International Journal of Food Microbiology 79 (2002) 131 141 www.elsevier.

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Application of cereals and cereal components in functional foods: a review

D. Charalampopoulos, R. Wang, S.S. Pandiella *, C. Webb
Department of Chemical Engineering, Satake Centre for Grain Process Engineering, UMIST, Manchester M60 1QD, UK Received 15 March 2002; received in revised form 23 April 2002; accepted 25 April 2002

Abstract The food industry is directing new product development towards the area of functional foods and functional food ingredients due to consumers demand for healthier foods. In this respect, probiotic dairy foods containing human-derived Lactobacillus and Bifidobacterium species and prebiotic food formulations containing ingredients that cannot be digested by the human host in the upper gastrointestinal tract and can selectively stimulate the growth of one or a limited number of colonic bacteria have been recently introduced into the market. The aim of these products is to affect beneficially the gut microbial composition and activities. Cereals offer another alternative for the production of functional foods. The multiple beneficial effects of cereals can be exploited in different ways leading to the design of novel cereal foods or cereal ingredients that can target specific populations. Cereals can be used as fermentable substrates for the growth of probiotic microorganisms. The main parameters that have to be considered are the composition and processing of the cereal grains, the substrate formulation, the growth capability and productivity of the starter culture, the stability of the probiotic strain during storage, the organoleptic properties and the nutritional value of the final product. Additionally, cereals can be used as sources of nondigestible carbohydrates that besides promoting several beneficial physiological effects can also selectively stimulate the growth of lactobacilli and bifidobacteria present in the colon and act as prebiotics. Cereals contain water-soluble fibre, such as h-glucan and arabinoxylan, oilgosaccharides, such as galacto- and fructo-oligosaccharides and resistant starch, which have been suggested to fulfil the prebiotic concept. Separation of specific fractions of fibre from different cereal varieties or cereal by-products, according to the knowledge of fibre distribution in cereal grains, could be achieved through processing technologies, such as milling, sieving, and debranning or pearling. Finally, cereal constituents, such as starch, can be used as encapsulation materials for probiotics in order to improve their stability during storage and enhance their viability during their passage through the adverse conditions of the gastrointestinal tract. It could be concluded that functional foods based on cereals is a challenging perspective, however, the development of new technologies of cereal processing that enhance their health potential and the acceptability of the food product are of primary importance. D 2002 Elsevier Science B.V. All rights reserved.
Keywords: Cereals; Probiotic; Prebiotic; Fermentation; Lactic acid bacteria; Bifidobacteria

1. Introduction The interest in developing functional foods is thriving, driven largely by the market potential for foods that can improve the health and well-being of consumers. The concept of functional foods includes foods or food

Corresponding author. Tel.: +44-161-200-4429; fax: +44-161200-4399. E-mail address: s.pandiella@umist.ac.uk (S.S. Pandiella).

0168-1605/02/$ - see front matter D 2002 Elsevier Science B.V. All rights reserved. PII: S 0 1 6 8 - 1 6 0 5 ( 0 2 ) 0 0 1 8 7 - 3

