Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Volume 66 , Issue 11
Showing 1-38 articles out of 38 articles from the selected issue
Analytical Chemistry Note
  • Shin-ichi INOMATA, Satoshi TSUCHIYA, Kazutaka IKEDA, Osamu SAITO, Tets ...
    2002 Volume 66 Issue 11 Pages 2461-2464
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The variegated cutworm, Peridroma saucia Hübner, is a lepidopteran pest to a large number of crops in Canada, the United States, and Europe. It was probably naturalized in Japan in the 1970s. The pheromone glands of the female moth include two components with electroantennographic activity in a ratio of 3:1. GC-MS analyses of pheromone extracts untreated and treated with dimethyl disulfide revealed the major component to be (Z)-11-hexadecenyl acetate and the minor component to be (Z)-9-tetradecenyl acetate. The synthetic pheromone was used to attract a large number of males in a vegetable field in Tokyo, which suggests that this species has already become a harmful pest in Japan.
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Organic Chemistry Regular Papers
  • Takeshi SASSA, Naoto TAJIMA, Mitsuyoshi SATO, Ai TAKAHASHI, Nobuo KATO
    2002 Volume 66 Issue 11 Pages 2356-2361
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Our search for new polar fusicoccins biosynthetically related to fusicoccin A from the culture filtrate of isolate Niigata 2-A of a peach Fusicoccum canker fungus resulted in the isolation of new fusicoccins named fusicoccins P and Q, and 3-epifusicoccins H and Q, together with 3′-deacetylfusicoccin A and 16-O-demethyl-3-epifusicoccin J. The structures of fusicoccins P and Q, and of 3-epifusicoccin Q were determined to be those of deisopentenylfusicoccin J, 12α-hydroxyfusicoccin H and 12α-hydroxy-3-epifusicoccin H, respectively, by NMR spectrometry and chemical derivation from known fusicoccins. 3-Epifusicoccin H was identified by comparing its 400 MHz NMR spectra with those of fusicoccin H. The lettuce seed germination-stimulating activity of these new fusicoccins was examined in the presence of ABA: fusicoccin P was highly active, while 3-epifusicoccins H and Q were slightly active, and fusicoccins H and Q were almost inactive. Possible biosynthetic pathways incorporating these new fusicoccins and 3-epifusicoccins from geranylgeranyl diphosphate to 3′-deacetlyfusicoccin A and 16-O-demethyl-3-epifusicoccin J are discussed.
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  • Mitsuru ITO, Hideshi OKUI, Harumi NAKAGAWA, Shigeru MIO, Ayako KINOSHI ...
    2002 Volume 66 Issue 11 Pages 2406-2414
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      A new series of N-oxydihydropyrrole derivatives was synthesized and evaluated for insecticidal activity against Nilaparvata lugens and Myzus persicae. Various substituents were introduced to the 1-position of the dihydropyrrole ring, and the derivatives obtained exhibited systemic and/or contact insecticidal activity. The structure-activity relationship revealed that small alkyoxy and alkoxyalkoxy groups were more favorable than alkylcarbonyloxy, alkoxycarbonyloxy, or sulfonyloxy groups as substituents at the 1-position.
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  • Jun MORISAWA, Chul-Sa KIM, Takehiro KASHIWAGI, Shin-ichi TEBAYASHI, Mi ...
    2002 Volume 66 Issue 11 Pages 2424-2428
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Sandaracopimarinol and (1S,6R)-2,7(14),10-bisabolatrien-1-ol-4-one were isolated and identified from Cryptomeria japonica as repellents against Armadillidium vulgare which is well known as an unpleasant pest in the house and as vegetable pest in Japan. These compounds strongly repelled A. vulgare when they were combined, although each compound alone did not show any activity.
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  • Kazutoshi SHINDO, Hidefumi SUZUKI, Toru OKUDA
    2002 Volume 66 Issue 11 Pages 2444-2448
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Paecilopeptin, a novel cathepsin S inhibitor, was produced and isolated from the culture supernatant of the fungal strain, Paecilomyces carneus. A spectroscopic analysis revealed the planar structure of paecilopeptin to be acetyl-Leu-Val-CHO. The stereochemistry of the constituent amino acids was analysed by chiral HPLC after oxidation and 6N HCl hydrolysis of paecilopeptin. The total synthesis of paecilopeptin was completed in six steps. Paecilopeptin inhibited human cathepsin S with an IC50 value of 2.1 nM in vitro.
