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AN INTRODUCTION TO THE APPLICATIONS OF INDUSTRIAL (WHITE) BIOTECHNOLOGY

EUROPABIO's BIOTECHNOLOGY INFORMATION KIT

1. Introduction:

Modern biotechnology is a powerful and versatile tool which can compete with chemical andphysical means of reducing energy and material consumption and minimising the generation ofwaste and emissions.There is general agreement that the use of biotechnology in industry does not simply remove pollutantsbut also will prevent pollution at the source.Efforts to achieve clean industrial products and processes will also bring great benefits to industry overthe next ten or twenty years.Industrial biotechnology, using microorganisms and biological catalysts (enzymes) to produce goods andservices, has come of age.

Biotechnology and CO2 emissions

The soya bean: an important renewable resourceThe soya bean has long been used to develop products ranging from foiod and diesel fuels to polymers,fabric softeners, solvents, adhesives, linoleum, rubber substitutes, printing inks, and plastics. Recent

advances in recombinant genetic biotechnology have made it possible to alter the lipid composition ofsoya beans to increase the variety of biohydrocarbons available for industrial applications. Amides, estersand acetates of biohydrocarbons are currently used as plasticisers, blocking/slip agents and mold-releaseagents for synthetic polymers. Biohydrocarbons linked to amines, alcohols, phosphates and sulfur groupsare used as fabric softeners, surfactants, emulsifiers, corrosion inhibitors, anti-static agents, hairconditioners, ink carriers, biodegradable solvents, cosmetic bases and perfumes. In combination withaluminium and magnesium, the soya bean is used to produce greases and marine lubricating materials.

2. Biotechnology in industrial sectors

Various parts of the industry are experimenting with the new tools offered by biotechnology. Of particularinterest is the possibility of using biobased resources as feedstocks in the larger volume sectors. While

biobased manufacturing will not necessarily always be cleaner, it is certain that wastes from biobasedmanufacturing will be more compatible with conventional wastewater treatment systems.

Pharmaceuticals

Today, many pharmaceuticals are semi-synthetic molecules, in that part of their structure is synthesisedby a living organism and later modified by chemical processing. Thanks to biocatalysis optimisedfermentation, and replacement of organic solvents by water, modern biotechnology contributes to cleanerproduction of such semi-synthetic antibiotics.

An enzymatic process for producing an antibioticThermostabilised enzymes and the development of a new bioreactor process by Kaneka Corporation areused to produce 2,000 metric tons a year of amoxicillin, an antibiotic. This all-enzymatic process hasdisplaced an older one in which part of the synthesis was carried out chemically but created problems,including coloring of the product, formation of by-products, and low energy efficiency.

Textiles and leather

The textiles industry is continuously seeking new sources of innovation, one of which is biotechnology. In1996 the global enzyme market for textiles amounted to $ 178 million. Moreover, textile and apparelcompanies are spending more time and money on environmentally relevant issues. Regulatory pressureis expected to intensify for both textiles and leather as less polluting technologies become available and itbecomes possible to generate less waste.Enzymes have been used in textile processing since the early part of this century to remove starch-basedsizing, but only in the past decade has serious attention been given to using enzymes for a wide range oftextile applications.

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Enzymes are expected to have an even greater impact on effluent quality as more fibre preparation, pre-treatment and value-added finishing processes convert to biotreatment. In addition, enzymes are veryeffective catalysts even under mild conditions and do not require the high energy input often associatedwith chemical processes.

Food

In the food sector, biotechnology has long played an accepted role in traditional processes, such ascheese making. Both modern and traditional biotechnology can be an important supportive tool for thefood industry and give considerable added value to food products. When evaluating the use ofbiotechnology "from the farm to the fork" it is necessary to balance the environmental impact ofcommercial agriculture with that of alternative production routes, such as growth of microorganisms infermentors or from fossil fuel feed stocks. The environmental benefits of producing food additives byfermentation or enzymatic routes instead of traditional organic synthesis are similar to those for otherspecialty chemicals. In the case of fermentation-derived preservatives, the effect is even more favorablewhen the fermentation broth is incorporated in the finished product. In the most desirable situation,bacteriocin-producing cultures are used in fermented foods (such as sauerkraut) where they consumecarbohydrates, naturally preserve the finished product and contribute nutritive value of their own. Abiotechnology application with very great potential environmental benefit would convert waste streamsfrom one process into raw materials for another, or upgrade underutilised raw materials into a morevaluable form.Ideas abound, including alternative uses for the grape pomace left over from wine-making, corn cobs as a

substrate for citric acid production, and cranberry waste as a substrate for fungal bioinoculants. Use ofthe large quantities of whey produced during cheese making also hold out great promise. One successfulapproach has been the production of lactose-fermenting yeasts as flavoring ingredients.

Sugars from starchesStarch processing involves the conversion of maize or another grain into dextrose and other syrups by ahydrolysis reaction. This was formerly done using acid at high temperature and pressure, but dextroseyields were limited to about 80 %, the process was hazardous and expensive and produced largequantities of salt as a by-product. The initial change to enzymatic hydrolysis in the 1960s increaseddextrose yields and eliminated the drawbacks of the acid process. In the 1970s, development ofimmobilised glucose isomerase enzymes enabled the production of high fructose corn syrup. In the1980s, thermostable alpha-amylases helped increase yields, and in the 1990s, recombinant thermostableamylases have helped reduce costs.

