Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Volume 73 , Issue 12
Showing 1-48 articles out of 48 articles from the selected issue
Award Review
  • Toshiaki KUDO
    2009 Volume 73 Issue 12 Pages 2561-2567
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    Termites thrive in the tropics and play an important role in lignocellulose degradation. This ability depends mainly on intestine microbes in the gut, but most of them are so-called unculturable microbes, which can not be cultivated by traditional culture methods. The recent development of molecular approaches such as the PCR method has made it possible to access the enormous numbers of unculturable microbes in the gut of termites.
    This review explains our research on the ecological role of the termite, the termite-microbe symbiotic system, and the functions of lignocellulose degradation using various molecular methods. In the future, new technologies such as genomics should make it possible to analyze and utilize unculturable microbial resources in natural environments.
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Analytical Chemistry Regular Paper
  • Shiro MAWATARI, Keita YUNOKI, Masaaki SUGIYAMA, Takehiko FUJINO
    2009 Volume 73 Issue 12 Pages 2621-2625
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    A method for the simultaneous purification of plasmalogens and sphingomyelin (SM) in human erythrocytes is described. Treatment of total lipids with n-hexane/acetone (1:1 v/v) resulted in selective precipitation of SM. Both the supernatant and the precipitate fractions were incubated with a phospholipase A1 (PLA1) from Aspergillus orizae for 3.5 h. The PLA1-treated lipids were extracted with n-hexane/isopropanol, the hexane layer was obtained using a Na2SO4 solution, and the hexane layer was further washed with water. At this step, the relative concentration of the plasmalogens was 92% of the total phospholipids in the supernatant fraction, and that of SM was 97.7% in the precipitate fraction. Each fraction was applied to high performance liquid chromatography (HPLC) for further purification. The plasmalogen and SM obtained were almost free of the other lipids. The purity of the plasmalogens and SM was monitored by HPLC, which can separate intact plasmalogens from their diacyl analogs.
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Organic Chemistry Regular Papers
  • Kenji MORI
    2009 Volume 73 Issue 12 Pages 2727-2730
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    (11Z,13Z)-11,13-Hexadecadienal, the female sex pheromone of the navel orangeworm (Amyelois transitella), was synthesized from commercially available 10-bromo-1-decanol in a 27% overall yield (8 steps). The synthesis was achieved by using 10-iododecanal, trimethylsilylacetylene and (Z)-1-bromo-1-butene as the key building blocks, employing Sonogashira-Hagihara coupling and Brown’s hydroboration-protonolysis as the key reactions. The terminal formyl group was installed in the earlier stage of the synthesis rather than in the final step. This procedure enabled the multi-gram-scale preparation of this economically important pheromone.
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  • Yasumasa SUGIYAMA, Akira HIROTA
    2009 Volume 73 Issue 12 Pages 2731-2734
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    Six compounds were isolated as radical scavengers from the culture broth of a marine-derived actinomycete strain USF-TC31. The structures of two novel compounds were determined to be those of N-carbamoyl-2,3-dihydroxybenzamide (5) and 2-acetamido-3-(2,3-dihydroxybenzoylthio)propanoic acid (6), and four known compounds were identified to be anthranilic acid (1), 2,3-dihydroxybenzoic acid (2), 2,3-dihydroxybenzamide (3) and benadrostin (4) on the basis of spectroscopic data. Compound 6 was characterized as a racemate by its specific rotation. Each of the obtained compounds was evaluated for DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, and compounds 2, 3, 5 and 6 each exhibited potent activity in comparison with the butylhydroxytoluene (BHT) positive control.
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Organic Chemistry Notes
Organic Chemistry Communication
  • Hyun-Sun PARK, Takeshi OHAMA
    2009 Volume 73 Issue 12 Pages 2803-2805
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    Supplementary material
    Atractylenolide III is an organic product of the herb plant Atractylodes ovata that is used as an anti-inflammatory. This compound has very limited solubility in water, but we succeeded in transforming it using a Chlamydomonas cell disruptant containing 9.1% dimethyl sulfoxide. Two types of metabolites were confirmed after 3 h of incubation by gas chromatography, besides the non-modified substrate. Nuclear magnetic resonance and gas chromatography-mass spectrometry analyses indicated that one of the metabolites had two hydroxyl groups whereas atractylenolide III had one hydroxyl group, but no metabolite was obtained when atractylenolide III was added directly to the Chlamydomonas culture medium for 10 d.
