Research Reports No.16 Abstract（2010）
16-01
Improvement of Starchy Foods by Acylated Brabched Oligosaccharide Phosphate
with Multi-Functional Properties
Koji Takahashi
Faculty of Agriculture, Tokyo University of Agriculture and Technology
Starchy foods easily retrograde, resulting in the reduced quality, and generally less interact with fat and oil giving good body texture. In the previous study, oleyl branched oligosaccharide phosphate (OA-BOS-P) was thus prepared from phosphate-rich potato starch by limitedly hydrolyzing with a-amylase followed by exhaustively hydrolyzing with glucoamylase, and then by acylating with lipase. In the present study, the multi-functionalities of OA-BOS-P were investigated in terms of exhibiting a high solubility, superior emulsifying ability, calcium-binding ability, and controlling ability to gelatinization and retrogradation of starch.
16-02
Development and Application of Purification of Polar Natural Food Colorants
using Spiral Countercurrent Chromatography
Koichi Inoue
School of Pharmacy, Kinjo Gakuin University
The multi-layered coils in high-speed countercurrent chromatography (HSCCC) are able to separate small molecules and natural products with organic-aqueous two-phase solvent system. However, it was found that with the heavy alcohol-aqueous two-phase solvent system, low stationary phase retention occurred that made it difficult to operate in HSCCC conditions. When the polar natural products such as polyphenols, antioxidant, and phenols were used by normal HSCCC, it is impossible that the separation and purification of the polar natural food colorants were applied. In the last few years new CCC rotors have been designed based on a spiral flow pathway. This name is spiral HSCCC. In this study, we apply the separation of antioxidants in anthocyanin colors such as red cabbage, red radish and perilla extracts by spiral HSCCC. These antioxidants were detected using HPLC with DPPH radical assay. Using these antioxidants, the K and a values were analyzed for two-phase solvent system. Two-phase solvent system composed of 1-butanol/0.1% TFA in water (50/50, V/V) for red cabbage, 1-butanol/acetic acid/water (40/10/50, V/V/V) for red radish, and tert-butyl methyl ether/1-butanol/acetonitrile/ water (20/20/10/50, V/V/V/V) for perilla extracts were used. In these results, the spiral HSCCC chromatograms of these antioxidants were shown in these figures.
16-03
Characteristics and source plant identification of cinnamon powder as a functional food
Michiho Ito
Graduate School of Pharmaceutical Sciences, Kyoto University
It was found that the stomach ulcers developed by water immersion restraint stress for mice could have been prevented by a continuous administration of cinnamon-containing food. Chinese or Vietnamese cinnamon powder was mixed with conventional mice foodstuff at doses of 2 to 100 mg/g per feed to make pellet chews. The chews were administered to mice according to the different design of experiments to reveal that the dose of cinnamon powder at 100 mg/g per feed significantly suppress development of stomach ulcers by water immersion restraint stress for mice. The better preventing effect was observed in the longer period administration groups. No difference in ulcer preventing effect was observed between the Vietnamese and the Chinese cinnamons although they have significant difference in essential oil content and taste. Cinnamaldehyde, the main component of essential oil of cinnamon powder, was examined for the ulcer preventing effect to be shown that it was significantly potent against ulcer development but less effective than the whole cinnamon powder. Food intake of cinnamon-administered mice was more than that of the control mice; however, their body weight increase was not proportional to the amount of intake. It might be interesting for the preventive medicine to avoid developing stomach ulcers by taking a kind of food additives or spices with conventional foodstuff. Further analyses are required for studying the functional compound contained in the cinnamon powders which prevents development of stomach ulcers.