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ingredients that exert a beneficial effect on host health and/or reduce the risk of chronic disease beyond basic nutritional functions (Huggett and Schliter, 1996). Successful types of functional products that have been designed to reduce high blood pressure, cholesterol blood sugar, and osteoporosis have been introduced into the market (Sanders, 1998). Recently, the functional food research has moved progressively towards the development of dietary supplementation, introducing the concept of probiotics and prebiotics, which may affect gut microbial composition and activities (Ziemer and Gibson, 1998). Probiotic foods are defined as those that contain a single or mixed culture of microorganisms that affect beneficially the consumers health by improving their intestinal microbial balance (Fuller, 1989). There is significant scientific evidence, based mainly on in vitro studies and on clinical trials using animals, suggesting the potentially beneficial effects of probiotic microorganisms. These include: metabolism of lactose, control of gastrointestinal infections, suppression of cancer, reduction of serum cholesterol, and immune stimulation (Gilliland, 1990; Salminen et al., 1998; Fooks et al., 1999). The necessity for epidemiological studies on healthy human populations to support the specific health promoting claims of a probiotic strain is generally highlighted (Sanders, 1998; Shortt, 1999; Saarela et al., 2000). Common microorganisms used in probiotic preparations are predominantly Lactobacillus species, such as Lactobacillus acidophilus, L. casei, L. reuteri, L. rhamnosus, L. johnsonii, and L. plantarum and Bifidobacterium species, such as Bifidobacterium longum, B. breve, B. lactis (Shortt, 1999). The incorporation of probiotic strains in traditional food products has been established in the dairy industry, leading to the production of novel types of fermented milks and cheeses (Gomes and Malcata, 1999). A prebiotic is a food ingredient that is not hydrolysed by the human digestive enzymes in the upper gastrointestinal tract and beneficially affects the host by selectively stimulating the growth and/or activity of one or a limited number of bacteria in the colon that can improve host health (Gibson and Roberfroid, 1995). Fibre is a general term of different types of carbohydrates derived from plant cell walls that are not hydro-

lysed by human digestive enzymes. Specific forms of dietary fibre are readily fermentable by specific colonic bacteria, such as bifidobacteria and lactobacilli species, increasing their cell population with the concomitant production of short-chain fatty acids (SCFA). These acids, especially butyrate, acetate, and propionate, provide metabolic energy for the host and acidification of the bowel (Sghir et al., 1998). Several clinical studies have also suggested that dietary fibre could promote beneficial physiological effects including laxation and blood cholesterol attenuation (Spiller, 1994), as well as blood glucose attenuation (Bijlani, 1985). It may also prevent cancer (McCann et al., 2001), diabetes (Wang et al., 2001), heart disease (Fernandez, 2001), and obesity (Iwata and Ishiwatari, 2001). However, epidemiological results have to be treated with great precaution due to the complexity of the possible mechanisms involved.

2. Cereal-based functional products The development of nondairy probiotic products is a challenge to the food industry in its effort to utilise the abundant natural resources by producing high quality functional products. In this respect, probiotic-containing baby foods or confectionery formulations have been developed by adding the strains as additives (Saarela et al., 2000). In recent years, cereals have also been investigated regarding their potential use in developing functional foods. Cereals are grown over 73% of the total world harvested area and contribute over 60% of the world food production providing dietary fibre, proteins, energy, minerals, and vitamins required for human health. The possible applications of cereals or cereal constituents in functional food formulations could be summarised:


as fermentable substrates for growth of probiotic microorganisms, especially lactobacilli and bifidobacteria  as dietary fibre promoting several beneficial physiological effects  as prebiotics due to their content of specific nondigestible carbohydrates  as encapsulation materials for probiotic in order to enhance their stability.

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3. Cereals as substrates for probiotics Lactic acid fermentation of cereals is a long-established processing method and is being used in Asia and Africa for the production of foods in various forms such as beverages, gruels, and porridge. Although differences exist between regions, the preparation procedure could be generalised. Cereal grains, mainly maize, sorghum, or millet grains, are soaked in clean water for 0.5 2 days. Soaking softens the grains and makes them easier to crash or wet-mill into slurry, from which hulls, bran particles, and germs can be removed by sieving procedures. During the slurring or doughing stage, which lasts for 1 3 days, mixed fermentations including lactic acid fermentation take place. During the fermentation, the pH decreases with a simultaneous increase in acidity, as lactic and other organic acids accumulate due to microbial activity. In Western countries, cereals, like wheat and rye, are used for sourdough production, which is traditionally prepared by adding a prefermented sourdough of good quality to the dough. These starter cultures can be characterised as mixed-strain cultures and are continuously propagated and distributed in small proportions in bakeries. The population of lactobacilli in fully fermented sourdoughs is more than 109 cfu g 1, while the lactic acid bacteria (LAB)/yeast ratio is generally 100:1 (Salovaara, 1998). The good growth of LAB in cereals suggests that the incorporation of a human-derived probiotic strain in a cereal substrate under controlled conditions would produce a fermented food with defined and consistent characteristics, and possibly health-promoting properties combining the probiotic and prebiotic concept. However, in designing such a novel fermentation food process, several technological aspects have to be considered such as the composition and processing of the cereal grains, the growth capability and productivity of the starter culture, the stability of the probiotic during storage, the organoleptic properties, and the nutritional value of the final product. 3.1. Effect of cereal composition on growth of probiotics Probiotic products are usually standardised based on the presumption that culture viability is a reasonable measure of probiotic activity, thus the ability of the