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Biochemistry & Molecular Biology Regular Papers
  • Se-Hwan LIM, Hye Won PARK, Chang-Hoon SHIN, Jai-Hyun KWON, Chan-Wha KI ...
    2009 Volume 73 Issue 12 Pages 2576-2582
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    Dry powder formulations for pulmonary delivery are attractive because many issues of solubility and stability can be minimized. Human insulin microcrystals with lactose carriers were produced for pulmonary delivery. The average particle diameter was 2.3 μm, with a narrow, monodispersed size distribution. The percentages of high molecular weight proteins (%HMWPs), other insulin-related compounds (%OIRCs), and A-21 desamido insulin (%Des) were very low throughout the microcrystal preparation process. Administration of the microcrystal powder by intratracheal insufflation significantly reduced the blood glucose levels of Sprague-Dawley rats. The percent minimum reductions of the blood glucose concentration (%MRBG) produced by the insulin microcrystal powder and by an insulin solution reached 40.4% and 33.4% of the initial glucose levels respectively, and their bioavailability relative to subcutaneous injection (F) was 15% and 10% respectively. These results confirm that the insulin microcrystal powder prepared is suitable for pulmonary delivery in an effective dosage form.
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  • Michiko YASUDA, Tsuyoshi ISAWA, Satoshi SHINOZAKI, Kiwamu MINAMISAWA, ...
    2009 Volume 73 Issue 12 Pages 2595-2599
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    Agriculturally important grasses contain numerous diazotrophic bacteria, the interactions of which are speculated to have some other benefits to the host plants. In this study, we analyzed the effects of a bacterial endophyte, Azospirillum sp. B510, on disease resistance in host rice plants. Rice plants (Oryza sativa cv. Nipponbare) were inoculated with B510 exhibited enhanced resistance against diseases caused by the virulent rice blast fungus Magnaporthe oryzae and by the virulent bacterial pathogen Xanthomonas oryzae. In the rice plants, neither salicylic acid (SA) accumulation nor expression of pathogenesis-related (PR) genes was induced by interaction with this bacterium, except for slight induction of PBZ1. These results indicate the possibility that strain B510 is able to induce disease resistance in rice by activating a novel type of resistance mechanism independent of SA-mediated defense signaling.
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  • Sung-Jin CHO, Ki Seok KOH, Eun LEE, Soon Cheol PARK
    2009 Volume 73 Issue 12 Pages 2609-2614
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    The earthworm provides an excellent model for investigating regeneration. Here we report the full-length cloning of three labial genes (Pex-lab01, Pex-lab02, and Pex-lab03) in the earthworm Perionyx excavatus. To analyze their expression pattern during head and tail regeneration, we used the reverse transcription-polymerase chain reaction. Our results indicate that the three labial genes were expressed only in the head-regenerating tissues. Also, we found that the expression of Pex-lab01 and Pex-lab02 is up-regulated, and this indicates their involvement in wound healing and the blastema formation processes during early head regeneration.
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  • Kei H. KOJO, Makoto T. FUJIWARA, Ryuuichi D. ITOH
    2009 Volume 73 Issue 12 Pages 2632-2639
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    While it has been established that binary fission of leaf chloroplasts requires the prokaryote-derived, division site determinant protein MinE, it remains to be clarified whether chloroplast division in non-leaf tissues and the division of non-colored plastids also involve the MinE protein. In an attempt to address this issue, plastids of cotyledons, floral organs, and roots were examined in the Arabidopsis thaliana mutant of the MinE (AtMinE1) gene, which was modified to express the plastid-targeted cyan fluorescent protein constitutively, and were quantitatively compared with those in the wild type. In the cotyledons, floral organs, and root columella, the plastid size in the atminE1 mutant was significantly larger than in the wild type, while the plastid number per cell in atminE1 appeared to be inversely smaller than that in the wild type. In addition, formation of the stroma-containing plastid protrusions (stromules) in the cotyledon epidermis, petal tip, and root cells was more active in atminE1 than in the wild type.
