Budapest, Hungary
April 18-19, 2002

Ministry of Education, Research and Development Division
H-1052 Budapest, Szervita tér 8.

Institute for Agrobotany
Tápiószele, Hungary

FVM Agrármarketing Centrum Kht
Budapest, Hungary

ELMA Rt.
First Hungarian Fruit Processing and Fruit Juice Manufacturer Share Company
Érsekhalma, Hungary

Influence of autumn herbicide treatments on the seasonal changes in carbohydrates in the roots of Cirsium arvense.
Wilson, Robert G.
University of Nebraska, 4502 Avenue I, Scottsbluff, Nebraska 69361, USA
rwilson1@unl.edu

Cirsium arvense is a troublesome weed found throughout the northern part of the USA. Each year land managers spend millions of dollars to try and control this weed. Results of several experiments have shown that herbicide applied in autumn is more effective than treatment made in the spring or summer. To examine this hypothesis in more detail, experiments were conducted near Scottsbluff, NE from 1999 to 2001 to examine the changes in glucose, fructose, sucrose, and fructans in the roots of Cirsium arvense following autumn application of two herbicides, dicamba and 2,4-D. Plant roots were exhumed from the soil and a root extract analyzed with a HPAEC-PAD with a Dionex 500 system. Plant shoot density was measured 7 months following treatment to determine herbicide effectiveness.

Dicamba applied at 0.56 kg/ha 4 to 11 days before the first frost (-2 C) was not as effective in controlling Cirsium arvense as treatments made 1 to 10 days after the first frost. Application of dicamba after the first frost resulted in a decrease in low- to mid-degree of polymerization fructans and increase in fructose in plant roots 30 days after treatment. Increases in Cirsium arvense control 7 months after treatment could be correlated with decreases in fructans and increases in fructose that were recorded in plant roots 30 days after herbicide treatment. In a similar experiment dicamba was applied at 0.3, 0.6, and 1.1 kg/ha and 2,4-D at 0.6, 1.1, and 2.2 kg/ha 4 days before the first autumn frost. Dicamba was more effective in controlling Cirsium arvense 7 months after treatment than 2,4-D and control with both herbicides increased as rate increased. Increases in weed control with herbicide observed 7 months after treatment could be correlated with decreases in low- to mid-degree of polymerization fructans that were recorded in plant roots 30 days after treatment.

Treatment of plants with 2,4-D was not as effective as dicamba in reducing fructans in plant roots. Quantities of fructan 1-exohydrolase (1-FEH) were increased in the roots of Cirsium arvense 30 days following treatment of the plant with dicamba.

(1) Ministry of Flemish Community, Centre of Agricultural Research, Department of Plant Genetics and Breeding, Caritasstraat 21, 9090 Melle, Belgium

(2) Chicoline, Research and Chicory Breeding, Rue de la Sucrerie 1, 7740 Warcoing, Belgium

The inulin yield of industrial chicory may be 30 % higher when sown before mid-April than at a sowing date one month later (Baert, 1997). Early sowing enhances the risk of bolting. There is a lot of genetic variation for bolting sensitivity at early sowing. The inheritance is mainly additive and the heritability is high (Baert and Van Bockstaele, 1999). In our selection programme we apply a strong selection for bolting resistance by selecting non bolted roots in very early sown trials. Does this selection affect important features as root yield and quality?

At the end of February 1998 and the beginning of March 1999 we sowed 4 chicory populations and selected not bolted roots in November. One year later we harvested seeds on the selected roots. We also produced a next generation of the original populations without selection for bolting resistance. In 2000 and 2001 the original populations and the seed lots of the next generation after and without selection were sown in February for bolting observation and in May for determination of fresh root yield, inulin content, inulin yield and inulin chain length ( as fructose/glucose ratio). We compared the performance of the generations after and without selection for bolting resistance with the correseponding original populations (relative value = 100).

The absolute average bolting % of the 8 original populations was 40. In the same trials the control varieties Orchies and Tilda had resp. 40 and 92 % bolters. Fig. 1. shows the effect of selection for bolting resistance. The selection reduced the number of bolters significantly by 31 % relative . Without selection the number of bolters increased nonsignificantly by 19 %. The effects of selection for bolting resistance on yield and inulin quality were slightly positive but non signifcant.

• Baert J. 1997. The effect of sowing and harvest date on inulin yield and composition of chicory roots. Industrial Crops and Products, 6, 195-199.
• Baert J. and Van Bockstaele E. 1999. Heritability of bolting sensitivity, fresh root weight, inulin content and inulin chain length of chicory (Cichorium intybus L.). Proceedings of the 4th European Symposium on Industrial Crops and Products, Schriftenreihe Nachwachsende Rohstoffe, Band 14, 188-194.

