Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Volume 71 , Issue 1
Showing 1-43 articles out of 43 articles from the selected issue
Award Review
  • Eiji MASAI, Yoshihiro KATAYAMA, Masao FUKUDA
    2007 Volume 71 Issue 1 Pages 1-15
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Lignins are the most abundant aromatic compounds in nature, and their decomposition is essential to the terrestrial carbon cycle. White rot fungi secreting phenol oxidases are assumed to be involved in the initial degradation of native lignin, whereas bacteria play a main role in the mineralization of lignin-derived low-molecular-weight compounds in soil. There are a number of reports on the degradation pathways for lignin-derived aromatic compounds, but their catabolism has not been enzymatically or genetically characterized. Sphingomonas paucimobilis SYK-6 is one of the best-characterized lignin-degrading bacteria. It can grow on a wide variety of lignin-related biaryls and monoaryls, including β-aryl ether, biphenyl, diarylpropane, and phenylpropane. These compounds are degraded via the protocatechuate (PCA) 4,5-cleavage pathway or multiple 3-O-methylgallate (3MGA) catabolic pathways. In this review, the enzyme systems for β-aryl ether and biphenyl degradation, O demethylation linked with one carbon metabolism, the PCA 4,5-cleavage pathway, and the multiple 3MGA catabolic pathways in SYK-6 are outlined.
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Organic Chemistry Regular Papers
  • Sun-Ju KIM, Chiami KAWAHARADA, Shigeki JIN, Makoto HASHIMOTO, Gensho I ...
    2007 Volume 71 Issue 1 Pages 114-121
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    The structurally unique glucosinolate (GSL), 4-(cystein-S-yl)butyl GSL, was identified in the leaves of hydroponically-grown rocket salad (Eruca sativa Mill.). Its electrospray ionization mass spectrometry (ESI-MS)/MS spectrum indicated that this unusual GSL had a molecular weight of 414 as a desulfo (DS)-GSL, and a molecular formula of C14H25N2O8S2 based on its negative ion matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) spectrum. For further confirmation, the 4-(cystein-S-yl)butyl DS-GSL was prepared with authentic L-Ser and purified dimeric 4-mercaptobutyl DS-GSL, and its chemical structure then confirmed by ESI-MS/MS data. It is named “glucorucolamine” as a trivial name from its ammonia sensitivity. This unique GSL was found to the greatest extent when rocket salad was grown in a 100% NH4+-N nutrient solution. Despite it clearly seems to reduce the detoxification of excess NH4+ in the leaves of rocket salad, present knowledge about the unique GSL is still far from being sufficient.
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  • Kensaku TAKARA, Keiko OTSUKA, Koji WADA, Hironori IWASAKI, Masatsugu Y ...
    2007 Volume 71 Issue 1 Pages 183-191
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    In the course of our work into the use of cane by-products, we have studied the isolation and structural determination of bioactive compounds in sugarcane molasses. In this study, three stereo isomers of syringyl glycerol 3′-O-β-D-glucopyranoside, three stereo isomers of guaiacyl glycerol 3′-O-β-D-glucopyranoside, a syringyl glycerol 2′-O-β-D-glucopyranoside, tachioside and a 2,3-dihydro-3,5-dihydroxy-6-methyl-4-(H)-pyran-4-one (DDMP) were isolated from the 25% methanol eluate by Amberlite XAD-2 column chromatography of sugarcane molasses. The structures of these compounds were determined on the basis of spectroscopic evidence. These isolated compounds were examined for their scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical species, and for their inhibitory activity against mushroom tyrosinase. All of the isolated compounds showed DPPH radical scavenging activity, while DDMP and tachioside showed mushroom tyrosinase inhibitory activity.
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Biochemistry & Molecular Biology Regular Papers
  • Kumi KIMURA, Satoshi TAWARA, Kiharu IGARASHI, Asako TAKENAKA
    2007 Volume 71 Issue 1 Pages 16-22
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Oxidative stress is recognized to be associated with the development of insulin resistance. Although free radicals are generated in various ways in vivo, very few radical generators have been used to investigate the effect of oxidative stress on cellular insulin signaling. In order to compare the effect of radical generators with different sites and durations of radical formation on liver insulin action, primary cultured rat hepatocytes were incubated with radical generators and insulin-dependent regulation of gene expression was examined. The hydrophobic 2,2′-azobis(2,4-dimethylvaleronitrile) (AMVN) radical and H2O2 increased plasma membrane damage, and the hydrophilic 2-2′-azobis(2-amidinopropane) dihydrochloride (AAPH) radical and buthionine sulfoxyimine (BSO) increased oxidation of intracellular substances. Paraquat (PQ) and H2O2 inhibited insulin-dependent repression of insulin-like growth factor-binding protein-1 (IGFBP-1) and phosphoenolpyruvate carboxykinase (PEPCK) gene expression. These results indicate that PQ and H2O2 impair insulin action effectively and are suitable for examining crosstalk between oxidative stress and insulin signaling in liver-cell culture systems.
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  • Yumiko ACHIWA, Kiyoshi HASEGAWA, Yasuhiro UDAGAWA
    2007 Volume 71 Issue 1 Pages 31-37
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    The phosphatidylinositol 3-Kinase-Akt (PI3K-Akt) pathway and the mitogen activated protein kinase (MAPK) pathway are important in the development and proliferation of various human cancers. It has been found recently that ursolic acid treatment affects growth and apoptosis in cancer cells. We sought to determine whether prominent signaling pathways, including the PI3K-Akt pathway and the MAPK (JNK, P38, and P44/42) pathway mediate these effects. Endometrial cancer cells often have high levels of phosphorylated Akt seen in conjunction with a PTEN mutation or deletion. Elevation in Akt protects the cancer cell from apoptosis. Ursolic acid treatment moderately decreased PI3K levels in SNG-II cells. Treatment also decreased phospho-Akt and phospho-P44/42 in a dose- and time-dependent fashion, dramatically in SNG-II cells and moderately in HEC108 cells. This effect was most pronounced following treatment with 50 μm ursolic acid for 72 h. Our study found inhibition of both the PI3K-Akt pathway and the MAPK pathway in two endometrial cancer cell lines, SNG-II and the poorly differentiated HEC108 cell line.
