(Vol.60 No.10 1996)
Extracellular Production of a Serratia marcescens Serine Protease
in Escherichia coli
Yasuo Ohnishi and Sueharu Horinouchi
Effect of Acetylation of Ovalbumin on Its Adsorption Behavior
at Solid/Liquid Interface
Anuradha Bhaduri, Naotoshi Matsudomi, and Kali Pada Das
Effects of Ascorbic Acid on the Deodorizing Activity of Polyphenols
against Methanethiol
Hideyuki Yasuda and Atsushi Onogi
Preparation of (})-2-(2,3-(>12<)1H(>22<)2)Jasmonic
Acid and Its Methyl Ester, Methyl (})-2-(2,3-(>12<)1H(>22<)2)Jasmonate
Hideharu Seto, Shozo Fujioka, Hiroshi Fujisawa, Kuniaki Goto, Hideaki Nojiri,
Hisakazu Yamane, and Shigeo Yoshida
New Oleanane Triterpene with Three Ketones Produced by Penicillium
simplicissimum ATCC 90288
Hideo Hayashi, Yoshihide Asabu, Katsushi Naito, Mitsuru Nakayama, Hiroshi
Nozaki, and Motoo Arai
Kojistatin A, a New Cysteine Protease Inhibitor Produced by Aspergillus
oryzae
Nobuyoshi Sato, Tatsuo Horiuchi, Mitsutoshi Hamano, Hiroshi Sekine, Satoru
Chiba, Hiroki Yamamoto, Takeya Yoshioka, Ikuo Kimura, Mikio Satake, and
Yoshiteru Ida
Acid Lipase Inhibitor in Chicken Plasma Identified as Apolipoprotein
A-I
Makoto Fujii, Tetsuro Higuchi, Satoru Mukai, Masami Yonekura, Takahiro
Yano, Hiroyuki Kawaguchi, Koh-ichi Nonaka, Takao Fukunaga, Yasushi Sugimoto,
and Shoji Yamada
Isolation and Analysis of Cinnamic Acid 4-Hydroxylase Homologous
Genes from a Hybrid Aspen, Populus kitakamiensis
Shinya Kawai, Ariko Mori, Takahiro Shiokawa, Shinya Kajita, Yoshihiro Katayama,
and Noriyuki Morohoshi
Purification and Characterization of Extracellular Chitin Deacetylase
from Colletotrichum lindemuthianum
Ken Tokuyasu, Mayumi Ohnishi-Kameyama, and Kiyoshi Hayashi
Physiological Study of Cysteine Synthase Isozymes in Spinacia
oleracea Seedlings
Sanae Kitamura, Takayuki Yamaguchi, Xia Zhu, and Masahiro Masada
Molecular Cloning and Nucleotide Sequence of the groEL Gene from
the Alkaliphilic Bacillus sp. Strain C-125 and Reactivation of Thermally Inactivated
Ώ-Glucosidase by Recombinant GroEL
Yi Xu, Tetsuo Kobayashi, and Toshiaki Kudo
Electrophoretic Analysis of Chemically Modified Taka-amylase
A Using a Slab Gel System with Multiple Lanes
Mikihiko Kobayashi and Yutaka Sasaki
Substrate Specificity of a Novel Alcohol Resistant Metalloproteinase,
Vimelysin, from Vibrio sp. T1800
Saori Takahashi, Kiyoaki Okayama, Shigeru Kunugi, and Kohei Oda
Nicardipine and Nifedipine Inhibit Fatty Acid Desaturases in
Rat Liver Microsomes
Hiroshi Kawashima, Kengo Akimoto, Saeree Jareonkitmongkol, Norifumi Shirasaka,
and Sakayu Shimizu
Effects of Mepanipyrim on Intracellular Trafficking: A Comparative
Study on Its Effects on Exocytic and Endocytic Trafficking of Proteins, Sphingolipids,
and Cholesterol
Ichiro Miura, Makoto Muroi, Nobue Shiragami, Isamu Yamaguchi, and Akira
Takatsuki
Chitinous Component of the Cell Wall of Fusarium oxysporum, Its
Structure Deduced from Chitosanase Digestion
Tamo Fukamizo, Yuji Honda, Hideyoshi Toyoda, Seiji Ouchi, and Sachio Goto
Simple Affinity Purification Method for Raw Starch-adsorbable
and -digesting Amylases with a Raw Starch Column
Hang Thi Mai, Setsuo Furuyoshi, Shinpei Yamamoto, and Toshiharu Yagi
Stereoconfiguration of Anteiso-fatty Acid Biosynthesized from
dl-Isoleucine in Rat Skin
Hirosuke Oku, Tomie Sakihama, and Isao Chinen
Identification of Low Molecular Weight Probes on Perforin- and
Fas-based Killing Mediated by Cytotoxic T Lymphocytes
Takao Kataoka, Masatoshi Taniguchi, Atsushi Yamada, Hidefumi Suzuki, Shin-ichi
Hamada, Junji Magae, and Kazuo Nagai
Estimation of pH and the Number of Lytic Granules in a CD8(>1+<)1
CTL Clone Treated with an Inhibitor of Vacuolar Type H(>1+<)1-ATPase,
Concanamycin A
Takao Kataoka, Michio Sato, Shunzo Kondo, and Kazuo Nagai
Analysis of Catalytic Action of Transglutaminase Induced in Human
Promyelocytic Leukemia (HL-60) and Human Hepatoblastoma (HepG2) Cells
Koji Ikura, Rika Shinagawa, Koji Hatano, Naoki Suto, and Ryuzo Sasaki
Purification and Properties of Adenosine 5'-Triphosphate Sulfurylase
from the Thermophilic Bacterium Bacillus stearothermophilus
Masaaki Onda, Akemi Morimoto, Ayako Simoide, Ken Iwata, and Hiroshi
Nakajimab
Purification and Characterization of an Endo Ώ-1,3-d-Mannanase
from Flavobacterium sp. AS-9
Tasuku Nakajima, Yutaka Maruyama, Asako Sato, Takehiko Matsumoto, Masahiro
Suenaga, and Eiji Ichishima
Detection of Protein A Produced by Staphylococcus aureus with
a Fiber-optic-based Biosensor
Ya Hsin Chang, Tsung Chain Chang, E-Fong Kao, and Chien Chou
Metabolism of Lysine, Threonine, and Leucine in Growing Rats on
Gluten or Zein Diets at Various Dietary Protein Levels
Changhyeug Kim, Hideyuki Tanaka, and Masaji Ogura
Analysis of the pH-Sensitive Swelling Rate of a Polyelectrolyte
Complex Gel Prepared from Xanthan and Chitosan by the Collective Diffusion Model
Hitoshi Kumagai, Chia-Hong Chu, Takaharu Sakiyama, Shinya Ikeda, and Kozo
Nakamura
Development of a Model for Analyzing the Swelling Rate of Ionic
Gels on the Basis of the Diffusion of Mobile Ions: Application to the pH-Sensitive
Swelling of a Polyelectrolyte Complex Gel Prepared from Xanthan and Chitosan
Chia-Hong Chu, Hitoshi Kumagai, Takaharu Sakiyama, Shinya Ikeda, and Kozo
Nakamura
Tastes Produced by Peptides Containing Ionic Groups and by Related
Compounds
Rie Kuramitsu, Masatoshi Takahashi, Kouji Tahara, Kozo Nakamura, and Hideo
Okai
Effects of Feeding Tryptophan-limiting Diets on the Conversion
Ratio of Tryptophan to Niacin in Rats
Katsumi Shibata, Hisako Shimada, and Takako Kondo
In Vivo Macrophage-stimulation Activity of the Enzyme-degraded
Water-soluble Polysaccharide Fraction from a Marine Alga (Gracilaria verrucosa)
Yasuko Yoshizawa, Jun Tsunehiro, Kazuyo Nomura, Masao Itoh, Fumio Fukui,
Akio Ametani, and Shuichi Kaminogawa
Antimutagenic Effect of Methanolic Extracts from Peanut Hulls
Gow-Chin Yen and Pin-Der Duh
Novel Benzacridine Derivative in the Green Pigment from Methyl
Caffeate and Butylamine
Goro Yabuta, Yukimichi Koizumi, Kazuko Namiki, Takatoshi Kawai, Tateki
Hayashi, and Mitsuo Namiki
Effect of Natural Food Colorings on Immunoglobulin Production
in Vitro by Rat Spleen Lymphocytes
Yuichiro Kuramoto, Koji Yamada, Osamu Tsuruta, and Michihiro Sugano
Antioxidative Compounds Isolated from Kokuto, Non-centrifugal
Cane Sugar
Yoko Nakasone, Kensaku Takara, Kouji Wada, Junichi Tanaka, Seiichi Yogi,
and Nobuji Nakatani
Effects of ΐ1¨4 Linked Galactooligosaccharides on Use of Magnesium
and Calcification of the Kidney and Heart in Rats Fed Excess Dietary Phosphorus
and Calcium
Osamu Chonan, Rie Takahashi, Hisako Yasui, and Masaaki Watanuki
Molecular Cloning of a Novel Gene, dtsR, Which Rescues the Detergent
Sensitivity of a Mutant Derived from Brevibacterium lactofermentum
Eiichiro Kimura, Chizu Abe, Yoshio Kawahara, and Tsuyoshi Nakamatsu
Efficient l-Serine Production from Methanol and Glycine by Resting
Cells of Methylobacterium sp. Strain MN43
Tairo Hagishita, Toyokazu Yoshida, Yoshikazu Izumi, and Toshio Mitsunaga
Control with Organic Solvents of Efficiency of Persolvent Cholesterol
Fermentation by Pseudomonas sp. Strain ST-200
Noriyuki Doukyu, Hideki Kobayashi, Harushi Nakajima, and Rikizo Aono
Biodegradation of Cellulose Acetate by Neisseria sicca
Kiyofumi Sakai, Tatsuo Yamauchi, Fumiko Nakasu, and Tatsuhiko Ohe
Molecular Cloning and Expression of the Pyrimidine Nucleoside
Phosphorylase Gene from Bacillus stearothermophilus TH 6-2
Kiyoshi Okuyama, Tomoki Hamamoto, Toshitada Noguchi, and Yuichiro Midorikawa
Production of Novel Branched Cyclofructans by Bacillus subtilis
MCI-2834
Sachiko Kushibe, Makoto Ueda, and Yuuki Morimoto
A Novel Gene That Interferes with the Phosphotransfer Signal
Transduction Mediated by the EnvZ Osmosensor in Escherichia coli
Kozo Hirokawa, Tomoaki Ogino, Hirofumi Aiba, and Takeshi Mizuno
Cloning of Stress Tolerance Gene in Torulaspora delbrueckii No.