The role of food has changed from the energy supply and sensory values to a health promoting character. According to some market surveys in Hungary 16% of consumer is oriented to buy healthy food. This is an upcoming trend in the new age of functional food. One of the best example for decreasing risk of a disease by selecting food fermented dairy products are known by their probiotic and prebiotic effects.

Probiotic microorganisms are able to survive in the human digestive system. Two groups of the human intestinal flora, Lactobacillus and Bifidobacterium are added (inoculated) to milk, which provides favorable carrier conditions to pass through the gastrointestinal tract. Microbial colonies of this two bacteria are able to be bind (immobilized) to the the enteric epithelium of the long bowel by the intake of prebiotic carbohydrates.

Fermented dairy products contain large number of cells and the public accepts the presence of living bacteria.

This products are palatable food items and their healthy effect is known by the consumer.

Prebiotics, like inulin and oligofructose are natoral carbohydrates (fructans) present in several fruits and vegetables like onion, garlic, banana, chicory and others. Chicory inulin and oligofructose are recognized as natural food ingredients and used as dietary fibres in European countries.

Inulin and oligofructose are practically indigestible in the small intestine, but fermented by bacteria colonising in the large bowel. The production of lactate and short-chain fatty acids leads to the selective stimulation of growth of the gut microflora.

The cooperation of prebiotics and probiotics provides excellent nutritional properties for functional food products. Regular consumption of fermented dairy products has a protective effect and decreases risk of intestinal diseases. Market value of functional food products is accepted by consumer science.

W. Praznik¹, E. Cieslik² and M.G. Lopez³

1Institute of Chemistry, University of Agricultural Sciences, Muthgasse 18, A-1190 Wien, Austria; Email:wpraznik@edv2.boku.ac.at

Szent István University, Faculty of Food Sciences, Department of Canning Technology, Budapest, Hungary
konz2@omega.kee.hu

Jerusalem artichoke is a plant that can serve as an alternative source of carbohydrate. The tubers contain 14-15 % inulin, 0.8-0.9 % minerals and 0.6-1.5 % raw fibre as basic components of healthy nutrition. Inulin is a polysaccharide, that consist from 80 % fructose and 20 % glucose. Since it is hard to digest for humans, it functions as dietary fibre. Carbohydrates, in addition to being our most important energy-giving nutrients, also cause “civilization illnesses” like diabetes, obesity and caries. Inulin, on the other hand, hydrolysed to fructose is better tolerated by diabetics; additionally, it is a more intensive sweetener , than saccharose and glucose. Other valuable nutrients of Jerusalem artichoke are the minerals. The high kalium content (0.4% w/w) of the tubers, for example, is of utmost importance.

Immunological adjuvants are of major importance as agents to potentiate the immune response. In humans, only aluminium-containing compounds are used as vaccines, since all the other known potent adjuvants - such as Freund's adjuvant - are too toxic. Since aluminium-hydroxids and phosphates give less intense and persistent response, it is of great interest to find alternative adjuvants that generate good immune response without inducing deleterious effects, both for human and veterinary use.

Presenting author: Vanesa Olivares-Illana, Instituto de Biotecnología UNAM, México.

Co-authors: Olvera -Carranza C., López-Munguía A.
vanesa@ibt.unam.mx

Fructosyltransferases are enzymes that catalyse the transfer of a fructosyl residue from sucrose or raffinose to a growing polysaccharide chain. The resulting polymer may be levan, when the fructose linkages are mainly  (2-6), as those occurring extensively in bacterias, or inulin, a (2-1) polymer found mainly in plants.

From peptides sequenced from the purified protein two primer were designed and a 3 Kpb fragment amplified by PCR, from which the whole sequence was obtained and expressed in E. coli DH5Some structure features of the enzyme deduced from the primary structure are discussed.

Applied Plant Research, Research Units Arable Farming, Field Production of Vegetables and Multifunctional Agriculture. P.O. Box 430, 8200 AK Lelystad, The Netherlands

l.van.den.brink@ppo.dlo.nl

In the Netherlands ca. 5000 ha of industrial chicory is grown for production of inulin and fructose. The chicory is harvested in general in the period beginning september untill the end of november. Later harvesting is riskfull because of wet soil conditions and frost. The capacity of the factory is not large enough to process the total harvest in this period. Medio february the last chicory is processed. So a large amount of chicory is stored in piles in the open air during ca. 8 weeks on the farms. Depending on the weatherconditions during storage and the quality of the chicory-roots considerable losses in yield can occur and also the quality can decrease.