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  • Jinhua DONG, Yutaka TAMARU, Toshiyoshi ARAKI
    2007 Volume 71 Issue 1 Pages 38-46
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    The β-agarase-d gene (agaD) from a marine bacterium, Vibrio sp. strain PO-303, was cloned and expressed in Escherichia coli. The gene consists of 1,362 bp and encodes a protein of 453 amino acids with a predicted molecular weight of 50,824. The full length of agarase-d consists of a signal peptide, a glycoside hydrolase family 16 catalytic module (CM), and a carbohydrate binding module (CBM). The full length of agarase-d without the signal peptide (rAgaDΔfull), the catalytic module (rAgaDCM), or the CBM (rAgaDCBM) was expressed in E. coli as recombinant proteins. rAgaDCM exhibited higher enzyme activity (63.6 units/mg) than rAgaDΔfull (1.20 units/mg) against agarose. rAgaDCM hydrolyzed agar and porphyran to several oligosaccharides and acted on neoagarohexaose to produce neoagarotetraose and neoagarobiose, but did not act on neoagarotetraose. rAgaDCBM bound to agarose.
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  • Masahide SUNAGAWA, Hitoshi MURATA, Yasumasa MIYAZAKI, Masaya NAKAMURA
    2007 Volume 71 Issue 1 Pages 47-50
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Mycorrhizal basidiomycetes, Suillus grevillei and S. bovinus, were transformed by particle bombardment. We isolated eight and four transformants from S. grevillei and S. bovinus respectively under three conditions differing in terms of target distance and helium pressure. The transformation frequencies were 1.2 and 0.6 transformants/μg DNA. Plasmid DNAs were integrated into the genomic DNA of the transformants and stably maintained. This is the first description of the transformation of S. grevillei and S. bovinus by particle bombardment.
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  • Kiyofumi SAKAI, Hayato YAMANAKA, Kunihiko MORIYOSHI, Takashi OHMOTO, T ...
    2007 Volume 71 Issue 1 Pages 51-57
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    A bacterium capable of assimilating 2,2-bis(4-hydroxyphenyl)propane (bisphenol A), strain BP-7, was isolated from offshore seawater samples on a medium containing bisphenol A as sole source of carbon and energy, and identified as Sphingomonas sp. strain BP-7. Other strains, Pseudomonas sp. strain BP-14, Pseudomonas sp. strain BP-15, and strain no. 24A, were also isolated from bisphenol A-enrichment culture of the seawater. These strains did not degrade bisphenol A, but accelerated the degradation of bisphenol A by Sphingomonas sp. strain BP-7. A mixed culture of Sphingomonas sp. strain BP-7 and Pseudomonas sp. strain BP-14 showed complete degradation of 100 ppm bisphenol A within 7 d in SSB-YE medium, while Sphingomonas sp. strain BP-7 alone took about 40 d for complete consumption of bisphenol A accompanied by accumulation of 4-hydroxyacetophenone. On a nutritional supplementary medium, Sphingomonas sp. strain BP-7 completely degraded bisphenol A and 4-hydroxyacetophenone within 20 h. The strain degraded a variety of bisphenols, such as 1,1-bis(4-hydroxyphenyl)ethane, 2,2-bis(4-hydroxy-3-methylphenyl)propane, 2,2-bis(4-hydroxyphenyl)butane, and 1,1-bis(4-hydroxyphenyl)cyclohexane, and hydroxy aromatic compounds such as 4-hydroxyacetophenone, 4-hydroxybenzoic acid, catechol, protocatechuic acid, and hydroquinone. The strain did not degrade bis(4-hydroxyphenyl)methane, bis(4-hydroxyphenyl)sulfone, or bis(4-hydroxyphenyl)sulfide.
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  • Yoshinobu KATO, Masahito SUGIURA, Takeshi MIZUNO, Hirofumi AIBA
    2007 Volume 71 Issue 1 Pages 77-83
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Flagella expression in Escherichia coli is controlled in a hierarchical manner, in which class-1 gene products, FlhDC, functions as a master regulator to control class-2 genes that encode motility-related genes. fliA, one of the class 2 genes, encodes flagellum-specific sigma factor (FliA/Sigma F/Sigma-28), which is necessary for the expression of class-3 genes. Previously, we carried out transcriptome analyses of all two-component regulatory systems of E. coli, and determined that the arcA mutant showed the motility-defective phenotype. In this study, we characterized the arcA mutant, and we present evidence that ArcA is necessary for the expression of FliA, but not for the master regulators, FlhDC. The phosphorylation site of ArcA is necessary for motility, while a cognate histidine kinase, ArcB, appears not to be involved in motility. This suggests that there must be regulatory factors other than ArcB interacting with ArcA to control flagella genes.
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  • Chih-Hung HUNG, Pei-Hua PENG, Chou-Chun HUANG, Hai-Lung WANG, Yu-Jen C ...