16-04
Prevention of metabolic syndrome by regulation of key molecules using natural food color
Takanori Tsuda
College of Bioscience and Biotechnology, Chubu University
Considerable attention has been focused on the health benefits of anthocyanins beyond their antioxidant content or their ability to improve vision. In this study, we tested the effect of dietary anthocyanin-rich bilberry extract (BBE) on hyperglycemia and insulin sensitivity in type 2 diabetic mice. We found that dietary BBE ameliorates hyperglycemia and insulin sensitivity via activation of AMP-activated protein kinase (AMPK). Dietary BBE significantly reduced blood glucose concentration and enhanced insulin sensitivity. AMPK was activated in white adipose tissue, skeletal muscle and the liver of diabetic mice fed BBE. This activation was accompanied by up-regulation of glucose transporter 4 in white adipose tissue and skeletal muscle, and suppression of glucose production and lipid content in the liver. At the same time, acetyl-CoA carboxylase was inactivated, and peroxisome proliferator-activated receptor-alpha, acyl-CoA oxidase and carnitine palmitoyltransferase-1A were up-regulated in the liver. These changes resulted in improved hyperglycemia and insulin sensitivity in type 2 diabetes. These findings provide a biochemical basis for the use of bilberry fruits and have important implications for the prevention and treatment of type 2 diabetes via activation of AMPK.
16-05
Development of quality assessment method for optically active food flavor compounds
and its biological behavior
(Enantiomeric analysis of food flavor compounds by multiple head-space SPME-GC/MS)
Koichi Saito
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Hoshi University
A quality assessment method for eight commercially available optically active flavor compounds: menthol, menthyl acetate, perillaldehyde, 1,8-cineol, a-pinene and limonene (both of which are impurities in 1,8-cineol), and neomenthol and neomenthyl acetate (structural isomers of menthol and menthyl acetate, respectively), was developed for commercial foods. The simultaneous determination of the eight flavor compounds and the optical purity test for the complete enantiomeric separation of each flavor compound were achieved with a gas chromatograph-mass spectrometer (GC/MS) equipped with a tandem capillary column of a fused silica column (DB-17MS) coupled with a cyclodextrin chiral column (b-DEX). The extraction of the flavor compounds from the food samples was carried out by the multiple head-space (MHS) solid-phase micro extraction (SPME) technique in order to avoid the matrix effect. The MHS technique improved quantification by SPME. Optimization of the MHS-SPME technique brought about excellent recoveries from the food samples. Of the twelve commercial samples subjected to the optical purity test, some were found to have undesirable enantiomers, which are not designated as flavor compounds by the food sanitation law in Japan. The results show that the developed method is effective and suggest that there is a need to conduct further optical purity tests for the quality assessment of flavor compounds in commercial foods.
16-06
Antioxidant enzymatically modified isoquercitrin suppresses the development of liver preneoplastic lesions
in rats induced by b-naphthoflavone
Kunitoshi Mitsumori
Graduate School, Institute of Symbiotic Science and Technology, Tokyo University of Agriculture and Technology
To investigate the modifying effect of enzymatically modified isoquercitrin (EMIQ) on hepatocellular tumor promotion induced by b-naphthoflavone (BNF) treatment, male rats were administered a single intraperitoneal injection of N-diethylnitrosamine (DEN) and were fed a diet containing BNF (0.5%) for 6 weeks with or without EMIQ (0.2%) in the drinking water after DEN initiation. One week after the commencement of the administration of BNF, rats were subjected to a two-thirds partial hepatectomy. The number and area of GST-P-positive foci, the number of COX2-positive cells and the area of elastica-van Gieson (EVG)-positive connective tissue fibers promoted by BNF were significantly suppressed by the administration of the antioxidant EMIQ. Real-time RT-PCR analysis revealed that EMIQ treatment decreased mRNA expression levels of Gstm1, Serpine1, Cox2 and Nfkbia and increased mRNA expression levels of Yc2 compared with those in the DEN-BNF group. These results suggest that co-administration of EMIQ suppresses the hepatocellular tumor-promoting activity of BNF in rats through the anti-inflammatory effects of EMIQ and restores the cellular redox balance altered by BNF.
16-07
Study on the relationship between the anthocyanin molecular species and color phenotype of roots
in the inbred lines of red radish (Raphanus sativus L.)
