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FAIR-CT95-0260
High quality oils, protein and bioactive products for food and non-food purposes based on biorefining of cruciferous oilseed crops |
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Proposal No: | FAIR-CT95-0260 |
| Date Prepared: | July 2001, September 1999, May 1999 | |
| Source: | Final Report
Abstract and
Executive Summary
Second Annual Progress report December 1997 Second Annual Progress Report - Bioraf, Denmark |
Final Report Abstract
Source: Final Report Abstract of June 2000
Consortium: The participants in this project were the Royal Veterinary and Agricultural University, Frederiksberg (Denmark), Instituto Sperimentale per le Colture Industriali, Bologna, (Italy), Institut National de la Recherche Agronomique, Nantes (France), The Scottish Agricultural College, Aberdeen (UK), Castrol A/S, Copenhagen (Denmark), Fraunhofer-lnstitut fuer Verfahrenstechnik und Verpackung, Freising (Germany) and the Bioraf Denmark Foundation, Aakirkeby (Denmark).
Abstract
Objectives - The overall project objective has been the development of "green chemistry" processes to produce new high quality, added value end-products from biorefined Cruciferous oilseeds, principally 00 rapeseed, and to evaluate:
For this purpose the project has been divided into four main areas:
Bioactives: To produce, characterise and evaluate bioactives from glucosinolates as potential biodegradable biocidal, antifungal, nematocidal and/or insecticidal agents and as health protective compounds and chiral intermediates for the pharmaceutical industry.
Protein Concentrates and Isolates: To produce, characterise and evaluate; protein concentrates and isolates as high quality/added value products as feed, food and non-food products; emulsifiers, surface active protein-lipid products, protein based hydrophobic biopolymers, thermoplastic materials as well as special protein based films and coatings including the biodegradability of moulded protein based polymers. In addition the aim was to formulate special high quality products for the food and feed industry, with the aim of optimal utilisation of all products resulting from the enzyme-based rapeseed refining process.
Lipids and Oils: To produce, characterise and evaluate; special edible- and technical oils, phospholipids, and other amphiphilic rapeseed constituents including investigation of the potential value of these products as natural technical oil additives.
Market Research and Economics: To find the optimal production set-up by market research and by economic and technical feasibility studies of the combined concept.
Results
The BOP processing technology has been developed as a unique method for agro-industry processing as supplement to traditional oil mill-extraction and processing of oilseed crops. Most important, and a prerequisite for recovery of the high quality oil, proteins and other constituents of cruciferous oilseeds, is an initial series of short time processing steps comprising inactivation of some seed enzymes - lipases, lipoxygenases and myrosinases. These processing steps need at the same time to be performed under gentle processing conditions protecting the native biomolecules - oils, proteins, lipids and glucosinolates - against transformation into artefacts. Gentle methods for separation of the aqueous emulsion have been developed to give a special oil free of unwanted amphiphilic compounds but with a high content of natural antioxidants. The process results initially in six fractions:
Biorefining of cruciferous oilseed crops by the processes considered in this project has been based on aqueous extraction or separation systems, following dehulling and cold pressing and/or enzyme catalysed cell wall degradation. The separations have comprised initial use of decanter centrifuges, ultrafiltration and flash chromatography techniques. These techniques enable unique opportunities for preparation of high quality oils, proteins and a number of other value added functional biomolecules for feed, food and non-food uses. These new biotechnological processes have been developed as "Green Chemistry" focused on product development of protein concentrates and isolates, protein based biopolymers, bioactives as biodegradable biocides, edible and technical oils and additives, produced as added value Biorefined Oilseed Products (BOP).
Efficient advanced, but still relatively simple and cheap, methods of analyses have been developed for analytical control of the processing, product quality and thereby optimised added value of the oilseed crops through added product value.
The results obtained covered:
Opportunities are now created for use of the biorefining techniques for:
Combined results of utilisation of the biorefining techniques and biorefined natural products will give the needed basis for:
Conclusion
In general 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.