    2007 Volume 71 Issue 1 Pages 98-103
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Degenerate primers were designed based on all possible sequences of the N-terminal and C-terminal regions of Delonix regia trypsin inhibitor (DrTI). Five hundred sixty-one bp of polymerase chain reaction (PCR) product was amplified using the above degenerate primers and genomic DNA and cDNA of Delonix regia as a template. The amplified PCR products were cloned and sequenced. DNA sequence analysis of cDNA and genomic clones of DrTI have the same nucleotide sequence in the coding region, and manifested a genomic clone without intervening sequences in the coding region. The amino acid sequence deduced from the DrTI genomic and cDNA clones agreed with that identified via amino acid sequencing analysis, except that two amino acid residues, Ser and Lys, existed between residues Lys141 and Ser142. DrTI open reading frame was then amplified and cloned in-frame with GST in pGEX4T-1 and overexpressed in Escherichia coli to yield a glutathione S-transferase (GST)-fusion protein with a calculated molecular mass of about 45 kDa. The recombinant DrTI (reDrTI) was derived by treating the GST-DrTI fusion protein with thrombin. Both the reDrTI and GST-DrTI fusion protein exhibited a strong identical inhibitory effect on trypsin activity.
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  • Hiromasa IMAISHI, Mariana PETKOVA-ANDONOVA
    2007 Volume 71 Issue 1 Pages 104-113
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    A cDNA encoding a cytochrome P450 (CYP76B9) was isolated from Petunia hybrida. Northern blot analysis revealed preferential expression of the gene in flowers and leaves. The recombinant yeast microsomes expressing CYP76B9 was allowed to react with capric acid and lauric acid as substrates. One major metabolite was produced from each fatty acid after incubation with yeast microsomes expressing CYP76B9. The metabolites were identified by gas chromatography–mass spectrometry (GC–MS) as ω-hydroxy capric acid and ω-hydroxy lauric acid. The kinetic parameters of the reactions were Km=9.4 μM and Vmax=13.6 mol min−1 per mol of P450 for capric acid, and Km=5.7 μM and Vmax=19.1 mol min−1 per mol of P450 for lauric acid. We found that the ω-hydroxy metabolites of capric acid and lauric acid can affect the plant growth of Arabidopsis thaliana. Plants grown in the presence of ω-hydroxy fatty acids exhibited shorter root length than control plants with the corresponding non-hydroxylated fatty acids.
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  • Hua SHAOFENG, Li SHUBEN, Xin JIAYIN, Niu JIANZHONG, Xia CHUNGU, Tan HA ...
    2007 Volume 71 Issue 1 Pages 122-129
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Methane monooxygenase hydroxylase was purified by chromatography and characterized by electrophoresis and spectroscopy. The molecular mass of hydroxylase was 201.3 KDa as determined by gel filtration, whereas the total molecular mass was 234 KDa as judged by SDS–PAGE. Structure study indicated that the enzyme is a homodimer structure, consisting of three subunits, designated α, β, and γ, with molecular masses of 58 KDa, 36 KDa, and 23 KDa respectively. IEF analysis indicated that the enzyme has a pI of 5.2. The UV–Vis spectrum of hydroxylase revealed an absorption peak near 281 nm and a weak shoulder peak around 395 nm–420 nm, and a fluorescence spectrum revealed an emission peak at 341.3 nm. Circular dichroism measurement indicated that hydroxylase mainly consists of α-helical regions. Finally, phylogenetic analysis indicated that this strain is very close to Methylosinus trichosporium OB3b.
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  • Masato EBIHARA, Makoto HATTORI, Tadashi YOSHIDA
    2007 Volume 71 Issue 1 Pages 130-137
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    T cell anergy is one of the mechanisms of immunological tolerance. We examined in this study the distinct responses of Th1 and Th2 cells to in vitro anergic stimulation using Th1 and Th2 cells from two strains of T cell receptor transgenic mice. Proliferation of the Th2 cells was difficult to suppress by anergic stimulation, while that of Th1 cells was significantly inhibited even by weak stimulation. However, IL-4 production by Th2 cells was definitely reduced by anergic stimulation, although the inhibition level of IL-4 was lower than that of IFN-γ production by Th1 cells. We also examined the reversal of anergy in both subsets. While both the anergized Th1 and Th2 cells responded to IL-2 stimulation, only the anergy of the Th2 cells could be reversed. This result indicates that progression of the cell cycle was not sufficient for anergy reversal in Th1 cells. Our findings indicate that the induction and reversal of T cell anergy might be affected by the distinct signaling features of Th1 and Th2 cells.
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  • Osamu YAMADA, Ryoko IKEDA, Yuka OHKITA, Risa HAYASHI, Kazutoshi SAKAMO ...
    2007 Volume 71 Issue 1 Pages 138-144
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    We found the orthologous genes required for RNA interference (RNAi) in the Aspergillus oryzae genome database, and constructed a set of tools for gene silencing using RNAi in A. oryzae. This system utilizes compatible restriction enzyme sites so that only a single target gene fragment is required to create the hairpin RNA cassette. For ease of handling, we also separated the construction of the hairpin RNA cassette for the target gene from its subsequent introduction into the expression vector. Using the brlA gene as a target for RNAi, we detected decreased mRNA levels and a delayed conidiation phenotype in the transformants. Furthermore, even though A. oryzae possesses three copies of the α-amylase gene, a single copy of an α-amylase RNAi construct was sufficient to downregulate the mRNA levels and decrease the enzymatic activity to 10% of control levels. Gene silencing by RNAi should provide a powerful genetic tool for post-genomic studies of the industrially important fungus A. oryzae.
