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World Journal of Dairy & Food Sciences 6 (1): 71-78, 2011 ISSN 1817-308X © IDOSI Publications, 2011

A Review Article on Edible Pigments Properties and Sources as Natural Biocolorants in Foodstuff and Food Industry

Ali Aberoumand

Department of Fisheries, Behbahan High Educational Complex, Behbahan, Iran

Abstract: Consumers are avoiding foods containing synthetic colourants, which lead food industries to replace them by natural pigments. Food colorants may be classified into synthetic, nature-identical, inorganic and natural colorants. Natural colorants for food are made from renewable sources. Most often, the colorants are extracted from plant material, but other sources such as insects, algae, cyanobacteria and fungi are used as well. Natural colorants are usually extracted and concentrated using either water or lower alcohols for water-soluble pigments and organic solvents for lipophilic pigments. Legislation restricts which colorants are allowed, what sources may be used for that particular colorant, what solvents may be used to extract it and the purity of the pigment. Colorants are formulated to make them more suitable for a variety of foods and drinks and to increase their stability.

Key words: Food colorants Natural colourants Pigments Properties

INTRODUCTION Examples of inorganic colors are titanium dioxide, gold and silver. Although structurally very diversified and Color is an important characteristic of food. Based on from a variety of sources, natural food colorants can be the color of the food, first impressions are made: Is the grouped into a few classes, the three most important of fruit immature, ripe, or overripe? Is the toast burnt? Is the which are: tetrapyrrols, tetraterpenoids and . food fresh? Based on these first impressions, a judgment The most important member of the tetrapyrrols is is made whether the food is safe to eat or not and , which is found in all higher plants. whether it can be expected to taste good or not. Since are tetraterpenoids that are as ubiquitous as color is closely associated with expectations, the addition chlorophyll, since they too are part of the photosynthetic of color to food is a way to fulfill these expectations. apparatus. They also give the yellow–orange–red color of Color is added to food for one or more of the following many fruits. are a group of flavonoids reasons: (1) to replace color lost during processing, (2) to that provide the red–purple shade of many fruits, in enhance color already present, (3) to minimize batch- particular berries (e.g., strawberries, elderberries and tobatch variations and (4) to color otherwise uncolored black currants). Other important classes of colorants are food. Food colors can be divided into four categories: (1) the (, lac, kermes and madder) natural colors, (2) nature-identical colors, (3) synthetic and the (). The terms “pigment” and colors and (4) inorganic colors. Natural colors are “dye” are often used interchangeably. Strictly speaking, pigments made by living organisms. Usually, pigments a pigment is insoluble in the given medium, whereas a made by modification of materials from living organisms, dye is soluble. Thus, carotenoids are dyes in oil but such as caramel, vegetable carbon and Cu-chlorophyllin pigments in water. This distinction may be difficult to (vide infra), are also considered natural though they are in maintain if nothing is assumed about the medium and in fact (except for carbon) not found in nature. Nature- the following the term “pigment” will be used for colored identical colors are man-made pigments which are also substances in general [1-8]. found in nature. Examples are , The development of foods with an attractive and riboflavin. Synthetic colors are man-made colors appearance is an important goal in the food industry. which are not found in nature, these are often azo-dyes. Increasingly, food producers are turning to natural food