In the years 1997 till 2002 research has been done on methods to reduce these losses. In uncovered piles and piles covered with different materials losses in yield and decrease in quality were measured. The occurance of frost during the storage period had a great influence on the losses. In years with frost during the first weeks of storage followed by a period with relative high temperatures high losses were measured in uncovered piles. With piles covered temporally only during the frost period less losses were measured. In other years without frost the losses in uncovered piles were lower than in covered piles with insufficient ventilation. Heatproduction of chicory is very high in comparison with sugarbeet. Without sufficient ventilation in the pile respiration losses were increasing and also diseases, especially Sclerotinia, were spreading quickly. In covered piles with forced ventilation it was possible to reach less losses and a higher inulin content. However the costs of these piles are very high.

Additional research was done in climate-rooms. The effect of temperature on inulin content, carbohydrate composition and losses of root yield was measured.

al.

29 Listopada 46, 31-425 Cracow, Poland: e-mail:rrciesli@cyf-kr.edu.pl

Besides their role as reserve carbohydrates, fructans may fullfil other more specific roles in plants which include drought or frost protectants and fructans acting as osmoregulators. Up to now no hard evidence has been provided that fructans act as drought or frost protectants in vivo. On the contrary, it was convincingly demonstrated that fructans indirectly (i.e. after hydrolysis by fructan degrading enzymes) contribute to osmoregulation during flower opening of the monocot species Hemerocallis [Bieleski et al., 1993].

Within dicots, fructan metabolism has been almost exclusively been studied within the Asteraceae. Here, we report on the study of fructan metabolism in Campanula rapunculoides, a representative of another dicot family (Campanulaceae). Inulin type fructans are detected in all organs of Campanula rapunculoides. Petals store inulin in the bud stage.

A rapid breakdown of inulin just before flower opening resulted in high concentrations of hexoses (especially fructose) and a temporal increase in the concentration of inulo-n-oses (Fn type fructans) which may contribute to the osmotic driving force involved in petal expansion.

A marked increase in 1-FEH activity was found together with the fructan degradation. The flower petal 1-FEH cDNA was cloned by 5' and 3' RACE RT-PCR starting with conserved primers. The translated cDNA groups together with chicory 1-FEH I, IIa and IIb, forming a distinct group within the cell-wall type glycosyl hydrolases because of their low iso-electic point. It was found that the Campanula flower cDNA was more homologous to chicory root 1-FEH IIa and IIb than to chicory 1-FEH I. The functionality of the clone has been confirmed by heterologous expression in Pichia pastoris.

Kerklaan 30, 9751 NN Haren, The Netherlands

Both polymers were synthesized by sucrase type of enzymes (glucosyl- and fructosyltransferases). These enzymes only need sucrose as substrate; the energy released by the cleavage of the glycosidic bond in sucrose is subsequently used for the polysaccharide synthesis reaction. During growth of L. reuteri on sucrose or maltose, the sucrases responsible for the synthesis of the glucan and the levan appeared to be completely bound to the cell wall, whereas during growth on sucrose part of the enzymes was released into the culture medium. The EPS production was not a stable characteristic in continuous cultures. Different spontaneous mutants appeared, such as the EPS negative mutant strain K24, lacking both the glucansucrase (a glucosyltransferase) and the levansucrase (a fructosyltransferase). Mutant 35-5, lacking levansucrase, appeared after a pH shift-down.

Using PCR techniques with degenerated primers based on known glucansucrase or fructosyltransferase amino acid sequences, chromosomal fragments containing a glucansucrase (gtfA) or a fructosyltransferase (ftfA) were amplified. Both fragments were sequenced and characterized at the amino acid level and phylogenetic trees of both types of sucrases were constructed.

Both the gtfA and the ftfA were cloned separately in Escherichia coli. Cell free extracts of the E. coli strain harboring the ftfA gene produced an inulosucrase, which synthesized inulin and fructose-oligosaccharides from sucrose. The recombinant glucansucrase and the L. reuteri glucansucrase synthesized the same unique glucan. These were the first examples of the isolation, characterization and cloning of Lactobacillus glucansucrase and fructosyltransferase genes.

We used a novel expression system for fructosyltransferases, namely tobacco BY2 suspension cultures. These cells can be transformed with the use of Agrobacterium and in both callus and suspension cultures of transformed cells enzymatic activity is high. 6G-FFT from onion expressed in BY2 cells can make a whole range of fructans, when 1-kestose is provided as a substrate. The Dionex profile of the fructans made by 6G-FFT resembles that of onion, showing that 6G-FFT is able to synthesize inulin neo-series of different DP’s and with different linkages. This strongly indicates that 1-SST and 6G-FFT are the enzymes responsible for the pattern of fructans found in onion, suggesting that these are the two fructosyltransferases present in onion.

In a collaboration with Advanta and Sensus, 1-SST and 6G-FFT from onion have been transformed to sugar beet. The beets containing 1-SST produce 1-kestose. The beets containing both 1-SST and 6G-FFT produce the onion-type inulin neo-series, showing that 1-SST and 6G-FFT are together sufficient to produce the fructans found in onion.