Yoshihiro Ozeki
Faculty of Technology, Tokyo University of Agriculture and Technology
Raphanus sativus cultivar 'Inuidani' bearing red roots had been established and hybridized with cultivar 'Miura'. All roots of the F1 progenies showed purple phenotype. Anthocyanins containing red roots of 'Inuidani' and purple roots of F1 were extracted using methanol containing 1% HCl, followed by partial purification using HP20 column chromatography. The eluted anthocyanins were hydrolyzed by 6 N HCl at 90°C for 60 min and purified using HP20. The aglycons were analyzed by high performance liquid chromatography (HPLC), indicated that main molecular species of anthocyanin aglycon were pelargonidin and cyanidin for red roots of 'Inuidani' and purple roots of F1, respectively. Because it was known that the hydroxyl group at the 3' position of B ring of anthocyanidin is introduced by flavonoid 3'-hydroxylase (F3'H), F3'H cDNA from the roots and F3'H gene from the leave of purple plants were isolated by PCR and their nucleotide sequence was determined. Beside F1 plants had one wild-type F3'H gene, two insertion sequences, Ty3/gypsy retrotransposon and its LTR sequence, were found in two independent F3'H genes of 'Inuidani' genome. All red F2 progenies accumulating pelargonidin had both two inserted F3'H genes in the genome, but all purple F2 progenies accumulating cyanidin had one wild-type F3'H gene without insertions. The genomic PCR for the other cultivar, 'Morning Breakfast', bearing the red roots containing pelargonidin, gave evidence of the disrupted F3'H gene by the other transposable elements, Helitron. It is showed that the PCR primers corresponding to these insertion sequences in F3'H genes might be useful to identify molecular phenotype in young plants at early stage of cultivation to predict the phenotype of root colors in adult plants.
16-08
Studies on the elicitation of sweetness of thaumatin
Tetsuya Masuda and Naofumi Kitabatake
pision of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University
Department of Natural Resources, Graduate School of Global and Environmental Studies, Kyoto University
Thaumatin is an intensely sweet-tasting protein and elicit a sweet taste at 50 nM. To clarify the structure-sweetness relationships in thaumatin, we attempted high-yields secretion of the recombinant thaumatin by Pichia pastoris to prepare high-quality crystals in the presence of cryoprotective glycerol for high resolution structural analysis. Approximately 100 mg L-1 of recombinant thaumatin I was obtained and preliminary crystallization analysis was performed for thaumatin mutants.
16-09
Identification analysis of major allergen proteins in cochineal dye
Hiroshi Akiyama
pision of Novel Foods and Immnochemistry, National Institute of Health Sciences
Carmine is a natural red pigment obtained from the dried gravid female cochineal insects (Dactilopius coccus or Coccus cacti). We identified a full-length cDNA encoding a protein, which we named CC38K, with 335 amino acids and a molecular mass calculated as 38 kDa. This amino acid sequence included all the partial amino acid sequences obtained from the purified proteins identified by IgE from allergic patients. The CC38K sequence showed homology to phospholipases. We could make the recombinant CC38K using a yeast expression system using a cold-inducible promoter. The recombinant CC38K protein was recognized by patients' sera indicating that this is a major allergen present in carmine. Four possible N-glycosylation sites were identified in the CC38K amino acid sequence. Therefore, we analyzed N-linked glycan on the major allergens in the cochineal insects. The results suggest that the allergens might have Mana1-6(Mana1-3)(Xylb1-2)Manb1-4GlcNAcb1-4(Fuca1-3)GlcNAc-Asn as N-linked glycan.
