciferous seeds – the Biorefined Oilseed Products (BOP), collected in the four groups (Figure 1; 1^st generation):

• bioactives in the “solubles” or “syrup” fraction
• protein concentrates
• lipids or oils
• carbohydrates, dietary fibres – feed pellets

These groups are again the basis for further separation and isolation of individual compounds according to the opportunities for an economical appropriate end-use (Figure 1; 2^nd generation). The combined project objective is production of new high quality added-value end-products from cruciferous oilseed crops, principally 00 rapeseed, avoid waste products from the process, and to evaluate:

• the product functionality
• the commercial potentials
• the combined technical and economical feasibility of the concept

Figure 1. Illustration of the fractionation of oilseed rape obtained with the different generations of biorefining compared to traditional oil mill processing.

Participants and their roles in the project

Co-ordinator: Ass. Professor Hilmer Sørensen, Chemistry Department, Royal Veterinary and Agricultural University, 40, Thorvaldsensvej, DK-1871 Frederiksberg C, Denmark and his research group. The task title is: “Basic Production” with sub-tasks (1) Pilot scale production control, (2) Glucosinolate isolation and conversion, (3) Characterisation of proteins, peptides, carbohydrates, and lipids in BOP products, (4) Edible and technical oils, (5) Production of concentrates and isolates from the HAC and LIPRO fractions, and (6) Special feed.

Dr. Sandro Palmieri, Instituto Sperimentale per le Colture Industriali, Via di Corticella, 133, IT40129 Bologna, Italy and his research group. The task title is: “Bioactive Basics” with the sub-tasks (1) Myrosinase purification, (2) Bioreactor optimization, (3) Bioactive molecules, production and characterisation, (4) Antitumor activity, and (5) Chiral intermediates.

Dr. Jacques Gueguen, Laboratory of Protein Biochemistry and Technology, Institut National de la Recherche Agronomique, 1627, Rue de la Géraudière, FR-44316 Nantes, and his research group, Dr. Claude Rabiller, Université de Nantes, Faculté des Sciences et des Techniques, 2, rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, and his research group, and Dr. Jacques Evrard, CETIOM, rue Monge, F-33600 Pessac, and his research group, all from France. The task title is “Biopolymers Basics” with sub-tasks (1) Protein composition and characterisation, (2) Lipid composition, extractions and characterisation, (3) Biomaterials: Preparation of films by casting, and (4) Lipophilization of proteins and peptides.

Dr. Kerr Walker, Scottish Agricultural College, Agronomy Department, Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland, and his research group. The task title is: “Biocide Functionality” with the sub-tasks (1) Formulation of biocides, (2) Antifungal activity, (3) Insecticidal activity, and (4) Nematocidal activity.

Mr. Kim Brinck, Castrol A/S, Esplanaden 7, DK-1263 København K, Denmark. The task title is: Oils and Additives” with the sub-tasks (1) Evaluation of the BOP-oil for technical applications and (2) Development and formulation of optimal combinations of natural additives.

Dr. Thomas Luck, Fraunhofer Institut für Verfahrenstechnik und Verpackung, Giggenhauser Str. 35, D-85345 Freising, Germany, and his research group. The task title is: ”Biopolymers – End-products” with the sub-tasks (1) Pilot scale protein extraction of 2S- and 12S fractions, (2) Tailoring into thermoplastic materials, and (3) Production of biodegradable hydrophobic films.

Mr. Christian Kjøller, Bioraf Denmark Foundation, Lykkesvej 11B, DK-3720 Aakirkeby, Denmark and his research group. The task title is: “Pilot Scale Production and Economics” with the sub-tasks (1) Pilot scale production of the BOP-products and (2) Market research and economics.

Expected deliverables and outcome

The expected deliverables and outcome are production in pilot plant scale of new high-quality and added-value products by use of biorefining techniques comprising aqueous extraction, sieving, decanting, centrifugations, ultrafiltrations, flash chromatography, and supercritical fluid techniques all as a “Green Chemistry” system.

