Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Volume 67 , Issue 7
Showing 1-33 articles out of 33 articles from the selected issue
Analytical Chemistry Regular Paper
  • Makoto KAWAHARA, Toshifumi YUI, Kunio OKA, Peter ZUGENMAIER, Shiho SUZ ...
    2003 Volume 67 Issue 7 Pages 1545-1550
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Chitosan salts with two medical organic acids having phenyl groups (salicylic and gentisic acids) exhibited fiber diffraction patterns of a new type of crystal which does not compare with known types I and II. The crystals, called type III salts, showed a fiber repeat of 2.550 nm and a meridional reflection at the 5th layer line. These results coupled with a conformational analysis indicate the chain conformation of chitosan with the salts to be a 5/3 helix, this helix differing from those of type I (an extended two-fold helix) and type II (a relaxed two-fold helix or a 4/1 helix). The fiber patterns of all the type III salts were similar. This observation has also been found with type II salts and is an indication that the acid ions are not arranged in regular positions in the crystals. A comparison of solid-state 13C-NMR spectra of the gentisic acid salt and the aspirin salt, which could not be crystallized, suggests that, in the latter salt, the chitosan molecules also formed a 5/3 helix.
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Organic Chemistry Regular Papers
  • Kentaro NAKAMINAMI, Yoshiaki SAWADA, Miwako SUZUKI, Hiromichi KENMOKU, ...
    2003 Volume 67 Issue 7 Pages 1551-1558
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Gibberellin (GA) plays an important role in the induction of germination of photoblastic lettuce (Lactuca sativa L. cv. Grand Rapids) seeds. We have previously shown that gene expression of a GA 3-oxidase (Ls3h1) increased after a red light treatment, resulting in an increase in the endogenous content of GA1, bioactive GA. Since the metabolism of GAs is also important for determining the endogenous levels of bioactive GAs, cDNAs encoding GA 2-oxidases (LsGA2ox1 and LsGA2ox2, for L. sativa GA 2-oxidase), which catalyze the deactivation of GAs, were isolated from lettuce seeds to investigate the regulation of these genes by light. An expression analysis shows that the mRNA levels of both enzymes was not markedly altered under different light conditions during germination. However, the amount of LsGA2ox2 transcripts was decreased to approximately half the level by red light. This reduction might play a role in the increase in GA1 level by red light in the lettuce seeds.
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  • Shin-etsu MUTO, Kenji MORI
    2003 Volume 67 Issue 7 Pages 1559-1567
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Four pheromone components of the female painted apple moth (Teia anartoides), an Australian insect pest, were synthesized. These were (Z)-6-henicosen-11-one (1), (6Z, 8E)-6,8-henicosadien-11-one (2), (Z)-cis-9,10-epoxy-6-henicosene (3), and (Z)-cis-9,10-epoxy-6-icosene (4). 2-Dodecanone was converted to 1 and 2, and both the enantiomers of 3 and 4 were synthesized from the enantiomers of 4-tert-butyldimethylsilyloxy-cis-2,3-epoxy-1-butanol.
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  • Makoto OJIKA, Yu ITOU, Youji SAKAGAMI
    2003 Volume 67 Issue 7 Pages 1568-1573
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Known polyene amide, clathrynamide A, and three novel related metabolites, debromoclathrynamide A, (4E,6E)-debromoclathrynamide A and (6E)-clathrynamide A, were isolated from an Okinawan marine sponge, Psammoclemma sp. The absolute stereochemistry of clathrynamide A was determined to be 3R,12R by the modified Mosher method. The structures of the three new compounds were elucidated by spectroscopic analyses as debromo derivatives and geometrical isomers. Their antifungal activities were also evaluated with a phytopathogenic fungus, indicating that both the bromine atom and double-bond geometry affected the activity.