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  • Yoshifumi MAEDA, Daiju DOUBAYASHI, Masaya OKI, Hiroaki NOSE, Akihiko S ...
    2009 Volume 73 Issue 12 Pages 2645-2649
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    An unnamed protein of Aspergillus oryzae RIB40 (accession no. XP_001727378), the amino acid sequence of which shows high similarity to those of formate oxidase isoforms produced by Debaryomyces vanjiriae MH201, was produced in Escherichia coli in C-His6-tagged form. The gene product, purified by affinity column chromatography, catalyzed the oxidation of formate to yield hydrogen peroxide but showed no evidence of activity on the other substrates tested. The Km and Vmax values at 30 °C at pH 4.5 were 7.9 mM and 26.3 μmole/min mg respectively. The purified enzyme showed UV-visible spectra atypical of ordinary flavoproteins. The UV-visible spectra of the enzyme and the UV-visible spectra, fluorescence spectra, and mass spectrometry of the extract obtained by boiling the purified enzyme suggested that the enzyme has a non-covalently bound FAD analog, which is expected to be 8-formyl-FAD.
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  • Seiichi FURUKAWA, Hiromitsu TANAKA, Jun ISHIBASHI, Shigeo IMANISHI, Mi ...
    2009 Volume 73 Issue 12 Pages 2665-2670
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    A cDNA encoding an IκB family protein was identified and the full nucleotide sequence was determined in the silkworm Bombyx mori. The IκB gene, designated BmCactus, was constitutively expressed mainly in the fat body and hemocytes. Transfection experiments on a B. mori cell line, NIAS-Bm-aff3, with expression vectors containing BmCactus, BmRelA, BmRelB, or the active portion of BmRelish1 showed that activation of the CecB1 gene promoter by either BmRelA or BmRelB, but not the active portion of BmRelish1, was strongly inhibited by BmCactus. In addition, activation of CecB1 gene by autoclaved E. coli in the cultured cells was observed regardless of the presence or absence of BmCactus. A gultathione S-transferase pull-down assay and analysis using a yeast two-hybrid system demonstrated that BmCactus interacted with the BmRel Rel homology domain, but not with the BmRelish Rel homology domain. These results suggest that BmCactus is involved in the Toll signal transduction pathway in B. mori.
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  • Sanhita ROY, Samir Kumar DUTTA
    2009 Volume 73 Issue 12 Pages 2671-2676
    Published: December 23, 2009
    Released: December 23, 2009
    JOURNALS FREE ACCESS
    A 516-bp winged bean chymotrypsin-trypsin inhibitor (WbCTI) gene was amplified from genomic DNA and cDNA isolated from winged bean using a pair of degenerate primers designed on the basis of the amino acid sequences of WbCTI. The amplified PCR products were cloned and sequenced to confirm their authenticity. DNA sequence analysis of the genomic and cDNA clones of WbCTI revealed the same nucleotide sequence in the coding region and showed WbCTI to be an intron less gene. WbCTI was subcloned into pTrc99A and expressed in Escherichia coli to yield a recombinant protein (rWbCTI). rWbCTI was purified by a rapid and single step immunoaffinity chromatography method, with an overall yield of 1.1 mg/g of wet cells. The homogeneity of the purified protein was checked by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which showed the presence of a single protein band. Functionally rWbCTI is indistinguishable from WbCTI, since both inhibit bovine trypsin and chymotrypsin in a 1:1 molar ratio. FPLC binding studies also confirmed that rWbCTI binds the proteases in a 1:1 molar ratio.
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  • Satoshi TATSUNO, Kenji ARAKAWA, Haruyasu KINASHI
    2009 Volume 73 Issue 12 Pages 2712-2719
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    Supplementary material
    Extensive mutations of lankacidin synthase genes were carried out to analyze the modular-iterative mixed polyketide biosynthesis of lankacidin. Three ketoreductase domains (lkcC-KR, lkcF-KR1, and lkcF-KR2) were inactivated by in-frame deletion and site-directed mutagenesis of their active sites. The mutants ceased or diminished lankacidin production, indicating that the three KR domains are functional in lankacidin biosynthesis. However, all of the KR mutants failed to accumulate the expected unreduced metabolites. Mutational analysis of two tandemly aligned acyl carrier protein domains (lkcC-ACP1 and lkcC-ACP2) revealed that either ACP is sufficient for lankacidin production. Disruption and complementation experiments on three unique genes/domain (lkcD for acyltransferase, lkcB for dehydratase, and lkcC-MT for a C-methyltransferase domain) suggested that their gene products function iteratively during lankacidin biosynthesis.