strain to attain high cell population is of primary importance. A concentration of approximately 107 cells ml 1 at the time of consumption is considered functional (Gomes and Malcata, 1999; Shortt, 1999). High cell growth rates and acidification rates would also result in reduction of fermentation times and enhance the viability of the specific strain by preventing growth of undesirable microorganisms present in the raw material (Marklinder and Lonner, 1992), which would lead to the formation of off-flavours (Svensson, 1999). Therefore, the adaptability of the probiotic in the substrate is a very important criterion in the selection procedure of a suitable strain (Oberman and Libudzisz, 1998). Lactobacilli and bifidobacteria have complex nutritional requirements such as carbohydrates, amino acids, peptides, fatty esters, salts, nucleic acid derivatives, and vitamins, which vary a lot from species to species (Severson, 1998). The principal carbohydrate constituents of cereal grains are starch, water-soluble or -insoluble components of dietary fibre, and several free sugars, such as glucose, glycerol, stachyose, xylose, fructose, maltose, sucrose, and arabinose. The contents of these components depend on the variety (Becker and Hanners, 1991), the processing, and the amount of water addition. Table 1 presents the composition of different varieties of cereals compared to that of milk. Cereals have higher content of some of the essential vitamins than milk, higher content of dietary fibre, and increased amount of minerals, especially phosphorus, but lower amount of fermentable carbohydrates, usually less than 1% in wheat dough. Information concerning the effects of cereal composition on the growth of probiotic microorganisms is limited. Marklinder and Lonner (1992) suggested the potential of fermented oatmeal soup (18.5%) containing viable LAB as a base for nutritive solution in enteral feeding. After testing several heterofermentative and homofermentative probiotic lactobacilli, it was concluded that oats are in general a suitable substrate for LAB growth, regardless of the differences between species and strains. Among the strains tested, L. acidophilus exhibited the slowest rates of pH reduction, and lowest levels of viable cells in the final product, due probably to its high requirements for several nutrients (Morishita et al., 1981). The highest viable cell counts, 3 109 and 1 109 cfu ml 1, were achieved using L. plantarum and L. reuteri, respectively. Addition of malted barley flour, proteases and amino acids

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Table 1 Composition of foods expressed as 100 g of edible portion Parameter Water (%) Protein (g) Fat (g) Carbohydrates (g) Fiber (g) Ash (g) Ca (mg) P (mg) Fe (mg) K (mg) Thiamin (mg) Riboflavin (mg) Niacin (mg) Mg (mg) Malt 8 13.1 1.9 77.4 5.7 2.4 40 330 4.0 400 0.49 0.31 900 140 Rice 12 7.5 1.9 77.4 0.9 1.2 32 221 1.6 214 0.34 0.05 1.7 88 Corn 13.8 8.9 3.9 72.2 2.0 1.2 22 268 2.1 284 0.37 0.12 2.2 147 Wheat 12 13.3 2.0 71.0 2.3 1.7 41 372 3.3 370 0.55 0.12 4.3 113 Sorghum 11 11 3.3 73.0 1.7 1.7 28 287 4.4 350 0.38 0.15 3.9 n.d. Millet 11.8 9.9 2.9 72.9 3.2 2.5 20 311 68 430 0.73 0.38 2.3 162 Milk (liquid) 87.4 3.5 3.5 4.9 n.d. 0.7 118 93 Trace 144 0.03 0.17 0.1 13

Source: Adapted from Severson (1998).