Corresponding Author: Ali Aberoumand, Department of Fisheries, Behbahan High Educational Complex, Behbahan, Iran. Postal address: No 22, second line of Ab va Barg, Zolfeghari Street, Behbahan, Khuzestan Province, Iran. Tel: +98-9166716540, Fax: +98-6712229969, E-mail: [email protected]. 71 World J. Dairy & Food Sci., 6 (1): 71-78, 2011 colors, since certain artificial color additives have this family plant displayed a broader color spectrum and demonstrated negative health issues following their were devoid of the mention drawbacks in red beet [18]. consumption [9]. Because of the deficiencies of existing Selected species of fruits recognized as natural food colorants, the demand for natural pigments purple-fleshed Hylocereus polyrhizus have very recently is repeatedly raised by the food industry. This demand been suggested as promising betacyanin source. The can be fuelled by research to offer a more natural-healthy primary findings on the pigment potential and the scarce way of coloring foods and provide a clean label knowledge about color characteristics encouraged more declaration. Therefore, part of plant pigment research is investigations. Preliminary data from different Hylocereus seeking new sources of pigments. This will not only genotype were promising and further investigations are directed in finding natural alternatives for synthetic dyes, needed for process optimisation with respect to color but also discover new procedures for the pigment shade and pigment yield. The main obstacle in these production. The diversity of tropical and subtropical fruits is the pectic substances. Method of degradation vegetation offers a promising range of unknown plant of pectic material is necessary to facilitate pigment release, compounds that might prove applicable for the needs of increasing yield and reducing processing wastes. In humans. The most common plant pigments are Malaysia fruits of Hylocereus polyrhizus (genus carotenoids, , anthocyanins and betalains. Hylocereus, Cactaceae) have been cultivated as fruit crop Most research has been focused on carotenoids and locally. Red color pigments from fruits of H. Polyrhizus anthocyanins but betalains have recently gained interest could be an interesting possibility as alternative to in food science. Commonly broad arrays of from red beet. In view of the fact that most natural containing extracts are used for purposes; food colorants are supplied as concentrated juices or so far, there is only one single betalainic source that has extracts and to get a better understanding of how pigment been extensively used in the food industry worldwide. profile and hue are related in making decision for Compared to anthocyanins, betalains are ideal for coloring commercial applicability, the present work aimed to: low-acid foodstuff as they maintain their color over a wide array of pH, from 3 to 7. The most important betalainic To extract betacyanin pigments from pulp and skin of sources for natural red coloring are selected varieties of Hylocereus polyrhizus. red beet, commercial preparations of which are mainly To identify the constituents of the betacyanin composed of the red-purple betanin and its C15-isomer pigments from pulp of Hylocereus polyrhizus. isobetanin. To evaluate the effect of enzyme treatment on yield Betacyanins are important constituents of of betacyanin constituent. pigments. They are the main compounds associated with To compare the color of extract from Hylocereus the red color exhibited by flowers, fruits and other plant polyrhizus to that of red beet [19-21]. tissues. Red beet has been established in the market as the oldest and most abundant red food colorant, called Natural Distribution of Pigments: Chlorophylls and betanin, that is known as E-162 in the European Union carotenoids are the most abundant pigments in nature. [10-13] and as 73.40 in the chapter 21 of the Code of They are involved in fundamental processes and life on Federal Regulations (CFR) section of the Food and Drug Earth depends on them. Plants, photosynthetic bacteria Administration (FDA) in the USA (Griffiths, 2005). and protozoa (plankton) are the main sources of the Practically it has been used to color foods such as organic materials that are required for the development yoghurt, confectionery, ice creams, syrups, sausages and of other living organisms such as vertebrate and processed meats. However, the typical earthy flavor invertebrate animals. Chlorophyll is not found in animals caused by geosmin and high nitrate concentrations but carotenoids accumulate in some organs (e.g., eyes) associated with the formation of carcinogenic and tissues (e.g., skin of fish, bird plumage). In general, nitrosamines [14, 15] may affect its commercial use. animal carotenoids are obtained from the common diet. Furthermore, the risk to carry-over earth-bound Other pigments are also found in animals; some have microorganisms from the raw material in red beet is a important functions (e.g., heme proteins, riboflavin), crucial point [16,17]. Thereby, there is a high demand for whereas the function of others is not yet completely clear other alternative compounds that can be substitute for red (e.g., melanins, flavonoids). Other organisms have beet. In recent years fruits from the Cactaceae have been interesting pigments that have been used or have introduced as a promising betalain source. The fruits of potential use. Lichens produce depsides, the ancient