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  • Ayako TAKADA, Genryou UMITSUKI, Kazuo NAGAI, Masaaki WACHI
    2007 Volume 71 Issue 1 Pages 158-164
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    The Escherichia coli RNase E is an essential endoribonuclease involved in processing and/or degradation of rRNAs, tRNAs, and non-coding small RNAs as well as many mRNAs. It is known that RNase E activity is somehow regulated by an RNA-binding protein Hfq, at least in some cases. We searched for proteins that showed changes in expression in both hfq::cat and rne-1 mutant cells as compared with the wild type, and found that a protein band of 49-kDa decreased in these mutant cells at 42 °C, the restrictive temperature for rne-1. N-terminal amino acid sequencing identified it as a mixture of GadA and GadB, two isozymes of glutamate decarboxylase involved in glutamate-dependent acid resistance. The rne-1 mutant as well as the hfq mutant showed decreased survival under acidic conditions (pH 2.5). Hfq is known to regulate the expression of GadA/B in RpoS- and GadY small RNA-dependent ways. We examined the expression of these two regulators in rne-1 mutant cells. In the mutant cells, the induction of GadY was defective at 42 °C, but the expression of RpoS was normal. These results indicate that RNase E is required for induction of the glutamate-dependent acid resistance system in a RpoS-independent manner.
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  • Shin-ichi HIRANO, Masaaki MORIKAWA, Kazufumi TAKANO, Tadayuki IMANAKA, ...
    2007 Volume 71 Issue 1 Pages 192-199
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    A putative gentisate 1,2-dioxygenase was encoded in the dibenzothiophene degradation gene cluster (dbd) from Xanthobacter polyaromaticivorans 127W. The deduced amino acid sequence showed high sequence similarity with gentisate dioxygenases from Pseudomonas alcaligenes (AAD49427, 65% identical), Bradyrhizobium japonicum (NP_766750, 64%), and P. aeruginosa (ZP_00135722, 54%), and moderate similarity with 1-hydroxy-2-naphthoate dioxygenase from Nocardioides sp. KP7 (BAA31235, 33%) and salicylate dioxygenase from Pseudaminobacter salicylatoxidans (AAQ91293, 33%). The enzyme, GDOxp, was heterologously produced in Escherichia coli and purified to homogeneity. GDOxp formed a tetramer and exhibited high dioxygenase activity against 1,4-dihydroxy 2-naphthoate as well as gentisate, suggesting unusually broad substrate specificity. GDOxp easily released ferrous ion under unfavorable temperature and pH conditions to become an inactive monomer protein. An inactive monomer protein can reconstitute a tetramer structure and restore enzyme activity in a cooperative manner upon the addition of ferrous ion. Chymotryptic digestion and protein truncation experiments suggested that the N-terminal region is important for the tetramerization of GDOxp.
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  • Ruchita SELOT, Virendra KUMAR, Sunita SHUKLA, Kumar CHANDRAKUNTAL, Mop ...
    2007 Volume 71 Issue 1 Pages 200-205
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Silkworms show high variability in silk quality and disease resistance. Attempts are on to combine the disease tolerance of multivoltine races and the silk quality of bivoltine races to generate new races with desirable phenotypic traits. We report the identification of a 26.5-kDa protein that is overexpressed in the gut juice of disease-resistant multivoltine races and that has anti-BmNPV activity. We have characterized this protein as a soluble NADH-oxidoreductase-like protein (BmNOX). Treatment of live BmNPV particles with BmNOX inhibited the capability of the viral particles to infect BmN cells in vitro.
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  • Gi-Young KIM, Ho-Jin CHAE, Ki-Hyung KIM, Man-Soo YOON, Kyu-Sub LEE, Ch ...
    2007 Volume 71 Issue 1 Pages 215-221
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Dendritic cells (DCs) are potent antigen presenting cells that are uniquely effective in generating primary immune responses. DCs that are manipulated to present tumor antigens induce antitumor immunity in animal models and preclinical human studies. A myriad of strategies have been developed to load tumor antigen effectively onto DCs. DC-tumor fusion presents a spectrum of tumor-associated antigens to helper T- and cytotoxic T-cell populations in the context of DC-mediated costimulatory signals. In this study, fusion cells (FCs) were generated with MCA-102 fibrosarcoma cells and murine bone marrow-derived myeloid DCs. The FCs coexpressed the DC-derived MHC class II and costimulatory molecules. The FCs also retained the functional properties of DCs and stimulated syngeneic T cell proliferation and interferon-γ (IFN-γ) production. Significantly, the results show that syngeneic T cells are primed by FCs to induce MHC class I-dependent lysis of MCA-102 fibrosarcoma. These findings indicate that fusions of tumor cells and DCs activate T-cell responses against syngeneic tumors.
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Biochemistry & Molecular Biology Notes
  • Yoshiko YAMAMOTO, Maki KURACHI, Kenichi YAMAGUCHI, Tatsuya ODA
    2007 Volume 71 Issue 1 Pages 238-241
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    We investigated the cytokine-inducing activities of guluronate (G3–G6) and mannuronate (M3–M6) oligomers on RAW264.7 cells with the Bio-Plex assay system. Relatively high levels of tumor necrosis factor-α (TNF-α), granulocyte colony-stimulating factor (G-CSF), monocyte chemoattractant protein-1 (MCP-1), regulated upon activation normal T cell expressed and secreted (RANTES), granulocyte macrophage (GM)-CSF, and eotaxin were induced by alginate oligomers to different extents depending on the oligomer structures, and low but significant levels of interleukin (IL)-1α, IL-1β, IL-6, IL-9, and IL-13 were also induced. Throughout all cytokines tested, M-oligomers tended to be more potent than G-oligomers in terms of cytokine induction, and this tendency was evident in differences between G3 and M3.