3110
Kuniho Nakata and Kazuhiko Okamura
Gene Analysis of Trehalose-producing Enzymes from Hyperthermophilic
Archaea in Sulfolobales
Kazuo Kobayashi, Masaru Kato, Yutaka Miura, Masako Kettoku, Toshihiro Komeda,
and Akihiro Iwamatsu
Syntheses and Biological Activities of Dihydro-5,6-dehydrokawain
Derivatives
Shinkichi Tawata, Shigehiko Taira, Naotada Kobamoto, Masanobu Ishihara,
and Seizen Toyama
@
Yasuo Ohnishi and Sueharu Horinouchi (>1υ<)1
Department of Biotechnology, Division of Agriculture and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113, Japan
@The Serratia marcescens serine protease (SSP) is one of the extracellular enzymes secreted from this Gram-negative bacterium. When the ssp gene, which encodes a SSP precursor (preproSSP) composed of a typical NH(>22<)2-terminal signal peptide, a mature enzyme domain, and a large COOH-terminal pro-region, is expressed in Escherichia coli, the mature protease is excreted through the outer membrane into the medium. The COOH-terminal pro-region, which is integrated into the outer membrane, provides the essential function for the export of the mature protein across the outer membrane. This is a very simple pathway, in contrast to the general secretory pathway exemplified by the secretion of a pullulanase from Klebsiella oxytoca, in which many separately encoded accessory proteins are required for the transport through the outer membrane. Moreover, the NH(>22<)2-terminal region of 71 amino acid residues of the COOH-terminal pro-sequence plays an essential role, as an eeintramolecular chaperone,'' in the folding of the mature enzyme in the medium. In addition to ssp, the S. marcescens strain contains two ssp homologues encoding proteins similar to SSP in amino acid sequence and size, but with no protease activity. Characterization of the homologue proteins and chimeric proteins between the homologues and SSP, all of which are produced in E. coli, has shown that they are membrane proteins that are localized in the outer membrane in the same manner as for SSP. By use of the COOH-terminal domain of SSP, pseudoazurin was exported to the cell surface of E. coli, which proves the usefulness of the SSP secretory system in the export of foreign proteins across the outer membrane.
Key words: protein secretion; pro-sequence; folding; serine protease; Serratia marcescens
@
Anuradha Bhaduri, Naotoshi Matsudomi,(>1*,υ<)1 and Kali Pada Das(>1**,υ<)1
Department of Chemistry, Bose Institute, 93/1 A.P.C. Road, Calcutta-700 009, India (>1* <)1Faculty of Agriculture, Yamaguchi University, Yamaguchi 753, Japan (>1** <)1Mason Institute of Ophthalmology, University of Missouri-Columbia, Columbia, MO 65212, U.S.A. Received September 18, 1995
@This paper reports the effect of modification of lysine residues on the adsorption of ovalbumin at alumina/water interface. It has been shown that the pH dependence of the adsorption changes on acetylation of lysine. Thus at pH 7.6 acetylated ovalbumin does not show any affinity for alumina surface although unmodified protein does. It seems that although electrostatic interactions are operative, surface unfolding of proteins and surface hydrophobicity of protein also control the adsorption of ovalbumin onto alumina.
Key words: acetylated protein; protein adsorption; solid/water interface; proteins at interfaces
@
Eiichiro Kimura,(>1υ<)1 Chizu Abe, Yoshio Kawahara, and Tsuyoshi Nakamatsu
Technology and Engineering Laboratories, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki-shi, Kanagawa 210, Japan Received November 1, 1995
@Several strains of Corynebacterium and Brevibacterium are known for their ability to secrete large amounts of amino acids, especially L-glutamate. We focused on the mechanism of L-glutamate secretion triggered by a detergent, namely polyoxyethylenesorbitan monopalmitate (PESP).
@A mutant strain, AJ11060, derived from Brevibacterium lactofermentum ATCC 13869 indicates the sensitivity to PESP. A multicopy suppresser gene that compliments the sensitivity of AJ11060 to the detergent was derived from a gene library of B. lactofermentum AJ12036. A 2855-bp DNA fragment was cloned and sequenced. An open reading frame was found that coded for the rescuer gene of the sensitivity to PESP of AJ11060 and was designated dtsR. The expression of the dtsR gene in B. lactofermentum was confirmed by using anti-DtsR antibody. The deduced DtsR protein indicated significant homology with some biotin enzymes such as the ΐ chain of propionyl-CoA carboxylase from rat (48.3%) and human (48.7%), or a 12S chain of methylmalonyl-CoA carboxyltransferase from Propionibacterium freudenreichii (43.1%).
Key words: Coryneform bacteria; Brevibacterium lactofermentum; l-glutamate fermentation; detergent sensitivity; dtsR gene
@
Ya Hsin Chang, Tsung Chain Chang,(>1*<)1 E-Fong Kao, and Chien Chou(>1υ<)1
School of Medical Technology, National Yang Ming University, Shi-Pai, Taipei 112, Taiwan, Republic of China (>1*<)1 Food Industry Research and Development Institute, P.O. Box 246, Hsinchu 300, Taiwan, Republic of China Received December 19, 1995
@Staphylococcus aureus is a pathogen important in causing human infections and intoxication. A sensitive fiber-optic that produces evanescent waves was developed for the detection of protein A, a product secreted only by S. aureus. In the immunosensor, a 40-mV argon-ion laser that generated laser light at 488 nm was used together with plastic optical fiber and antibodies to protein A were physically adsorbed onto the fiber. The principle of the detection involved a sandwich immunoassay with fluorescein isothiocyanate conjugated with anti-(protein A) immunoglobulin G to produce signals of the antigen-antibody reaction. The detection limit was 1 ng of protein A per milliliter. The fiber-optic immunosensor could be used for rapid and specific detection of S. aureus in clinical specimens and foods.