72 World J. Dairy & Food Sci., 6 (1): 71-78, 2011 and most extensively used dyes which were used as is mentioned in the Bible [26]. Use of natural biocolarants textile dyeing agents. In addition, they have application in food is known from Japan in the shosoin text of the as sunlight filters, as chemical indicators (litmus paper, Nara period (8th century) contains references to colored pH indicators) and as cytological stains. Some of the soybean and adzuki-bean cakes, so it appears that pigments obtained by treatment of lichen substances colored processed foods had been taken at least by are orcein and : More than 1000 pigments people of some sections. Study of color intensified, since have been identified in fungi. Consequently, the late 19th century, to understand: diversity of fungi pigments is the second in importance, after plant flavonoids. Fungi are not photosynthetic The phenomenon for survival of animals and plants, and do not contain chlorophyll. The relation between color and evolution theories; distribution in fungi is restricted to some orders (e.g., and Pharagmobasidiomycetidae, Discomycetes). In addition, The role imparting in comparative physiology. flavonoids are scarce in fungi whereas riboflavin imparts the yellow color in the genera Use of natural colorant in Thus, studies on biocolorant are greatly impulsed by food industry appears to have multidimensional potential their multiple functions [27]. The art of coloring spread [22]. For example, in addition to coloring property,- widely with the advancement of civilization [28]. carotene may be used in food as an essential vitamin source or betalains as source of essential amino acid or Source of Biocolorants: Plants, animals and anthocyanins as quality control marker of food stuffs. microorganisms are the sources of natural biocolorants, Flavonoids are colorants with high pharmacological but few of them are available in sufficient quantities for promise. Sometimes, diets with carotenoid mixtures are commercial use as food colorant and mostly are plant recommended instead of having just one particular origin. For biotechnological production of such colorants, carotenoid, because great variability of radicals and plants and microorganisms are more suitable due to their microenvironments take place in vivo [23]. Process understanding of proper cultural techniques and development and use of new and available technologies processing. Natural colorants obtained from plant origin for production optimization are the important area of are pepper, red beet, grapes, saffron [29, 30]. Nowadays, research on natural food grade colors. Therefore, it fermentative production of food grade pigments are requires a thorough review of the biotechnological available in the market, for example; color from Monascus potentials of ‘natural food grade biocolorants’. sp., from Xanthophyllomyces dendrorhous, Arpink red color from Penicillium oxalicum, riboflavin History of Biocolorants: Man has always been interested from Ashbya gossypii and -carotene from Blakeslea in colors. The art of dyeing is of long past and many of trispora. Also a number of microorganisms produce the dyes go back into prehistory. In Europe, it was biocolors in good amount that includes Serratia and practiced during the Bronze Age. The earliest written Streptomyces [31]. record of the use of natural dyes was found in China dated 2600 BC. In Indian subcontinent, dyeing was known Biocolorants in Food Industry: To consider the natural as early as in the Indus Valleyperiod (2500 BC) and has biocolors as the colorant, stability, yield and price are been substantiated by findings of colored garments of mostly the constrains. Most of them are sensitive to pH, cloth and traces of madder dye in the ruins of heat and sunlight [32]. Inspite of such factors, the natural Mohenjodaro and Harappa civilization (3500 BC). In biocolorants are gaining importance because of health Egypt, mummies have been found wrapped in colored and hygiene, nutrition, pharmaceutical activities, fashion cloth. Chemical tests of red fabrics found in the tomb of and environmental consciousness, indicates relative King Tutankhamen in Egypt showed the presence of dependency on natural products. Colors derived from , a pigment extracted from madder. minerals (lead chromate, copper sulphate) may cause The dye was used by the people of Aztec serious health problems [33]. However, in last few and Maya culture period of Central and North America. decades, synthetic additives are severely criticized and By the4th century AD, dyes such as woad, madder, weld, consumers show inhibition toward these products, Brazilwood, indigo and a dark reddish-purple were consequently they prefer to use the natural colorants known. Brazil was named after the woad found there [24]. [35]. In the 1960s in US, the environmental activists Henna was used even before 2500 BC, [25] while saffron demonstrated against the use of such food additives