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  • Nobukazu SHITAN, Miyako TANAKA, Koichiro TERAI, Kazumitsu UEDA, Kazufu ...
    2007 Volume 71 Issue 1 Pages 242-245
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    To examine whether human ATP-binding cassette (ABC) transporters play a role in the detoxification of plant alkaloid berberine, we investigated berberine transport using multidrug resistance protein1 (MDR1) and multidrug resistance-associated protein1 (MRP1). Cells expressing MDR1 or MRP1 accumulated less berberine. Berberine accumulation depended on the cellular ATP level, and was reversed by typical inhibitors of MDR1, suggesting that human MDR1 and MRP1 directly efflux berberine as their substrate.
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  • Noritaka IWAI, Takuya FUJII, Hirokatsu NAGURA, Masaaki WACHI, Tomoya K ...
    2007 Volume 71 Issue 1 Pages 246-248
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Supplementary material
    We comprehensively investigated the effects of substitution of the benzyl group in S-benzylisothiourea derivatives on antibacterial activity, because we found previously that some substitutions enhanced it. A 2,4-Cl2-derivative was found to be the most effective compound, it was stronger than the original one in Gram-negative rod shaped-bacteria such as Escherichia coli and Salmonella typhimurium.
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  • Keita IMAI, Yoichi NODA, Hiroyuki ADACHI, Koji YODA
    2007 Volume 71 Issue 1 Pages 249-252
    Published: January 23, 2007
    Released: January 23, 2007
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    Overproduction of the ER membrane protein Rcr1 makes Saccharomyces cerevisiae resistant to Congo red by reducing the chitin content through a unknown mechanism. By both co-immunoprecipitation and yeast two-hybrid experiments, specific interaction between Rcr1 and the ubiquitin ligase Rsp5 was found. This binding was largely mediated by a singular VPEY sequence in Rcr1 in addition to PPSY, the consensus ligand motif of the WW domains. Mutant analysis indicated that Rsp5 and other Rcr1-interacting proteins discovered in the current screen were not engaged in Congo red resistance.
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  • Tsutomu NAKAGAWA, Kazuhiro NISHIUCHI, Jyunji AKAKI, Hideyuki IWATA, Ry ...
    2007 Volume 71 Issue 1 Pages 256-260
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Human prorenin attached by a decahistidine tag at the C-terminus was produced in Chinese hamster ovary cells. The tagged protein secreted into the culture medium was in the inactive prorenin form, and was activated to mature renin by proteolytic removal of its prosegment by trypsin in the same manner as native prorenin. The tagged (pro)renin was efficiently purified by metal-chelate affinity chromatography. The enzymatic properties of mature renin carrying the tag were similar to native renin. These results indicate that the introduction of a decahistidine tag at the C-terminus does not interfere with either the correct folding of prorenin or the catalytic activity of mature renin.
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  • Takashi SUZUKI, Masaaki ITO, Toru EZURE, Masamitsu SHIKATA, Eiji ANDO, ...
    2007 Volume 71 Issue 1 Pages 261-264
    Published: January 23, 2007
    Released: January 23, 2007
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    To establish a strategy to generate N-acylated proteins modified with fatty acids having a specific chain length, tGelsolin-streptag, an epitope-tagged model protein having an N-myristoylation motif, was synthesized using an insect cell-free protein synthesis system in the presence of acyl-CoA with various fatty acid chain lengths. It was found that the fatty acid species attached to the N-termini fully depended on the acyl-CoA species added to the reaction mixture. N-Acylated proteins with fatty acid chain lengths of 8, 10, 12, and 14 were generated successfully.
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  • Mochammad Amin ALAMSJAH, Keiko ISHIBE, Daekyung KIM, Kenichi YAMAGUCHI ...
    2007 Volume 71 Issue 1 Pages 265-268
    Published: January 23, 2007
    Released: January 23, 2007
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    α-linolenic acid and linoleic acid isolated from Ulva fasciata showed toxic effects on red tide phytoplankters in a concentration-dependent manner. Among six species tested, raphidophycean flagellate Heterosigma akashiwo was the most susceptible to these fatty acids, and 50% lethal concentrations (LC50) of α-linolenic acid and linoleic acid were estimated to be 0.58 and 1.91 μg/ml respectively, whereas dinoflagellate Gymnodinium impudicum and Heterocapsa circularisquama were highly resistant and no significant toxic effects were observed up to 1,000 μg/ml. Both fatty acids were less toxic to fish (devil stinger), zooplankters (brine shrimp and rotifer), and mammalian cell lines (U937, HeLa, Vero, and CHO cells) than H. akashiwo.
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Biochemistry & Molecular Biology Communications
  • Yasumasa MIYAZAKI, Masakazu HIRAIDE, Hajime SHIBUYA
    2007 Volume 71 Issue 1 Pages 273-278
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Several functional genes encoding putative proteins, heat shock protein 70, sphingosine phosphate lyase, and Na+/H+ antiporter, were cloned from the basidiomycete Fomitopsis pinicola, a wood-rotting fungus isolated in the tropical mangrove forest of Pohnpei Island of the Federated States of Micronesia. The deduced amino acid sequences of the obtained genes involved in heat shock resistance, lipid synthesis, and salt tolerance showed diverse similarities to other homologous proteins. Molecular phylogenetic trees of these proteins suggested that encoded proteins of the cloned genes of F. pinicola differed remarkably from other homologs in various organisms, even fungal proteins. Putative candidates for other genes related to several cellular metabolisms were also amplified, implying the possible existence of those genes in F. pinicola. This is the first report of possibly functional genes derived from a basidiomycetous mushroom growing in tropical islands such as Micronesia. The genes found in this study might play important roles in the cellular survival of the basidiomycete F. pinicola under severe environmental conditions.