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  • Yukihiro KANATO, Sayaka ONO, Ken KITAJIMA, Chihiro SATO
    2009 Volume 73 Issue 12 Pages 2735-2741
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    Glycosaminoglycans (GAGs), large anionic glycopolymers, are the glycan portion of proteoglycans and are important components of the extracellular matrix. Recently we reported that polysialic acid, a polyanionic glycopolymer specific to the brain, binds neurotrophic factors to form a large complex. It is not clear whether GAGs also bind neurotrophic factors to form a large complex. In the present study, we demonstrate that a brain-derived neurotrophic factor (BDNF) dimer directly binds GAGs other than chondroitin and hyaluronic acid to form a large complex. Neurotrophin-3 showed similar GAG binding properties. Furthermore, BDNF, after forming a large complex with GAG, bound to the BDNF receptors tropomyosin-related kinase (Trk) B and p75 neurotrophin receptor (NTR). These findings suggest that GAGs function to produce a reservoir of BDNF and other neurotrophic factors, and may serve to regulate their local concentrations on the cell surface.
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Biochemistry & Molecular Biology Notes
Biochemistry & Molecular Biology Communication
  • Noriyuki YANAKA, Yurika KASEDA, Anna TANAKA, Yoshihito NOGUSA, Naoki S ...
    2009 Volume 73 Issue 12 Pages 2809-2811
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    Zinc finger protein ZPR1 (ZPR1) binds to eukaryotic translation elongation factor 1α (eEF1α) in response to growth stimuli, and is also involved in transcription and cell cycle regulation. In this study, we characterized the interaction of ZPR1 and eEF1α and generated a ZPR1 mutant that constitutively interacted with eEF1α. ZPR1-ΔA (Δ193–246) bound to eEF1α independently of Zn2+ in vivo. This study indicates that ZPR1-ΔA (Δ193–246) is a useful tool to provide structural insights into ZPR1 and to investigate the biological significance of the interaction between ZPR1 and eEF1α.
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Food & Nutrition Science Regular Papers
  • Jun LI, Shinya MATSUMOTO, Akihiro NAKAMURA, Hirokazu MAEDA, Yasuki MAT ...
    2009 Volume 73 Issue 12 Pages 2568-2575
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    Soluble soybean polysaccharide (SSPS) was fractionated into two sub-fractions, a high-molecular-weight fraction (HMF) and a low-molecular-weight fraction (LMF) by the ethanol-extraction method. Characterization of the sub-fractions, that is, analysis of chemical composition, gel filtration, and SDS–PAGE, revealed that the main component of HMF was a large polysaccharide molecule with covalently-attached peptides, possibly corresponding to the intact SSPS molecule. LMF consisted of free peptides and saccharides of small size, which might have occurred as by-products during the production process of SSPS. HMF exhibited high ability to emulsify oil droplets and stabilize α-casein dispersions in an acidic pH region, but this ability of LMF was inferior to HMF. On the other hand, LMF had higher activity to prevent the oxidation of emulsified lipids than HMF. These results suggest that HMF and LMF had different characteristics and functional properties, and that the combination of the two sub-fractions generates the multi-functions of commercial SSPS.
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  • Meng-Kwoon SIM, Yong-Chiat WONG, Xiao-Guang XU, Sai-Zhen SIM, Daniel T ...
    2009 Volume 73 Issue 12 Pages 2583-2588
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    This study researched the effects of chicken meat extract on blood glucose and insulin level, membrane glucose transporter-4 (GLUT4), and tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) in type 2 diabetic KKAy mice and GK rats. Eight-week-old KKAy mice and GK rats and euglycemic control animals, C57BL/6J mice and Wistar rats, were orally administered a liquid commercial chicken meat extract, BRAND’S Essence of Chicken (BEC), for up to 8 weeks. BEC (1.5 ml/kg) had no effect on blood insulin levels, but significantly lessened the hyperglycemia in the diabetic animals. In the BEC-treated diabetic animals, insulin induced a significant increase in plasma membrane GLUT4 and cytosolic tyrosine-phosphorylated IRS-1, indicating that it attenuates insulin resistance. The present findings are the first demonstration of the hypoglycemic action of a dietary protein, and they lend credence to the reported benefits of using chicken meat as a source of protein in the dietary management of diabetic individuals.