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Organic Chemistry Notes
Biochemistry & Molecular Biology Regular Papers
  • Akira KAWAKAMI, Midori YOSHIDA
    2002 Volume 66 Issue 11 Pages 2297-2305
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      We isolated two cDNAs of winter wheat (Triticum aestivum L.), designated wft1 and wft2, which encoded sucrose:fructan 6-fructosyltransferase (6-SFT) and sucrose:sucrose 1-fructosyltransferase (1-SST; EC 2.4.1.99), respectively, which are involved in the synthesis of fructan in wheat. wft1 and wft2 were cloned by screening of a cDNA library with probed-cDNA fragments corresponding to plant fructosyltransferase and invertase. The identity of the clones was verified by functional characterization of recombinant proteins expressed in methylotrophic yeast, Pichia pastoris. Northern blotting showed that the level of wft2 transcripts increased from autumn to early winter in the crown tissues of all field-grown wheat cultivars examined. Higher levels of wft1 and wft2 transcripts were found in leaf tissues of snow mold-resistant cultivars, which accumulated more fructan than other cultivars. Our results showed that Wft1 and Wft2 were important in fructan accumulation during cold hardening of winter wheat.
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  • Masako SHINJOH, Noribumi TOMIYAMA, Taro MIYAZAKI, Tatsuo HOSHINO
    2002 Volume 66 Issue 11 Pages 2314-2322
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The D-sorbitol dehydrogenase gene, sldA, and an upstream gene, sldB, encoding a hydrophobic polypeptide, SldB, of Gluconobacter suboxydans IFO 3255 were disrupted in a check of their biological functions. The bacterial cells with the sldA gene disrupted did not produce L-sorbose by oxidation of D-sorbitol in resting-cell reactions at pHs 4.5 and 7.0, indicating that the dehydrogenase was the main D-sorbitol-oxidizing enzyme in this bacterium. The cells did not produce D-fructose from D-mannitol or dihydroxyacetone from glycerol. The disruption of the sldB gene resulted in undetectable oxidation of D-sorbitol, D-mannitol, or glycerol, although the cells produced the dehydrogenase. The cells with the sldB gene disrupted produced more of what might be signal-unprocessed SldA than the wild-type cells did. SldB may be a chaperone-like component that assists signal processing and folding of the SldA polypeptide to form active D-sorbitol dehydrogenase.
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  • Kozo HIROKAWA, Naoki KAJIYAMA
    2002 Volume 66 Issue 11 Pages 2323-2329
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Agrobacterium tumefaciens AgaE-like protein had a similar sequence to that of a fructosyl amino acid oxidase from Corynebacterium sp. strain 2-4-1. To characterize the AgaE-like protein, we produced the enzyme in Escherichia coli, and purified it to homogeneity. The molecular mass of recombinant AgaE-like protein was 42 kDa on SDS-PAGE and 85 kDa on gel filtration. The protein acted on N-fructosyl valine and N-fructosyl glycine as substrates, but not on glycated protein or Nε-fructosyl lysine. Apparent Km for N-fructosyl valine and N-fructosyl glycine were 1.64 and 0.31 mM, respectively. The AgaE-like protein had maximum activity at pH 7.8 and 35°C in 0.1 M potassium phosphate, but more than 80% of its activity was lost at 40°C or more. In contrast to eukaryotic fructosyl amino acid oxidases, the AgaE-like protein contained noncovalently bound FAD as a cofactor and was inactive against Nε-fructosyl Nα-Z(benzyloxycarbonyl)-lysine. These characteristics were similar to a fructosyl amino acid oxidase from Corynebacterium sp. strain 2-4-1, suggesting that these prokaryotic enzymes comprise a new family of fructosyl amino acid oxidases.
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  • Akio SUGIHARA, Yuji SHIMADA, Atsuo NOMURA, Tadamasa TERAI, Masaki IMAY ...
    2002 Volume 66 Issue 11 Pages 2347-2355
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      With the aim of developing a new cholesterol esterase for eliminating lipids on used contact lenses, microorganisms were screened for the enzyme activity. A Pseudomonas aeruginosa isolated from soil was found to produce a desirable enzyme. The enzyme had an isoelectric point of 3.2, and molecular mass of 58 kDa. The optimal temperature was around 53°C at pH 7.0, and the optimal pH was from 5.5 to 9.5. The enzyme was stable between pH 5 and 10 for 19 h at 25°C, and retained its activity up to 53°C on 30 min of incubation at pH 7.0. The rates of hydrolysis of cholesteryl esters of different fatty acids were in the following order: linoleate>oleate>stearate>palmitate>caprylate>myristate> laurate, caprate>caproate>butyrate, acetate. Addition of (tauro)cholate to a final concentration of 100 mM markedly promoted the hydrolysis of triglycerides of short-, medium-, and long-chain fatty acids. When used with taurocholate, the enzyme acted as an effective cleaner for contact lenses stained with lipids consisting of cholesteryl oleate, tripalmitin, and stearyl stearate.