Key words: biosensor; fiber optic; protein A; Staphylococcus aureus
@
Makoto Fujii,(>1υ<)1 Tetsuro Higuchi, Satoru Mukai, Masami Yonekura,(>1*<)1 Takahiro Yano,Hiroyuki Kawaguchi, Koh-ichi Nonaka, Takao Fukunaga, Yasushi Sugimoto, and Shoji Yamada(>1**<)1
Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, Korimoto 1-21-24, Kagoshima 890, Japan (>1* <)1Faculty of Agriculture, Ibaraki University, 3-21-1 Amimachi-chuo, Inashiki-gun, Ibaraki 300-03, Japan (>1** <)1Laboratory of Marine Biochemistry, Faculty of Fisheries, Kagoshima University, Kagoshima 890, Japan Received January 10, 1996
@We have reported a inhibitor of acid lipases in liver lysosomes and erythrocytes from chickens [M. Fujii et al., Int. J. Biochem., 22, 895-898 (1990)]. In this paper, the properties of the inhibitor were described in comparison with those of apo A-I of chicken.
@The purified inhibitor migrated with the same mobility on SDS-PAGE as apo A-I, and had a molecular weight of 27,000. The peptide map from the lipase inhibitor was similar to that of apo A-I. Antibodies to the acid lipase inhibitor also reacted with apo A-I. Apo A-I inhibited the acid lipase activities of liver lysosomes and erythrocytes from chickens as strongly as the lipase inhibitor. The N-terminal amino acid sequence of lipase inhibitor was identical to that of apo A-I as far as residue 20. The amino acid sequence of peptides obtained from the inhibitor by cleavage with CNBr corresponded to internal sequence of apo A-I, and so the CNBr-peptides were derived by cleavage after the methionine residues in apo A-I. The findings showed that the inhibitor of the acid lipases in liver lysosomes and erythrocytes from chickens was identical to apo A-I.
Key words: apo A-I; lipase inhibitor; acid lipase; lysosome; chicken
@
Changhyeug Kim,(>1*<)1 Hideyuki Tanaka,(>1υ<)1 and Masaji Ogura
Department of Applied Biological Chemistry, Faculty of Agriculture, Utsunomiya University, Mine-350, Utsunomiya 321, Japan (>1*<)1 Department of Applied Biochemistry, United Graduate School, Tokyo University of Agriculture and Technology, 3-8-1 Harumi-cho, Fuchu-shi, Tokyo 183, Japan Received January 22, 1996
@The metabolic fates of the carbon skeletons of leucine, lysine, and threonine were studied in growing rats on the diets containing graded levels of protein calorie percentages (10, 20, 30, and 40 PC%) by use of either gluten or zein at 4100 kcal of metabolizable energy per kg of diets. In growth experiment for 21 days, body weight gain, food intake, and body fat increased at higher PC% in the gluten diets, but rats given zein did not maintain their initial weight even at 40 PC%. The concentration of plasma free lysine remained low with the zein diets, but plasma threonine increased at 10 and 20 PC% in the gluten and zein diets, respectively. Plasma leucine increased as the protein level increased either dietary protein. More than 70% of (>114<)1C was incorporated into body protein 12 h after an intraperitoneal injection of labeled lysine in all groups, but little (>114<)1CO(>22<)2 was expired in rats on the gluten and zein diets. About 79% of (>114<)1C-threonine was incorporated into body protein in rats given the gluten and zein diets at 10 PC%, but the values were gradually decreased with increasing the dietary protein levels. Some 40-50% of (>114<)1C-leucine was incorporated into the body protein in rats given the gluten diets, and the values for the zein diets were extensively decreased in the higher PC% groups where the expired (>114<)1CO(>22<)2 was inversely increased to a great extent. These results showed that, when a specific amino acid was limiting or deficient in the diet, the major portion of the labeled amino acid was utilized for body protein synthesis and little was oxidized to carbon dioxide, whereas the oxidative degradation of essential amino acid other than limiting one was increased and the efficiency of the amino acid utilization was relatively decreased.
Key words: essential amino acid; amino acid metabolism; limiting amino acids; gluten; zein
@
Shinya Kawai,(>1υυ<)1 Ariko Mori,(>1*<)1 Takahiro Shiokawa, Shinya Kajita, Yoshihiro Katayama,(>1*<)1 and Noriyuki Morohoshi(>1*<)1
Laboratory of Plant Technology, Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Saiwai-cho 3-5-8, Fuchu, Tokyo 183, Japan (>1*<)1 Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, Saiwai-cho 3-5-8, Fuchu, Tokyo 183, Japan Received January 24, 1996
@Cinnamic acid 4-hydroxylase (CA4H) is the second enzyme involved in phenylpropanoid biosynthesis and is a member of the cytochrome P-450 superfamily. Three CA4H homologous genes, cyp73a, cyp73b, and cyp73c, and a cDNA clone of cyp73a were isolated from a genomic library and a cDNA library of a hybrid aspen; Populus kitakamiensis, and were characterized. They might be interrupted by two introns each. cyp73a and cyp73b were very similar to each other not only in coding regions but also in non-coding regions. Southern blot analysis showed that four homologous genes for CA4H constructed a small gene family in the diploid genome of P. kitakamiensis. In the promoter regions, there were many common cis-element-like sequences in phenylpropanoid biosynthesis genes.
Key words: CYP73; cytochrome P-450; cinnamic acid 4-hydroxylase; phenylpropanoid; Populus kitakamiensis
@
Ken Tokuyasu,(>1υ<)1 Mayumi Ohnishi-Kameyama,(>1*<)1 and Kiyoshi Hayashi (>1**<)1
Biomaterials Conversion Laboratory, National Food Research Institute, 2-1-2 Kannondai, Tsukuba, Ibaraki 305, Japan (>1* <)1Speciation Laboratory, National Food Research Institute, 2-1-2 Kannondai, Tsukuba, Ibaraki 305, Japan (>1** <)1Enzyme Applications Laboratory, National Food Research Institute, 2-1-2 Kannondai, Tsukuba, Ibaraki 305, Japan Received January 26, 1996
@Chitin deacetylase, active in the presence of acetate (96% of the enzymatic activity was retained in the presence of 100 mM sodium acetate), was purified to electrophoretic homogeneity from a culture filtrate of Colletotrichum lindemuthianum (944-fold with a recovery of 4.05%). The enzyme was induced in the medium after the eighth day of incubation simultaneously with the blackening of the medium. The molecular mass of the enzyme was 31.5 kDa and 33 kDa as judged by SDS-PAGE and gel filtration, respectively, suggesting that the enzyme is a single polypeptide. The optimum temperature was 60C and the optimum pH was 11.5-12.0 when glycol chitin was used as substrate. The enzyme was active toward glycol chitin, partially N-deacetylated water soluble chitin, and chitin oligomers the degrees of polymerization of which were more than four, but was less active with chitin trimer and dimer, and inactive with N-acetylglucosamine. The K(>2m<)2 and k(>3(/)cat<)3 for glycol chitin were 2.55 mM and 27.1 s(>1-1<)1, respectively, and those for chitin pentamer were 414 ΚM and 83.2 s(>1-1<)1, respectively. The reaction rates of the enzyme toward glycol chitin and chitin oligomers seemed to follow the Michaelis-Menten kinetics.
Key words: chitin; chitin deacetylase; Colletotrichum lindemuthianum
@
Tairo Hagishita, Toyokazu Yoshida,(>1υ<)1 Yoshikazu Izumi,(>1*<)1 and Toshio Mitsunaga
Department of Food and Nutrition, Faculty of Agriculture, Kinki University, Nara 631, Japan (>1*<)1 Department of Biotechnology, Faculty of Engineering, Tottori University, Tottori 680, Japan Received February 9, 1996
@Resting cells of methanol-utilizing microorganisms isolated from soils were examined for L-serine production under conditions in which L-serine-degradation was suppressed. Strain MN43, a facultative methylotrophic bacterium identified as a Methylobacterium sp., was selected for further studies. Under the optimal conditions, 65 mg/ml L-serine was produced by this bacterium from 50 mg/ml glycine and 104 mg/ml methanol in 5 days, with a molar conversion ratio from glycine to L-serine of 93%. This production is the highest so far reported for microbes producing l-serine.