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  • Satoru WATANABE, Masumi SATO, Kaori NIMURA-MATSUNE, Taku CHIBAZAKURA, ...
    2007 Volume 71 Issue 1 Pages 279-282
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Three dnaK and four dnaJ genes have been identified in the genome of cyanobacterium Synechococcus elongatus PCC 7942. Our comprehensive analysis of yeast two-hybrid screening revealed a specific interaction among DnaK2, DnaJ2, and RNase E, an essential endoribonuclease. We examined the effects of DnaK2 and DnaJ2 on RNase E activity by monitoring the digestion of psbAII transcript in vitro. The addition of DnaK2 and DnaJ2 obviously inhibited RNase E activity in an ATP-dependent manner. These results suggest that DnaK2 and DnaJ2 are involved in RNA degradation through interaction with RNase E.
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Food & Nutrition Science Regular Papers
  • Der-Zen LIU, Yu-Chih LIANG, Shyr-Yi LIN, Yin-Shiou LIN, Wen-Chun WU, W ...
    2007 Volume 71 Issue 1 Pages 23-30
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Wild and solid-state cultures (SSC) of Taiwanofungus camphoratus (aka Antrodia camphorata and Chang-chih [CC]) were sequentially extracted with cold water, methanol, and hot water to get cold-water-soluble (CWS), methanol-soluble (MS), and hot-water-soluble (HWS) extracts, respectively. Only the MS extract exhibited angiotensin-converting enzyme (ACE) inhibitory activities. The antihypertensive effects of the MS extract (10 mg/kg BW) were measured in spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKY) rats. MS extract of the SSC type was able to effectively lower the systolic blood pressure (SBP) and diastolic blood pressure (DBP) of SHR, but not of WKY rats, the results being significantly different from those for distilled water only (the blank). However, wild CC and its MS extract were not as effective as the SSC type in reducing SHR blood pressure and had no effect on WKY rats. SSC-type CC might be developed into a health food with the ability to regulate blood pressure.
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  • Truong Tuyet MAI, Nguyen Van CHUYEN
    2007 Volume 71 Issue 1 Pages 69-76
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    A screening of 5 plants used for making drinks in Vietnam revealed a Cleistocalyx operculatus (Roxb.) Merr and Perry flower bud extract to have the highest inhibitory activity against the α-glucosidase enzyme. The anti-hyperglycemic effects of an aqueous extract from flower buds of Cleistocalyx operculatus (CO), a commonly used material for drink preparation in Vietnam, were therefore investigated in vitro and in vivo. In vitro, the CO extract inhibited the rat-intestinal maltase and sucrase activities, with IC50 values of 0.70 and 0.47 mg/ml, respectively. These values are lower than those for a guava leaf extract (GE; IC50 0.97 and 1.28 mg/ml, respectively). Postprandial blood glucose testing of normal mice and STZ-induced diabetic rats by maltose loading (2 g/kg body weight (bw)) showed that the blood glucose reduction with CO (500 mg/kg bw) was slightly less than that with acarbose (25 mg/kg bw) but was more potent than that with GE (500 mg/kg bw). In an 8-week experiment, the blood glucose level of STZ diabetic rats treated with 500 mg of CO/kg bw/day was markedly decreased in comparison with that of non-treated diabetic rats. Consequently, CO is considered to be a promising material for preventing and treating diabetes.
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  • Hideharu FUKAO, Hideki YOSHIDA, Yoh-ichi TAZAWA, Takahiko HADA
    2007 Volume 71 Issue 1 Pages 84-90
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Antithrombotic activities of odorless garlic powder were demonstrated in blood fibrinolytic and coagulation systems. Though the odorless garlic preparation did not influence tissue-type plasminogen activator (t-PA) or its inhibitor secretions from human umbilical vein endothelial cells, it enhanced plasmin generation by t-PA on fibrin film and in chromogenic assays by 1.8-fold and 8.7-fold respectively. The coagulation system was considerably reduced after the administration of the garlic in a rat in situ loop model, indicating that increased levels of thrombin-antithrombin III (TAT) complex in the control group were significantly reduced to normal (sham) in the garlic group (p<0.05), which was associated with decreasing tendencies towards prolonged or increased values of coagulation parameters in the control group. These findings suggest that odorless garlic not only activates fibrinolytic activity by accelerating t-PA-mediated plasminogen activation, but also suppresses the coagulation system by downregulating thrombin formation, suggesting a beneficial role in preventing pathological thrombus formation in such cardiovascular disorders.
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  • Hiroshi OKAWA, Tatsuya MORITA, Kimio SUGIYAMA
    2007 Volume 71 Issue 1 Pages 91-97
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    The effect of dietary supplementation with cysteine on the plasma homocysteine concentration was investigated in rats fed on 10% casein (10C) and 30% casein (30C) diets. The 10C diet significantly increased the plasma homocysteine concentration as compared with the 30C diet. The hyperhomocysteinemia induced by the 10C diet was significantly suppressed by cysteine supplementation even at a 0.3% level, whereas cysteine did not decrease the plasma homocysteine concentration when added to the 30C diet. In contrast, 0.3% methionine supplementation of the 10C diet tended to increase the plasma homocysteine concentration. Cysteine supplementation to rats fed on the 10C diet did not alter the plasma cysteine concentration and the hepatic activities of cystathionine β-synthase and betaine:homocysteine S-methyltransferase, whereas it significantly decreased the hepatic concentrations of S-adenosylmethionine and betaine. These results suggest that cysteine supplementation might be effective for suppressing the hyperhomocysteinemia induced by a low-protein diet.