Influence of xxxx short-chain fructo-oligosaccharides (sc-FOS) on bioavailability of magnesium and calcium in human

(1) Nutri-Health SA – 3, avenue Paul Doumer F-92500 Rueil-Malmaison

(2) Béghin Meiji – 14 boulevard du Général Leclerc F-92 572.Neuilly sur Seine

Numerous studies conducted in rats suggest that various Non-Digestible Oligosaccharides (NDO’s) may improved mineral absorption in human. The physiological mechanisms which controlled intestinal magnesium and calcium absorption in human are different (passive vs active and down-regulated) and explain the contradictive results observed in recent clinical studies.

Magnesium, as a co-factor of numerous metabolic reactions, is a pivotal nutrient for many body functions. An optimal food daily intake contribute to bone health. In European countries, magnesium food consumption is often below the Recommended Daily Intake (RDI: 6 mg/kg body weight). The SUVIMAX’s study¹ conducted in France has demonstrated that more than 70% of French population does not fit the RDI. Moreover, 20% of the French population has a magnesium daily intake below two third of the RDI which states a magnesium deficiency. Two recent clinical studies^{2 3} conducted in human have demonstrated that 7-10g/day sc-FOS ACTILIGHT over a long period improve dietary magnesium absorption from 12% in post-menopausal women to 18% in adolescents.

It is well established than an optimal dietary calcium intakes (RDI: 900-1300 mg/day) contributes to reduction of risk of osteoporosis. As a limited amount of ingested calcium is absorbed (30-45%) a nutritional challenge is to improved food calcium bioavailability. Several human studies have recently reported an improvement of calcium absorption with NDO’s. Positive effect of NDO’s on calcium absorption have only been observed when NDO’s where given over a short period of time (7-9 days) or in combination with high dietary calcium intakes (1500 mg/day). On the other hand, results observed with NDO’s (especially fructans) over long periods (4-5 weeks) and low or normal dietary calcium intakes (650-950 mg/day) are negative. These clinical data suggest a down-regulation of the active calcium absorption mechanisms which would limit the potential beneficial effects of NDO’s on calcium absorption. As long as a scientific consensus on the NDO’s effect on calcium absorption is not achieved, related functional claim is out of place. Further studies are ongoing.

Feed Research Institute, Chinese Academy of Agricultural Sciences;

Address:12 Zhongguan Nandajie, Beijing100081,CHINA;

E-mail: wangjianhua@mail.caas.net.cn; Phone: 0086-10-62145735; Fax: 0086-10-68975127

* The first author Dr Wang Jian Hua is the responsible and corresponding person of this paper.

An yeast strain Kluyveromyces marxianus IW9801 (CGMCC0360) with high inulinase yielding was screened. The genome DNA from the strain was extracted as template DNA of PCR; The special primers on purpose of cloning for whole exo-inulinase via PCR amplification were designed and synthesized; The expression vectors in Pichia pastoris GS115 was constructed, and the host strain was transformed via the vector, recombiants were screened; The expression of the target protein in shaking flask and fermentor were initially optimized, respectively. The highest exo-inulinase activity 577U/ml for the recombinant Pichia pastoris and 289U/ml for original strain Kluyveromyces marxianus reached when a high cell density cultivation was developed for inulinase production at the 96th hour and 120th hour of fermentation, respectively. It is of great importance of enzyme industry and application in food industry.

Key words: Kluyveromyces marxianus, Pichia pastoris, cloning, DNA recombination, inulinase, high density cell cultivation, and high fructose syrups.

(1) Agricultural Research Institute of the Hungarian Academy of Sciences, Martonvásár, Hungary; (2) Dept. of Genetics and Molecular Biology, University of Pécs, Hungary
galibag@pengiun.mbki.hu

Frost tolerance of wheat is determined by the interactions of plant genotype and environment. To achieve the genetically determined frost resistance the plant must be cold hardened. During cold treatment the metabolism of plant is altered due to the temperature dependent regulation of the activity of certain enzymes and changes in the pattern of gene expression. To verify the genetic relationship between the frost tolerance and the cold induced changes in carbohydrate metabolism, chromosome substitution, recombinant and deletion lines were studied. Major genes influencing frost tolerance (Fr) and vernalisation requirement (Vrn) were localised on the long arm of 5A chromosome. The mechanism whereby the Vrn-A1-Fr1 interval affects freezing tolerance remains to be determined. One possible approach to solve this puzzle is the localization of genes affecting metabolic processes during cold acclimation. The effect of cold hardening on the dynamics of frost tolerance and on water-soluble carbohydrate content, including fructan, sucrose, fructose and glucose, was studied in substitution lines. Significant correlation between these parameters was detected during the 51-day hardening period. A study using 5A chromosome recombinant lines showed that the gene regulating sucrose accumulation seemed to be closely associated with the Vrn-A1 locus but separable from the Fr1. This result was verified using 5A deletion lines.

