Journal of Applied Glycoscience
Online ISSN : 1880-7291
Print ISSN : 1344-7882
ISSN-L : 1344-7882
Volume 50 , Issue 1
Showing 1-16 articles out of 16 articles from the selected issue
  • Masanori Kobayashi, Michio Kubota, Yoshiki Matsuura
    2003 Volume 50 Issue 1 Pages 1-8
    Published: January 20, 2003
    Released: February 23, 2011
    JOURNALS FREE ACCESS
    The three-dimensional structure of maltooligosyl trehalose synthase has been determined by the multiple isomorphous replacement method and refined at 1.9 Å resolution. The structure of the enzyme consists of three domains, A, B and C. The catalytic site resides between the C-terminal end of the central-barrel in the (β/α)8-structure of domains A and B, as in the usual α-amylase family enzymes. Domain C is composed of an eight-stranded β-sandwich structure. Domain A contains two large extra structural frameworks excursed from the barrel. These extra structures are shown to be indispensable to form the pocket at the active site where intramolecular transglycosylation takes place. A possible mechanism of catalysis is also discussed based on the refined structure.
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  • Naoko Matsunaga, Setsuko Takahashi, Keiji Kainuma
    2003 Volume 50 Issue 1 Pages 9-13
    Published: January 20, 2003
    Released: June 28, 2010
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    A rapid and efficient rice starch isolation method was tested on two varieties of newly developed rice, Hoshinishiki (Kanto181) and Saikaimochi227 (a high-amylose and a high-yield waxy rice), and a reference rice variety, Nipponbare. Rice flour was stirred in 0.05 or 0.075 M aqueous sodium hydroxide for 3 h once or three times, neutralized with 1 M hydrochloric acid and washed with water. Isolated rice starch was compared with the starch isolated by the conventional method in terms of chemical and physical properties. The improved method reduced processing time to two days, while the conventional method required one to three weeks. The improved method lowered damaged starch and protein contents, but slightly lowered starch recovery. Non-waxy rice starches isolated by the two methods differed in physical properties upon testing by photopastegraphy and Rapid Visco Analyser (RVA), whereas no difference was observed in waxy rice. The phosphorus content in Nipponbare starch decreased during the isolation process due to degradation of the lysophospholipid, and the phosphorus content was the lowest in the starch isolated by the conventional method. Sodium content was high in the starch isolated by the improved method, although the influence of residual sodium on the viscosity of the paste was small. To reduce residual sodium in the starch isolated by the improved method, washing with water eight times after neutralization was needed. Thus, we propose the improved procedure for rice starch preparation. Rice flour is washed with 0.05 M aqueous sodium hydroxide three times, neutralized and then washed eight times with water. In the case of high-amylose or waxy rice flours, alkali washing can be reduced to once.
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  • Hiroki Takata, Kohji Ohdan, Takeshi Takaha, Takashi Kuriki, Shigetaka ...
    2003 Volume 50 Issue 1 Pages 15-20
    Published: January 20, 2003
    Released: February 23, 2011
    JOURNALS FREE ACCESS
    By sequencing the whole genome of the hyperthermophilic bacterium, Aquifex aeolicus, a gene (Aq722) encoding a protein with high similarity to the branching enzyme (BE, EC 2.4.1.18) has been found. Based on the putative amino acid sequence, a polynucleotide encoding the BE gene was chemically synthesized and the gene was expressed in Escherichia coli. More than 95% of the recombinant enzyme was present within the cells as insoluble but catalytically active aggregates. Heat treatment of the aggregate suspension at 70°C resulted in about 30% solubilization of the BE activity. Enzymatic properties of both soluble and insoluble forms of the BE were analyzed. Both forms exhibited maximum activities at 75°C and pH 7.5-8.0, and were stable at temperatures up to 85°C. The insolubleform BE was less stable than the soluble form in neutral and acidic pH regions; after 30-min incubation at 70°C and pH 7.0, 90 and 50% activities of soluble andinsoluble forms remained, respectively. When amylopectin was used as a substrate, the A. aeolicus BE cyclized the B-chains, which connect the cluster structures of amylopectin. The product (highly-branched cyclic dextrin), with weight-average degree of polymerization (DPw) 1200, has a ring structure with DPw 50 and noncyclic chains with an average unit chain length of 16 connected to the ring.