16-10
New function of plant polyphenols in abnormal vascular smooth muscle contraction
Katsuko Kajiya
Department of Molecular Physiology and Medical Bioregulation, Yamaguchi University Graduate School of Medicine
Although hypercholesterolemia is a major risk factor of vascular diseases, the relationship between cholesterol (Chol) and abnormal vascular smooth muscle (VSM) contraction has not been clarified yet. A Rho-kinase (ROK)-mediated Ca2+-sensitization of VSM contraction contributes to abnormal VSM contractions such as vasospasm. We previously identified sphingosylphosphorylcholine (SPC) as an upstream pathogenic factor for this abnormal pathway. Interestingly, the degrees of SPC-induced Ca2+-sensitization in humans correlated well with serum total Chol and LDL-Chol, but inversely correlated with HDL-Chol levels. Since Chol is highly enriched in membrane microdomains, lipid rafts, we focused on the roles of rafts in the VSM Ca2+-sensitization. SPC induced the translocation of cytosolic ROK to rafts. The depletion of Chol by b-cyclodextrin destroyed rafts and abolished the SPC-induced ROK translocation and Ca2+-sensitization. Furthermore, we have succeeded in developing the model membrane of lipid rafts, and the surface plasmon resonance measurements on the model membranes provided the first direct evidence for higher affinity of SPC for the rafts membrane than for normal (non-raft) membrane. In addition, our finding of selective inhibition of VSM Ca2+-sensitization by natural products stimulated us to examine the effects of many functional components from plants on abnormal VSM contraction induced by SPC, providing some hints of structure-activity correlation.
16-11
Pharmacokinetis and mechanisms for intestinal absorption of several flavonoid glycosides -role of transporters
Toshiaki Makino
Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Nagoya City University
Quercetin, a flavonol contained in various vegetables and herbal medicines, has various biological activities including anti-cancer, anti-allergic and anti-oxidative activities. However, its low oral bioavailability due to insolubility in water has limited its use as a food additive or dietary supplement. In previous reports, we found that the elongation of a-linkage of glucose moiety in quercetin-3-O-glucoside (isoquercitrin, IQC) enhances the bioavailability of quercetin. In order to clarify the mechanisms for the intestinal absorption of quercetin glycosides, in vitro hydrolysis reactions of quercetin glycosides by the homogenate of rat intestinal epithelium were evaluated. IQC, quercetin-3-O-maltoside (Q3M) and enzymatically modified isoquercitrin (a-oligoglucosyl isoquercitrin, EMIQ) were rapidly hydrolyzed to quercetin by rat intestinal epithelium, but quercetin-3-O-rutinoside (rutin) was not. a-Monoglucosyl rutin (aMR) and a-oligoglucosyl rutin (aOR) were hydrolyzed to rutin. Acarbose inhibited the hydrolysis of EMIQ to quercetin via IQC, and phrolizin and lactose inhibited the hydrolysis of IQC to quercetin, suggesting that intestinal epithelial enzymes such as mucosal maltase-glucoamylase (MGM) and lactase phrolizin hydrolase (LPH) play important roles in the hydrolysis and absorption of quercetin glycosides.
16-12
Short-chain fatty acid production from Gum arabic and its effect on the adipose and musclar tissues
Kazunari Ushida
Animal Science, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University
Acacia spp. produce gum exudates, traditionally called as gum arabic (GA) or gum acacia which are widely used in the food industry such as emulsifiers, adhesives, and stabilizers. The literature indicates that GA can be fermented by the intestinal bacteria to short chain fatty acid, particularly propionate. Propionate may be used as a precursor of gluconegenesis in the liver which can reduce glucogenic amino acid utilization. That reduction may reduce the muscle protein turnover. As a dietary fiber, GA should have a potential to control lipid metabolism. In this experiment, a long term effect of GA on muscle weight and adipose tissue weight of female mice at 13 weeks old which received a drinking water containing GA (1.0%) for 180 days. Cecal bacterial composition was significantly affected by GA and as a result, cecal propionate was increased (P<0.1). Weight of the Mesenteric fat and the Visceral fat tended to decrease by GA (P<0.1). At the same time, Gastrocnemius muscle weight tended to be bigger by GA (P<0.1). Area of adipocyte was significantly decreased by GA (P<0.01). While serum triglyceride was unchanged, free fatty acid was higher and total cholesterol was lower in GA mice (P<0.1). Blood urea nitrogen was also decreased by GA(P<0.1). Gene expression in the Visceral adipose tissue was changed particularly in lipid utilization as energy such as Adrb3, HSL, ATGL, Plin and Fiaf.