Key words: l-serine production; methylotroph; Methylobacterium; serine pathway; enzymatic production
@
Sanae Kitamura, Takayuki Yamaguchi, Xia Zhu, and Masahiro Masada(>1υ<)1
Department of Bioresources Chemistry, Faculty of Horticulture, Chiba University, Matsudo 648, Chiba 271, Japan Received February 16, 1996
@A polyclonal antiserum against cysteine synthase (CSase, EC 4.2.99.8) isozymes (CSases 1, 2, and 3) was prepared for physiological study of the isozymes. From the results of immunotitration and double immunodiffusion analyses, the polyclonal antiserum was proved to have the same degree of reactivity toward each isozyme. The accumulation of the isozymes during germination of spinach seeds was investigated by Western blot analysis. Although CSase 1 was not detected in the cotyledons of dark-grown seedlings, it was found in the green cotyledons of seedlings that had been grown in the light. In the extract of roots, CSase 1 was not detected in any developmental stage. On the other hand, CSases 2 and 3 were detected in all developmental stages and tissues. These results indicate that the expression of CSase 1 is regulated by light, and that the regulatory mechanism of CSase 1 expression differs from those of CSase 2 and 3 expression. Western blot analysis showed that the hydrated seeds contained a new CSase band, which was different from known CSases in terms of electrophoretic mobility.
Key words: cysteine biosynthesis; cysteine synthase; O-acetylserine(thiol)lyase; O-acetyl-l-serine; sulfate assimilation
@
Noriyuki Doukyu, Hideki Kobayashi, Harushi Nakajima, and Rikizo Aono(>1υ<)1
Department of Bioengineering, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta 4259, Yokohama 226, Japan Received February 19, 1996
@Pseudomonas sp. strain ST-200 oxidizes cholesterol dissolved in an organic solvent overlying the medium. Major conversion products are cholest-4-en-3-one (C4EO), 6ΐ-hydroxycholest-4-en-3-one (HCEO), and cholest-4-ene-3,6-dione (CEDO). Productivity of each conversion product was altered by changing organic solvents used to dissolve the cholesterol. Generally, HCEO was predominant among the products. HCEO was produced even by cells grown without cholesterol and then killed with harmful organic solvents. The yield of the most oxidized product, CEDO, was improved when the cells were grown in the presence of cholesterol dissolved in a less toxic solvent, cyclooctane.
Key words: bioconversion of cholesterol; cholesterol conversion; Pseudomonas sp.; persolvent fermentation; organic solvents
@
Kiyofumi Sakai,(>1υ<)1 Tatsuo Yamauchi,(>1*<)1 Fumiko Nakasu,(>1*<)1 and Tatsuhiko Ohe
Department of Biochemistry, Osaka Municipal Technical Research Institute, 1-6-50 Morinomiya, Joto-ku, Osaka 536, Japan (>1*<)1 Fiber and Textile Research Laboratories, Teijin Ltd., 3-4-1 Minohara, Ibaraki, Osaka 567, Japan Received February 22, 1996
@Bacteria capable of assimilating cellulose acetate, strains SB and SC, were isolated from soil on a medium containing cellulose acetate as a carbon source, and identified as Neisseria sicca. Both strains degraded cellulose acetate membrane filters (degree of substitution, DS, mixture of 2.8 and 2.0) and textiles (DS, 2.34) in a medium containing cellulose acetate (DS, 2.34) or its oligomer, but were not able to degrade these materials in a medium containing cellobiose octaacetate. Biodegradation of cellulose acetate (DS, 1.81 and 2.34) on the basis of biochemical oxygen demand reached 51 and 40% in the culture of N. sicca SB and 60 and 45% in the culture of N. sicca SC within 20 days. A decrease in the acetyl content of degraded cellulose acetate films and powder was confirmed by infrared and nuclear magnetic resonance analyses. After 10-day cultivation of N. sicca SB and SC, the number-average molecular weight of residual cellulose acetate decreased by 9 and 5%, respectively. Activities of enzymes that released acetic acid and produced reducing sugars from cellulose acetate were mainly present in the culture supernatant. Reactivity of enzymes for cellulose acetate (DS, 1.81) was higher than that for cellulose acetate (DS, 2.34).
Key words: cellulose acetate; biodegradation; Neisseria sicca; esterase; cellulase
@
Hitoshi Kumagai,(>1υ<)1 Chia-Hong Chu, Takaharu Sakiyama,(>1υυ<)1 Shinya Ikeda, and Kozo Nakamura
Department of Applied Biological Chemistry, Division of Agriculture and Agricultural Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan Received February 26, 1996
@The pH-sensitive swelling of a xanthan/chitosan complex gel, which swelled in NaOH solutions of pH 9-12, was analyzed by the collective diffusion model developed by Tanaka and Fillmore, which is based on the diffusion of a polymer network. The time-course for swelling of the gel beads under a change in pH from 11 to 10 fitted well with the equation derived by Tanaka and Fillmore. The collective diffusion coefficient characteristic of the gel was also obtained, although the value of this coefficient was higher than that of the cooperative diffusion coefficient determined from dynamic light scattering experiments by about 4 orders of magnitude. This result indicates that the swelling rate of the gel due to pH change was not controlled by cooperative diffusion of the polymer networks.
Key words: swelling rate; collective diffusion model; dynamic light scattering; ionic gel; chitosan
@
Chia-Hong Chu, Hitoshi Kumagai,(>1υ<)1 Takaharu Sakiyama,(>1υυ<)1 Shinya Ikeda, and Kozo Nakamura
Department of Applied Biological Chemistry, Division of Agriculture and Agricultural Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan Received February 26, 1996
@A model for analyzing the swelling rate of ionic gels was developed on the basis of the diffusion of a species of mobile ion. This model was applied to the analysis of pH-sensitive swelling of a xanthan/chitosan complex gel in NaOH solutions of pH 9-12, using the sodium ion as the reference mobile ion. The time-course for swelling of gel beads with a pH change from 11 to 10 was successfully described by the developed model. The values for the diffusion coefficient obtained by fitting the model to the data were of the same order as those for the diffusion coefficient of the sodium ion measured for a membrane of the complex gel. Thus, it was confirmed that the swelling rate of the gel due to pH change was mainly controlled by the diffusion of mobile ions. However, the time-course for swelling of the gel at pH values below 10 was not satisfactorily explained by the model developed, suggesting that the change in the degree of ionization during swelling also affected the swelling rate of the xanthan/chitosan complex gel.
Key words: swelling; kinetics; diffusion coefficient; polyelectrolyte complex gel; mass coordinate
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Yi Xu,(>1*<)1 Tetsuo Kobayashi,(>1*,**,υ<)1 and Toshiaki Kudo(>1*<)1
(>1*<)1 Microbiology Laboratory, The Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, Wako, Saitama 351-01, Japan (>1**<)1 Department of Applied Biological Sciences, School of Agricultural Sciences, Nagoya University, Chikusa, Nagoya, Aichi 464-01, Japan Received February 28, 1996
@The groEL gene of the alkaliphilic Bacillus sp. strain C-125 was cloned in Escherichia coli and sequenced. The groEL gene encoded a polypeptide of 544 amino acids and was preceded by the incomplete groES gene, lacking its 5'-end. The sequence of the derived amino acids was 87.5% identical to that of B. subtilis, 85.4% identical to that of B. stearothemophilus, and 60.9% identical to that of E. coli. The GroEL protein was expressed in E. coli. Purified GroEL protected yeast Ώ-glucosidase from irreversible aggregation at a high temperature and the addition of Mg-ATP was essential for reactivation of the Ώ-glucosidase. The addition of E. coli GroES increased recovery of the enzyme activity, indicating that C-125 GroEL could function in coordination with E. coli GroES.
Key words: alkaliphilic Bacillus; GroES; GroEL
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Rie Kuramitsu,(>1υυ<)1 Masatoshi Takahashi,(>1*<)1 Kouji Tahara,(>1*<)1 Kozo Nakamura,(>1*<)1 and Hideo Okai (>1*<)1
Department of Chemistry, Faculty of General Education, Akashi College of Technology, Uozumi, Akashi, Hyogo 674, Japan (>1* <)1Department of Fermentation Technology, Faculty of Engineering, Hiroshima University, 1-4-1 Kagamiyama, Higashihiroshima 724, Japan Received March 11, 1996
@The typical tastes imparted by ionic groups are salty-like of sodium chloride and umami-like of monosodium glutamate, but the relationship between the taste and chemical structure has not previously been elucidated. One of the reason for the difficulty in understanding the taste-structure relationship is the presence of the ambiguous and unfavorable tastes of neutral salts. We define the strange tastes of neutral salts (TNS) collectively in a specific category, and then the tastes due to ionic groups. Sour, salty, and umami tastes and TNS were studied for different acidic and basic groups and various combinations of both groups to elucidate the taste characteristics of the ionic groups. The results reveal that the tastes due to ionic groups have common characteristics, being different from bitter and sweet tastes.