increased the rate of pH-decrease and the total amount of lactobacilli in the final product (Marklinder and Lonner, 1994). In a similar study, L. acidophilus was successfully cultivated in an enzymatically hydrolysed oat mash reaching 109 cfu ml 1 (Bekers et al., 2001). In our study (Charalampopoulos et al., 2002), human-derived strains of L. reuteri, L. plantarum, L. acidophilus, and a L. fermentum strain isolated from cereals were cultured in malt, barley, and wheat extracts formulated without the addition of any supplements. The growth parameters are presented in Table 2. The malt medium supported better cell growth than barley and wheat due to the increased

amounts of maltose, sucrose, glucose, and fructose (approximately 15 g l 1 of total fermentable sugars) and free amino nitrogen (approximately 80 mg l 1). It must be emphasised that each strain demonstrated a specific preference for one or more sugars, which has been reported for LAB isolated from fermented cereal products (Gobbetti and Corsetti, 1997). The similar fermentation patterns observed in wheat and barley for all the strains tested could be attributed to the low total fermentable sugar (3 4 g l 1) and the low free amino nitrogen concentration (15.3 26.6 mg l 1). L. plantarum exhibited the highest cell population owing to its unique ability to tolerate low pH values by

Table 2 Numerical values of estimated microbial growth parameters in sterile malt, barley, and wheat media Medium Malt Microorganism L. L. L. L. L. L. L. L. L. L. L. L. fermentum plantarum reuteri acidophilus fermentum plantarum reuteri acidophilus fermentum plantarum reuteri acidophilus lmax (h 1) 0.62 F 0.04 0.41 F 0.03 0.38 F 0.02 0.19 F 0.02 0.43 F 0.05 0.20 F 0.02 0.13 F 0.01 0.18 F 0.03 0.53 F 0.05 0.23 F 0.02 0.13 F 0.01 0.15 F 0.01 X0 (log10cfu ml 1) 6.85 F 0.08 6.90 F 0.10 6.20 F 0.07 6.89 F 0.06 6.90 F 0.11 6.71 F 0.13 6.14 F 0.04 7.02 F 0.04 6.93 F 0.09 7.21 F 0.05 6.22 F 0.03 7.02 F 0.02 Xmax (log10 cfu ml 1) 9.68 F 0.03 10.11 F 0.18 8.86 F 0.06 8.10 F 0.06 9.12 F 0.05 9.43 F 0.10 7.28 F 0.05 7.73 F 0.03 9.28 F 0.04 9.29 F 0.06 7.20 F 0.04 7.71 F 0.03 pH 3.77 F 0.09 3.40 F 0.09 3.72 F 0.09 3.73 F 0.09 4.61 F 0.09 3.92 F 0.09 4.88 F 0.09 3.93 F 0.09 4.50 F 0.09 3.83 F 0.09 4.40 F 0.09 3.73 F 0.09

Barley

Wheat

lmax = maximum specific growth rate, X0 = initial cell population, X max = maximum cell population at the end of the exponential phase, pH = pH at the end of the exponential phase. The maximum specific growth rate was estimated using a logistic-type equation, applied to the data obtained during growth. Source: Adapted from Charalampopoulos et al. (2002).