The effect of drought and salt stresses on the water-soluble carbohydrate content in wheat seedlings was examined to characterize the involvement of major sugar components in the adaptive processes. Plants differing in drought and salt tolerance were grown hydroponically, and they were exposed to consecutive water (polyethylene glycol) and salinity (NaCl) stresses. The rate of sucrose and fructan accumulation in stems proved to be reliable marker either for drought or salinity tolerance.

Plant Research International 6700 AA Wageningen The Netherlands

The objective of our research is to understand the mechanism of regulation in order to increase the quality and length of the fructans in the agricultural crop chicory (Cichorium intybus L.).

Previous studies on the localization of fructan synthesizing enzymes have been performed using antibodies against the proteins. Here we report the localization of 1-fructan: fructan fructosyltransferase (1-fft ) expression in chicory taproots.

At first we analysed the expression driven by the 1-fft promoter regions by expressing GUS in transgenic chicory plants under control of two different 5'upstream regulatory sequences of 1-fft.

We also analyzed the expression, by Northern analysis, of the native 1-fft in the different rings of the root.

In transversal root slices, when GUS was driven by an 1100 bp. promoter fragment, GUS activity was visualized in tissue located around the vascular cambium, in young xylem and phloem. The Northern analyses showed highest 1-fft expression around the vascular cambium. This agreed with the finding that the 1-FFT protein was located around the phloem, but expression was also found around young xylem tissue. These results showed that the cloned 1100bp. promoter fragment contained the regulator sequences needed for 1-fft regulation.

The plants harbouring the construct containing the 500bp promoter-GUS fusion did not show any GUS activity, unless the root slices were incubated in a high sucrose-solution. This shorter fragment seemed to have a stress or sugar response element.

There are many evidences suggesting that fructans, low-molecular-weight non-digestible carbohydrates which are naturally occuring dietary constituents may improve the quality of life and increase disease resistance in both humans and animals. The potential health benefits of theese compounds are: prebiotic effect and better resistance to pathogens, reduction in blood lipids and the risk for coronary heart disease, antitumor properties and immune stimulation. The stimulatory effects of fructooligosaccharides on intestinal calcium and magnesium absorption have been well studied and the results are extremely promising.

The aim of this experiment was to examine whether the FOS consumption affected bioavailability of Ca and Mg in rats.

The calcium and magnesium levels in serum and urine of rats were determined in Medical Laboratory Centre S.C. ”Diagnostyka”. The contents of these elements in diets and faeces were analyzed by the flame method, with a Philips PU9100X AAS, after ashing of the samples in the muffle furnace (450 ^oC, 12 h).

Data were analyzed by two-factorial ANOVA.

Growth of rats was not affected by fructooligosaccharides. As compared to the control groups (grups I, III), the addition of FOS (group II, IV) to the diet with normal and deficient amount of Ca increased the concentration of calcium and magnesium in blood and decreased the level of these elements in urine and faeces. The absorption ratio of calcium increased from 34,9 to 59,5% and magnesium – from 66,2 to 78,1% at normal level calcium in diet, and in condition of 50% calcium deficiency these values increased – for calcium (z 73,3 do 77,4%) and for magnesium (z 67,9 do 82,8%). In contrast, rats fed the diet with 30% excess of calcium and 5% FOS had lower absorption of these elements ( from 47,8 to 35,9% for calcium and from 73,7 to 69,2% for magnesium) despite higher values of Ca and Mg in blood and dry mass of faeces.

FOS consumption enhances calcium and magnesium absorption. These stimulatory effects of FOS have been explained by their fermentation in the large intestine to short-chain fatty acids, which in turn reduce luminal pH.

In this study , FOS had similar effects on mineral absorption.

Departament of Human Nutrition Agricultural University of Cracow

al. 29 Listopada 46, 31-425 Cracow, Poland: e-mail: rrciesli@cyf-kr.edu.pl

Fruit juices are attractive and popular food products. The rich range of components – fruits, vegetables, herbs and spices and especially attractive tropical plants – creates the possibility of numerous new compositions of drinks. The use of Jerusalem artichoke is a good example. As the bulbs of the plant contain inulin and fructooligosaccharides, they become an important component in the production of functional food products.