Summary
Objectives
The overall project objective is to produce new end-products of high added-value from biorefined cruciferous oilseeds - principally rapeseed - and to evaluate the product functionality, the commercial potentials and the combined technical and economic feasibility of the concept. For this purpose the project is broken down into four main objectives as follows.
Bioactives To produce, characterise and evaluate bioactives from glucosinolates as potential biodegradable biocides - antifungal, nematocidal or insecticidal agents - and as chiral intermediates for the pharmaceutical industry.
Protein concentrates and isolates To produce, characterise and evaluate high-quality or added-value protein concentrates and isolates for use in:
Investigation of the biodegradability of moulded protein-based polymers is also required. Special high quality products for the food and feed industry will be formulated with the purpose of optimal utilisation of all products resulting from the enzyme-based rapeseed process.
Lipids and Oils To produce, characterise and evaluate special edible and technical oils, phospholipids, dolicholes and other amphiphilic rapeseed constituents, including the potential value of these products as natural technical oil additives.
Market Research and Economics To find the optimal production set-up by market research and by economic and technical feasibility studies of the combined concept.
Activities
Biorefining of cruciferous oilseed crops is based on aqueous extraction or separation systems, following enzyme catalysed cell wall degradation. The separations comprise initial use of decanter, centrifuges, ultrafiltration and flash chromatography techniques. These techniques enable unique opportunities for preparation of high quality oils, proteins and a number of other value-added functional biomolecules for feed, food and non-food uses. These new biotechnological processes have been developed as "Green Chemistry" focused on product development of protein concentrates and isolates, protein based biopolymers, bioactives as biodegradable biocides, edible and technical oils and additives, produced as added value Biorefined Oilseed Products (BOP).
Progress
The BOP processing technology has been developed as a unique method for agro industry processing as supplement to traditional oil mill-extraction and processing of oilseed crops. Most important, and a prerequisite for recovery of the high quality oil, proteins and other constituents of cruciferous oilseeds, is an initial series of short time processing steps comprising inactivation of some seed enzymes - lipases, lipoxygenases and myrosinases. These processing steps need at the same time to be performed at gentle processing conditions protecting the native biomolecules (oils, proteins, lipids and glucosinolates) against transformation into artefacts. Efficient methods of analyses for control of the processing conditions and for control of the product composition and quality are thus needed and have been developed. After inactivation of the above mentioned enzymes at optimised conditions the process with cell wall degrading enzymes is carried out at 50ºC to facilitate an easy and mild release and recovery of the rapeseed constituents. The gentle separations of the aqueous emulsion have been developed to give a special oil without unwanted amphiphilic compounds but with a high content of natural antioxidants. The process has resulted initially in six fractions:
Achievements
The process has been developed to a level where only limited glucosinolate degradation occur, and with these compounds concentrated in the HAC fraction, which give the optimal basis for glucosinolate isolation. FC procedures have now been developed for isolation of glucosinolates, sinapine and soluble proteins present in the HAC fraction. A cheap and simple technique for isolation of large amounts of pure myrosinase has been developed and bioreactors with immobilised myrosinase for production of biocides have been constructed.
With the intact glucosinolates concentrated in the HAC fraction, this gives also opportunities for using glucosinolate rich syrup as water soluble biocide precursors. Great amounts of glucosinolates and syrup rich in glucosinolates have been produced and these products are now used as water solutions applied prior to application of water solutions of myrosinase which give in situ release of biocides, thereby avoiding otherwise serious problems with safety requirements, which have been well documented. Comprehensive studies of the biocidal effects of glucosinolates/glucosinolate products toward insects, fungi and nematodes have been performed and promising results have been obtained. In addition negative effects from glucosinolates/glucosinolate products in the protein-, oil- and lipid fractions have been avoided.