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maintaining a proton (pH) and charge gradient between the inside and the outside of the cells even in the presence of high amounts of lactate and protons (Giraud et al., 1998). L. acidophilus exhibited the poorest growth probably because of substrate deficiency in specific nutrients, confirming the importance of substrate composition in conjunction with the nutritional requirements of the specific strain. 3.2. Survival of probiotics A key factor in the selection of suitable probiotic starter is its ability to survive the acidic environment of the final fermented product (in vitro) and the adverse conditions of the gastrointestinal tract (in vivo). The survival of the probiotic bacteria in vitro might be influenced by the metabolites formed by the starter such as lactic acid and acetic acid, hydrogen peroxide, and bacteriocins (Saarela et al., 2000). Although differences exist between species and specific strains, lactobacilli are generally considered to be intrinsically resistant (Kashket, 1987), especially at pH values higher than 3.0 (Hood and Zottola, 1988; Jin et al., 1998). Of the various probiotic bacteria, L. casei and L. plantarum appear to have longer shelf lives than L. acidophilus, L. reuteri, and bifidobacteria in cultured milk (Lee and Salminen, 1995). Based on the above, the optimum final pH and the concentrations of lactic acid and acetic acid in fermented cereal product in relation to the properties of each specific probiotic strain have to be investigated in order to maximise the viability during storage. Besides the intrinsic stability of each strain, the inclusion of slow-metabolising energy sources, such as arginine, fructose, citric acid, and malic acid, which could be present in cereals, has been reported to enhance the viability by providing energy (Lee and Salminen, 1995). Survival of the probiotic strains during gastric transit is also influenced by the physicochemical properties of the food carrier used for delivery. The buffering capacity and the pH of the carrier medium are significant factors, since food formulations with pH ranging from 3.5 to 4.5 and high buffering capacity would increase the pH of the gastric tract and thus enhance the stability of the probiotic strain (Kailasapathy and Chin 2000; Zarate et al., 2000). In addition, it has been shown that malt, wheat, and barley extracts exhibited a significant protective effect

on the viability of human-derived L. plantarum and L. acidophilus strains under acidic conditions mimicking the stomach, which based on supporting experiments with dietary constituents could be mainly attributed to the presence of soluble sugars in the cereal extracts and to a less extent to the free amino nitrogen content, depending on the strain. (Charalampopoulos et al., data not shown). 3.3. Organoleptic properties The grains of corn, sorghum, millet, barley, rye, and oats contain appreciable amount of crude fibre and lack gluten-like proteins of wheat. The traditional foods made from these grains usually lack flavour and aroma (Chavan and Kadam, 1989). Lactic acid fermentation improves the sensorial value, which is very much dependent on the amounts of lactic acid, acetic acid and several aromatic volatiles, such as higher alcohols and aldehydes, ethyl acetate and diacetyl, produced via the homofermentative or heterofermentative metabolic pathways. Consequently, an appropriate selection of the strain is necessary to efficiently control the distribution of the metabolic end products (Lonner and Preve-Akesson, 1988; Damiani et al., 1996; Hansen et al., 1989). Knowledge of the biochemical pathways leading to flavour production can help in making the right choice of starter. However, the end product distribution of lactic acid fermentations depends also on the chemical composition of the substrate (carbohydrate content, presence of electron acceptors, nitrogen availability) and the environmental conditions (pH, temperature, aeorbiosis/anaerobiosis), controlling of which would allow specific fermentations to be channelled towards a more desirable product (Hansen and Hansen, 1994). In general, probiotic products obtained using a single strain probiotic starter are hardly acceptable to consumers, lacking sensory appeal due to a rather sour and acidic taste. For milk-based products, the probiotic strains are often mixed with Streptococcus thermophilus and L. delbrueckii (Saarela et al., 2000). In this respect, another alternative in enhancing the aromatic profile of the final product would be the incorporation of supporting strains being able to bring out the preferred flavour. It is important that the supporting strains grow in the cereal substrate and do not act antagonistically towards the probiotic strain. In indus-

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trial cereal fermentations L. sanfransisco, the most important sourdough bacteria, is usually mixed with Sacharomyces exiguus or Candida milleri, improving the overall organoleptic properties (Gobbetti, 1998). 3.4. Nutritional value Lactic acid fermentation improves usually the nutritional value and digestibility of cereals. Cereals are limited in essential amino acids such as threonine, lysine, and tryptophan, thus making their protein quality poorer compared with animals and milk (Chavan and Kadam, 1989). Their protein digestibility is also lower than that of animals, due partially to the presence of phytic acid, tannins, and polyphenols which bind to protein thus making them indigestible (Oyewole, 1997). Lactic acid fermentation of different cereals, such as maize, sorghum, finger millet, has been found effectively to reduce the amount of phytic acid, tannins and improve protein availability (Chavan et al., 1988; Lorri and Svanberg, 1993). Increased amounts of riboflavin, thiamine, niacin, and lysine due to the action of LAB in fermented blends of cereals were also reported (Hamad and Fields, 1979; Sanni et al., 1999). Khetarpaul and Chauhan (1990) reported improved minerals availability of pearl millet fermented with pure cultures of lactobacilli and yeasts.