73 World J. Dairy & Food Sci., 6 (1): 71-78, 2011 and this attitude was spread out widely. Activists also in cosmeticsand foods, particularly in dairy products campaigned for the natural colorants highlighting their (cheese), confectionary (soft and hard candy), fish and nutritional characteristics as a sales tool. A strategy that meat products, fruit products, beverages, snacks, beer has failed earlier, later converted it into a total success, and wine. It is more stable in photo degradation than that resulting from changes in social attitude. Thus, a of carotene [42]. However, canthaxanthin is not worldwide tendency to use natural colorants is generated. considered as food additive under EU regulation. Currently, people interpret the content of synthetic Cochineal insect ( coccus), best cultivated products as contaminant and the tendency has been on ficusindica, extract is known to have carmine reinforced [36]. There are number of advantages of using or carminic acid, which appears as magenta-red upon natural biocolorants, over synthetic colorants, further application. Water insoluble forms of carmine exhibit a boosted because of their pharmacological properties. color range from pink to purple. It is resistant to light, Products are of good market value if they are colored with heat and chemical oxidation, often it is more stable than natural compounds, for example; annatto in Cheddar synthetic food colorants but unstable at low pH. The cheese [37]. water soluble form is used in alcoholic drinks with calcium carmine; but water insoluble form is used in a Food Grade Natural Biocolorants: Natural biocolorants variety of products. Together with ammonium, carmine is are under ‘exempt from certification’category of FDA and used inmeat, sausages, processed poultry products, EU for food use and are: annatto extract, dehydrated red alcoholic drinks, beverages, bakery and dairy products, beet, canthaxanthin, carotene, Dactylopuis coccus extract, including desserts and sweets. On an average, people cotton seed meal, grape skin extract, fruit and vegetable consume one or two drops of carminic acid annually with juices, tagetes extract, carrot oil, oil of corn endosperm, food [43]. Cotton seed meal is the by-product after oil paprika and paprika oleoresin, riboflavin, saffron, turmeric extraction from cotton seed. Gossypol is a fat soluble and turmeric oleoresin and (flavoxanthins, yellow pigment that occurs in cotton seed in bound and rubiaxanthins, ). Chlorophylls, in spite of its free forms. The bound form of gossypol is combined with large intake amount per day. Yellow to orange color of free amino acids which constitute a medium quality Annatto comes from the outer layer of seeds of the protein feed [44]. Grape skin extract (enocianina) imparts tropical tree Bixa orellana. The carotenoids, bixin and a reddish purple color to beverages [45]. Some fruits norbixin, are responsible for appearance of yellow to contain a single type of anthocyanin ( in apple, orange color. The pH and solubility affect the color hue; cherry, fig, etc), some contain two major types (cyaniding the greater the solubility in oil, the brighter is the color. and peonidin in ascherry, canberry); or some with several Water soluble, oil soluble and oil/water dispersible forms anthocyanins (grape). As colorant, grape juice is available of annatto are available. Since it precipitates at low pH, is in a variety of colors: red (shades of cherry, raspberry or also available as emulsion, an acid proof state [38]. strawberry), purple and yellow. It gives a strawberryred Annatto has been used for over two centuries as a food shade in application. It is used to color a number color especially in cheese and in various other food ofnon-beverage foods, including gelatin desserts, fruit products [39]. Red beet () extract shows fillings and in certain confectionaries. Anthocyanins from variety of colors, depending on their content of yellow banana bract and Oxalis triangularis are found as compound and may have a good flavor. Also a potential source of food colorants [56]. Acylated bluish-red color produced by a compound known as anthocyanins from different edible sources (black carrot betanin and is stable at higher pH range than red cabbage cultivars) are also used in food industry [47]. Vegetable extract [40]. There is no limit on its upper usage level. It juice is actually fermentable liquid product, either has wide application in different food commodity from unfermented or lactic acid fermented, obtained from the beverages to candy and from dairy to cattle products [41]. edible part of one or more vegetables for direct Canthaxanthin, a carotenoid, is commercially produced consumption and preserved exclusively by physical from the algae Haematococcus lacustris. Canthaxanthin means. The juice shall be free from skins, seeds and other is known mainly as the natural pigment of the coarse parts of the source vegetables. Tomato juice and orangeyellow chanterelle mushroom. Canthaxanthin has blends based on tomato have long been popular and various physiological functions and can be converted account for over 90% of the non-fruit juice trade. into vitamin A under stress. Canthaxanthin is used in is the principal compound derived from tomato, poultry for the appearance of color shade of the yolk, enlisted under US system. Under EU legislation,