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  • Toshiyuki NAKAMURA, Takayuki SASAKI, Minoru FUJIMORI, Kazuyuki YAZAWA, ...
    2002 Volume 66 Issue 11 Pages 2362-2366
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Bifidobacterium longum is a nonpathogenic anaerobic bacterium among normal bacterial flora. Recently, it was reported that B. longum accumulated in hypoxic solid tumors. The gene of interest was expressed in transfected B. longum by the shuttle vector pBLES100 in solid tumors. In this report, we constructed pBLES100-S-eCD, which included the cytosine deaminase gene. We confirmed by western blotting that transfected B. longum produced cytosine deaminase. In addition, transfected B. longum produced cytosine deaminase that converted 5-fluorocytosine into 5-fluorouracil. B. longum could be useful for enzyme/pro-drug therapy of hypoxic solid tumors.
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  • Sakihito KITAJIMA, Masami UEDA, Satoshi SANO, Chikahiro MIYAKE, Takayu ...
    2002 Volume 66 Issue 11 Pages 2367-2375
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Depletion of the electron donor ascorbate causes rapid inactivation of chloroplastic ascorbate peroxidase (APX) of higher plants, while cytosolic APX is stable under such conditions. Here we report the cloning of cDNA from Galdieria partita, a unicellular red alga, encoding a novel type of APX (APX-B). The electrophoretic mobility, Km values, kcat and absorption spectra of recombinant APX-B produced in Escherichia coli were measured. Recombinant APX-B remained active for at least 180 min after depletion of ascorbate. The amino-terminal half of APX-B, which forms the distal pocket of the active site, was richer in amino acid residues conserved in chloroplastic APXs of higher plants rather than cytosolic APXs. In contrast, the sequence of the carboxyl-terminal half, which forms the proximal pocket, was similar to that of the cytosolic isoform. The stability of APX-B might be due to its cytosolic isoform-like structure of the carboxyl-terminal half.
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  • Koji YUASA, Masayoshi MAESHIMA
    2002 Volume 66 Issue 11 Pages 2382-2387
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Vacuoles of radish (Raphanus sativus) contained a Ca2+-binding protein (RVCaB) of 43 kDa. We investigated the Ca2+-binding properties of the protein. RVCaB was expressed in Escherichia coli and was purified from an extract by ion-exchange chromatography, nitrocellulose membrane filtration, and gel-filtration column chromatography. Ca2+-binding properties of the recombinant protein were examined by equilibrium dialysis with 45Ca2+ and small dialysis buttons. The protein was estimated to bind 19Ca2+ ions per molecule with a Kd for Ca2+ of 3.4 mM. Ca2+ was bound to the protein even in the presence of high concentrations of Mg2+ or K+. The results suggested that the protein bound Ca2+ with high ion selectivity, high capacity, and low affinity.
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  • Isao TAKAHASHI, Sayoko SUGIURA, Hirotoshi OHTA, Kazuo OZAWA, Tadashi K ...
    2002 Volume 66 Issue 11 Pages 2402-2405
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Serological detection of antibodies specific to human cytomegalovirus (HCMV) is not reliable because the assay uses the whole HCMV protein fraction. Antigenic materials composed of well-characterized viral proteins are being tried for serodiagnosis in Europe. Epitopes of antibodies to HCMV phosphoprotein 150 (pp150) encoded by UL32 in Japanese individuals were investigated for comparison with the results in Europe. The major epitopes on amino acid residues 496 to 652 of HCMV pp150 were identified and the detection of antibodies with an enzyme-linked immunosorbent assay (ELISA) of synthetic peptides against the main epitopes was established. Fifteen seropositive and five seronegative serum samples for the epitope mapping and 131 seropositive and 50 seronegative samples for ELISA were investigated. Overlapping 15-mer peptides moving by two amino acids through V496-H652 were synthesized. The main epitope regions were V508-D530, L526-Q544, S536-D554, T616-G634, S624-P642, and L632-H652. When each peptide was conjugated with bovine serum albumin for ELISA, 80.9% of the seropositive samples were judged to be positive. The results of this study are the same as those for European sera, so the antigenic materials developed in Europe might be used to replace the whole HCMV protein fraction in Japanese.
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  • Yoshio TANAKA, Tsunehiro AKI, Yûki HIDAKA, Yûji FURUYA, Se ...