blood glucose, and insulin contents in human body. Water-insoluble fibre contains lignin, cellulose, hemicelluloses (Bingham, 1987; Marlett, 1990), and nonstarchy polysaccharides such as water-unextractable arabinoxylan. Between cereal grains, the content of dietary fibre varies (Table 3) (Nelson, 2001; Herrera et al., 1998). In cereal botanical components, the majority of dietary fibres generally occur in decreasing amounts from the outer pericarp to the endosperm, except arabinoxylan, which is also a major component of endosperm cell wall materials. The procedures for the isolation and purification of dietary fibre and the techniques involved in their quantitative and structural analysis were developed for the isolation of dietary fibre from conventional milling streams. The combination of the debranning or pearling technology and the subsequent simplified milling process might produce processing streams of more specified botanical components, such as the outer pericarp, the inner pericarp, the seed coat, the aleurone cells, the embryo, and the starchy endosperm. Targeting at particular dietary fibres in each of these streams according to the knowledge of fibre distribution in cereal grains, these isolation procedures would be simplified and their products more purified. 4.2. b-Glucan One of the most important members of the dietary fibre family is h-glucan. It is unbranched polysaccharides composed of (1 ! 4) and (1 ! 3) linked h-Dglucopyranosyl units in varying proportions. Various forms of h-glucan have been recognised as having

4. Dietary fibre from cereal grains and their prebiotic and physiological effects 4.1. Definition Dietary fibre is the edible part of plants or analogous carbohydrates, which resists the hydrolysis by alimentary tract enzymes. In addition, fibre is not totally unavailable either, because a portion of dietary fibre is metabolised to volatile fatty acids in the gastrointestinal tract. Dietary fibre can be divided into two categories according to their water solubility. Each category provides different therapeutic effects. Water-soluble fibre consists mainly of nonstarchy polysaccharides, mainly h-glucan and arabinoxylan. By forming viscous solution, soluble fibre slows intestinal transit, delays gastric emptying, and reduces glucose and sterol absorption by the intestine. Soluble fibre also decreases serum cholesterol, prostprandial

Table 3 Comparison of total dietary fibre content in cereal grains Cereals Legumes Rye Corn Triticale Oats Wheat Sorghum Barley Finger millet Rice Total dietary fibre (%, db) 13.6 28.9 15.5 15 14.5 14 12 10.7 10 6.2 7.2 3.9 F 0.2

Source: Compiled from Herrera et al. (1998) and Nelson (2001).

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important positive therapeutic effects on coronary heart disease, on the reductions of cholesterol and glycemic response (Wood, 1993; Beer et al., 1995). In addition, oat h-glucan has been reported to selectively support the growth of lactobacilli and bifidobactera in rat experiments (Ryhanen et al., 1996) and in in vitro studies (Jaskari et al., 1993). High molecular weight h-glucans, up to 3 million Da (Wood et al., 1991), are viscous due to labile cooperative associations. Low molecular weight h-glucans, as low as 9000 Da (Gomez et al., 1997), can form soft gels as the chains are easier to rearrange to maximise linkages. When exposed to physical forces and chemical or enzymatic hydrolysis, molecular size of h-glucan reduces to achieve molecular weights of 0.4 2 million Da in typical food preparations (Beer et al., 1997). Hydrolysates of oat h-glucan have been reported to stimulate the growth of three Bifidobacterium strains and L. rhamosus GG (Kontula et al., 1998). Among all the cereal grains, barley and oats contain the highest level of h-glucan, covering the ranges of 3 11% and 3 7% on a dry basis, respectively. It is usually concentrated in the inner aleurone cell walls and subaleurone endosperm cell walls of barley (Koksel, 1999), oats (Wood, 1993), and wheat (Wood, 1997). Considerable amount of h-glucan is also found in the crease area of wheat (Wood, 1984) and possibly other grains. Wheat is not recognised as a source of hglucan because of its much lower content, usually below 1% on a dry basis. The physical properties of wheat grain, however, allow the development of pearling technology to separate the aleurone layer as potential source of h-glucan. The pearling process applies friction and abrasion to debran grains for the improvement of milling performance. Developments such as the PeriTec process from Satake Engineers and the Tkac process from Tkac and Timm Enterprises provide the opportunity to individually collect wheat botanical components. The combination of the pericarp, the seed coat, and the nucellus forms a rich source of both arabinoxylan and lignin. The bran section in the crease area and the subaleurone section might be separated by the subsequent milling process and mixed into the aleurone section from debranning, as a source of h-glucan. The successful application of the pearling technology to other grains would rely on factors such as proper conditioning of the grains prior to debranning and special design of the debranner.