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  • Takanori Nihira, Supannee Kansarn, Toshiaki Kono, Gentaro Okada
    2003 Volume 50 Issue 1 Pages 21-25
    Published: January 20, 2003
    Released: July 01, 2011
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    A distinct endo-Cellulase component derived from Acremonium cellulase, a commercial Cellulase material from Acremonium cellulolyticus, was extensively purified by consecutive column chromatography and was designated as cellulase I. Cellulase I was homogeneous on both Native-and SDS-PAGE, and was completely free from β-glucosidase. The enzyme showed high specific activity for Avicel and extremely low specific activity for CMC, 0.69 and 0.40 U/mg of protein, respectively. The molecular mass (SDS-PAGE) and p1 value of cellulase I were about 61 kDa and 5.0, respectively. The purified enzyme contained 19.8% carbohydrate as glucose. The N-terminal amino acid sequence from the 2nd up to the 20th residue of the enzyme wasQ-*-V-W-G-Q-*-G-G-Q-G-W-S-G-A-T-(S)-*-A-. The optimum pH and temperature for cellulase I were 5.0and 55°C, respectively. Cellulase I was completely stable over the range of pH 3.0-6.0 at 4°C for 24 h and at temperatures below 55°C. The activity of cellulase I was partially inactivated by 5 mM Mn2+, Fe2+, Hg2+ and KMn04 to various inhibition extents. Cellulase I split various cellulosic substrates to produce predominant cellobiose and a small amount of glucose as the final hydrolysis products. The enzyme was characterized as an exceedingly low endo-type cellulase on the basis of its actions on both CMC and cellooligosaccharides.
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  • Masahiro Kojima, Koji Ogawa, Hiroko Ito, Kazuya Kohayakawa
    2003 Volume 50 Issue 1 Pages 27-31
    Published: January 20, 2003
    Released: July 01, 2011
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    Maximum expansion ratio and minimum bulk density and water solubility of cornstarch-poly (lactic acid) (PLA) extrudates were predicted using response surface methodology. An increase in temperature decreased the expansion ratio markedly at low moisture content (around 12%). Maximum expansion ratio was 30.2 at 110°C barrel temperature, with 12% moisture content and 19.4% PLA content. Although bulk density of the extrudates was inversely related to the expansion ratio (r = - 0.652), the conditions for minimum bulk density were different. Minimum bulk density was 0.011 (g/cm3) at 170°C barrel temperature, with 15% moisture content and 20.5% PLA content. Water solubility was significantly dependent on PLA contents, minimum water solubility was 24.3% at 110°C barrel temperature, with 12.1% moisture content and 22.8% PLA content. The conditions for closest responses to the maximum or minimum values were predicted using MINITAB response optimizer software. Thirty point one of the expansion ratio, 0.020 (g/cm3) of the bulk density and 24.7% of water solubility were obtained at 110°C barrel temperature, 12% moisture contents and 21.9% PLA contents. For biodegradability of starch-PLA extrudates, an increase in PLA content decreased the biodegradability rate significantly.
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  • Kazumi Funane, Shuryo Nakai, Kazue Terasawa, Tetsuya Oguma, Hiroshi Ka ...
    2003 Volume 50 Issue 1 Pages 33-35
    Published: January 20, 2003
    Released: June 28, 2010
    JOURNALS FREE ACCESS
    Amino acid residues of Cyclodextran glucanotransferase from Bacillus circulans T-3040 were replaced using site-directed mutagenesis to improve the productivity of cyclodextrans. The computer program of random-centeroid optimization for genetics was applied for selecting the positions of mutations to change hydrophobicity or bulkiness of the enzyme protein. Two mutants, namely A452N and V744L, produced larger amounts of CIs per hour than the wild-type enzyme. A452N and V744L mutant enzymes increased the reaction velocities 3- and 2-fold, and Km values 9- and 3-fold, respectively. Activation by Ca2+ and inactivation by Cu2+ were reduced in both mutants. Optimum temperature and pH, and stability against changes in heat and pH of the two mutants were not affected. Wild-type and mutant enzymes were all inactivated with 1-ethyl-3-[3-(dimethylamino) propyl] carbodiimide in the absence of Ca2+, however, inactivation was reduced by an additional 10 mM of Ca2+.