Key words: umami peptide; salty peptide; neutral salt; taste
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Shinkichi Tawata, Shigehiko Taira, Naotada Kobamoto, Masanobu Ishihara, and Seizen Toyama
Department of Bioscience and Biotechnology, College of Agriculture, University of the Ryukyus, 1 Senbaru, Nishihara-cho, Okinawa 903-01, Japan Received March 14, 1996
@The syntheses and biological activities of dihydro-5,6-dehydrokawain derivatives against plant pathogenic fungi and termites were investigated. Dihydro-5,6-dehydrokawain was isolated by a simple method without chromatography from the leaves of Alpinia speciosa K. SCHUM. The white crystalline compound obtained was identified as dihydro-5,6-dehydrokawain (1) by instrumental analyses. 4-Hydroxy-6-(2-phenylethyl)-2H-pyran-2-one (3) was prepared by hydrolyzing dihydro-5,6-dehydrokawain. Three dihydro-5,6-dehydrokawain derivatives were synthesized by reacting 3 with phosphoric agents.
Among the synthesized compounds, dimethyl [6-(2-phenylethyl)-2-oxo-2H-pyran-4-yl]phosphorothionate (4) had the strongest antifungal activity of 91% at 100 ppm against Corticium rolfsii.
Key words: dihydro-5,6-dehydrokawain; dimethyl [6-(2-phenylethyl)-2-oxo-2H-pyran-4-yl]phosphorothionate; 4-hydroxy-6-(2-phenylethyl)-2H-pyran-2-one; antifungal activity; Corticium rolfsii; Alpinia speciosa K. Schum
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Mikihiko Kobayashi and Yutaka Sasaki
National Food Research Institute, Kannondai, Tsukuba 305, Japan Received April 8, 1996
@The limitation of slab gel electrophoresis is in the fixed gel concentration. We devised a slab gel plate to which six different gel concentrations could be applied. Various molecular forms of Taka-amylase A (TAA) prepared by chemical modification were analyzed using this multiple lane system. Modification of Lys, Trp, and carboxyl groups of TAA gave charge isomer forms having different affinities for the substrate, soluble starch. Cross-linked oligomeric forms of TAA were resolved in monomer to tetramer forms in 6.3-12.3% gels. The affinity of TAA for various substrates was also evaluated by the multiple lanes containing different concentrations and various substrates. Moreover, the rapid and simple handling of stained gels by ethanol dehydration and subsequent lapping is described.
Key words: affinity gel electrophoresis; gel electrophoresis; multiple lane slab gel; Taka-amylase A
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Saori Takahashi, Kiyoaki Okayama, Shigeru Kunugi,(>1*<)1 and Kohei Oda
Department of Applied Biology, and (>1*<)1 Department of Polymer Science and Engineering, Faculty of Textile Science, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606, Japan Received April 8, 1996
@Vimelysin is a novel alcohol resistant metalloproteinase from Vibrio sp. T1800. The substrate specificity of vimelysin was studied by using natural and furylacryloyl dipeptide substrates. Vimelysin cleaved mainly Pro(>17<)1-Phe(>18<)1 bond and slightly Tyr(>14<)1-Ile(>15<)1 bond in human angiotensin I. Vimelysin also cleaved mainly Phe(>124<)1-Phe(>125<)1 and Tyr(>116<)1-Leu(>117<)1 bonds, and slightly His(>15<)1-Leu(>16<)1, His(>110<)1-Leu(>111<)1, Ala(>114<)1-Leu(>115<)1, and Gly(>123<)1-Phe(>124<)1 bonds in oxidized insulin B-chain. The substrate specificity of vimelysin, by using furylacryloyl (Fua) dipeptides were also studied. The ratio of k(>3(/)cat<)3 /K(>2m<)2 for Fua-Gly-Phe-NH(>22 <)2/Fua-Gly-Leu-NH(>22<)2, Fua-Phe-Leu-NH(>22 <)2/Fua-Gly-Leu-NH(>22<)2, and Fua-Phe-Phe-NH(>22 <)2/Fua-Gly-Leu-NH(>22<)2 were 15.9, 27.8, and 59.0, respectively. These results indicate that vimelysin easily recognizes phenylalanine in P1' positions, which is different from thermolysin.
Key words: vimelysin; alcohol resistant; metallo-proteinase; substrate specificity; Vibrio sp.
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Kiyoshi Okuyama, Tomoki Hamamoto, Toshitada Noguchi, and Yuichiro Midorikawa
Research & Development Division, Yamasa Corporation, Choshi, Chiba 288, Japan Received April 18, 1996
@The pyrimidine nucleoside phosphorylase (Py-NPase) of Bacillus stearothermophilus TH 6-2 is a dimer of 46-kDa subunits and catalyzes the reversible phosphorolysis of uridine and thymidine. The gene encoding this pyrimidine nucleoside phosphorylase ( pyn gene) has been cloned and sequenced from B. stearothermophilus TH 6-2. The pyn gene corresponded to an open reading frame of 1299 nucleotides that translates into a putative 433 amino acid protein with a molecular weight of 46,271. The deduced amino terminal sequence of Py-NPase coincided with that previously found for the purified enzyme. The deduced amino acid sequence of Py-NPase shared significant similarity with those of human and Escherichia coli thymidine phosphorylases. The cloned pyn gene was overexpressed in E. coli cells to produce an active enzyme in large quantities that accounted for approximately 20% of the total protein.
Key words: Bacillus stearothermophilus ; pyrimidine nucleoside phosphorylase; gene cloning; sequence similarity
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Katsumi Shibata, Hisako Shimada, and Takako Kondo
Department of Human Health Science, Faculty of Human Sciences, Osaka International University for Women, Moriguchi, Osaka 570, Japan Received April 18, 1996
@We investigated the effects of feeding various types of nicotinic acid-free, tryptophan-limiting diets on the conversion ratio of tryptophan to niacin in rats. Various tryptophan-limiting diets were made by adding zein, gelatin, glycine, threonine, methionine, or glycine+threonine+methionine to a nicotinic acid-free, 9% casein diet. When the rats were fed with the tryptophan-limiting diets, the conversion ratio of tryptophan to niacin was markedly decreased. However, the ratio recovered after the addition of tryptophan to the tryptophan-limiting diets. These results clearly prove that the conversion was lowest when the rats were fed with the tryptophan-limiting diets. Therefore, we think that the pellagragenic factor of corn is simply due to a low content of tryptophan, but the adverse effect is due to a low conversion ratio of tryptophan to niacin.
Key words: tryptophan conversion to niacin; amino acid; tryptophan-limiting diet; nicotinamide; rat
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Yasuko Yoshizawa, Jun Tsunehiro, Kazuyo Nomura,(>1*<)1 Masao Itoh,(>1*<)1 Fumio Fukui, Akio Ametani,(>1**<)1 and Shuichi Kaminogawa(>1**<)1
Research and Development Center, Showa Sangyo Co., Ltd., 2-20-2 Hinode, Funabashi, Chiba 273, Japan (>1*<)1 R & D Department, Shin Nihon Chemical Co., Ltd., 19-10 Showa-cho, Anjo, Aichi 446, Japan (>1**<)1 Department of Applied Biological Chemistry, The University of Tokyo, Bunkyo-ku, Tokyo 113, Japan Received April 22, 1996
@The water-soluble polysaccharide fraction from Gracilaria verrucosa (GWS) has been reported to increase the phagocytic activity of mice [Yoshizawa et al., Nippon Shokuhin Kogyo Gakkaishi, 41, 557-560 (1994)]. In this study, the macrophage-stimulation activity of enzyme-degraded GWS (GWS-E) was investigated by intraperitoneally and orally administering GWS-E to mice. The intraperitoneal administration of GWS-E increased the number of peritoneal exudate cells (PEC), and increased the phagocytic activity and oxygen radical-secreting activity (spontaneous chemiluminescence) of PEC. This administration could also stimulate splenic macrophages (SPM), increasing radical-secreting activity. When GWS-E was administered orally, the radical-secreting activity of PEC and SPM increased. In this case of oral administration, the activity of SPM increased in a dose-dependent manner, while that of PEC had an optimum dose. These results indicate that GWS-E had macrophage-stimulation activity in vivo and would be suitable as a source for a physiologically functional food with protective and immunopotentiating activity.