    2002 Volume 66 Issue 11 Pages 2415-2423
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The fungal strain Mortierella alliacea YN-15 is an arachidonic acid producer that assimilates soluble starch despite having undetectable α-amylase activity. Here, a α-glucosidase responsible for the starch hydrolysis was purified from the culture broth through four-step column chromatography. Maltose and other oligosaccharides were less preferentially hydrolyzed and were used as a glucosyl donor for transglucosylation by the enzyme, demonstrating distinct substrate specificity as a fungal α-glucosidase. The purified enzyme consisted of two heterosubunits of 61 and 31 kDa that were not linked by a covalent bond but stably aggregated to each other even at a high salt concentration (0.5 M), and behaved like a single 92-kDa component in gel-filtration chromatography. The hydrolytic activity on maltose reached a maximum at 55°C and in a pH range of 5.0-6.0, and in the presence of ethanol, the transglucosylation reaction to form ethyl-α-D-glucoside was optimal at pH 5.0 and a temperature range of 45-50°C.
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  • Norihito NAKAMICHI, Masaya MURAKAMI-KOJIMA, Eriko SATO, Yasuko KISHI, ...
    2002 Volume 66 Issue 11 Pages 2429-2436
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The complete genome sequence of Arabidopsis thaliana revealed that this higher plant has a tremendous number of protein kinases. We recently isolated a novel type of protein kinase, named AtWNK1, which shows an in vitro ability to phosphorylate the APRR3 member of the APRR1/TOC1 quintet that has been implicated in a mechanism underlying circadian rhythms in Arabidopsis. We here address two issues, one general and one specific, as to this novel protein kinase. We first asked the general question of how many WNK family members are present in this higher plant, then whether or not other members are also relevant to circadian rhythms. The results of our analyses showed that Arabidopsis has at least 9 members of the WNK1 family of protein kinases (designated here as WNK1 to WNK9), the structural design of which is clearly distinct from those of other known protein kinases, such as receptor-like kinases and mitogen-activated protein kinases. They were examined with special reference to the circadian-related APRR1/TOC1 quintet. It was found that not only the transcription of the WNK1 gene, but also those of three other members (WNK2, WNK4, and WNK6) are under the control of circadian rhythms. These results suggested that certain members of the WNK family of protein kinases might play roles in a mechanism that generates circadian rhythms in Arabidopsis.
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  • Takashi MORIMOTO, Yoshihito SUZUKI, Isomaro YAMAGUCHI
    2002 Volume 66 Issue 11 Pages 2437-2443
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      An expression library of Arabidopsis thaliana cDNAs was randomly introduced into A. thaliana. The transformant pool was used to obtain a line, c105, with reduced apical dominance and irregular positioning of leaves and flowers. The inserted DNA was a 3′-fragment of the ribosomal protein S6 gene with antisense orientation. The transcriptional level of the ribosomal protein S6 was lower in c105 than in the wild-type plant. Introduction of the same fragment into the wild-type plant gave phenotypes similar to those of c105, so the phenotypes of c105 were due to the S6 antisense expression. The phenotypes suggest selectively reduced function of specific proteins rather than an overall decrease in protein function caused by defective ribosomal biogenesis.
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Biochemistry & Molecular Biology Notes
  • Koji SHIGETA, Keisuke OOTAKI, Hideki TATEMOTO, Tsutomu NAKANISHI, Akir ...
    2002 Volume 66 Issue 11 Pages 2491-2494
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      A methanol extract of Coptidis Rhizoma effectively enhanced the outgrowth of neurite in PC12 cells induced by nerve growth factor (NGF). Following solvent partition and preparative HPLC, berberine was isolated as the major active compound. Berberine enhanced the proportion of neurite-bearing cells in a dose-dependent manner without cytotoxicity. Its structural relatives, palmatine and coptisine, showed a slightly weaker NGF-enhancing effect than berberine. These three alkaloids inhibited acetylcholinesterase activity at a level comparable to that of physostigmine, but this inhibition was not responsible for the potentiation of NGF-induced neurite outgrowth. It is demonstrated for the first time that protoberberine alkaloids potentiated the NGF-induced differentiation of neural cells.
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  • Norihiro FUTAMURA, Yuzuru MUKAI, Masahiro SAKAGUCHI, Hiroshi YASUEDA, ...