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  • Masatsugu Yamashita, Hideya Adachi, Takeshi Nakamura, Hajime Taniguchi ...
    2003 Volume 50 Issue 1 Pages 37-39
    Published: January 20, 2003
    Released: June 28, 2010
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    The formation of amylose complexes with glycerol monooleate (MG-C18:1) or glycerol monostearate(MG-C18:0) was conducted in aqueous solution using a commercial amylose with an average degree of polymerization (DP) of 18. The chain distributions of amylose precipitated by forming a complex in aqueous solution (20°C) were analyzed by high-performance anion exchange chromatography with a pulsed amperometric detector (HPAEC-PAD). The critical DPs of amylose forming complexes with MG-C18:1 and MG-C18:0 were DP 31 and 29, respectively. The principal chains of amylose forming complexes with MG-C18:1 and MGC18: 0 were DP 35-36 and DP 33-34, respectively.
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  • Mee Son, Haruhide Mori, Masayuki Okuyama, Atsuo Kimura, Seiya Chiba
    2003 Volume 50 Issue 1 Pages 41-44
    Published: January 20, 2003
    Released: February 23, 2011
    JOURNALS FREE ACCESS
    The hydrolytic reaction of carbohydrate-hydrolase is essentially accompanied by a reverse reaction (the condensation reaction), meaning that only the substrate capable of being hydrolyzed is produced by the reverse reaction. Honeybee α-glucosidase I can't hydrolyze isomaltose, but is capable of hydrolyzing maltose, kojibiose and slightly nigerose. Nevertheless, the enzyme catalyzes the formation and accumulation of isomaltose from glucose together with α-glucobioses such as maltose, kojibiose and nigerose. This finding is in conflict with the data that the enzyme has no hydrolytic activity toward isomaltose. However, the conflict for the peculiar phenomenon on the reaction was rationally explained by the evidence that isomaltose might be formed by the intramolecular transglucosylation via other α-glucobioses that are easily produced from glucose by the condensation reaction. It is suggested that the usual transglycosylation of carbohydrate-hydrolase may be accompanied by an intramolecular transfer reaction.
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  • Yoshimi Sugimoto, Yotaro Konishi, Hidetsugu Fuwa
    2003 Volume 50 Issue 1 Pages 45-49
    Published: January 20, 2003
    Released: June 28, 2010
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    Starch granules were prepared from three kinds of Azuki beans (Vigna angularis ): Akanedainagon (dainagon), Erimoazuki (Erimo) and Shiroazuki. The properties of the starch granules were examined. The results obtained were as follows: 1) The mean particle sizes of the Dainagon, Erimo and Shiroazuki starches as determined by videomicrometer were 39.3, 38.0 and 37.5, um for long axis, and 32.4, 31.8 and 29.5, um for short axis, respectively. 2) The initiation temperature for gelatinization of Erimo starch by differential scanning calorimetry, photopastegraphy and Brabender's amylography (6 and 8 % starch suspensions) tended to be higher than the respective values of the other two starches. 3) The breakdown of Shiroazuki starch as determined by Brabender's amylography (8%) was 271 BU, which was relatively lower than the respective values of the other two starches. 4) The apparent amylose contents of the Dainagon, Erimo and Shiroazuki starches as determined by amperometric titration were 29.0, 27.4 and 28.7%, respectively, and those by the enzyme-chromatographic method were 30.4, 28.8 and 31.2%, respectively. The amylose content of Erimo starch was lower than that of the other two starches. 5) X-ray diffractograms of the Azuki bean starches showed Cb type patterns. 6) The susceptibility of Dainagon and Shiroazuki starch granules to a-amylase of the porcine pancreas was similar to that of normal maize starch granules, having relatively high susceptibility, and that of Erimo starch granules was lower than the respective values of the other two starches. By scanning electron microscopy (SEM), a stripe structure on the Azuki bean starch granules was observed following attack by pancreatin.