The work was aimed to evaluate the quality of fruit juices (cherry and black currant juices) with various admixtures of the juice obtained from the bulbs of Polish species of Jerusalem artichoke, i.e. Albik and Rubik. The bulbs were harvested twice, in the fall and in spring, after storing them in the soil over the winter. The Jerusalem artichoke juice was obtained by means of a laboratory periodic juice extractor. Then, the juice was closed in hermetic containers, pasteurised and stored in the temperature of 2-4^oC. The material prepared in this manner was used for further research. The evaluation procedure covered mixtures of cherry and black currant juices with an admixture of topinambur juice in the following ratios: 1:1 and 1:3. Organoleptic assessment was carried out according to a 5-grade scale. The following parameters were considered: clarity, colour, aroma and taste.

The quality of standard juices was high and it reached 4.4 for cherry juice and 4.7 for black currant juice. Each admixture of Jerusalem artichoke to the juices reduced the number of points a mixture received. The juice of Rubik species harvested in the fall mixed in the 1:1 ratio with black currant juice was assessed to be fairly satisfactory, while the juice of Albik species was found to be unsatisfactory. The 1:1 mixtures of juices extracted from Albik and Rubik bulbs gathered in spring were found to be fairly satisfactory. The change in the ratio to 1:3 resulted in an improvement of the assessment results. Irrespective of the plant species and the time of bulb gathering, the juices were found to be good.

1. Central Food Research Institute Budapest Hungary

2. Szent István University, Faculty of Veterinary Sciences, Department of animal

Breeding Budapest, Hungary

b.czukor@cfri.hu

Lactic acid bacteria are generally considered as beneficial agents for human and animal health. Several strains of Lactic acid bacteria and their combinations with different prebiotics already used as feed additive to improve growth performances and digestic hygiene

Our investigations were aiming to compare the effects of diet,. containing JAP and Bb-12 in Rats. For these experiments thermophilic culture as Bifidobacterium Bb 12 stains was fermented on DSMZ_58 medium with 40 g/l Jerusalem Artichoke supplementation. The ferment broth was concentrated and separated by DDS membran filtration with RO membranes

The other aim of our study was to investigate the effects of Jerusalem Artichoke powder and Bifidobacterium Bb-12 supplemented diets (fed for 14 days) on the fallowing parameters :1, feed intake, 2. body weight, 3. feed/gain, 4. activity of pancreatic hydrolyses(amylase, lipase and trypsin) in the pancreatic tissue homogenate and small intestinal content, 5. large intestinal flora (Lactic acid producers, Coliforms and Sulfid reducing Clostridia in Rats.

1. The supplementation of the rat diet with JAP (2%) and JAP (2%) and Bb. 12 slightly but not significantly increased the weight grain and improved the feed conversion rate of the rats, however the other investigated JAP concentrations and also the Bifidobacterium supplementation exerted no beneficial effects on these parameters.
2. The supplementation of rat diet with JAP and/ or Bb.12 did not influenced significantly the activity of pancreatic hydrolyses.
3. The supplementation of the rat diet with JAP and/or Bb. 12 significantly and dose dependently decreased the number of coliform germs. JAP 2% increased, JAP 6 %, Bb. 12 and JAP + Bb. 12 decreased the number of lactic acid producers.

Ukrainian State University of Food Technologies

In this work were tested different methods of inulin isolation from vegetable raw material: pressing, extraction, centrifugion etc.

Also was tested different kind of raw material: fresh roots and bulbs and roots dried by different methods.

In the result of investigation was founded that in depending from final products quality can be choosed the method of inulin isolation.

Ukrainian State University of Food Technologies

A functional claims refers to the positive consequences of the interactions between a food component and specific genomic, biochemical, cellular or physiological functions without direct reference to any health effect or disease prevention.

A health claim refers to the prevention of a patology or a disease by consuming a spesific food component or food ingredient.

In this work we discuss of different inulins properties which give it possibilities to use it with claims «functional» and «health» food.

* Centro de Estudos de Produçăo e Tecnologia Agrícolas, Instituto de Investigaçăo Científica Tropical, Apartado 3014, 1301 Lisboa Codex, Portugal

Chicory (Cichorium intybus L.) has been cultivated in Portugal (Ribatejo region and Azores) as a coffee substitute. Considering the increased value of this crop amongst inulin-rich materials, namely for the production of syrups with a high content of fructose, studies on industrial up-grading of this raw-material were developed. The carbohydrates composition of the fresh roots showed a very high potential of the portuguese chicory (19,9-21,3 % inulin and 21,2-22,9 % total fructose); inulin and total fructose yields reached values of 7,9-10,8 t ha^-1 and 8,5-11,5 t ha^-1, respectively.

Following earlier work, a technological process to obtain fructose-rich concentrates from chicory was applied to chicory roots harvested at different dates.

Max-Planck-Institute of Molecular Plant Physiology, D-14424 Potsdam, Germany;

Fructans have for a long time been considered as protective agents in the drought and freezing tolerance of plants. We have shown before that low concentrations (up to 10 mg/ml) of inulin isolated from chicory roots and dahlia tubers stabilize egg phosphatidylcholine (EPC) large unilamellar vesicles during freeze-drying by hydrogen bonding to the phosphate in the lipid headgroups (Hincha et al., 2000, Eur. J. Biochem. 267, 535-540).