Key words: macrophage activation; radical secretion; sulfated polysaccharide; oral administration; red alga
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Hiroshi Kawashima,(>1*,υ<)1 Kengo Akimoto,(>1υ<)1 Saeree Jareonkitmongkol, Norifumi Shirasaka, and Sakayu Shimizu
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Kyoto 606, Japan Received April 22, 1996
@Nicardipine and nifedipine, Ca channel blockers, inhibited rat liver microsomal desaturases, though verapamil, methoxyverapamil, cinnarizine, flunarizine, and diltiazem did not. However, nicardipine and nifedipine apparently did not inhibit the fungal desaturation in Mortierella alpina 1S-4. Nicardipine inhibited rat liver microsomal ’5 desaturase specifically (50% inhibitory concentration, 170 ΚM), and nifedipine inhibited ’6 desaturase specifically (78 ΚM). The inhibition by nicardipine and nifedipine is uncompetitive, the K(>2i<)2 values for ’5 and ’6 desaturases being 62 and 44 ΚM, respectively.
Key words: nicardipine; nifedipine; desaturase; polyunsaturated fatty acid; Ca channel blocker
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Sachiko Kushibe, Makoto Ueda,(>1υ<)1 and Yuuki Morimoto(>1*<)1
Mitsubishi Chemical Co. Research and Development Division Yokohama Research Center, 1000, Kamoshida, Aoba-ku, Yokohama 227, Japan (>1* <)1Kashima Plant, Mitsubishi Chemical Co., Ltd., 14 Sunayama Hasaki-machi, Kashima-gun, Ibaraki 314-02, Japan Received May 2, 1996
@Transfructosylated derivatives of cyclofructans (CFs) were synthesized using enzymes from Bacillus subtilis MCI-2834. The structures of the transfer products were analyzed by FBMS spectrometry and (>113<)1C-NMR, and HPLC with acid- and enzymatic hydrolysis products. It has been proven that these products are novel branched-chain oligosaccharides composed of cyclohexaose and fructose residues.
Key words: cyclofructan; branched-cyclofructan; Bacillus subtilis
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Kozo Hirokawa, Tomoaki Ogino, Hirofumi Aiba, and Takeshi Mizuno
Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464, Japan Received May 2, 1996
@In Escherichia coli, expression of the major outer membrane proteins, OmpC and OmpF, is regulated in response to the medium osmolarity and other environmental stimuli. A two-component signal transduction system, mediated by EnvZ and OmpR, is crucially responsible for this osmotic regulation of the ompC and ompF genes. In this study, an E. coli gene was cloned, which interferes with expression of both the ompC and ompF genes at the level of transcription, provided that the cloned gene was introduced in E. coli cells by a multicopy plasmid. The gene product was identified as F107, which was previously characterized as a hypothetical protein in E. coli genome databases. F107 containing 107 amino acids appears to be highly hydrophobic, and has a sequence similarity to the eukaryotic type of cytochrome-c oxidase subunit III. The mechanism by which F107 inhibits transcription of ompC and ompF was examined extensively, mainly by using a set of envZ and ompR mutants. These results suggested that F107 interferes specifically with a function of the EnvZ osmosensory kinase. Possible mechanisms by which F107 affect the EnvZ function are discussed.
Key words: Escherichia coli ; EnvZ osmosensor; phosphotransfer signal transduction; OmpC and OmpF
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Kuniho Nakata(>1υ<)1 and Kazuhiko Okamura(>1υυ<)1
Central Research Laboratories, Mercian Corporation, 9-1 Johnan 4-chome, Fujisawa, Kanagawa 251, Japan Received May 13, 1996
A cryptic plasmid (pSRY1) was found in Torulaspora delbrueckii No. 3110. A plasmid vector (pSRY21) was constructed by ligating it to pPR1-RIM-C ORF with a cycloheximide resistance gene (RIM-C). The transformation efficiency of the strain No. 3110 with pSRY21 by the protoplast-PEG method was increased by 3000-fold by treatment of the protoplasts with polylysine [poly(Γ-L-lysine)].
@Previously we have reported a mutant of No. 3110, T1, which can neither assimilate nor accumulate trehalose. The mutant was salt- and temperature-sensitive. We cloned a DNA fragment from No. 3110, which, when introduced, complemented these mutant characters of T1. These results suggested an important role of metabolism of trehalose for the stress tolerance.
Key words: polylysine; cycloheximide; stress tolerance; trehalose; transformation
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Ichiro Miura,(>1*,υ<)1 Makoto Muroi,(>1**<)1 Nobue Shiragami,(>1**<)1 Isamu Yamaguchi,(>1*<)1 and Akira Takatsuki(>1**,υυ<)1
(>1*<)1 Microbial Toxicology Laboratory, and (>1**<)1 Animal and Cellular Systems Laboratory, The Institute of Physical and Chemical Research (RIKEN ), Wako-shi, Saitama 351-01, Japan Received May 23, 1996
@Mepanipyrim, N-(4-methyl-6-prop-1-ynylpyrimidin-2-yl)aniline, diminished the cell surface expression of envelope glycoproteins of Newcastle disease and vesicular stomatitis viruses at concentrations where their synthesis was not profoundly affected. Intoxication by diphtheria toxin and ricin and recycling of transferrin were not affected even when cells were treated with mepanipyrim for 2 h before the addition of these probes, indicating that mepanipyrim does not act on the endocytic and recycling pathways of these proteins. Metabolic conversion of C(>26<)2-NBD-ceramide to sphingomyelin and its back-exchange to the medium was also not affected, but synthesis and back-exchange of C(>26<)2-NBD-glucosylceramide were greatly influenced, and an accumulation of LDL-derived, unesterified cholesterol was induced by the drug. These results are discussed relating to the site(s) of action of mepanipyrim.
Key words: mepanipyrim; glycoprotein trafficking; sphingolipid; cholesterol; brefeldin A
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Gow-Chin Yen(>1υ<)1 and Pin-Der Duh (>1*<)1
Department of Food Science, National Chung Hsing University, 250 Kuokuang Road, Taichung 402, Taiwan, Republic of China (>1*<)1 Department of Food Science, Chia-Nan College of Pharmacy and Science, Tainan 717, Taiwan, Republic of China Received October 30, 1995
@The antimutagenic effects of methanolic extracts of peanut hulls (MEPH) were evaluated by the Ames test. MEPH inhibited the mutagenicity of 4-nitroquinoline-N-oxide (NQNO), a direct-acting mutagen. MEPH also inhibited the mutagenicity of some indirect-acting mutagens and decreased in the order of 2-amino-3-methyl-imidazo(4,5-f )quinoline (IQ)>aflatoxin B(>21<)2 (AFB(>21<)2)>2-amino-6-methyldipyrido(1,2-a : 3',2'-d)imidazole (Glu-P-1)>3-amino-1,4-dimethyl-5H-pyridol(4,3-b)indole (Trp-P-1)>benzo(a)pyrene (B(a)P) for S. typhimurium TA98, and IQ>Trp-P-1>Glu-P-1> AFB(>21<)2>B(a)P for S. typhimurium TA100.
Key words: antimutagenicity; mutagen; peanut hull
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Goro Yabuta,(>1υ<)1 Yukimichi Koizumi,(>1a<)1 Kazuko Namiki,(>1b<)1 Takatoshi Kawai,(>1c<)1 Tateki Hayashi,(>1d<)1 and Mitsuo Namiki (>1d<)1
NODAI Research Institute (NRI ), Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156, Japan (>1a <)1Department of Brewing and Fermentation, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156, Japan (>1b <)1Department of Food and Nutrition, Sugiyama Jogakuen University, Nagoya 465, Japan (>1c <)1Eisai Co., Ltd., Tsukuba, Ibaraki 300-25, Japan (>1d <)1Department of Food Science and Technology, Nagoya University, Chikusa-ku, Nagoya 464, Japan Received February 6, 1996
@A crystalline product resulted from the reduction and acetylation of the green pigment formed from the reaction of methyl caffeate and butylamine. Its structure was determined to be that of a novel benzacridine, i.e., dimethyl-7-butyl-6,9,10-triacetoxybenz[k, l ]-acridine-1,2-dicarboxylate.
Key words: green pigment; benzacridine derivative; caffeate and amine; X-ray crystallographic analysis
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Hideyuki Yasuda and Atsushi Onogi
Lotte Central Laboratory Co., Ltd., 3-1-1 Numakage, Urawa, Saitama 336, Japan Received February 26, 1996
@The effects of ascorbic acid on the deodorizing activity of polyphenols against methanethiol (CH(>23<)2SH) was investigated. The addition of ascorbic acid caused loss of the deodorizing activity of polyphenols against CH(>23<)2SH. It was assumed that ascorbic acid prevented the oxidation of polyphenols, which is the essential step for the deodorizing action against CH(>23<)2SH and so suppressed the deodorizing activity.