The mild process condition results in preservation of native antioxidants, which give the oil and protein fractions pronounced resistance towards auto-oxidation. Added value protein concentrates, and unique oil with high levels of valuable native antioxidants, without residues of organic solvents and phospholipids, reduce/eliminate the need for degumming and other comprehensive oil refining processes. The protein concentrates have been evaluated by use of balance trials, and fiber pellets produced in special formulation of the hulls and a concentrated syrup remaining after protein, oil and glucosinolate isolation have been tested by use of in vitro determination of enzyme digestible organic matter. Although not yet optimal, protein based films have been produced, and proteins are now produced in products which are concentrates with a level of protein close to the requirement for protein isolates (90% of DM). Various protein products have also been tested for nutritive value in balance trials. Special fiber mix pellets have been produced from hulls and HAC-syrup unused after glucosinolate and protein isolation. These pellets have been developed as ruminant feed and they have been evaluated in vitro in a test for enzyme digestible organic matter (EDOM).
Future actions
The process gives opportunities of continued development, with new generations of the biorefinery process and thereby increased values of the products. Supercritical Fluid Extraction (SFE) is a technique that is on the way to be introduced for production of high quality, added value products (BOP) as special lipids and amphiphilic compounds, natural products as antioxidants, phenolics and other valuable compounds. Another important technique under development for production of pure individual proteins and protein isolates is the IsoPrime procedure, which has resulted in promising initial results. In co-operation with Castrol A/S a filtration and a low-pressure column steam- refining system is engineered for pilot scale production of biorefined oil, and oil produced therefrom will be evaluated, as will special protein and peptide fractions for potential value as emulsifiers.
The future actions will be directed towards isolation of greater amounts of various glucosinolates, development of the procedures and products used as biocides, further development of the techniques for production of protein isolates, production of protein based film, surfactants and emulsifiers. These actions will also comprise new input in the BOP process, which gives opportunities of continued development, with special focus placed on fine chemicals, valuable natural products, lipids and proteins. SFT (SFE+SFC), IsoPrime and FC techniques will be used both for preparative purposes and for analytical control of the products produced. In co-operation with Castrol A/S the new filtration and low-pressure column steam-refining systems engineered for pilot plant products of biorefined oil will be used to optimise the quality of BOP oil, and correspondingly focus will be placed on optimising the properties of proteins/peptides for use as emulsifiers.
Discussion
The work performed up to now has fulfilled expectation for all tasks and in several cases additional progress has been obtained. This is thus the case with respect to production of high quality oils, production of different types of protein concentrates with expected added value and with the opportunities for production of protein isolates, e.g. myrosinases, in the pilot p] ant scale. The development of new advanced supercritical fluid techniques (SFT; SFE and SFC) and IsoPrime techniques for protein isolation are considered as important tools both for production of added value products and documentation of the high quality products obtainable by use of the BOP process. Difficulties have been met with respect to the solubility of the proteins in the PRM fractions which create problems in relation to productions of some of the required non-food products as e.g. "films".
Appreciable progress has, however, been made also in this part of the project, even though the "film" produced up to now are of lower quality than required. In the area of biocide trials several glucosinolate-derived products have the desired biocide effects, significant progress has been made and a considerable number of unexplored opportunities for an increased utilisation of biodegradable nature friendly biocides appear to exist. The results of the economic and technical feasibility studies carried out indicates that a combined BOP-production can be a viable investment with the present actual processing costs and calculated investments, when the yield and product functionality can be fully realised. The most economically viable lines in the process are those that produce refined oil and emulsifiers as well as the production line that results in protein isolates, biocides and fine agrochemicals. From an economic point of view further research should be emphasised on these two BOP processes.
The process lines which uses PRM and produce protein concentrates, and dietary fibres creates a significant lower economic surplus due to a lower value of these products. Focus should still be placed on this task with respect to development of hydrophobic protein products, films and emulsifiers with a higher market value. At this stage the combined BOP-process shall still contain all sub-processes, but later on the total process will be reduced in complexity. Each sub-process will be analysed to determine whether the intermediate products should be sold directly to market or go into the next sub-process.
A further effort has still to be made within the studied process area focusing on development of the new biotechnology based on Green Chemistry and added value products, which have to be seen in context with the broad bio-diversity of the primary products and opportunities for synergy in combined production. The development of applied biotechnology based 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 in food, feed, non-food and high tech agroindustries.