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Organic Chemistry Notes
Biochemistry & Molecular Biology Regular Papers
  • Nariaki WAKIUCHI, Ryohei SHIOMI, Hajime TAMAKI
    2003 Volume 67 Issue 7 Pages 1465-1471
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Galactinol, 1-O-(α-D-galactopyranosyl)-myo-inositol, was produced from sucrose as a starting material.
      UDP-Glc was prepared with sucrose and UDP using sucrose synthase partially purified from sweet potato roots. Then, the UDP-Glc was converted to UDP-Gal using yeast UDP-Gal 4-epimerase from a commercial source. Finally, galactinol was produced from the UDP-Gal and myo-inositol using galactinol synthase partially purified from cucumber leaves. The product was identified as galactinol by the retention times of HPLC, α-galactosidase digestion, and NMR spectrometry.
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  • Sunyo JUNG, Jung Sung CHUNG, Sun Mi JANG, Ja Ock GUH, Hee Jae LEE, San ...
    2003 Volume 67 Issue 7 Pages 1472-1478
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Protoporphyrinogen oxidase (Protox) is the last shared enzyme of the porphyrin pathway. As a continuation of our previous work in which the transgenic rice plants expressing the Bacillus subtilis Protox in the cytoplasm or the plastid showed resistance to diphenyl ether herbicide, this study was undertaken to identify the effects of tertapyrrole biosynthesis in these transgenic rice plants. The transgenic plants either targeted into plastids or expressed in cytoplasm showed higher Protox activity than wild-type plants did. Photosynthetic activity, measured as a quantum yield of photosystem II, was slightly higher in transgenic plants than in wild-type plants, but chlorophyll contents were not significantly different between transgenic and wild-type plants. As for porphyrin biosynthesis, both cytoplasm-expressed and plastid-targeted transgenic plants showed increased synthesis of aminolevulinic acid, Mg-Proto IX, and protoheme in comparison to wild-type plants whereas synthesis of protoporphyrin IX was similar for wild-type and transgenic plants. These results indicate that either cytoplasm or plastid expression of B. subtilis Protox in rice can upregulate the porphyrin pathway leading to increase in photosynthetic efficiency in plants.
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  • Thida SRIPUAN, Kazuhiro AOKI, Kenji YAMAMOTO, Dararat TONGKAO, Hidehik ...
    2003 Volume 67 Issue 7 Pages 1485-1491
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      α-Galactosidase was purified from a fresh fruiting body of Ganoderma lucidum by precipitation with ammonium sulfate and column chromatographies with DEAE-Sephadex and Con A-Sepharose. The purified enzyme was homogeneous on polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was similar to that of Mortierella vinacea α-galactosidase. The molecular mass of the enzyme was about 56 kDa by SDS-polyacrylamide gel electrophoresis, and about 249 kDa by gel filtration column chromatography. The optimum pH and temperature were 6.0 and 70°C, respectively. The enzyme was fully stable to heating at 70°C for 30 min. It hydrolyzed p-nitrophenyl-α-D-galactopyranoside (Km=0.4 mM) but hydrolyzed little o-nitrophenyl-α-D-galactopyranoside. It also hydrolyzed melibiose, raffinose, and stachyose. The enzyme catalyzed the transgalactosylation reaction which synthesized melibiose. The product was confirmed by various analyses.
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  • Takashi FUKADA, Masanobu ONO, Shohei SAKATA, Noriyuki KIOKA, Hiroshi S ...
    2003 Volume 67 Issue 7 Pages 1515-1521
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      It has been known that mouse, rat, and human N-acetylglucosaminyltransferase I (GnT-I) genes produce at least two transcripts, which differ in their 5′-untranslated region (5′-UTR) length, and the longer transcript is preferentially expressed in brains. However, the physiological meaning of this brain-specific expression pattern was unknown. We cloned the rat GnT-I gene and analyzed its structure. It consisted of five exons, and four of them coded only 5′-UTRs. A putative NF-κB binding site was found in the 5′-flanking sequence for the transcript that was previously shown to be induced by inflammation. The unusually long 5′-UTR of the major GnT-I transcript in rat brain was shown to inhibit protein production from the following coding sequence in COS7 cells. Comparison of the GnT-I protein/mRNA ratio in rat brain and liver showed that GnT-I mRNA in the brain was translated 3.8-times less efficiently than in the liver. Implications are discussed of these results in regulation of GnT-I expression in rat brain.