    2002 Volume 66 Issue 11 Pages 2495-2500
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Many plant pathogenesis-related (PR) proteins are allergenic. We isolated three cDNAs, Cry j 3.1, Cry j 3.2, and Cry j 3.3, that encoded homologs of Jun a 3, a PR protein allergen in Juniperus ashei, from a cDNA library derived from the pollen of Cryptomeria japonica. The predicted amino acid sequences encoded by the three cDNAs were more than 85% identical to each other and about 57% identical to the sequence of Jun a 3. The Cry j 3 genes seemed to form a small multigene family in the genome of C. japonica. Expression of Cry j 3 was strong in roots and in female and male strobili; expression was weaker in cotyledons, leaves, stems, and pollen grains.
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  • Yoshiyuki ASHIDA, Masaki NISHIMOTO, Akihito MATSUSHIMA, Junko WATANABE ...
    2002 Volume 66 Issue 11 Pages 2511-2514
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Genes for two geraniol-responsive factors, designated McEREBP1 and McWRKY1, from cultured shoot primordia of Matricaria chamomilla were cloned. The deduced amino acid sequences of these genes were highly similar to those of the family of ethylene-responsive element binding proteins and elicitor-induced DNA-binding proteins containing a WRKY domain, respectively. The levels of McEREBP1 and McWRKY1 mRNAs were maximum when measured 1 h after treatment of the cultured cells with geraniol.
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  • Hong WU, Hitoshi SHIMOI, Kiyoshi ITO
    2002 Volume 66 Issue 11 Pages 2515-2519
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      A β-1,6-glucanase was purified to apparent homogeneity from a commercial yeast digestive enzyme prepared from Streptomyces rochei by a series of column chromatographies. The molecular mass of the purified enzyme was 60 kDa by SDS-PAGE. The purified enzyme had an optimum pH range from 4.0 to 6.0 and was stable in the same pH range. The enzyme was stable under 50°C but lost almost all activity at 60°C. The enzyme was specific to β-1,6-glucan and had little activity towards β-1, 3-glucan and β-1, 4-glucan. When the β-1,6-glucan was hydrolyzed with the purified enzyme for 5 h, the reaction products contained 20% glucose, 36% gentiobiose, and 44% other oligosaccharides, suggesting that the enzyme is an endo-type glucanase. When the purified enzyme was used for the digestion of the cell wall of Saccharomyces cerevisiae, cell-wall proteins covalently bound to the cell-wall glucan were recovered as soluble forms, suggesting that this enzyme is useful for analysis of yeast-cell wall proteins.
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Biochemistry & Molecular Biology Preliminary Communication
Food & Nutrition Science Regular Papers
  • Kunimasa MORITA, Kazuhiro TOBIISHI
    2002 Volume 66 Issue 11 Pages 2306-2313
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The effects of nori (Porphyra yezoensis), a kind of red alga, on the gastrointestinal absorption and reabsorption of 17 types of dioxin were investigated in male Wistar rats. The rats were fed with 4 g of the control diet or 4 g of the nori diet containing a standard dioxin solution (233 ngTEQ/kg of body weight) for five consecutive days. In the group fed with the 10% nori diet, the fecal excretion of dioxin from days 1 to 5 was higher (p<0.01) than that of the control group by 5.5-fold for 2,3,7,8-TCDD, 6.6-fold for 1,2,3,7,8-pentaCDD, and 6.0-fold for 2,3,4,7,8-pentaCDF. In another experiment, the rats were fed with 4 g of the control diet containing a standard dioxin solution (2991 ngTEQ/kg of body weight) on the first day of the experiment and then given the control diet for 7 consecutive days, before being given either the control diet or the nori diet for 28 consecutive days more. In the group fed with the 10% Nori diet, the fecal excretion of dioxin during the period from days 8 to 35 was higher (p<0.01 or p<0.05) than that of the control group by 2.4-fold for 2,3,7,8-TCDD, 2.3-fold for 1,2,3,7,8-pentaCDD, and 2.4-fold for 2,3,4,7,8-pentaCDF. These results suggest that the administration of nori prevented dioxin from being efficiently absorbed and reabsorbed from the gastrointestinal tract, and might be useful for protecting humans exposed to dioxin from ill effects.