Animal feed

Since the common protein sources used in animal feeds (e.g. soya, fishmeal, wheat and maize ) aredeficient in methionine, lysine, threonine and tryptophan, these essential amino acids are added assupplements to monogastric diets, e.g. for poultry and pigs. Whereas methionine is produced by chemicalsynthesis (300,000 tons in 1996) lysine, threonine and tryptophan are produced by industrialfermentation, using mutants of Corynebacterium glutamicum and recombinant strains of E.coli.

Feed enzymes are designed to degrade components of raw materials that limit digestibility and/or lead tohigher levels of excretion of manure, nitrogen and phosphorus. Endoxylanases and phytases are thebest-known feed-enzyme products. Endoxylanase enzymes hydrolyse phytic acid and release inorganicphosphate, thereby avoiding the need to add inorganic phosphates to the diet and reducing phosphorusexcretion. If phytase is added to feeds for pigs to liberate phosphate in the feed, phosphate release inmanure is reduced by 30 %. In a country like the Netherlands, this would reduce the phosphate releasedinto the environment by 20,000 tons a year.The marginal price increase in the feed cost to farmers (about 2 %) would be compensated for by areduced levy on discharge of phosphate.

Pulp and Paper

The pulp and paper industry is very capital-intensive with small profit-margins. It must meet increasingdemand for pulp and paper and, at the same time, comply with increasingly stringent environmentalregulations. Driven by market and environmental demands for less chlorinated products and by-products,it is the fastest growing market for industrial enzymes. In the United States, this market is projected togrow by 15 % a year for the next ten years.

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In paper-making, various processes are used to separate the cellulose fibres from the lignin in wood toform a slurry (pulp) that is then processed into paper and board. Existing chemical pulping operationscreate a great deal of pollution. Biopulping, which involves the treatment of lignocellulosic materials withlignin-degrading fungi, has been shown to result in energy savings and strength improvements. Enzymesare now also being incorporated into the pulping process, where they offer a number of advantages.The structure and chemical chemical composition of pulp fibres are of paramount importance for paperstrength and other properties. Enzymes can be used to reduce fibre coarseness, increase paper densityand smoothness, and improve appearance. Most pulp is produced using the kraft process. Kraft pulpshave a characteristic brown colour, which must be removed by bleaching before manufacturing paper forwriting or other products for which appearance is important. Chlorination is traditionally used, but pulpmanufacturers are turning to other techniques because of consumer resistance and environmentalregulations. According to studies conducted in Finaland, hemicellulases (mainly xylanases) improvebleaching. They are now being used commercially in Scandinavia, Canada, the United States, and Chile.Treating kraft pulps with xylanases significantly reduces chemical consumption with almost no loss in pulpyiels or quality. A new enzyme that is better suited to the temperatures and pH found in pulp processinghas also been developed in Israel and successfully tested in a large-scale trial.

Using bacteria to remove by-productsAdding polymers to paper stops fibres from becoming waterlogged and gives the paper wet strength.However, the polymer production process creates contaminants which reduce its effectiveness. CarburyHerne Limited and Hercules Inc. have developed a bioprocess for removing these by-products.

Two strains of bacteria are used to digest the by-products which are then washed out of the polymerbefore it is applied to the paper. This treatment is considered not only more environmentally acceptable,but it is also less expensive than developing a new product or a new manufacturing process to do thesame job.The process has now been adopted at production scale at two plants that make packaging paper for foodliquids. As the bioreactors were built into existing production lines, costly redesign of the productionprocess was avoided.

Energy

Biotechnology is having a major effect on the economics and the environmental impact of the energy

sector. Biotechnology can produce cleaner coal and petroleum, chiefly by removing sulfur and thusreducing the environmental contaminants released during combustion. Production of low-sulfur fuels willextend fossil fuel reserves and reduce levels of air contaminants. Biotechnology also has the potential forproducing equivalents to petroleum distillates, such as biodiesel. Ethanol, methane, and molecularhydrogen are even cleaner fuels, all of which would, if produced biologically, greatly lower levels ofgreenhouse gases.

The bioconversion of synthesis gas to liquid fuels such as methanol is also being investigated. Synthesisgas is a mixture of CO, H2 and CO2 made by the partial oxidation of any carbon-based material. Feedsfor the production of synthesis gas include agricultural, municipal, and paper wastes and biomass grownspecifically for this purpose. The range of feeds for synthesis gas make it a particularly versatile source offuels. With potentially lower processing costs and greater carbon yield, fuels derived from synthesis gasare an attractive alternative to fuels produced by fermenting biomass-derived sugars.

BioethanolBioethanol is a liquid transportation fuel. Currently, most bioethanol is made from sugar cane, maize andother starch crops. In the United States, close to a billion gallons of ethanol are produced annually, and inBrazil production may be four times that. However, a tax credit is needed to achieve a competitive marketprice. To be economically competitive with fossil fuels, the technology for producing ethanol frombiomass-derived sugars will require using high-yield low-cost crops and more efficient methods ofconverting lignocellulosic waste material into fermentable sugars. These two areas are the focus ofcurrent research. In studies sponsored by the Department of Energy, US scientists are investigating asimultaneous saccharification and fermentation procedure for converting cellulose to ethanol. Theprocess combines cellulose hydrolysis and fermentation steps in one vessel to produce high yields. Theobjective is to develop, by the year 2000, technologies for producing ethanol from biomass at a cost thatwill be competitive, without tax incentives, with the cost of gasoline.

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