Introduction
This item is based on the individual progress
report for participant number seven (Bioraf), that is responsible for the pilot
scale production and economic determinations within this project. The main
objectives are to:
These objectives are being met through implementation of two tasks under the overall heading of Pilot Scale Production and Economics. Within this sub-task 7.1 covers Pilot scale production and technical feasibility, including aspects of:
The economics are dealt with by a sub-contractor to this participant, the Danish Institute of Agricultural and Fisheries Economics, that are fulfilling the following objectives:
Based on the technical findings, market research and economic assessment will be carried out concerning the economic viability of the integrated production system from agricultural raw material (seeds) to intermediate/finished products. As sub-task 7.2 (Market research and economics)
Results
The required fractions are produced using aqueous
enzymatic extraction of Cruciferous oilseeds, in pilot scale, obtaining
data on production costs and market value. The processes include dry
milling, continuous cooking, wet milling, enzyme reaction in tanks, heat
exchangers, decanters, centrifuges, wet sieves, ion-exchangers,
evaporators, fine grinder/dryer, spray dryer, oil refining system, freeze
drier, super fluid extraction, which can be combined in different ways and
to optimise the processes. Seeds used include high erucic acid and other
rape varieties as well as mustard (Sinapis alba), B. juncea,
B. campestris, Tropaeolum majus and Crambe abyssinica.
The composition was found to vary as follows:
| Crop/variety | Dry matter w/w% | Protein w/w% | Oil w/w% |
| Rape 00 Apex | 91.4 | 20.1 | 42.1 |
| Rape 00 Jazz | 91.0 | 21.7 | 43.6 |
| Rape HEAR | 93.4 | 22.5 | 45.2 |
| S. Alba | 92.4 | 29.6 | 23.6 |
| B. campestris | 93.4 | 26.9 | 30.8 |
| B. juncea | 93.0 | 24.7 | |
| Crambe abyssinica | 92.9 | 22.9 | 28.2 |
Tests on the oil refining system have indicated that some reconstruction is required. A low pressure pre-stripping column is needed in order to remove residues of water before heating to avoid unnecessary oxidation and pressure drop. Further, low pressure measurement equipment and automatic process control systems has been proven necessary to obtain reproducible results. Reconstruction has been initiated and a demonstration production of up to 8 tons oil is planned for 1999.
In various experiments batches of protein rich meal were obtained with from 41.5 to 64.5 percent protein. Other partners have evaluated these in bio-polymer and binding applications, which indicated that further optimisation is required in respect to solubility and the level of colour. Trials were initiated using cold pressing, substituting the enzyme process. A small pre-pressing unit has been installed for this purpose, with trials planned for spring 1999.
Prepared proteins have been used for preparation of enzymatic protein hydrolysates. Pilot scale trials are planned for preparation of hydrolyses oilseed proteins in early 1999.
Six Cruciferous (non 00 rape) varieties have been processed to produce glucosinolates and desulfoglucosinoolates, in order to evaluate their mixed and individual biological activity.
The economic analysis has focused on the price trends for the various output products of the process. In addition, the economic viability has been estimated at different price levels for the input and output commodities. It was observed that the market price of rape rose by 70 percent since early 1996. However, the future price will depend on world market price for other oil crops such as soya. The protein fraction is the most valuable component, it is comparable to casein, which has shown some increase in price, but in now fairly stable. Refined rapeseed oil is the second most important product. Having fallen in the early part of the product, a marked increase in price has occurred over the last two years. Other products, such as molasses and wheat bran, are fairly low in value and remain fairly stable.
The value of the biocide components is difficult to predict. There are no similar products on the market and it is not yet known how these might substitute for other pesticide products in the market.
An economic model has been run applying the updated prices. Over the last reporting period, the profitability has slightly reduced; due to higher raw material prices and a decrease in value of some products. The technical process in not fully developed yet and it is possible that costs will increase beyond the estimate. Hence, a sensitivity analysis has been carried out, that indicated that the process could be economically viable even if the overall investment costs doubled.
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