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  • Katsuko KAJIYA, Shigenori KUMAZAWA, Tsutomu NAKAYAMA
    2002 Volume 66 Issue 11 Pages 2330-2335
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Green tea contains a high concentration of such catechins as (−)-epicatechin (EC), (−)-epigallocatechin (EGC), (−)-epicatechin gallate (ECg), and (−)-epigallocatechin gallate (EGCg). Their biological activities have been evaluated by in vitro experiments using cultured cells or bacteria, but the order of activity of the various catechins differed with the study. We have been studying the interaction of tea catechins with lipid bilayers, and clarified that the number of hydroxyl groups on the B-ring, the presence of the galloyl moiety, and the stereochemical structure of each catechin govern their affinity for lipid bilayers. We investigated in this present study the effects of various external factors on the affinity of tea catechins for lipid bilayers by using liposomes as model membranes. The amount of tea catechins incorporated into the lipid bilayers increased with increasing salt concentration in an aqueous medium and decreased with increasing negative electric charge of the lipid bilayers. Furthermore, the amount of EGCg or ECg incorporated into the lipid bilayers increased with increasing EC concentration. These results reveal that the salt concentration in an aqueous medium, the electric charge of the membrane, and the presence of other catechins governed the affinity of tea catechins for the lipid bilayers.
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  • Makoto YOSHIMOTO, Shoji YAHARA, Shigenori OKUNO, Md. Shahidul ISLAM, K ...
    2002 Volume 66 Issue 11 Pages 2336-2341
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The caffeoylquinic acid derivatives, 3-mono-O-caffeoylquinic acid (chlorogenic acid, ChA), 3,4-di-O-caffeoylquinic acid (3,4-diCQA), 3,5-di-O-caffeoylquinic acid (3,5-diCQA), 4,5-di-O-caffeoylquinic acid (4,5-diCQA) and 3,4,5-tri-O-caffeoylquinic acid (3,4,5-triCQA), and caffeic acid (CA) were isolated from the sweetpotato (Ipomoea batatas L.) leaf. We examined the antimutagenicity of these caffeoylquinic acid compounds to promote new uses of the sweetpotato leaf. These caffeoylquinic acid derivatives effectively inhibited the reverse mutation induced by Trp-P-1 on Salmonella typhimurium TA 98. The antimutagenicity of these derivatives was 3,4,5-triCQA>3,4-diCQA=3,5-diCQA=4,5-diCQA>ChA in this order. There was no difference in the antimutagenicity of all dicaffeoylquinic acid derivatives. A comparison of the activities and structures of these compounds suggested that the number of caffeoyl groups bound to quinic acid played a role in the antimutagenicity of the caffeoylquinic acid derivatives. The sweetpotato leaves contained distinctive polyphenolic components with a high content of mono-, di-, and tricaffeoylquinic acid derivatives and could be a source of physiological functions.
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  • Ken KATO, Yukihiro TAKADA, Hiroaki MATSUYAMA, Yoshihiro KAWASAKI, Seii ...
    2002 Volume 66 Issue 11 Pages 2342-2346
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      We investigated the calcium bioavailability of milk calcium, taken with or without cheese. Twenty-four 6-week-old male rats for a meal-feeding experiment were trained to consume an AIN-76 diet within 2 h (2 times per day) for 2 weeks. The rats were then divided into three experimental groups, each fed 2 types of experimental diets: Control group, Cheese group, and Ca-Cheese group. The rats were each alternately given 2 types of experimental diets at 2-h meal-feeding for 31 days. The breaking force and energy of the femur in the Ca-Cheese group were significantly higher than in the control group. The bone mineral density (BMD) of the lumbar spine and the femur in the Ca-Cheese group was also significantly higher than in the other two groups. These results indicate that milk calcium taken with cheese increases bone strength and BMD efficiently, results that may be useful for the prevention of osteoporosis.
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  • Seon-Kang CHOI, Motoyasu ADACHI, Shigeru UTSUMI
    2002 Volume 66 Issue 11 Pages 2395-2401
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Soy glycinin has five major subunits which are classified into two groups according to their homology in amino acid sequences (group I, A1aB1b, A1bB2 and A2B1a; group II, A3B4 and A5A4B3). It has been reported that the peptide fragments derived from the A1a and A2 chains of the A1aB1b and A2B1a subunits had bile acid-binding ability and that the region of 114-161 residues of the A1a chain was responsible for this bile acid-binding ability. In this study, we constructed A1a, A3 and 9 deletion mutants of A1a lacking various numbers of residues at the C-terminus, and evaluated their bile acid-binding ability by a cholic acid-conjugated column and fluorescence analysis. The bile acid-binding ability of A1a was higher than that of A3 and there was a remarkable decrease in the bile acid-binding ability between the Δ[138-291] and Δ[130-291] mutants. The 130-138 region is rich in hydrophobic residues. In this regard, when we constructed the Δ[129-134] mutant lacking six contiguous hydrophobic residues (VAWWMY) and evaluated its bile acid-binding ability, a similar remarkable decrease in the bile acid-binding ability was observed. These results indicate that the 129-134 residue region (VAWWMY) with high hydrophobicity was important for bile acid-binding of A1a.