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  • Katsuyoshi KAMIIE, Tetsuro YAMASHITA, Hideharu TAIRA, Shin-ichiro KIDO ...
    2003 Volume 67 Issue 7 Pages 1522-1529
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Elongation factor 1 (EF-1) from the silk gland of Bombyx mori consists of four subunits: α (51 kDa), β (26 kDa), γ (49 kDa), and δ (33 kDa). The EF-1α subunit catalyzes the binding of aminoacyl-tRNA to the ribosome concomitant with the hydrolysis of GTP. The EF-1α-bound GDP is then exchanged for GTP by the EF-1βγδ complex. To facilitate analysis of the roles of the individual EF-1β, γ, and δ subunits in GDP/GTP exchange on EF-1α, we cloned the cDNAs for these subunits and expressed them in Escherichia coli. EF-1β, EF-1γ, and the carboxyl-terminal half of EF-1δ were expressed, purified, and examined for protein:protein interactions by gel filtration chromatography and by a quartz-crystal microbalance method. An 80-kDa species containing EF-1β and γ subunits in a 1:1 molar ratio was detected by gel filtration. A higher molecular weight species containing an excess of EF-1γ relative to EF-1β was also detected. The amino-terminal region of EF-1β (amino acid residues 1-129) was sufficient for binding to EF-1γ. The carboxyl-terminal half of EF-1δ did not appear to form a complex with EF-1γ.
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  • Nozomu YASUTAKE, Kazuhide TOTANI, Yoichiro HARADA, Shinobu HARAGUCHI, ...
    2003 Volume 67 Issue 7 Pages 1530-1536
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      A condensation reaction between N-acetyllactosamine and glycerol was directly catalyzed by using a commercially available cellulase preparation from Trichoderma reesei. 1-O-β-N-Acetyllactosaminyl-(R, S)-glycerols (1) were readily synthesized in a 5% yield based on the N-acetyllactosamine added and conveniently isolated by two-step column chromatographies. The use of a partially purified enzyme increased 2.3-fold the yield of 1, compared to that of the crude enzyme containing β-D-galactosidase activity. When various alkanols (n:2-4) were used in the condensation reaction, the corresponding alkyl β-N-acetyllactosaminides were obtained in yields of 0.3-1.1% of the desired compounds.
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Biochemistry & Molecular Biology Notes
Food & Nutrition Science Regular Papers
  • Yoshiaki MIYAKE, Kenichiro MINATO, Syuichi FUKUMOTO, Kanefumi YAMAMOTO ...
    2003 Volume 67 Issue 7 Pages 1443-1450
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Potent antioxidative hydroxyflavanones were produced with Aspergillus saitoi from hesperidin or naringin, which are flavanone glycosides in citrus fruit with weak antioxidative activity. The hydroxyflavanone produced from hesperidin was identified as 8-hydroxyhesperetin (8-HHE), a novel substance, and those from naringin were identified as carthamidin (6-hydroxynaringenin) and isocarthamidin (8-hydroxynaringenin) by FAB-MS, 1H-NMR and 13C-NMR analyses. The antioxidative activity of these hydroxyflavanones was examined by using the free radical-scavenging system of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the methyl linoleate oxidation system. The hydroxyflavanones (8-HHE, carthamidin, and isocarthamidin) exhibited stronger activity than the flavanone glycosides (hesperidin or naringin) and their aglycones (hesperetin or naringenin). The activity of 8-HHE and isocarthamidin was comparable to that of α-tocopherol, and that of carthamidin was weaker than that of isocarthamidin. The hydroxyflavanones, which were hydroxylated on A ring of flavanone by Aspergillus saitoi, were obtained as potent antioxidants.