We have now investigated the effect of chain length of polysaccharides on membrane interaction. Using Fourier transform infrared (FTIR) spectroscopy, we found that fructans with increasing chain length (DP=degree of polymerization 2-5) are progressively more effective in lowering the gel to liquid-crystalline phase transition temperature (T_m) of freeze-dried EPC, while in the case of glucans an inverse correlation exists between DP and reduction of T_m. For the DP 5-sugars, T_m was 9°C lower for the fructan than for the glucan, indicating reduced sugar-membrane interactions for the glucan as compared to the fructan.

A reduced interaction of the higher DP glucans and an increased interaction of the respective fructans with the phospholipid headgroups in the dry state was also indicated by dramatic differences between samples with glucans and fructans in the phosphate asymmetric stretch region of the infrared spectrum.

The capacity to interact with membranes correlates with stabilizing effects on liposomes against leakage of aqueous content after air-drying and rehydration, indicating that fructans are good candidates for the protection of cellular membranes against damage during freezing or periods of drought. As an in vivo test system, we have transformed Arabidopsis thaliana with the fructosyl transferase genes of globe artichoke (Cynara scolymus) and have analyzed the leaf carbohydrate content of the transgenic plants. Expression of only 1-SST leads to the formation of small amounts of inulin type oligofructans with a DP of up to 6, while

combined expression of 1-SST and 1-FFT causes a reduction in the fructan content of leaves and lowers the maximal DP of fructans. We conclude that in the presence of vacuolar invertases, the reversibility of the fructosyltransfer reaction of 1-FFT prevents fructan accumulation by promoting a reversion of chain elongation. This is further substantiated by the finding that cold exposure of the plants, which leads to a reduction of vacuolar invertase activity, results in an increased accumulation of fructans in 1-SST/1-FFT expressing plants.

(1) Department of Agroenvironmental Science and Technologies,

University of Bologna, Via Filippo Re 6/8, 40126 Bologna, Italy.

Eleven varieties of chicory (Cichorium intybus, L.) were compared in Bologna (44° 33’ N, 11° 21’ E, 32 a.s.l.) in the year 2000. Photosynthetic capacity and other physiological parameters were measured in plants with more then 20 fully expanded leaves. Inulin content, fresh and dry weights and nitrogen content (Kjeldahl method) of roots and leaves were determined. Root fresh weight ranged from 54.1 t ha^-1 (Fedora nova) and 70.4 t ha^-1 (Candi) with significant difference among varieties. Leaf fresh weight was also significant different: Tilda (26.1 t ha^-1), Inula (24.5 t ha^-1) and Candi (24.4 t ha^-1) did show the best performances. Tilda and Candi had also the highest root fresh weights. Root dry matter percentages were slightly negatively correlated to root fresh weights, therefore varieties with a significant higher fresh weight have not ever had a significant higher dry weight. Average root nitrogen content did not differ among varieties ranging from 0.78% (Inula) to 0.62% (Tilda and FD9708). Nitrogen content was statistically lower in roots than leaves that removed 47% average total nitrogen from the soil. Also root inulin percentages and productions were not significant different among varieties. Average inulin content (d.m.) was 54.1 ± 2.8%; Regalo showed the highest value (58.9%), whereas Fedora nova the lowest (49.7%). The highest inulin production was obtained by Tilda (9.2 t ha^-1), the lowest by Fedora nova and FD9607 (both 6.9 t ha^-1). Net assimilation rate (A) changed among varieties: Regalo (17.6 mol CO₂ m^-2 s^-1) and Tilda (15.0 mol CO₂ m^-2 s^-1) did show the highest net photosynthesis, Hicor and Fedora nova the lowest (9.8 and 8.0 mol CO₂ m^-2 s^-1). Significant correlation between net assimilation rate, root dry weight (r = 0.47*), inulin percentage (r = 0.74**) and inulin production (r = 0.65**) were found. Therefore net photosynthesis measured at this stage did show fairly representative of the whole season inulin accumulation rate. Transpiration rate (E) was very closely correlated to net assimilation rate (r = 0.75**). The instantaneous water use efficiency (WUEi) was not correlated to photosynthesis but was statistically different among varieties. FD9708 had both the highest WUEi and the intrinsic water use efficiency (A/gs). WUEi was also negatively correlated (r = - 0.60**) to root dry weight. Stomatal conductance was positively correlated to photosynthesis (r = 0.66**) but not to the intercellular CO₂ concentrations. This probably mean that stomatal limitation was mostly relevant to limit photosynthetic activity.