I. Direct results

Lipids and oil as high quality edible and technical products, as well as evaluation of natural technical oil additives. The oil quality is controlled by SFT (Bjergegaard et al., 1999b).

Protein concentrates and isolates as high quality and added-value products in feed, food and non-food products, emulsifiers, surfactants, protein-lipid products, and protein based hydrophobic biopolymers.

Bioactives as potential biodegradable biocides against herbivores, micro-organisms, weeds, as potential antitumor compounds and chiral intermediates of the pharmaceutical industry.

II. Indirect results

Development of applied biotechnology; chromatography, ultrafiltration, supercritical fluid extractions, and enzyme technology, which also will be applicable to non-cruciferous plants.

Development of applied methods of analyses for characterisation of starting materials, products and for production control (Sørensen et al., 1999).

Applications

The development of applied biotechnology based on aqueous extraction systems and techniques used in this project is expected to be applicable in processing of a broad spectrum of agricultural crops increasing their value and utilisation by creating new outlet in feed, food and renewable non-food products.

Progress summary

Aqueous enzymatic based extraction of cruciferous oilseeds has been developed as a unique method for processing as supplement to traditional extraction and processing methods (Bagger et al., 1998). Thereby, glucosinolate degradation is avoided and the intact glucosinolates are isolated and evaluated as potential plant protection agents. In addition negative effects from these compounds in the protein, oil and lipid fractions have been avoided (Bjergegaard et al., 1996). Also the mild process gives a preservation of the native antioxidants, which give the oil and protein fractions pronounced resistance towards autooxidation. The biorefining process thus produces a unique vegetable oil from the seeds, without residues of organic solvents or other unwanted constituents, including phospholipids, thus eliminating a need for degumming or other comprehensive oil refining processes. Protein concentrate, individual biologically active proteins, as well as glucosinolates and phenolics are products isolated from the process. Patent application has been submitted concerning production of high value proteins. Bioreactors with immobilised enzymes for production of biocides have been constructed. Surplus products from the process are combined in a pellet suitable for cattle feeding. The process is thus without waste products, as all parts of the seeds will be separated into usable products.

Exploitation of the results

Advanced but relatively simple and cheap methods of analyses as MECC, SFT, ELISA, and chemometrics are developed and used to efficient quality control of starting materials, processes and end-products (Andersen et al., 1999; Bjergegaard et al., 1999ab; Sørensen et al., 1999). Moreover, exploitation of the results comprise:

• Developed techniques in lab-scale for production/isolation of various natural products of high
• value
• Transfer of lab-scale developed techniques to the Pilot Plant scale
• Pilot Plant scale isolation of enzyme used in biocide production
• Pilot Plant scale isolation of glucosinolates used as biocide precursors
• Production of added-value rapeseed oil and other lipids-amphiphilic compounds for use in feed-
• food-, and non-food industry
• Production of protein concentrates and isolates for use as special feed, food, and non-food

Visions

Opportunities for use of the biorefining techniques to:

• Nature-friendly “Green Chemistry” processing of oil and protein rich agricultural crops
• Production of high-quality vegetable oil as food without residues of organic solvents and with high content of the plants native antioxidants
• Production of added-value proteins for food and other special purposes where high-quality is necessary (Bagger et al., 1998; Figure 2)
• Production of biodegradable-nature friendly biocides
• Isolation of various natural products for use as added-value renewable fine chemicals; chiral-, labile- and structurally complex molecules

Combined results of utilization of the biorefining techniques and biorefined natural products will give the needed basis for:

• Increased value of agricultural crops
• Optimal utilization of renewable resources
• Benefits to producers, end-users and the nature.

Figure 2. Developments in the aqueous based enzyme extraction processing of oilseed crops (compare to Figure 1) give an increased number of added-value products and thereby the potential of increased added-value of oil and protein rich crops.

Acknowledgements

The Commision of European Union (Contract FAIR CT 95-0260) is gratefully acknowledged for financial support to this work.

References

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