4.3. Oligosaccharides Oligosaccharides, such as lactulose, fructo-oligosaccharides, transgalacto-oligosaccharides (Gibson et al., 1995; Bouhnik et al., 1997) have received increased attention, especially because they have been shown to be effective in stimulating the growth of bifidobacteria and lactobacilli in human large intestine. These oligosaccharides can be isolated from plant materials or can be synthesised enzymatically (Crittenden and Playne, 1996). In the food industry, simple oligasaccarides are used as bifidogenic substances and some infant products contain them in the hope that this might provide some of the benefits attributed to oligosaccharides in human milk (RiveroUrgell and Santamaria-Orleans, 2001). At least two types of oligosaccarides exist in cereal grains. They are galactosyl derivatives of sucrose, stachyose and raffinose, and fructosyl derivatives of sucrose, fructooligosaccharides (Henry and Saini, 1989). The exact distributions of these polymers within cereal grain have not been fully established. In respect of wheat, reported values suggest their distributions in all milling products, including bran (Yamada et al., 1993), germ (Pomeranz, 1988), and flour (Nakazawa et al., 2000). Wheat germ is particularly rich in raffinose family oligosaccharides, 7.2% on a dry basis. Total sugar content in the aleurone cells have been approximated as 11.1% (Mizuochi, 1999) on a dry basis, while the reported values for milling flour fall in the range of 1.2 1.6% (Pomeranz, 1988). Extraction of oligosaccharides from natural resources has not been fully developed due to the complexity of these substances and their connections with other macromolecules, particularly proteins. Oligosaccharide concentrate might be obtained from cereal botanical constituents by exploring their water solubility. Taking wheat as example, the water washing process commonly applied for gluten separation would retain watersoluble oligosaccharides in the starch slurry. After the removal of starch by centrifugation or vibrating screen, the residual paste might be centrifuged to obtain a coloured fibrous substance, starch tailings. Oligosaccharides can be released from other cell wall components by the treatment of cellulase to these tailings. Separation of oligosaccharides from both wheat germ residue, usually after oil extraction, and from the aleurone layer might be developed based on the sol-