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  • Jun YAMAKOSHI, Satoshi FUKUDA, Takuya SATOH, Ryouhei TSUJI, Makoto SAI ...
    2007 Volume 71 Issue 1 Pages 165-173
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    We investigated the mechanism of the antihypertensive effect of less-sodium soy sauce containing γ-aminobutyric acid (GABA) in spontaneously hypertensive rats (SHRs). When SHRs were given a diet with less-sodium soy sauce containing GABA (GABA-rich soy sauce group) for 6 weeks, the systolic blood pressure decreased as compared with that in rats fed diets with less-sodium soy sauce or a solution of salt. Renal sympathetic nerve activity (RSNA) and positive Na balance were reduced, and the urinary Na excretion tended to increase in the GABA-rich soy sauce group. Vascular hypertrophy of the thoracic aorta and the coronary and renal interlobular arteries tended to reduce in the GABA-rich soy sauce group. These results suggest that inhibition of Na retention by natriuresis, as a result of inhibition of RSNA by the GABA in the soy sauce contributed to the antihypertensive effect of GABA in the SHRs. Intake of less-sodium soy sauce containing GABA might help to reduce overall cardiovascular risk.
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  • Motoko OARADA, Tohru GONOI, Tsuyoshi TSUZUKI, Miki IGARASHI, Katsuya H ...
    2007 Volume 71 Issue 1 Pages 174-182
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Psychological stress has been shown to modulate immune functions. In this study, we investigated the effect of dietary oils (olive oil, soybean oil, and fish oil) on the social isolation stress-induced modulation of lymphocyte immunological activities in mice. In olive oil-fed, but not soybean oil- or fish oil-fed, mice, a 2-week isolation stress decreased the lymphocyte proliferative response, reduced the interferon-γ and interleukin (IL)-10 secretions and increased the IL-4 secretion by lymphocytes. The isolation stress reduced the arachidonic acid content of lymphocytes markedly, moderately, and not at all in the olive oil-, soybean oil-, and fish oil-fed mice, respectively. In the olive oil-fed, but not soybean oil- or fish oil-fed, mice, the isolation stress up-regulated the expression level of mRNA for splenic heat-shock protein 70 and increased lymphocyte sensitivity to the antiproliferative effect of corticosterone. This is the first demonstration that effect of psychological stress on lymphocyte immunological activities can vary depending upon the dietary fatty acid composition.
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  • Fumiki AOKI, Shinichi HONDA, Hideyuki KISHIDA, Mitsuaki KITANO, Naoki ...
    2007 Volume 71 Issue 1 Pages 206-214
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    We applied licorice flavonoid oil (LFO) to high-fat diet-induced obese C57BL/6J mice and investigated its effect. LFO contains hydrophobic flavonoids obtained from licorice by extraction with ethanol. The oil is a mixture of medium-chain triglycerides, having glabridin, a major flavonoid of licorice, concentrated to 1.2% (w/w). Obese mice were fed on a high-fat diet containing LFO at 0 (control), 0.5%, 1.0%, or 2.0% for 8 weeks. Compared with mice in the control group, those in the 1% and 2% LFO groups efficiently reduced the weight of abdominal white adipose tissues and body weight gain. A histological examination revealed that the adipocytes became smaller and the fatty degenerative state of the hepatocytes was improved in the 2% LFO group. A DNA microarray analysis of the liver showed up-regulation of those genes for beta-oxidation and down-regulation of those for fatty acid synthesis in the 2% LFO group. These findings suggest that LFO prevented and ameliorated diet-induced obesity via the regulation of lipid metabolism-related gene expression in the liver.
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Food & Nutrition Science Notes
  • Mi Jeong KIM, Soo Jung CHOI, Hye Kyung KIM, Chang-Ju KIM, Bumshik HONG ...
    2007 Volume 71 Issue 1 Pages 226-230
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    An ethanolic extract of Allium tuberosum Rottl. demonstrated potent choline acetytrasferase (ChAT) activating activity (45%) among 30 screened Korean edible plants. The ChAT activator of A. tuberoum Rottl. was purified and identified as ferulic acid [4-hydroxy-3-methoxycinnamic acid] via silica-gel open-column chromatography, HPLC, EI-MS, and 1H/13C-NMR. The ferulic acid from A. tuberoum Rottl. exhibited ChAT activity in a dose-dependent manner and showed notoxicity in a cell viability assay (MTT assay). We suggest that the ferulic acid from A. tuberoum Rottl. was the strong ChAT activator.
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  • Yasushi ENDO, Yuko OSADA, Fumiko KIMURA, Hitoshi SHIRAKAWA, Kenshiro F ...
    2007 Volume 71 Issue 1 Pages 231-233
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    The mechanism for the plasma and liver triacylglycerol-reducing effects of Japanese torreya (Torreya nucifera) seed oil containing sciadonic acid (all-cis-5, 11, 14-eicosatrienoic acid) is reported. Male SD rats were fed experimental diets containing 10% torreya, corn, or soybean oil for 4 weeks, and the activities and mRNA expression of the enzymes involved in lipid metabolism were measured in the liver. The activities of some hepatic enzymes involved in fatty acid synthesis were lower in the rats fed torreya oil.
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  • Hirofumi HIRAYASU, Yumiko YOSHIKAWA, Satoshi TSUZUKI, Tohru FUSHIKI
    2007 Volume 71 Issue 1 Pages 234-237
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    In the large-intestinal mucosae of rats orally administered dextran sulfate sodium, which induces an enteritis resembling ulcerative colitis (UC), the activity for granzyme A, a lymphocyte tryptase, increased at an earlier stage than that at which UC markers (growth-regulated gene product/cytokine-induced neutrophil chemoattractant-1 and caspase-3) increased. This suggests involvement of the enzyme in the exacerbation and perpetuation of enteritis.