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  • Hiroshi Takii, Minoru Yoshiyama, Michiyo Yanase, Hiroshi Kamasaka, Tak ...
    2003 Volume 50 Issue 1 Pages 51-54
    Published: January 20, 2003
    Released: February 23, 2011
    JOURNALS FREE ACCESS
    We developed a sugarless chewing gum that promotes dental re-calcification using phosphoryloligosaccharides (POs). Since the gum contains 79% (w/w) sugar alcohol, it was apprehensive that a gastrointestinal disorder would happen when an excess amount of gum is consumed at once. Sixteen women and 19 men ingested 14 tablets of gum with POs for consecutive 14 days, and the influence on gastrointestinal condition was investigated. On the first day of the experiment, the average times of the transient feces for women and men were 0.9 and 0.2 times, respectively. On the final day, transient feces for women and men were 0.4 and 0.1 times, respectively. The volunteers' gastrointestinal condition was not influenced even after ingesting an excessive amount of POs at once, which was equal to that contained in 70 tablets. These results suggested that the POs did not affect the volunteers' gastrointestinal condition, and that the increase in times of evacuation was caused by sugar alcohol.
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  • Toshiaki Komaki
    2003 Volume 50 Issue 1 Pages 55-60
    Published: January 20, 2003
    Released: February 23, 2011
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    This review summarizes three main topics from my work on enzyme application for starch processing, which is a major field of research in the development of industrial applications for enzymes. I have engaged in such research for more than 50 years, beginning in 1951 while I was with the enzyme production department of Nagase Co., Ltd. The first problem was the complete liquefaction of sweet potato suspension at high concentration using an enzyme process. With the “gradual heating method, ” a conventional method of liquefaction, the saccharified solution became turbid and it was impossible for the solution to be filtered and purified. The turbid liquid was generated by means of rearrangement of starch molecular chains that were not digested by enzymes in the process of gradual warm-up liquefaction. For the insoluble starch particles to be digisted, they had to be treated by heat at more than 120°C and have enzymes added again. As a liquefaction method to prevent generating insoluble starch particles, I developed the “Instantaneous heating method.” With this method, a starch suspension is instantaneously heated to 88-92°C with a-amylase. Since the method was inadequate for corn starch, double liquefaction was employed for the corn starch so that α-amylase could be added again after heating the starch solution at over 120°C. As thermostable α-amylase was produced and available from B. subtilis MK 385, the problem with corn starch liquefaction was solved by instantaneously heating at 100-110°C with a jet cooker. The second topic was the industrial production of glucose using an enzyme process. I started industrial production of glucoamylase originating from Aspergillus or Rhizopus and developed a saccharification method for starch to obtain 95-96% yield of glucose. The third problem was the development of isomerization from glucose to fructose. Glucose isomerase originating from Streptmyces phaeocromogenus was accumulated within a strain pellet. The cell with the enzyme was immobilized by an anion exchange resin modified with quaternary pyridine. It formed granulations, and cross-linked to boost its physical strength. Thus, continuous isomerization was developed with a tubular flow bioreactor packing the immobilized cell and production of glucose/fructose syrup as an alternative sweetening to sucrose was started.
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  • Shigetaka Okada
    2003 Volume 50 Issue 1 Pages 61-65
    Published: January 20, 2003
    Released: February 23, 2011
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    The main topics from the results of the studies by the author and collaboraters during the four decades are reviewed. The studies are related to glucanotransfer activity of amylases and related enzymescyclodextrin glucanotransferase, α-amylase, D-enzyme and branching enzyme-and included the following subjects. 1) Coupling sugar (maltooligosyl sucrose) was produced from the mixture of dextrin and sucrose by Cyclodextrin glucanotransferase. The cariogenicity of the coupling sugar was studied by a group at the Japanese National Institute of Health and several universities, and coupling sugar was proved to be an anticariogenic sweetener. It was the first example of a so-called functional oligosaccharide which had a physiological property in addition to the conventional function of sweeteners. 2) Synthesis of some glucosides: hesperdin dihydrochalcone glucoside, glucosyl stebioside, glucosyl hesperidin and hydroquinone glucoside. Among them, stevioside is about 140 fold as sweet as sucrose, but has a slightly bitter taste and after taste. The taste was improved by enzymic glucosidation. Hydroquinone glucoside inhibited tyrosinase activity, and consequently the melanin accumulation of human skin. Applications for hydroquinoe glucoside in the cosmetic field have started recently. 3) Discovery of cycloamylose containing 17 or more glucose residues. Potato D-enzyme catalyzes an intramolecular transglycosylation reaction on amylose to produce cycloamylose, a novel α-1, 4 glucan composed of 17 or more glucose residues. X-ray structure of a cycloamylose with 26 glucose residues was explained. 4) Development of cyclic clusterdextrin, a new dextrin containing cyclic parts. Branching enzymes catalyze the cyclization of amylose and amylopectin. The product from amylopectin had a narrow molecular weight distribution compared with those of commercial dextrins. The cyclic clusterdextrin was soluble in water to give a highly stable and clear solution. The solution had a low propensity for retrogradation.