16-13
Evaluation of commercially available antimicrobial compounds
for controlling Listeria monocytogenes in minced tuna and salmon roe
Hajime Takahashi
Department of Food Science and Technology, Faculty of Marine Science, Tokyo University of Marine Science and Technology
Listeria monocytogenes found in minced tuna and fish roe has the potential to cause listeriosis. Although refrigeration is critical for these seafoods, enforcing strict temperature regulations on distributors, as well as consumers, is difficult. Alternatively, L. monocytogenes growth could be controlled using antimicrobials. In this study, we investigated the effect of nisin and other commercially available antimicrobial compounds (lysozyme, ɛ-polylysine, and chitosan) on L. monocytogenes in raw minced tuna and salmon roe to determine their effectiveness. The L. monocytogenes-inoculated food samples were incubated with each antimicrobial at 10°C for 7 days or 25°C for 12 h. NisaplinTM (nisin-containing antimicrobial) effectively inhibited L. monocytogenes growth in minced tuna at 500 ppm and salmon roe at 250 ppm, within the commonly selected shelf life. The optimal concentration of each antimicrobial was determined. These results will be useful for selection of the most effective antimicrobial(s) at the optimal concentration to prevent pathogenic growth in raw minced tuna and salmon roe.
16-14
The differences in chitinase activity in anatomies of various plant species
Yukio Takii
Department of Food Science and Nutrition, School of Environmental Sciences, Mukogawa Women's University
Research regarding the distribution of intracellular chitinase activity, which is found in the skin and flesh of 46 plants from 24 families. Examples which have high chitinase activity in live anatomy weight are persimmon (Ebenaceae Diospyros kaki) skin and flesh, Cucurbitaceae hurdies melon flesh, muskmelon (Cantaloupes) flesh, mango (Anacardiaceae Mangifera indica) skin and flesh, avocado (Lauraceous Persea Americana) skin, mube (Lardizabalaceae Stauntonia hexaphylla) flesh, and kiwifruit (Actinidiaceae Actinidia deliciosa) flesh. As the result of examination using the skin of 5 items from Ebenaceae which family has the highest chitinase activity, Fuji-gaki (from Ehime Prefecture) has enzyme activities, which are 1.080U per 100g of live skin weight, and 68.2U/mg protein per the protein content of the skin.
16-15
Development of Recyclable Catalyst towards Syntheses of Macrocyclic Perfumery Ingredients
Hisahiro Hagiwara
Graduate School of Science and Technology, Niigata University
Thirteen- to eighteen-membered lactones were synthesized by ring-closing olefin metathesis reactions of bis-olefins with heterogeneous Grubbs Ru-supported ionic liquid catalysts (Ru-SILC), in which homogeneous Grubbs catalysts were confined in pores of alumina with the aid of an ionic liquid. The Grubbs-SILC exhibited higher catalytic performance than their homogeneous counterparts and could be repeatedly used several times without any pre-treatment after simple filtration.
16-16
Study on a refreshment and relaxation effects of a life space perfume component
Akiyoshi Sawabe
Department of Applied Biological Chemistry, Faculty of Agriculture, Kinki University
Environmental stress is present in casual everyday life variously. How to feel about the stress is various so as to say so many men, so many minds. In this study, we investigated the building materials used in an office and life space. We reviewed a refreshment effect and aromatherapy indication by stress marker evaluation as environmental harmony evaluation for essential oils of building materials and the isolation products. In this study, for cortisol and amylase in saliva of human, the essential oil of hiba (Thujopsis dolabrata) indicated suppressive effect on stress.
16-17
Synthesis of chiral lactones as aroma chemicals employing environmentally friendly organocatalyst
Kunihiko Takabe and Nobuyuki Mase
Department of Molecular Science, Faculty of Engineering, Shizuoka University
Organocatalytic asymmetric a-hydroxymethylations of cycloalkanones with aqueous formaldehyde solution have been developed to synthesize chiral lactones that are important synthons in aroma chemicals as well as pharmaceuticals. L-Threonine-catalyzed a-hydroxymethylation of 5- to 8-membered cycloalkanones afforded the desired aldol products in 31-63% yields with 82-93% ee. Enantioselectivities were increased up to 96-99% ee employing lipase-catalyzed kinetic resolution of the aldol products. Further chemical transformations of the aldols gave various bioactive chiral lactones. In addition, the transitional state of organocatalytic asymmetric a-hydroxymethylations of cyclopentanone was proposed.