Szent István University, Faculty of Food Sciences, Department of Canning Technology, Budapest, Hungary

Jerusalem artichoke is a plant that can serve as an alternative source of carbohydrate. The tubers contain 14-15 % inulin, 0.8-0.9 % minerals and 0.6-1.5 % raw fibre as basic components of healthy nutrition. Inulin is a polysaccharide, that consist from 80 % fructose and 20 % glucose. Since it is hard to digest for humans, it functions as dietary fibre. Carbohydrates, in addition to being our most important energy-giving nutrients, also cause “civilization illnesses” like diabetes, obesity and caries. Inulin, on the other hand, hydrolysed to fructose is better tolerated by diabetics; additionally, it is a more intensive sweetener , than saccharose and glucose. Other valuable nutrients of Jerusalem artichoke are the minerals. The high kalium content (0.4% w/w) of the tubers, for example, is of utmost importance.

1^F-fructosyltransferase (1^F-FTase), an enzyme produced in intracellular and extracellular forms during aerobic cultivation of Aureobasidium pullulans CCY 27-1-1194, has been shown to have a high activity, stability and regioselectivity for the production of 1^F-type fructooligosaccharides . In this study, we examined the effect of the initial sucrose concentration, c_s0, (50, 200 and 350 g/L) and time of duration (1, 2 and 4 days) on the FTase production in shaked flask cultivations. After each cultivation run the following quantities were measured and calculated: the mass of wet and dry cells produced, the intra- and extracellular fructosyltransferase activity obtained, the final sugar and total organic acid content in the cultivation medium, sucrose conversion, specific enzyme activity per mass of cells and the yield factor of biomass on substrate, Y_XS.

It was observed that sucrose was not only consumed by the reactions of intracellular metabolism but a significant part was converted by enzymatic reactions into glucose, fructose and fructooligosaccharides. We further found that the cell growth was strongly inhibited by the sucrose concentration when, during the first two days, it was fastest at c_s0 = 50 g/L. After four days, the cell concentration reached approximately the same value in all runs when the sucrose conversion was complete only at c_s0 = 50 g/L. The overall FTase production partly followed the trend of biomass growth at individual sucrose concentrations but the highest values of enzyme activity were achieved at c_s0 = 350 g/L since the very first day. The specific activity per cell mass was also the highest in this case and was decreasing with the time of cultivation in all cases. This observation may be explained by that high sucrose concentrations promote FTase production which was demonstrated by the high fructooligosaccharide concentrations in the medium after the first day of cultivation. The amount of organic acids produced was growing both with the initial sucrose concentration and cultivation time. The Y_XS values were essentially independent of cultivation time but differed significantly with regard to the initial sucrose concentration. The values of about 0.25 at c_s0 = 50 g/L witness an efficient coupling of fermentative and respiratory pathways. At other cases, the Y_XS values were below 0.1 which indicated that part of the substrate might be transformed into acids in non-growth associated processes.

References

Madlová A., M. Antošová, M. Baráthová, et al. (1999). Chem. Pap. 53(6): 366-369.

1Institute of Chemistry, University of Agricultural Sciences, Muthgasse 18, A-1190 Wien, Austria;

* Author to whom correspondence should be addressed; e-mail:dario@um.es

Many researches have hypothetized that one reason to explain the enhanced absorption of some dietary minerals in rats fed with non-digestible oligosaccharides is the increase of the epithelial cell layer of cecum and colon (Ohta, et al., 1994; Tulung et al., 1987; Younes et al., 1994; Campbell et al., 1997; Lopez et al., 1999; Rémésy et al., 1992, 1993; Scharrer and Lutz, 1992; Levrat et al., 1991; Chonan et al., 1996), although it could not be the only factor to estimulate mineral absorption (Chonan and Takahashi, 1999; Chonan et al., 1997).

The effect of eight different experimental infant formulas on colonic epithelial cell proliferation was tested. Infant formulas were administered to rats during 30 days to evaluate the prebiotic (GOS at 1,2; 5 and 10 %), probiotic (Bifidobacterium longum and Bifidobacterium infantis) and synbiotic (a combination of both). A diet of infant formula without pre- and probiotics was used control, meanwhile a maintenance commercial diet for rats was used to monitorize growing of them.

Crypt depth, cell density (number/crypt) and the relationship between both, were the parameters estimated to study the epithelial cell proliferation.

The addition of GOS (prebiotic diets) and together with probiotics (synbiotic diet) to infant formulas had a significant effect (p<0,05), higher than obtained in control group on proximal and distal colon portion crypt proliferation, independently of the GOS dose. However, the use of probiotics only had stimulatory effect on distal colon portion. Nor prebiotics, probiotics or synbiotics showed significant effect on cecum epithelial cell proliferation respecting to the control group.