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  • Setsuko Takahashi
    2003 Volume 50 Issue 1 Pages 67-75
    Published: January 20, 2003
    Released: July 01, 2011
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    1. The cooking and processing properties of newly developed rice. Newly developed rice was studied for its cooking and processing properties as milled rice, flour, and starch. To clarify the cooking, taste and physicochemical properties of rice is important in order to use it in cooked and processed foods. For the purpose of clarifying the factors influencing the taste of rice, the structure of starch was analyzed by measuring the amylose content and amylopectin unit-chain distribution using gel filtration. The average chain-length of amylopectins was determined by the rapid Smith degradation method and non-reducing residue measurement. The cooked rice, whose starch was low in apparent amylose content measured by gel chromatography, high in longer chain length fraction (Fr. II) and short in average chain-length of amylopectin, had a soft and sticky texture, and it was preferred in sensory evaluation. Thus, the relationship between the taste of rice and the rice starch structure was observed. In addition, a rapid, convenient and accurate method of evaluating rice texture is essential because the firmness and adhesiveness of cooked rice is important in the evaluation of sensory attributes. Using the new model of tensipresser at low and high compressibilities, the new texture evaluation method for cooked rice was applied to measure the firmness, adhesiveness, strength, and flexibility of cooked rice. The relationship between the texture and sensory attributes was clarified. II. Cooking and processing properties of starch derivatives. Laboratory samples of starch derivatives from corn and wheat starches were studied for the influences of chemical treatment and lipid on their gelatinization, swelling, and gelforming properties and the experiment was carried out in cooperation with Dr. Seib. In addition, the effect of starch derivatives on Japanese foods was examined. A commercial sample of hydroxypropylated wheat starch, obtained from Midwest Grain Products Inc., was used for Udon noodles, pie fillings, and oriental dumplings, and the texture and sensory attributes were compared with those of hydroxypropylated waxy maize, which isthe most popular starch derivative in United States and was obtained from National Starch and Chemical Co. Those Japanese foods prepared with hydroxypropylated wheat starch showed improved softness of the gel, texture and preferences, reduced preparation time and increased storage stability at low temperature, compared with those prepared with wheat starch. The result indicated that starch derivatives had the effect of improving the quality of cooked foods.
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  • Takahisa Kanda
    2003 Volume 50 Issue 1 Pages 77-81
    Published: January 20, 2003
    Released: July 01, 2011
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    The mode of degradation of native cellulose has not been fully established. The mode of hydrolysis of highly purified cellulases, exo- and endo-type cellulases from fungi (Irpex lacteus, Trichoderma reesei and Aspergillus niger) were investigated by using pure cellulosic materials with different crystallinities of cellulose I type in addition to bacterial celluloses of two type different types (cellulose I and II) and valonia cellulose. At the same time, we also investigated the hydrolysis action of exo- and endo-type cellulases by using water soluble substrates such as cellooligosaccharide and barley glucan. From these results, it was found that the characteristic mode of action of each cellulase can be clearly understood by using only pure crystalline cellulosic substrates. Furthermore, we will describe the synergistic actions of cellulase components in addition to related enzyme on cellulose degradation in this paper.