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  • Yung-Hsien CHANG, Jai-Sing YANG, Jiun-Long YANG, Chang-Lin WU, Shu-Jen ...
    2009 Volume 73 Issue 12 Pages 2589-2594
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    Ganoderma lucidum (G. lucidum) is a medicinal mushroom having biological effects such as immunomodulation and anti-tumor actions. In China and many other Asian countries, G. lucidum is used as a folk remedy to promote health and longevity. Although many studies have shown that G. lucidum modulates the immune system, including, for example, antigen-presenting cells, natural killer (NK) cells, and the T and B lymphocytes, the effects of G. lucidum on the WEHI-3 leukemic BALB/c mice are unclear. We attempted to determine whether G. lucidum would promote immune responses in BALB/c mice injected with WEHI-3 leukemia cells. The effects of G. lucidum on the survival rate of WEHI-3 leukemia cells injected into BALB/c mice were examined. It increased the percentages of CD3 and CD19, but decreased the percentages of Mac-3 and CD11b markers, suggesting that differentiation of the precursor of T and B cells was promoted but macrophages were inhibited. It decreased the weight of spleens as compared with control mice. It also promoted phagocytosis by macrophage from peripheral blood mononuclear cell (PBMC) and it also promoted natural killer cell activity. It decreased the percentage of leukemia cells in the spleens of mice before they were injected with WEHI-3 cells. Apparently, G. lucidum affects murine leukemia WEHI-3 cells in vivo.
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  • Shizue SAEGUSA, Mamoru TOTSUKA, Shuichi KAMINOGAWA, Tomohiro HOSOI
    2009 Volume 73 Issue 12 Pages 2600-2608
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    We investigated whether non-pathogenic Saccharomyces cerevisiae and human commensal opportunistic pathogenic Candida albicans stimulate cytokine responses of human neutrophil-like HL-60 cells pre-treated with either 1 μM retinoic acid or 1.25% dimethyl sulfoxide (DMSO). Intact and heat-killed S. cerevisiae enhanced secretion of interleukin (IL)-1β, IL-6, IL-8, IL-12, IL-18, MCP-1/CCL2 and TNF-α from retinoic acid-treated HL-60 cells, accompanied by alterations in mRNA expression of the cytokines. Heat-killed C. albicans promoted secretion of IL-6, IL-8, IL-12, MCP-1 and TNF-α, while intact C. albicans slightly enhanced secretion of IL-1β, IL-8 and IL-18. In response to yeast stimuli, retinoic acid-treated HL-60 cells generally secreted cytokines more strongly than DMSO-treated HL-60 cells. Gene expression levels of Toll-like receptor (TLR)1, TLR2, TLR4, TLR6 and dectin-1 in HL-60 cells were additionally affected by retinoic acid or DMSO and by co-culturing with S. cerevisiae or C. albicans. Our results suggest that both intact and heat-killed S. cerevisiae and C. albicans induce cytokine responses of neutrophils in the intestine, and stimulate host immune function.
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  • Celia CONESA, Coloma POCOVÍ, María-Dolores PÉREZ, ...
    2009 Volume 73 Issue 12 Pages 2615-2620
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    The possibility of using recombinant human lactoferrin from rice (rhLF) makes it necessary to study its differences from the protein of milk. In this work, the binding of different iron-saturated forms of rhLF to Caco-2 cells was studied. Iron-saturated rhLF bound in higher proportion than the apo-form, but, the data obtained for specific binding were not compatible with receptor-mediated binding. Competition assays showed the same binding capacity for human milk lactoferrin as for rhLF to Caco-2 cells. Another basic protein of milk, lactoperoxidase, was found to compete with rhLF for binding to Caco-2 cell membranes, suggesting an electrostatic interaction. The transport of iron (59Fe) bound to rhLF and to citrate and the transport of rhLF (125I-labeled) were studied on Caco-2 monolayers. Transport of iron was found to be significantly greater when bound to citrate than to rhLF. The amount of intact lactoferrin that traversed the Caco-2 monolayers was very low, suggesting degradation of it across these cells.