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ubility of oligosaccharides in 80% ethanol solution (Henry and Saini, 1989). Purification of these materials will require the application of cellulose column chromatography (Mizuochi, 1999) gel filtration (Palmacci et al., 2001), high-performance liquid affinity chromatography (Zopf et al., 1989). Cereal bioprocessing through enzymatic reactions or through fermentation can also produce a large range of oligosaccharides with potential prebiotic properties. The a-amylase present in the cereal grain can hydrolyse the gelatinised starch granules, and the extent of the hydrolysis could be regulated through temperature control. The different fractions of the oligosaccharides obtained could then be separated and their functionality could be tested. Another alternative for the hydrolysis of the starch would be through fungal fermentation of the use of solid state fermentation technology. The processing steps prior to the starch hydrolysis (e.g., milling) could also have an effect in the biotransformation and should be taken into account. 4.4. Resistant starch Resistant starch has been recognised as a functional fibre performing an important role in digestive physiology. Similar to oligosaccharides, especially fructooligosaccharides, it escapes digestion and provides fermentable carbohydrates for colonic bacteria. Resistant starch has also been shown to provide benefits such as the production of desirable metabolites including short-chain fatty acids in the colon. In addition to its therapeutic effects, resistant starch provides better appearance, texture, and mouthfeel than conventional fibres (Martinez Flores et al., 1999). Opportunities exist for the development of ingredients from resistant starch as prebiotics for decreasing the risk of bowel diseases. Resistant starch might be classified into four categories (Yue and Waring, 1998), but the natural types are frequently destroyed when subjected to modern food processes. Resistant starch is naturally found in cereal grains and in heated starch or starch-containing foods. The manufacture of resistant starch usually involves partial acid hydrolysis and hydrothermal treatments (Brumovsky and Thompson, 2001), heating, retrogradation (Schmiedel et al., 2000), extrusion cooking (Gebhardt et al., 2001), chemical modification (Wolf et al., 1999), and repolymerisation (Yue and Waring, 1998).

5. Encapsulation of probiotic strains using cereal fractions In the last years, several encapsulation techniques using cereal fractions have been tested in order to improve the viability of the probiotic strains in functional foods. The possibility of using high amylose maize (amylomaize) starch granules as a delivery system for probiotic bacteria has been investigated by Wang et al. (1999). In this case, Bifidobacterium strains isolated from a healthy human were used adhered to amylomaize starch granules. In vitro studies showed that growth in these conditions led to enhanced survival of the probiotic strains. Survival in vivo was also monitored by measuring the faecal level of Bifidobacterium after oral administration of the strain to mice. A sixfold better recovery of the strains was noted for cells grown in amylose-containing medium compared with the control. A modified method using calcium alginate for the microencapsulation of probiotic bacteria in yoghurt has also been reported in the literature (Sultana et al., 2000). Incorporation of maize starch (a prebiotic) with alginate improved the encapsulation of viable bacteria as compared to when the bacteria were encapsulated without the starch. The survival of encapsulated cultures was in all cases higher than with the free cells. Techniques such as spray drying could be used to produce small uniformly coated microspheres containing the viable probiotic bacteria (ORiordan et al., 2001). Other encapsulation techniques have been used by other authors. Jankowski et al. (1997) used capsules of a liquid starch core with calcium alginate membranes, while Selmer-Olsen et al. (1999) and Shah and Ravula (2000) used pure Ca-alginate gel beads. Adhikari et al. (2000) used kappa-carrageenan for the encapsulation of bifidobacteria in yogurt. In all these cases, the encapsulation technique could be used to transmit probiotic bacteria via fermented products provided that the sensory characteristics of the product are improved or maintained.

6. Future perspectives Cereals are generally suitable substrates for the growth of human-derived probiotic strains. Regardless

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of the relatively big differences in performance between species and the complexity of cereal substrates, a systematic approach is needed in order to identify the intrinsic and processing factors that could enhance the growth and, more importantly, the survival of the probiotic strain in vitro and in vivo. The possible improvement of the organoleptic properties should also be investigated by using supporting cultures that act synergistically on the probiotic strains. Additionally, the functionality of colonic strains could be improved by the presence of specific nondigestible components of the cereal matrix that could act as prebiotics. The possibility of separating specific fractions of nondigestible soluble fibre from different types of cereals or cereal by-products, either through primary processing technologies, such as pearling and sieving, or through enzymatic modifications, looks very promising. The development of new functional ingredients has the advantage that food manufacturers can add extra value to products the consumer is already familiar with. Developing new foods involves larger marketing campaigns and often the consumer needs an adaptation time to the new product. By either developing new and innovative products or just reformulating existing ones, nutritional food ingredients enable manufacturers to meet and exceed the expectations of todays health-conscious consumer. Cereals not only have the ability to grow and deliver probiotic lactic acid bacteria to the human gut, but also contain potentially prebiotic compounds whose functionality should be explored. References
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