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  • Yutaka KONISHI, Keiko HAGIWARA, Makoto SHIMIZU
    2002 Volume 66 Issue 11 Pages 2449-2457
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Fluorescein is a marker-dye customary applied to the evaluation of tight-junctional permeability of epithelial cell monolayers. However, the true mechanism for the permeation has not been elucidated. Transepithelial transport of fluorescein in Caco-2 cell monolayers was therefore examined. Fluorescein transport was dependent on pH, and in a vectorical way in the apical-basolateral direction, but it was independent of the tight-junctional permeability of monolayers of these human intestinal cells. The permeation of fluorescein was concentration-dependent and saturable; the Michaelis constant was 7.7 mM and the maximum velocity was 40.3 nmol min−1 (mg protein)−1. Benzoic acid competitively inhibited fluorescein transport, suggesting that fluorescein is transported by a monocarboxylic acid transporter (MCT). Antioxidative polyphenolic compounds such as ferulic acid from dietary sources, competitively inhibited the permeation of fluorescein. These compounds probably share a transport carrier with fluorescein. Measurement of the effects of phenolic acids on fluorescein transport across Caco-2 monolayers would be a useful way to evaluate the intestinal absorption or bioavailability of dietary phenolic acids.
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Food & Nutrition Science Notes
  • Mitsuo MIYAZAWA, Atsushi SUGIE, Masaki SHINDO
    2002 Volume 66 Issue 11 Pages 2458-2460
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The biotransformation of (−)-verbenone was investigated with human liver microsomes by using GC-MS. Regioselective biotransformation was observed when (−)-verbenone was incubated with the liver microsomes. (−)-10-Hydroxyverbenone was formed from (−)-verbenone of kinetic analysis showed that the Km and Vmax values for the hydroxylation of (−)-verbenone by liver microsomes from three human samples, HG-70, HG-56 and HG-23, were 1.1 mM and 4.8 nmol/min/nmol P450, 0.6 mM and 2.1 nmol/min/nmol P450, and 2.8 mM and 4.6 nmol/min/nmol P450, respectively.
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  • Komang Ayu NOCIANITRI, Shoji SAKAKIBARA, Takashi KANNO, Hiroto KIKUCHI ...
    2002 Volume 66 Issue 11 Pages 2465-2470
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The effects of some methyl-containing compounds added to a choline-deficient diet on the metallothionein mRNA level in the rat liver were studied. The addition of choline or carnitine to the choline-deficient diet did not induce a gain in body weight, while the addition of either betaine or methionine to the choline-deficient diet, or of methionine to the choline-deficient diet with choline significantly increased the body weight. The metallothionein mRNA level in the liver of rats fed on the choline-deficient diet was similar to that of rats fed on the choline-deficient diet with choline, betaine or carnitine. However, the addition of methionine to the choline-deficient diet with or without choline caused a marked suppression in the metallothionein mRNA level in the liver. It is thus surmised that the metallothionein mRNA level in the liver might be regulated by the dietary content of methionine.
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  • Hiromitsu AOKI, Nguyen Thi Minh KIEU, Noriko KUZE, Kazue TOMISAKA, Ngu ...
    2002 Volume 66 Issue 11 Pages 2479-2482
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The carotenoids in Gac fruit (Momordica Cochinchinensis spreng) were analysed by high-performance liquid chromatography (HPLC), and the concentrations of β-carotene, lycopene, zeaxanthin and β-cryptoxanthin were measured. Lycopene was found to be predominantly present in the Gac seed membrane at a concentration of up to 380 μg/g of seed membrane. The concentration of lycopene in the Gac seed membrane was about ten-fold higher than that in known lycopene-rich fruit and vegetables, indicating that Gac fruit could be a new and potentially valuable source of lycopene.
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  • Md. Shahidul ISLAM, Makoto YOSHIMOTO, Norihiko TERAHARA, Osamu YAMAKAW ...
    2002 Volume 66 Issue 11 Pages 2483-2486
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      The anthocyanin composition of three varieties, Simon No. 1, Kyushu No. 119, and Elegant Summer, in sweetpotato (Ipomoea batatas L.) leaves was examined for promoting new uses. Fifteen anthocyanin compounds were identified and measured. HPLC clearly showed quantitative differences, but not qualitative ones. The anthocyanins were acylated cyanidin and peonidin type. The result suggests that the major anthocyanin composition of sweetpotato leaves is cyanidin type.
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  • Hiromi YAMASHITA, Yoko NANBA, Miki ONISHI, Masumi KIMOTO, Miki HIEMORI ...