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  • Takanori HASEGAWA, Kozue HIRAKAWA, Takashi MATSUMOTO, Shinji TOKI, Yos ...
    2009 Volume 73 Issue 12 Pages 2626-2631
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    To determine the effects of Lactobacillus plantarum strain HSK201 on Japanese cedar pollinosis, a single-blind, placebo-controlled study was conducted in 2008. The HSK201 group was administered fermented milk prepared with HSK201 for 8 weeks, and the placebo group was administered non-fermented milk adjusted to the same acidity and taste. We found HSK201 strain intake to suppress both helper T cell type 1/2 ratio reduction and serum Japanese cedar pollen-specific IgE elevation at the peak of pollen dispersion. In addition, the nasal and ocular symptom scores in the HSK201 group were also lower than those in the placebo group during the early phase of the pollen season. Although this was a preliminary study with 19 employees of our own company serving as subjects, the results suggest that ingestion of the HSK201 strain alleviates pollinosis symptoms during the period when pollen exposure is low and the symptoms are mild.
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  • Yuko NAGATA, Kenji HASHIGUCHI, Yuya KAMIMURA, Mutsuko YOSHIDA, Toshiha ...
    2009 Volume 73 Issue 12 Pages 2650-2655
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    This study aimed to assess variations in the human gastrointestinal transit tolerance of Lactobacillus plantarum strain No. 14 after culture with glucose and with fructose. Transit tolerance was determined at 37 °C against simulated gastric juices at pH values of 2.5, 3.0, and 3.5, and against simulated small intestinal juices containing 0%, 0.2%, or 0.4% oxgall. The simulated gastrointestinal transit tolerance of Lactobacillus plantarum strain No. 14 varied with the carbon source. Hence we compared the amounts of exopolysaccharide from Lactobacillus plantarum strain No. 14 cultured in various carbon sources. The exopolysaccharide levels were 146.5±8.1 mg/l (culture) with glucose, and 20.1±17.0 mg/l (culture) with fructose. When exopolysaccharide was removed by centrifugation, the simulated gastric tolerance of Lactobacillus plantarum strain No. 14 cultured with glucose decreased markedly, but that with fructoce did not decrease. These results suggest that the gastrointestinal transit tolerance of Lactobacillus plantarum strain No. 14 is related to exopolysaccharide contents.
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  • Chaluntorn VICHASILP, Kiyotaka NAKAGAWA, Phumon SOOKWONG, Yasuyo SUZUK ...
    2009 Volume 73 Issue 12 Pages 2684-2689
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    Mulberry 1-deoxynojirimycin (DNJ, a potent α-glycosidase inhibitor) has therapeutic potency against diabetes mellitus. However, the amount of DNJ in mulberry leaves is low (about 0.1%), and a more effective extraction method is needed. Ultrasound-assisted extraction (UAE) was applied in this study for mulberry DNJ extraction, and five factors, the percentage of ethanol in the extraction solvent (x1), ratio of the extraction solvent to mulberry sample (x2), ultrasonic power (x3), extraction temperature (x4) and extraction time (x5), were investigated by fractional factorial 2(5-1) design (FFD) to obtain the optimum extraction efficiency (DNJ yield, Y1) and extraction productivity (total yield, Y2). The results showed that x2, x3 and x5 had significant impact on Y1 and Y2, and were further optimized by response surface methodology (RSM). Under the optimized conditions (x2, x3 and x5 of 7 ml/g, 180 W and 260 s, respectively), DNJ-enriched powder (0.8%) was produced with high extraction efficiency (98%) and productivity (20%), enabling this product to be used for nutraceutical purposes.
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  • Fuminori KAWABATA, Naohiko INOUE, Yukiko MASAMOTO, Shigenobu MATSUMURA ...