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  • Mako NAKAYA, Hiroyuki ONDA, Kyoko SASAKI, Akiko YUKIYOSHI, Hirofumi TA ...
    2007 Volume 71 Issue 1 Pages 253-255
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Royal jelly is known as a functional food containing many useful minerals. In this study, we found an anti-environmental estrogen activity of royal jelly. Bisphenol A (BPA) is an environmental estrogen that stimulates proliferation of human breast cancer MCF-7 cells. Royal jelly inhibited the growth-promoting effect of BPA on MCF-7 cells, even though it did not affect the proliferation of cells in the absence of BPA. In addition, this inhibiting effect of royal jelly was heat-stable.
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  • Hirohito YAMAKAWA, Hiroshi AKIYAMA, Yumi ENDO, Kiyoko MIYATAKE, Kozue ...
    2007 Volume 71 Issue 1 Pages 269-272
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    A sensitive qualitative detection method for soybeans in foods by using the polymerase chain reaction (PCR) was developed. For specific detection of soybeans with high specificity, the primer pair of Gym 81/Gym 82 was designed on the gene encoding the Glycine max repetitive sequence. The trace amount of soybeans in commercial food products could be qualitatively detected by this method.
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Microbiology & Fermentation Technology Regular Papers
  • Tomoyuki KOSAKA, Shunichi NAKAYAMA, Keizo NAKAYA, Sadazo YOSHINO, Kens ...
    2007 Volume 71 Issue 1 Pages 58-68
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    The solventogenic sol operon consisting of bld, ctfA, ctfB, and adc was cloned from Clostridium saccharoperbutylacetonicum strain N1-4. These genes share as high as 95–98% similarity with the corresponding sol genes of Clostridium beijerinckii NCIMB 8052. The N1-4 sol gene cluster was transcribed in a polycistronic manner under the control of two promoters, and its transcription was highly induced during solventogenesis. Strain DGN3-4, the degenerated strain derived from N1-4, maintained the sol genes, but transcription of the DGN3-4 sol operon was hardly induced during solventogenesis. A substance extracted from the culture supernatants of wild-type N1-4 allowed us to induce transcription of the sol operon in DGN3-4. These results suggest that the degeneration is caused by the incompetence of the induction mechanism of the sol operon, and that transcription might be under the control of a quorum-sensing mechanism.
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  • Hiroaki IWAKI, Yoshie HASEGAWA
    2007 Volume 71 Issue 1 Pages 145-151
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Bacterial strain KU-15, identified as a Burkholderia terrae by 16S rRNA gene sequence analysis, was one of 11 new isolates that grew on 2-nitrobenzoate as sole source of carbon and nitrogen. Strain KU-15 was also found to grow on anthranilate, 4-nitrobenzoate, and 4-aminobenzoate. Whole cells of strain KU-15 were found to accumulate ammonia in the medium, indicating that the degradation of 2-nitrobenzoate proceeds through a reductive route. Metabolite analyses by high-performance liquid chromatography indicated that 3-hydroxyanthranilate, anthranilate, and catechol are intermediates of 2-nitrobenzoate metabolism in strain KU-15. Enzyme studies suggested that 2-nitrobenzoate degradation occurs via the formation of 2-hydroxylaminobenzoate and that the pathway branches at this point to form two different aromatic intermediates: anthranilate and 3-hydroxyanthranilate. PCR amplifications and DNA sequencing revealed DNA fragments encoding a polypeptide homologous to 2-amino-3-carboxymuconate 6-semialdehyde decarboxylase and anthranilate 1,2-dioxygenase.
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  • Kenichi HOJO, Chinami MIZOGUCHI, Naoki TAKETOMO, Tomoko OHSHIMA, Kazuh ...
    2007 Volume 71 Issue 1 Pages 152-157
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    We surveyed the distribution of salivary Lactobacillus and Bifidobacterium species in periodontitis patients and healthy subjects. Approximately 700 lactobacilli and 300 bifidobacterial isolates were obtained from 16 young, orally healthy subjects (mean age ± standard deviation: 21.0±2.0 y), 16 periodontitis patients (51.6±13.8 y), and 14 well-maintained former periodontitis patients (60.2±9.6 y). Among eleven Lactobacillus species detected in saliva, L. salivarius, L. gasseri, and L. fermentum were prevalent, but no species was specifically associated with periodontal health. In contrast, of four Bifidobacterium species, B. adolescentis was specifically (P<0.05) prevalent in young healthy subjects compared with the other two groups. Furthermore, the bifidobacterial count of the well-maintained subjects was the highest (P<0.05) among the groups. These results suggest that bifidobacterial count and species might be associated with periodontal health status and/or age.
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Microbiology & Fermentation Technology Note
  • Masahiro HAYASHI, Tsugiyo YUKINO, Fumio WATANABE, Emi MIYAMOTO, Yoshih ...
    2007 Volume 71 Issue 1 Pages 222-225
    Published: January 23, 2007
    Released: January 23, 2007
    [Advance publication] Released: January 07, 2007
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    Newly isolated thraustochytrids showed uptake of vitamin B12 from the culture into the cells. Cultivation of thraustochytrids in a medium containing 1 μg/ml of vitamin B12 greatly increased the contents of vitamin B12 in the cells. Similarly to Schizochytrium limacinum, odd numbered fatty acids decreased in the cells of new isolates cultivated with vitamin B12. Vitamin B12-enriched thraustochytrids, strain mh0186, enhanced the population growth of rotifers fed on the cells as sole feed.
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