74 World J. Dairy & Food Sci., 6 (1): 71-78, 2011 lycopene (E 160d) is considered as food additives. It is Carotenoids: Carotenoids are one of the most important highly stable under a wide range of temperature and groups of natural pigments. It has been estimated that pH, hence used as common food colorant. It is available nature produces about 100 million tons of thesepigments in liquid form or as cold-water dispersible powder. Carrot per year. They give brilliant yellow and red colours to has long been a component of tomato blends [13]. many fruits, vegetables, roots, flowers and autumn leaves; Red-cabbage juice produces a bright pink to red color they produce the colour of the egg yolk, of many algae, to a product with a pH less than 4.0 and is soluble in yeasts and mushrooms, Crustaceae as well as of the water, but not in oil. Use of Marigold (Tagetes erecta) feathers and skins of many birds. The carotenoids have flowers as source of food colorants is known from attracted the curiosity of scientists since the beginning Aztec civilization. Marigold flowers are by far the most of organic chemistry, actually since 1818. Numerous abundant natural source for commercial [48]. The outstanding chemists made valuable contributions and antimutagenic activity of carotenoids of Aztec marigold many tools and methods have been introduced into was evaluated. The presence of some essential amino organic chemistry via the carotenoids'. Imention the early and fatty acid increases the nutritional quality of carrot analyses (by Willstätter and Zechmeister), the seed oil. Corn endosperm oil is a reddish-brown liquid rediscovery of column chromatography by Kuhn and composed chiefly of glycerides, fatty acids, sitosterols Karrer and of the thin-layer chromatography by Callebaut and carotenoid pigments obtained by isopropyl alcohol [l2], the elucidation of the rule by Ruzicka and and hexane extraction from the gluten fraction of yellow of the symmetrical structures of squalene, /3-carotene corn grain [49]. It is used in chicken feed as color and lycopene by Karrer, the studies on comparative additive.Paprika (Capsicum annuum) is the pioneer and biochemistry by Goodwin3, the first rules on widely [50] used carotenoid as food colorant. In paprika, u.v.absorption of polyenes by Richard Kuhn, the red carotenoidsare dominated by canthaxanthin, investigation of cis-trans isomerism [54] and the first capsorubin where as yellow xanthophylls includes syntheses by Karrer and Inhoffen. The application of cryptoxanthin, zeaxanthin, antheraxanthin and -carotene n.m.r. spectroscopy by Jackman and Weedon5 proved to [51]. Paprika oleoresin is mainly extracted from the pods. be a valuable tool for the identification of carotenoids, It contains three main naturally occurring pigments: the synthesis of which was especially advanced by the capsanthin, capsorubin and carotene. This combination development of condensation reactions for the buildup produces a bright orange to red-orange in food products. of the conjugated chain (Wittig6, Homer7). We aimed at The oleoresin is oil soluble, when emulsified becomes the development of commercial syntheses of some water dispersible [52]. Riboflavin (vitamin B2) has a selected carotenoids arid the investigation of their use as variety of applications as a yellow food colorant. Its use food colourants. By substituting natural colouring matters is permitted in most countries. Applications include for artificial dyes we fulfil a physiological desideratum that dressings, sherbet, beverages, instant desserts, ice good food should not contain unnatural pigments [54]. creams, tablets and other products. Riboflavin has a special affinity for cerealbased products, but its use in Production of Carotenoids from Natural Sources: The these applications is somewhat limited due to its slight classical method for producing /3-carotene and other odour and naturally bitter taste. carotenoids is solvent extraction of the plant material. The natural source, the main pigments, possible Other Application in Food Industry provitamin A activity of the extract and the degree of Food Preservatives: Several natural biocolorants, pigmentation of eggs and broilers, after feeding, are including anthocyanin, additionally show antagonistic indicated. activity to certain bacteria, viruses and fungi thus to Carrots, palm oil, alfalfa, dried grass and leaves are protect food from microbial spoilage (Bridle and [53]. the starting materials for practically all preparations of Some are also active against protozoa (Leishmania natural carotene with provitamin A activity. The extracts brasiliensis) and insects (Calliphora erythrrocephala). from alfalfa, grass, leaves, Tagetes and yellow maize, Sometimes, carotenoids can act as sun screen to containing either lutein or zeaxanthin, are valuable maintain the qualityof food by protecting from intense supplements to chicken feed because allxanthophylls light. It has beenreported that corn carotenoids inhibit colour the egg yolk in contrast to the . Besides the synthesis of aflatoxin by Aspergillus flavus (90%) /3-carotene, annatto which is an extract of the seeds of and by most of the A. parasiticus (30%) strains [54]. Bixa Orellana is also used for colouring dairy products.