    2009 Volume 73 Issue 12 Pages 2690-2697
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    Capsinoids are non-pungent capsaicin analogs which increase energy expenditure like capsaicin. However, the mechanisms underlying the enhancement of their energy expenditure despite their non-pungency are poorly understood. We suggest here that capsinoids increase energy expenditure in wild-type mice, but not in transient receptor potential vanilloid 1 (TRPV1) knockout mice, implying that capsinoids increase energy expenditure via TRPV1. The jejunal administration of capsinoids to anesthetized mice raised the temperature of the colon and intrascapular brown adipose tissue. Denervation of the extrinsic nerves connected to the jejunum inhibited this temperature elevation. These findings suggest that capsinoids increase energy expenditure by activating the intestinal extrinsic nerves. Although the jejunal administration of capsinoids did not raise the tail skin temperature, an intravenous injection of capsinoids did, indicating that capsinoids could barely pass through the intestinal wall into the blood. Taken together, gastrointestinal TRPV1 may be a critical target for capsinoids to enhance energy expenditure.
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  • Ken-ichi YAMAZAKI, Noriko KATO-NAGAOKA, Tomomi SUZUKI, Kan SHIDA, Masa ...
    2009 Volume 73 Issue 12 Pages 2705-2711
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
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    We examined cytokine production and allergic reactions in mice fed ad libitum (AL) and subjected to dietary restriction (DR). DR retarded the increase in body weight, and peripheral blood T cells in the DR mice produced less IFN-γ and more IL-4 in response to immobilized anti-CD3 mAb. Systemic immunization and intranasal challenge with ovalbumin (OVA) induced accumulation of leukocytes into the lung, increase in IL-4 level in bronchoalveolar lavage fluid (BALF), and rise in serum IgE in the AL mice. In contrast, these allergic symptoms were alleviated in the DR mice. Furthermore, the relative proportion of IL-4-producing T cells responsive to OVA was less in the DR mice than the AL mice. DR tended to decrease the proportion and cytolytic activity of NK cells in the spleen, especially in younger mice. These results indicate that DR can prevent the expansion of allergen-specific IL-4-producing T cells followed by suppression of the allergic reaction, but might dampen NK cell activity.
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Food & Nutrition Science Notes
Microbiology & Fermentation Technology Regular Papers
  • Mark GAFFNEY, Sean DOYLE, Richard MURPHY
    2009 Volume 73 Issue 12 Pages 2640-2644
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    Extracellular xylanase production by the thermophilic fungus Thermomyces lanuginosus 195 in solid state fermentation (SSF) was found to be significantly affected by fermentation temperature, duration, and inoculum volume (p≤0.001). Optimization of these parameters corresponded to a 21.7% increase in xylanase yield. Maximum activity (2,335 U/g of wheat bran) was obtained when 10 g of wheat bran was inoculated with 10 ml of liquid culture and cultivated at 45 °C for 40 h. The influence of supplemental carbon and nitrogen sources (3% w/v) on xylanase production was also assessed. Wheat bran, supplemented with glucose and cellulose, facilitated 10% and 7% increases in relative activity respectively. Ammonium based salts, nitrates, and a number of organic nitrogen sources served only to reduce xylanase production (p≤0.005) significantly. The enhanced xylanase titers achieved in the present study emphasize the need for optimizing growth conditions for maximum enzyme production in SSF.
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  • Tong DAN, Kenji FUKUDA, Michiko SUGAI-BANNAI, Naoya TAKAKUWA, Hidemasa ...
    2009 Volume 73 Issue 12 Pages 2656-2664
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    Part of the exopolysaccharide gene cluster of Lactobacillus fermentum TDS030603 was characterized. It consists of 11,890 base pairs and is located in the chromosomal DNA, 13 open reading frames of which were encoded. Out of the 13 open reading frames, six were found to be involved in exopolysaccharide synthesis; however, five were similar to transposase genes of other lactobacilli, and two were functionally unrelated. Expression analysis revealed that the exopolysaccharide synthesis-related genes were expressed during cultivation. Southern analysis using specific primers for the exopolysaccharide genes indicated that duplication of the gene cluster did not occur. The plasmid-cured strain maintained its capacity for exopolysaccharide production, confirming that the exopolysaccharide gene cluster of this strain is located in the chromosomal DNA, similarly to thermophilic lactic acid bacteria. Our results indicate that this exopolysaccharide gene cluster is likely to be functional, although extensive gene rearrangement occurs.