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  • Motomitsu Kitaoka
    2003 Volume 50 Issue 1 Pages 83-87
    Published: January 20, 2003
    Released: February 23, 2011
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    Phosphorolytic enzymes that act on carbohydrates are interesting enzymes not only in terms of their basic properties but also in regard to their various applications. However, they have not been studied in as much detail as other types of enzymes such as the hydrolases and the synthases. Phosphorolytic enzymes have attracted attention because of their application in the synthesis of carbohydrate chains, exploiting their strict regiospecificities. In addition, a number of new phosphorolytic enzymes have been discovered recently. We have been investigating the phosphorolytic enzymes and have discovered their unique characters and applications. In this paper, the following are described: 1. Analyses of the reaction characterized by celobiose phosphorylase from Cellvibrio gilvus; 2. Characterization of laminaribiose phosphorylase from Euglena gracilis; 3. A specific colorimetric quantification of cellobiose employing cellobiose phosphorylase; 4. Synthesis of laminarioligosaccharides; 5. Conversion of sucrose into cellobiose using two phosphorolytic enzymes.
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  • Akira Yamazaki, Miwako Kinefuchi
    2003 Volume 50 Issue 1 Pages 89-96
    Published: January 20, 2003
    Released: June 28, 2010
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    Until recently, the factor that has played a predominant role in food processing was not "pressure" but "heat, " although both factors are independently responsible for transforming the state of a substance. In food processing, high pressure treatment produces no aberrant matter that puts the safety of food at risk because the food processing can be achieved without any cleavage of covalent bond contained in the ingredients of food. Moreover, high pressure treatment is considered to be very promising for food processing of the future. This is because decomposition of nutrients and production of stench can be minimized more effectively and energy consumption can be reduced more efficiently when compared with heat treatment. Further, whena food product in a container is subjected to high pressure treatment, uniform processing throughout the food can be guaranteed. It has been confirmed that pressure treatment exhibits the following effects, which are useful to improve the quality of food to create new added value . 1) Removal of air bubbles: Air bubbles trapped in a solid substance can be removed therefrom by placing it in a solvent . Then a relatively low pressure ranging between about 10 to 100 MPa is applied and then rapidly reduced . By using this method, we have succeeded in developing food products, for example "rice cakes whose dispersion rate is decreased to one sixth" and "low-salt pickles prepared in 10 min." 2) Impregnation of a solid substance with a liquid and extraction of an ingredient from the solid substance: The sequential operation of applying a pressure of about 50 to 300 MPa to a solid substance and reducing the applied pressure can form a flow path for liquids in the solid substance. The solid substance can be thus impregnated with a liquid and a predetermined ingredient can be extracted from the solid substance effectively through the path. More specifically, we have completed the development of "low allergenic rice" by extracting an allergen-containing protein from rice, and "unpolished rice that can be cooked by normal procedures" by improving the water absorption properties of unpolished rice. 3) Control of enzyme reaction and destruction of cell wall and cell membrane: By the application of a pressure of 200 to 600 MPa to a living organism, the cell wall and the nuclear membrane in the cell are destroyed to cause the leakage of cell sap and halt the activity of some enzymes . By taking advantage of the above-mentioned effect, we have succeeded in the extraction of trehalose from the yeast cell with an enzyme by splitting the trehalose, i.e., the trehalase being deactivated . In addition, it has been found that excess sludge occurring from industrial waste can be made more degradable through self-digestion by the application of high pressure. 4) Denaturation of starch: A pressure treatment of about 200 to 600 MPa destroys the crystal structure of starch so that the centric crosses under polarized light are lost . Consequently, the starch exhibits the same viscosity as obtained by gelatinization after heat treatment and the increased amylase digestibility. Using a variety of starches, the change in the state of water in the starches caused by the high pressure treatment was observed to obtain basic data. We have searched for reasons why the rice subjected to pressure treatment can become tastier after cooking than non-treated rice, and finally clarified the texture characteristics of cooked rice. In particular, it has been found that even though the cooked high-pressure processed rice suffers from retrogradation during storage, it can be restored by microwave heating to such an extent that the degree of gelatinization can surpass that obtained immediately after cooking. Moreover, we have succeeded in developing production lines capable of achieving high pressure application and establishing an energy-saving manufacturing system for producing the cooked high-pressure processed rice on an industrial sca
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