Key words: deodorizing activity; methanethiol; polyphenols; (-)-epigallocatechin gallate; ascorbic acid
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Tamo Fukamizo,(>1*,υ<)1 Yuji Honda,(>1*<)1 Hideyoshi Toyoda,(>1**<)1 Seiji Ouchi,(>1**<)1 and Sachio Goto(>1*<)1
(>1* <)1Laboratory of Biophysical Chemistry, and (>1** <)1Laboratory of Plant Pathology, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara 631, Japan Received March 8, 1996
@The cell wall of Fusarium oxysporum was digested with commercial Bacillus pumilus chitosanase. The chitosanase produced low molecular weight heterooligosaccharides consisting of GlcN and GlcNAc from the cell wall. A main component of the digestion products was identified as 2-amino-2-deoxy-ΐ-D-glucopyranosyl-(1¨4)-2-acetamido-2-deoxy-D-glucopyranose. The chitosanase appeared to be more effective than Streptomyces griseus chitinase for cell wall digestion. Moreover, maltose was unexpectedly found in the digestion products, indicating that the cell wall contains Ώ-1,4-linked glucan chain as a polysaccharide component.
Key words: Fusarium oxysporum; cell wall structure; chitosanase; chitooligosaccharide; (>113<)1C-NMR
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Hideharu Seto,(>1υ<)1 Shozo Fujioka, Hiroshi Fujisawa,(>1*<)1 Kuniaki Goto,(>1*<)1 Hideaki Nojiri,(>1**<)1 Hisakazu Yamane,(>1**<)1 and Shigeo Yoshida
The Institute of Physical and Chemical Research (RIKEN), Hirosawa, Wako, Saitama 351-01, Japan (>1*<)1 Research and Development Center, Nippon Zeon Co., Ltd., Kawasaki, Kanagawa 210, Japan (>1**<)1 Biotechnology Research Center, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan Received March 14, 1996
@For use as the internal standards in a quantitative analysis of natural jasmonic acid (JA) and methyl jasmonate (JAMe) by gas chromatography-mass spectrometry-selected ion monitoring, (})-2-(2,3-(>12<)1H(>22<)2)JA and its methyl ester, (})-2-(2,3-(>12<)1H(>22<)2)JAMe, were efficiently prepared from 2-(2-pentyl)-2-cyclopentenone through catalytic semi-deuteriogenation of acetylenic intermediates with deuterium gas in pyridine.
Key words: jasmonic acid; methyl jasmonate; (})-2-(2,3-(>12<)1H(>22<)2)jasmonic acid; methyl (})-2-(2,3-(>12<)1H(>22<)2)jasmonate
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Yuichiro Kuramoto,(>1υ<)1 Koji Yamada, Osamu Tsuruta, and Michihiro Sugano
Laboratory of Food Science, Department of Food Science and Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-81, Japan Received March 14, 1996
@Water-soluble (cacao pigment, cochineal pigment, corn pigment, betanin, carthamus yellow, and monascus pigment) and water-insoluble (gardenia yellow, laccaic acid, bixin, and curcumin) natural colorings inhibited IgE production by rat spleen lymphocytes at 10 and 1 ΚM, respectively. Although many of these colorings only inhibited the production of IgG and IgM at high concentrations, the water-insoluble colorings enhanced IgM production even at 1 ΚM. These results suggest that natural colorings have immunoglobulin production-regulating activity.
Key words: food additive; immunoglobulin; spleen lymphocytes
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Yoko Nakasone,(>1υ<)1 Kensaku Takara, Kouji Wada, Junichi Tanaka,(>1*<)1 Seiichi Yogi,(>1*<)1 and Nobuji Nakatani (>1**<)1
Department of Bioscience and Biotechnology, College of Agriculture, University of the Ryukyus, Nishihara-cho, Okinawa 903-01, Japan (>1* <)1College of Science, University of the Ryukyus, Nishihara-cho, Okinawa 903-01, Japan (>1** <)1Faculty of Science of Living, Osaka City University, Sumiyoshi-ku, Osaka 558, Japan Received March 25, 1996
@Five antioxidative compounds were isolated from dichloromethane extracts of kokuto, a kind of non-centrifugal sugar, and identified by spectral analysis as syringaresinol (1), medioresinol (2), coniferyl alcohol (3), sinapyl alcohol (4), and 3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone (5). The antioxidative activities of 1 as determined by the ferric thiocyanate method and by the thiobarbituric acid (TBA) method were respectively superior to and similar to those of butylated hydroxyanisole (BHA). 2 and 5 had stronger activities than BHA, while 3 and 4 respectively had almost the same and weaker activities than BHA.
Key words: antioxidant; non-centrifugal sugar; kokuto; phenyl propanoid; lignan
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Hang Thi Mai, Setsuo Furuyoshi,(>1*<)1 Shinpei Yamamoto, and Toshiharu Yagi(>1υ<)1
Department of Bioresources Science, Faculty of Agriculture, and (>1*<)1 Research Institute of Molecular Genetics, Kochi University, Monobe, Nankoku, Kochi 783, Japan Received April 8, 1996
@A simple purification procedure for raw starch-adsorbable and -digesting amylases (RSAs) was devised. The method depended on an affinity column, which was prepared by mixing raw corn starch and Hyflo Super-Cel. RSAs were specifically adsorbed on the matrix, and eluted with a buffer containing 1% ΐ-cyclodextrin. This column could be used to purify RSAs from Streptomyces thermocyaneoviolaceus and a recombinant strain of E. coli.
Key words: amylase; raw starch; affinity chromatography; raw starch-digesting amylase
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Kazuo Kobayashi,(>1υυ<)1 Masaru Kato, Yutaka Miura, Masako Kettoku, Toshihiro Komeda,(>1*<)1 and Akihiro Iwamatsu (>1*<)1
Applied Bioresearch Center, Kirin Brewery Co., Ltd., 3 Miyaharacho, Takasaki-shi, Gunma 370-12, Japan (>1*<)1 Central Laboratories for Key Technology, Kirin Brewery Co., Ltd., 1-13-5 Fukuura, Kanazawa-ku, Yokohama-shi, Kanagawa 236, Japan Received April 9, 1996
@The genes encoding new trehalose-producing enzymes from S. acidocaldarius ATCC33909 were cloned to analyze the distribution of these genes in Sulfolobales. Comparison of the amino acid sequences with S. solfataricus KM1 showed approximately 50% similarity. Southern analysis suggests that homologues of the trehalose-producing enzyme genes exist widely in Sulfolobales and strains in Sulfolobales were classified into three kinds of genotypes.
Key words: trehalose; Sulfolobus ; trehalose-producing enzyme; Southern analysis; genotype
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Hirosuke Oku,(>1υ<)1 Tomie Sakihama, and Isao Chinen
Laboratory of Applied Biochemistry, Faculty of Agriculture, University of Ryukyus, Nishihara-cho, Okinawa, 903-01, Japan Received April 15, 1996
@Isoleucine (Ile) is a precursor for the biosynthesis of anteiso-fatty acids in rat skin, and among the four possible stereoisomers of Ile, L-Ile, and L-allo-Ile were selectively used for biosynthesis of anteiso-fatty acids. This study examined the optical rotation of anteiso-fatty acid derived from DL-Ile to ascertain its stereoconfiguration. Specific rotation of anteiso-fatty acid derived from DL-Ile favorably compared with that derived from L-Ile, suggesting the selective biosynthesis of the (S )-enantiomer of anteiso-fatty acid in rat skin.
Key words: stereoconfiguration; anteiso-fatty acid; branched-chain; stereoselectivity; rat skin
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Takao Kataoka,(>1υ<)1 Masatoshi Taniguchi, Atsushi Yamada, Hidefumi Suzuki, Shin-ichi Hamada, Junji Magae, and Kazuo Nagai
Department of Bioengineering, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226, Japan Received April 16, 1996
@Perforin- and Fas-based killing pathways are two major mechanisms of cytotoxic T lymphocytes (CTL)-mediated cytotoxicity. In this paper, we have reported the identification of low molecular weight probes on CTL-mediated cytolysis. In addition to inhibitors of acidification so far reported, three other groups of compounds have been identified to block perforin-based cytolysis by the CD8(>1+<)1 CTL clone: (1) an inhibitor of actin polymerization (cytochalasin D), (2) respiratory inhibitors (antimycin A and oligomycin A), and (3) protein kinase inhibitors (calphostin C, herbimycin A, K252a, and staurosporine). Since Fas-based cytolysis by CD4(>1+<)1 CTL clone was inhibitable or rather increased by these agents, only vacuolar type H(>1+<)1-ATPase inhibitors such as concanamycin A have been shown to be highly specific probes to block perforin-based CTL-mediated cytotoxicity.