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  • Koichiro MIYAKE, Shingo KAKITA
    2009 Volume 73 Issue 12 Pages 2677-2683
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    γ-Glutamylcysteine synthetase (γGCS, EC 6.3.2.2) catalyzes the formation of γ-glutamylcysteine from L-glutamic acid (Glu) and L-cysteine (Cys) in an ATP-dependent manner. While γGCS can use various amino acids as substrate, little is known about whether it can use non-amino acid compounds in place of Cys. We determined that γGCS from Escherichia coli has the ability to combine Glu and amines to form γ-glutamylamides. The reaction rate depended on the length of the methylene chain of the amines in the following order: n-propylamine > butylamine > ethylamine >> methylamine. The optimal pH for the reaction was narrower and more alkaline than for the reaction with an amino acid. The newly found catalytic ability of γGCS was used in the production of theanine (γ-glutamylethylamine). The resting cells of E. coli expressing γGCS, in which ATP was regenerated through glycolysis, synthesized 12.1 mM theanine (18 h) from 429 mM ethylamine.
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  • Tomokazu ITO, Nobuyuki UOZUMI, Tatsunosuke NAKAMURA, Sayuri TAKAYAMA, ...
    2009 Volume 73 Issue 12 Pages 2698-2704
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    An Escherichia coli mutant lacking three major K+ uptake systems, Trk, Kup, and Kdp, did not grow under low K+and high Na+ concentrations. The introduction of fkuA and of fkuB of a marine bacterium, Vibrio alginolyticus, has been reported to compensate for the growth defect by accelerating the rate of K+ uptake (Nakamura, Katoh, Shimizu, Matsuba, and Unemoto, Biochim. Biophys. Acta, 1277, 201–208 (1996)). We investigated the function of unknown genes of E. coli, yggS and yggT, homologs of fkuA and fkuB respectively. E. coli TK2420 cells, which lack the three K+ uptake systems, did not grow under high Na+ or mannitol concentrations. The growth defect was compensated by the introduction of the yggT gene alone: yggS was not required. Here we found that YggT endowed E. coli cells with a tolerance for osmotic shock, and discuss a possible mechanism.
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  • Noppon LERTWATTANASAKUL, Emi SHIGEMOTO, Nadchanok RODRUSSAMEE, Savitre ...
    2009 Volume 73 Issue 12 Pages 2720-2726
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    The function of mitochondrial Adh3 in the thermotolerant yeast Kluyveromyces marxianus was investigated. An ADH3-disrupted mutant exhibited growth retardation on non-fermentable carbon sources, except for ethanol, and this was suppressed by supplementation with antioxidants. Detailed analysis of the phenotype revealed that the mutant showed an increase in the activity of NADH dehydrogenase, sensitivity to H2O2, and accumulation of reactive oxygen species (ROS), and that these carbon sources increased the activity of succinate dehydrogenase. The increase in both activities may reflect enhanced expression of both dehydrogenases by elevation of their substrate levels. The ROS level became low when antioxidants were added. These findings suggest that the ADH3 mutation and such carbon sources cause an elevation of the substrate level of the respiratory chain and eventually of the ROS level via increased expression of primary dehydrogenases, which in turn causes cell growth retardation. Adh3 might thus play a crucial role in the control of the NADH/NAD+ balance in mitochondria.
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Microbiology & Fermentation Technology Notes
Microbiology & Fermentation Technology Communication
  • Masato IKEDA, Masahiro BABA, Nobuyoshi TSUKAMOTO, Tomoha KOMATSU, Sato ...
    2009 Volume 73 Issue 12 Pages 2806-2808
    Published: December 23, 2009
    Released: December 23, 2009
    [Advance publication] Released: December 07, 2009
    JOURNALS FREE ACCESS
    Mutagenized cell libraries of Corynebacterium glutamicum were screened for mutants that lost the ability to grow under low oxygen concentrations. The resulting high-oxygen-requiring mutants were used to clone wild-type DNA fragments that could complement the phenotype. Sequencing and subcloning analyses identified six genes, Cgl0807, Cgl1102, Cgl0600, Cgl1427, Cgl2857, and Cgl2859, as the genes responsible for complementation. Some of these genes showed cross-complementation of the mutants in oxygen-limiting static culture, suggesting the utility of these genes for improved growth and production under oxygen limitation.
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