75 World J. Dairy & Food Sci., 6 (1): 71-78, 2011

For red carotenoids large quantities of paprika are (e.g., turmeric) could be made in this way. Colorants extracted. Torula yeast and certain algae are other sources made in this way will probably face some hurdles, at least of red polyene pigments used as colourants for oil and in Europe where genetically modified food is generally cheese. The market for natural carotenes has declined viewed with a large degree of skepticism by the since the introduction of synthetic carotene. Today, only consumers. Finally, new colorants would have to be one crystalline carotene preparation extracted from approved by the authorities, which is very costly because dehydrated carrots is still on the market. Recently it was of the various toxicological studies needed to confirm announced that /3-carotene and further carotenoids will the safety of a new food additive. The above gives a fairly be produced in Brazil by fermentation from mated strains compressed overview of the most important natural of Blakeslea trispora [54]. food colorants. The reader is referred to a number of excellent textbooks for more detailed information on plant CONCLUSION pigments and food colorants [12-16].

Research on carotenoids has included all branches REFERENCES of natural sciences has led the way to pure natural food colouring matters. It is my firm belief that for food 1. Alexandra, P.E., G.M. Monica, R.E. Wrolstad and colouring it is desirable to substitute artificial dyes by M.B.A. Gloria, 2001. Anthocyanins from Oxalis pure natural pigments. Furthermore I believe that pure triangularis as potential food colorants. Food Chem., synthetic carotenoids will gradually replace the natural 75: 211-216. extracts in the same way as the pure synthetic vitamins 2. Andrews, A.G., H.G. Phaff and M.P. Starr, 1976. have practically eliminated the impurevitamin preparations Carotenoids of Phaffia rhodozyma, a red-pigment obtained from natural sources. fermenting yeast. Phytochem., 15: 1003-1007. 3. Baranyovits, F.L.C., 1978. Cochineal carmine: an Future Prospects: Development of “new” food colorants ancient dye with a modern role. Endeavor, 2: 85-92. can be divided into two categories: those covered by 4. Barth, M.M., C. Zhou, K.M. Kute and G.A. existing legislation and those that are not. Colorants Rosenthal, 1995. Determination of optimum covered by existing legislation are not really new, but conditions for supercritical fluid extraction of encompass subjects such as new sources of known carotenoids from carrot (Daucus carota L.) tissue. J. pigments (e.g., a newly discovered fruit rich in Agric. Food Chem., 43: 2876-2878. carotenoids), new ways of formulating existing pigments 5. Bartley, G.E. and P.A. Scolnik, 1995. Plant (e.g., blue anthocyanin) and improvement of existing carotenoids: pigments for photoprotection, visual sources (e.g., by breeding). Breeding and selection of attraction and human health. Plant Cell., 7: 1027-1038. high-yield strains will most likely only lead to minor 6. Beyer, P., S. Al-Balli, X. Ye, P. Lucca, P. Schenb, R. improvements. Major improvements could be achieved Welsch and I. Potmyllus, 2002. Golden Rice: by genetic manipulation, but this is not allowed in the Introducing the Carotene biosynthesis pathway into EU and the naturalness of the colorant would be (partly) rice endosperm by genetic engineering to defeet lost. Vitamin A deficiency. J. Nutr., 132: 5065-5105. Production of colors by fermentation has a number 7. Bosevska, M., D. Karlovic, J. Turkulov and D. Pericin, of advantages: cheaper production, possibly easier 1993. Quality of corn germ oil obtained by extraction, higher yields, no lack of raw materials and no aqueous enzymatic extraction. J. Am. Oil Chem. Soc., seasonal variations. Fermented colors are already used 70: 1273-1277. today: D. salina, B. trispora, spirulina and monascus. It 8. Boyles, M.J. and R.E. Wrolstad, 1993. Anthocyanin is not unlikely that new, fermented colors such as composition of red raspberry juice: influences of lycopene from B. trispora will become allowed in the near cultivar, processing and environmental factors. J. future. A giant leap forward in color production could be Food Sci., 58: 1135-1141. achieved by combining genetic manipulation and 9. Bridle, P. and C.F. Timberlake, 1997. Anthocyanins as fermentation. natural food colourselected aspects. Food Chem., Microorganisms could be made to produce colorants 58: 103-109. in high yield by inserting genes coding for the colorant 10. Britton, G., 1991. Carotenoids. Method. Plant even colorants not naturally produced by microorganisms Biochem., 7: 473-518.

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