Key words: CTL; cytotoxicity; Fas; perforin
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Takao Kataoka,(>1υ<)1 Michio Sato,(>1*<)1 Shunzo Kondo,(>1*<)1 and Kazuo Nagai Department of Bioengineering, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226, Japan (>1*<)1 Electron Microscopy Section, Mitsubishi Kasei Institute of Life Sciences, 11 Minamiooya, Machida 194, Japan Received May 1, 1996
@A vacuolar type H(>1+<)1-ATPase inhibitor, concanamycin A (CMA), raised the pH and induced morphologic changes such as vacuolation in lytic granules in the CD8(>1+<)1 cytotoxic T lymphocyte clone OE4. Here we measured the pH in the lytic granules present in OE4 by immunoelectron microscopic observation using 3-(2,4-dinitroanilino)-3'-amino-N-methyldipropylamine (DAMP) as a pH probe. In control OE4, only the peripheral region, but not cores, were heavily stained with DAMP. Exposure to 100 nM CMA for 60 min raised the pH in the lytic granules from 5.7}0.2 to 6.8}0.2. Furthermore, the number of lytic granules in OE4 was estimated under phase contrast microscopy after Wright staining. About 20 lytic granules in average were detected in control cells, while CMA induced vacuolation and decreased their number by 20 to 30%.
Key words: acidic pH; concanamycin A; cytotoxic T lymphocytes; lytic granules; vacuolar type H(>1+<)1-ATPase.
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Hideo Hayashi,(>1υ<)1 Yoshihide Asabu, Katsushi Naito, Mitsuru Nakayama, Hiroshi Nozaki,(>1*<)1 and Motoo Arai
Department of Applied Biological Chemistry, College of Agriculture, Osaka Prefecture University, Sakai, Osaka 593, Japan (>1* <)1Department of Biological Chemistry, Faculty of Science, Okayama University of Science, Ridai-cho, Okayama 700, Japan Received April 22, 1996
@A new oleanane triterpene was isolated from okara fermented with Penicillium simplicissimum ATCC 90288. Its structure was established to be 7ΐ,15Ώ,24-trihydroxyolean-12-en-3,11,22-trione by spectroscopic techniques and an X-ray crystallographic analysis.
Key words: oleanane triterpene; Penicillium simplicissimum
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Osamu Chonan, Rie Takahashi, Hisako Yasui, and Masaaki Watanuki Yakult Central Institute for Microbiological Research, 1796 Yaho, Kunitachi, Tokyo 186, Japan Received May 17, 1996
@Magnesium deficiency was induced in male Wistar rats by adding an excess of phosphorus and calcium to the diet (1.195 g of phosphorus and 1.04 g of calcium/100 g of diet). Feeding of these animals with a diet containing ΐ1¨4 linked galactooligosaccharides (4'-GOS) (5 g of 4'-GOS/100 g of diet) increased the apparent magnesium absorption ratios and the concentrations of magnesium in the serum and femur, and reduced accumulation of calcium in the kidney and heart. We speculate that the use of magnesium increased by feeding 4'-GOS to a limited extent prevented the lower magnesium status and the severity of calcification of the kidney and heart caused by excess dietary phosphorus and calcium.
Key words: ΐ1¨4 galactooligosaccharides; magnesium; calcification; kidney; heart
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Koji Ikura,(>1*,υ<)1 Rika Shinagawa, Koji Hatano, Naoki Suto, and Ryuzo Sasaki
Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Kyoto 606-01, Japan (>1*<)1 Department of Chemistry and Materials Technology, Faculty of Engineering and Design, Kyoto Institute of Technology, Matsugasaki, Kyoto 606, Japan Received May 17, 1996
@Transglutaminase is a calcium-dependent enzyme that catalyzes an amine incorporation and a cross-linking of proteins. Intracellular transglutaminase is induced when human promyelocytic leukemia HL-60 cells are treated with retinoic acid and human hepatoblastoma HepG2 cells, with interleukin-6. To find whether the intracellular reaction catalyzed by transglutaminase increases when the enzyme is induced in these cells, the transglutaminase-catalyzed incorportation of (>114<)1C-labeled methylamine into cellular proteins was measured. The incorporation level of the labeled methylamine into proteins of HL-60 and HepG2 cells did not increase after the transglutaminase had been induced. The presence of the calcium ionophore A23187 did not affect these results. These findings suggested that even after the enzyme induction the catalytic action of intracellular transglutaminase is maintained at a constant level in these cells by unknown regulatory mechanism(s).
Key word: transglutaminase
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Masaaki Onda, Akemi Morimoto, Ayako Simoide, Ken Iwata, and Hiroshi Nakajima
Department of Biochemistry, Unitika Research and Development Center, 23 Kozakura, Uji, Kyoto 611, Japan Received May 21, 1996
@ATP sulfurylase, which catalyzes the first step on the sulfate activation, was purified to apparent homogeneity from Bacillus stearothermophilus, with ammonium sulfate fractionation and successive column chromatography. The enzyme had an apparent molecular mass of 100 kDa, consisting of two equal-sized-44 kDa subunits. The optimum pH was about 8.5-8.7. Divalent cations such as Mn(>12+<)1, Mg(>12+<)1, and Co(>12+<)1 were required for its activity. Apparent K(>2m<)2 values for ATP and SO(>32-(/)4<)3 were 0.045 mM and 0.2 mM, respectively. The enzyme can use deoxyadenosine 5'-triphosphate as a substrate. The enzyme was thermostable and did not show significant loss of activity for 15 min of incubation up to 70C, suggesting a practical use for a continuous reaction.
Key words: ATP sulfurylase; thermophilic bacterium; thermostable; sulfate activation; adenosine 5'-phosphosulfate
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Tasuku Nakajima,(>1*<)1 Yutaka Maruyama, Asako Sato, Takehiko Matsumoto, Masahiro Suenaga, and Eiji Ichishima
Department of Applied Biological Chemistry, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981, Japan Received June 18, 1996
@A novel mannanase that has a strict specificity for the Ώ-1,3-mannosidic linkage has been isolated and characterized. The enzyme was purified to homogeneity from the cultural supernatant of a soil bacterium, Flavobacterium sp. AS-9 isolated by enrichment culture on Auricularia mannan of which the structure was an Ώ-1,3-linked D-mannan main chain branched with short segments of xylose or glucuronic acid. Purified mannanase was optimally active between pH 6 and 8. The apparent molecular mass of the enzyme was 94 kDa by SDS gel electrophoresis under reducing conditions and 100 kDa by gel filtration HPLC. The enzyme only hydrolyzed Ώ-1,3-D-mannan with endo-type action to produce Ώ-1,3-mannooligosaccharides of various sizes. The smallest product was Ώ-1,3-mannobiose.
Key words: fungal Ώ-d-mannan; Ώ-1,3-d-mannanase
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Nobuyoshi Sato, Tatsuo Horiuchi, Mitsutoshi Hamano, Hiroshi Sekine,(>1*<)1 Satoru Chiba,(>1**<)1 Hiroki Yamamoto,(>1**<)1 Takeya Yoshioka,(>1**<)1 Ikuo Kimura,(>1**<)1 Mikio Satake,(>1**<)1 and Yoshiteru Ida(>1***<)1
Research and Development Division, Kikkoman Corporation, 399 Noda, Noda-shi, Chiba 278, Japan (>1*<)1 Noda Institute for Scientific Research, 399 Noda, Noda-shi, Chiba 278, Japan (>1**<)1 Central Research Laboratory, Nippon Suisan Corporation, 559-6 Kitano-machi, Hachioji-shi, Tokyo 192, Japan (>1***<)1 School of Pharmaceutical Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142, Japan Received March 25, 1996
@A new cysteine protease inhibitor, named kojistatin A, was found in the extract of a solid culture of Aspergillus oryzae ATCC 20386. The structure of kojistatin A was determined on the basis of chemical and spectroscopic evidence. Kojistatin A specifically inhibited cysteine proteases such as papain, ficin, and bromelain.
Key words: Aspergillus oryzae; kojistatin A; cysteine protease inhibitor