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  • Chong-Suk KWON, Ho Yong SOHN, Sung Hee KIM, Ji Hyun KIM, Kun Ho SON, J ...
    2003 Volume 67 Issue 7 Pages 1451-1456
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      In the process of screening for pancreatic lipase inhibitors, which could be used as an anti-obesity measure, the methanol extract of Dioscorea nipponica Makino powder (DP) appeared to have potent inhibitory activity against porcine pancreatic lipase with an IC50 value of 5-10 μg/ml, where the enzyme activity was assayed by using 4-methylumbelliferyl oleate as a substrate. Further purification of active components present in the herb generated dioscin that belongs to the saponin family. Dioscin and its aglycone, diosgenin, both suppressed the time-dependent increase of blood triacylglycerol level when orally injected with corn oil to mice, suggesting their inhibitory potential against fat absorption. Sprague-Dawley rats fed on a high-fat diet containing 5% Dioscorea nipponica Makino and 40% beef tallow gained significantly less body weight and adipose tissue than control animals fed on a high-fat diet alone during an 8-week experimental period (P<0.05).
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  • Shiro MAWATARI, Yasushi OHNISHI, Yoshikazu KAJI, Tohru MARUYAMA, Kaori ...
    2003 Volume 67 Issue 7 Pages 1457-1464
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Effects of high dietary cholesterol on erythrocyte membrane lipids were studied. Feeding rats with a diet containing 0.5% cholesterol and 0.15% sodium cholate for two weeks induced changes in erythrocyte membrane lipids including a decrease in cholesterol, an increase in α-tocopherol (α-Toc) and changes in the fatty acid composition of phospholipids. Oleic acid and linoleic acid increased, while arachidonic acid decreased in phosphatidylcholine. Saturated fatty acids decreased and unsaturated fatty acids increased in phosphatidylethanolamine. Almost the same changes in membrane lipids were also noted after six weeks of feeding rats with the diet. A diet containing 0.5% cholesterol but without sodium cholate caused a decrease in erythrocyte cholesterol and an increase in erythrocyte α-Toc after two weeks of feeding, as compared to the basal diet, indicating that high dietary cholesterol, but not sodium cholate, was responsible for these changes in the erythrocyte membrane.
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  • Kaoru KOHYAMA, Tomoko SASAKI, Haruka DAN
    2003 Volume 67 Issue 7 Pages 1492-1498
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Using a multiple-point sheet sensor (MSS), load and contact area were directly measured for compression of four different foods. The MSS provided temporal and spatial changes in stress applied on the sample surface during the testing. The sum of load value detected by the MSS corresponded to the load measured by a universal testing machine during the compression. The contact area between a flat probe and food surface varied with the variety of foods even though under a small strain, and increased as compression strain increased. The active stress, that is, the load divided by the contact area, was different from conventional stress, that is, the load divided by the initial cross-sectional area. The value of active stress provided a better explanation of textural characteristics of food, because texture is often sensed under a large deformation and mixed assessment of mechanical and geometrical properties.
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  • Hisako TAKIGAWA-IMAMURA, Takumi SEKINE, Mitsuo MURATA, Kiyoshi TAKAYAM ...