16-18
Quality evaluation of essential oils by a taste-sensing system
Nobuo Kawahara
Research Center for Medicinal Plant Resources, National Institute of Biomedical Innovation
Recently, it has been recognized effectiveness and functionality of aromatherapy, a natural holistic approach to therapy using essential oils and other plant extracts. Many common essential oils have medicinal properties that have been applied in folk medicine since ancient times and are still widely used today. Essential oils are registered in "The Japan's Specifications and Standards for Food Additives" mainly used as bitter substances and anti-oxidants, and also 7 essential oils (cinnamon oil, clove oil, fennel oil, eucalyptus oil, mentha oil, orange oil and turpentine oil) are registered in "The Japanese Pharmacopoeia Fifteenth Edition".
In the course of our study for development of a new method for the quality evaluation of essential oils, we investigated the profile analysis of 17 kinds of essential oils by a taste-sensing system last year. As the results, 17 kinds of essential oils were classified into 7 types by the taste distributions. Furthermore, the taste pattern of clove oil was almost same as eugenol, a main component of clove oil and showed good correlation between the taste intensity of anionic bitterness and the content of eugenol.
In this year, we investigated the quantitative analysis of 11 kinds of commercial clove oils and 12 kinds of commercial thyme oils, both of which the taste patterns of oils are similar to these main components, by a taste-sensing system and HPLC. As the results, the content of eugenol in clove oils observed by a taste-sensing system showed equivalence to that of HPLC, whereas, the content of tymol in thyme oils showed a little difference between these two methods. Through these studies, we showed possibility of the quality evaluation of essential oils by a taste-sensing system, although the quantitative analysis by this system might be suitable for some kind of essential oils but not for some others.
16-19
The evaluation of food texture using biomedical measurement during gel swallowing
Kazuhiro Hori, Sumiko Yokoyama*, Takahiro Ono*, Kenichi Tamine*, Jugo Kondo*,
Sato Hamanaka*, Yoshinobu Maeda*, Rika Yahagi, Hiroshige Taniguchi, Makoto Inoue
pision of Dysphagia Rehabilitation, Niigata University Graduate School of Medical and Dental Sciences
*pision of Oromaxillofacial Regeneration, Osaka University Graduate School of Dentistry
As the bolus conditions are known to modify the swallowing behaviors, it should be important to evaluate the effects of the food texture on tongue function. The purpose of this study is to clarify the influence of gel consistency on tongue pressure during swallowing.
At first, to determine experimental condition, the tongue pressure during saliva, 5mL, 10mL swallowing in normal and chin-down posture were measured. The tongue pressure was recorded by our original sensor sheet with five measuring points attached on the hard palate directly with denture adhesive. In the results, the larger magnitude and longer duration were recorded during saliva swallowing and in chin-down posture. On the other hands, the magnitude and duration during 5mL and 15mL swallowing showed no significant difference.
Secondly, the influence of gel consistency on tongue pressure during swallowing was measured with tongue pressure sensor sheet. To distinguish between squeezing and swallowing periods, swallowing sound was recorded with a small microphone attached at the neck surface of the level of the cricoids cartilage. The 5 ml of water and gel agent at different concentrations (1.0%, 1.8%, 2.8% Gelup J-3843; 0.075%, 0.2%, 0.35% Gellan gum; San-Ei Gen F.F.I., Inc.) were chosen as a test material. The duration and magnitude of tongue pressure during squeezing and swallowing were compared among these conditions. In the results, the duration and magnitude of tongue pressure during squeezing were increased with increasing the concentrations of materials as well as the number of squeezing in one trial. On the other hand, they were not affected by the bolus conditions, i.e., concentrations, during swallowing.
In conclusion, the tongue pressure production during squeezing may be regulated by the consistency of gel agent for forming the bolus with adequate texture to be swallowed.