    2002 Volume 66 Issue 11 Pages 2487-2490
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      A 36-kDa allergen, Tri a Bd 36K, was purified from wheat albumin and characterized. The protein was similar to barley peroxidase BP-1 both in its amino acid sequence and peroxidase activity. The enzyme seemed to contain L-fucose and D-mannose and the glycan moiety reacted with IgE antibodies in a patient's serum.
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Microbiology & Fermentation Technology Regular Papers
  • Jun OGAWA, Woro Triarsi SULISTYANINGDYAH, Hiromi TANAKA, Kenji KANO, T ...
    2002 Volume 66 Issue 11 Pages 2376-2381
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Bilirubin dehydrogenase, a membrane-bound enzyme that catalyzes the one-step oxidation of ditaurobilirubin and bilirubin to ditaurobiliverdin and biliverdin, respectively, in the presence of an electron acceptor, was found in Aspergillus ochraceus IB-3, and purified from the membrane fraction through solubilization by Triton X-100. Phenazine and quinone derivatives acted as electron acceptors. Accumulation of ditaurobiliverdin and biliverdin by enzyme catalysis increased the absorbance at 660 nm, which is far from the range of wavelengths affected by serum ingredients. The enzyme selectively oxidized ditaurobilirubin at low pH, so changes in the reaction pH enable the enzyme to discriminate between the bilirubin fractions ditaurobilirubin (an example of conjugated bilirubin) and bilirubin (an example of unconjugated bilirubin). Using the enzyme, 2 to 80 μM of ditaurobilirubin were measured accurately by monitoring the changes in absorbance at 660 nm.
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  • Toyokazu YOSHIDA, Kohei FUJITA, Toru NAGASAWA
    2002 Volume 66 Issue 11 Pages 2388-2394
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      After enrichment culture with indole-3-carboxylate in static culture, a novel reversible decarboxylase, indole-3-carboxylate decarboxylase, was found in Arthrobacter nicotianae FI1612 and several molds. The enzyme reaction was examined in resting-cell reactions with A. nicotianae FI1612. The enzyme activity was induced specifically by indole-3-carboxylate, but not by indole. The indole-3-carboxylate decarboxylase of A. nicotianae FI1612 catalyzed the nonoxidative decarboxylation of indole-3-carboxylate into indole, and efficiently carboxylated indole and 2-methylindole by the reverse reaction. In the presence of 1 mM dithiothreitol, 50 mM Na2S2O3, and 20% (v/v) glycerol, indole-3-carboxylate decarboxylase was partially purified from A. nicotianae FI1612. The purified enzyme had a molecular mass of approximately 258 kDa. The enzyme did not need any cofactor for the decarboxylating and carboxylating reactions.
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Microbiology & Fermentation Technology Notes
  • Hiroshi TERANO, Kyosuke NOMOTO, Shigehiro TAKASE
    2002 Volume 66 Issue 11 Pages 2471-2473
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      This study provides new information on the Fe uptake system capable of supporting growth of the organism. Pseudomonas fluorescens isolated from the rhizosphere of barley, a gramineous plant, produced a siderophore under iron-limiting conditions. Its chemical structure was identified as pyochelin, on the basis of 1H and 13C NMR data of a stable methyl ester derivative. The same iron-limiting conditions induced a new set of outer membrane proteins (75 and 55 kDa), consistent with a siderophore-mediated iron-uptake system.
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  • Yasuyuki HASHIDOKO, Eriko ITOH, Kentaro YOKOTA, Tadashi YOSHIDA, Satos ...
    2002 Volume 66 Issue 11 Pages 2474-2478
    Published: 2002
    Released: June 19, 2003
    JOURNALS FREE ACCESS
      Five Gram-negative bacteria, all of which were Enterobacteriaceae, were isolated from the phyllosphere of green or senescing leaves of Rosa rugosa, and their phenotypic and physiological characteristics were examined. Partial 16S rDNA sequences led to identification of these isolates as Pantoea agglomerans, Klebsiella terrigena, Erwinia rhapontici, and two strains of Rahnella aquatilis. Interestingly, these phyllosphere bacteria had certain phenotypic and physiological convergences, while they showed their own metabolic properties toward phenolic compounds of plant origin. In particular, the two Ra. aquatilis isolates from the green leaves had a substrate-inducible gallate decarboxylase activity in the resting cells that had been cultured in 1 mM gallic acid- or protocatechuic acid-containing medium. The other three isolates from the senescing leaves did not have this enzyme activity. Simple phenolics that the Ra. aquatilis decarboxylatively produced from benzoic acid derivatives had better antimicrobial activities than those of the substrates.
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