    2003 Volume 67 Issue 7 Pages 1499-1506
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      Glucose incorporation is regulated mainly by GLUT4 in skeletal muscles. Here we report that treatment of L6 myotubes with scriptide, a hydroxamic acid-based histone deacetylase (HDAC) inhibitor, stimulated 2-deoxyglucose uptake. The effect appeared only after 24 hr, resulting in 2.4-fold glucose uptake at treatment day 6. Scriptide acted synergistically with insulin, indicating it stimulated a distinct pathway from the insulin signaling pathway. It was not observed in undifferentiated myoblasts or 3T3-L1 adipocytes, suggesting a muscle-specific effect of scriptide. A five-carbon chain and hydroxamic acid, essential for histone deacetylase inhibition, were indispensable for this effect, and trichostatin A stimulated glucose uptake as well. Scriptide increased the cellular content of GLUT4, and induced GLUT4 translocation, but GLUT4 mRNA level did not change, indicating scriptide functions posttranslationally. Our results indicated a novel function for HDAC inhibitors of increasing GLUT4 content and its translocation in muscle cells, resulting in stimulation of glucose uptake.
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  • Akio AMETANI, Toshio SAKURAI, Yoshinori KATAKURA, Satoru KUHARA, Hidek ...
    2003 Volume 67 Issue 7 Pages 1507-1514
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      We compared T-cell responses to regions in residues 21-40 of A and B variants of bovine milk β-lactoglobulin (β-LG) that vary by two different amino acid residues at 64 and 118. Results showed that T cells from C57/BL6 and C3H/HeN mice immunized with peptide 21-40 or BALB/c mice immunized with peptide 21-32 or 25-40 responded more vigorously to β-LG B than to β-LG A. This difference in response to 25-40 in BALB/c mice was not observed when β-LGs B and A were denatured, suggesting that the conformation difference affects display of the determinant 25-40. Reactivity of anti-β-LG monoclonal antibodies and molecular modeling using molecular dynamics calculations revealed subtle differences in the three-dimensional structure of these two variants. Furthermore, substitution of two amino acid residues at sites distant from the T-cell determinant induced differential determinant display on antigen-presenting cells, possibly due to subtle conformational changes in β-LG.
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Food & Nutrition Science Note
  • Hirosuke OKU, Sawitree WONGTANGTINTHARN, Hironori IWASAKI, Takayoshi T ...
    2003 Volume 67 Issue 7 Pages 1584-1586
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      This paper describes the in vitro effect of conjugated linoleic acid (CLA) on fatty acid biosynthesis. Among the rat liver enzymes involved in fatty acid biosynthesis, fatty acid synthetase (FAS) showed the largest activity fluctuation with the types of fatty acids. Of the fatty acids, CLA was the most potent inhibitor of FAS, and the 9c, 11t-rather than the 10t, 12c-isomer showed greater inhibition. CLA also significantly lowered the incorporation of [14C]-acetate into phospholipid in breast cancer cells, supporting the view that CLA inhibits fatty acid biosynthesis through the interaction with FAS.
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Food & Nutrition Science Communications
  • Yasuyuki TAKENAKA, Futoshi NAKAMURA, Taichi YAMAMOTO, Masaaki YOSHIKAW ...
    2003 Volume 67 Issue 7 Pages 1620-1622
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      We found that enterostatin (VPDPR), an anorexigenic peptide for a high-fat diet, significantly reduces serum cholesterol levels after oral administration of 100 mg/kg for 3 days in mice fed a high cholesterol-cholic acid diet. DPR, a peptide fragment of VPDPR, also had hypocholesterolemic activity at a dose of 50 mg/kg. Food intake was not suppressed under these dietary conditions. Fecal excretion of cholesterol and bile acids was increased significantly by both VPDPR and DPR. Interestingly, DPR induced hypocholesterolemic effects just two hours after a single oral administration at a dose of 100 mg/kg.
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  • Hitoshi INOKUCHI, Hisako HIROKANE, Tsuyoshi TSUZUKI, Kiyotaka NAKAGAWA ...
    2003 Volume 67 Issue 7 Pages 1623-1627
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      The anti-angiogenic property of vitamin E compounds, with particular emphasis on tocotrienol, has been investigated in vitro. Tocotrienol, but not tocopherol, inhibited both the proliferation and tube formation of bovine aortic endothelial cells, with δ-tocotrienol appearing the highest activity. Also, δ-tocotrienol reduced the vascular endothelial growth factor-stimulated tube formation by human umbilical vein endothelial cells. Our findings suggest that tocotrienol has potential use as a therapeutic dietary supplement for minimizing tumor angiogenesis.
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Microbiology & Fermentation Technology Regular Papers
  • Masao SAKAI, Hiroyuki FUTAMATA, Shinjiro KANAZAWA
    2003 Volume 67 Issue 7 Pages 1479-1484
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      We did tests using swarm plates, to examine the effects of various salts and their concentrations on the chemotaxis of fluorescent Pseudomonas strains. As a result, we found that the swarming ability of the Pseudomonas strains was inhibited by high concentrations of Ca2+. The growth of the strains was not affected at the high concentration of Ca2+, but the cells grown in swarm agar under the condition were extended in the filaments. Most of the cells had reached 10 μm to 40 μm in length. Such cell elongation was not observed with salts other than calcium salts. A significant correlation between the cell elongation and the decrease of swarming ability by the high concentration of Ca2+ was observed.
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  • Tsuyoshi SUGIO, Mitsuko FUJII, Fumiaki TAKEUCHI, Atsunori NEGISHI, Ter ...
    2003 Volume 67 Issue 7 Pages 1537-1544
    Published: 2003
    Released: August 08, 2003
    JOURNALS FREE ACCESS
      A highly mercury-resistant strain Acidithiobacillus ferrooxidans MON-1, was isolated from a culture of a moderately mercury-resistant strain, A. ferrooxidans SUG 2-2 (previously described as Thiobacillus ferrooxidans SUG 2-2), by successive cultivation and isolation of the latter strain in a Fe2+ medium with increased amounts of Hg2+ from 6 μM to 20 μM. The original stain SUG 2-2 grew in a Fe2+ medium containing 6 μM Hg2+ with a lag time of 22 days, but could not grow in a Fe2+ medium containing 10 μM Hg2+. In contrast, strain MON-1 could grow in a Fe2+ medium containing 20 μM Hg2+ with a lag time of 2 days and the ability of strain MON-1 to grow rapidly in a Fe2+ medium containing 20 μM Hg2+ was maintained stably after the strain was cultured many times in a Fe2+ medium without Hg2+. A similar level of NADPH-dependent mercury reductase activity was observed in cell extracts from strains SUG 2-2 and MON-1. By contrast, the amounts of mercury volatilized for 3 h from the reaction mixture containing 7 μM Hg2+ using a Fe2+-dependent mercury volatilization enzyme system were 5.6 nmol for SUG 2-2 and 67.5 nmol for MON-1, respectively, indicating that a marked increase of Fe2+-dependent mercury volatilization activity conferred on strain MON-1 the ability to grow rapidly in a Fe2+ medium containing 20 μM Hg2+. Iron oxidizing activities, 2,3,5,6-tetramethyl-p-phenylenediamine (TMPD) oxidizing activities and cytochrome c oxidase activities of strains SUG 2-2 and MON-1 were 26.3 and 41.9 μl O2 uptake/mg/min, 15.6 and 25.0 μl O2 uptake/mg/min, and 2.1 and 6.1 mU/mg, respectively. These results indicate that among components of the iron oxidation enzyme system, especially cytochrome c oxidase activity, increased by the acquisition of further mercury resistance in strain MON-1. Mercury volatilized by the Fe2+-dependent mercury volatilization enzyme system of strain MON-1 was strongly inhibited by 1.0 mM sodium cyanide, but was not by 50 nM rotenone, 5 μM 2-n-heptyl-4-hydroxy-quinoline-N-oxide (HQNO), 0.5 μM antimycin A, or 0.5 μM myxothiazol, indicating that cytochrome c oxidase plays a crucial role in mercury volatilization of strain MON-1 in the presence of Fe2+.
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Microbiology & Fermentation Technology Note
Microbiology & Fermentation Technology Communication
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