Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Volume 75 , Issue 4
Showing 1-42 articles out of 42 articles from the selected issue
Award Reviews
  • Yoshichika KAWAI
    2011 Volume 75 Issue 4 Pages 609-617
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    It has been suggested that polyphenol-rich diets decrease the risk of cardiovascular diseases. Although studies of the bioavailability of polyphenols, particularly their absorption and metabolism, have been reported recently, the tissue and cellular distributions underlying their biological mechanisms remain unknown. It is difficult to evaluate the specific localization of tissue and/or cellular polyphenols, because the method is limited to chromatography. To overcome these difficulties, we have developed anti-polyphenol antibodies to characterize immunohistochemically the localization of polyphenols and their metabolites in vivo. Two novel monoclonal antibodies were raised against quercetin and tea catechins, which represent flavonoid-type polyphenols distributed in foods and beverages, and are expected to exhibit anti-oxidative and anti-inflammatory activities in vivo. Using these antibodies, we identified activated macrophages as a specific target of these flavonoids during the development of atherosclerotic lesions. This review describes recent findings on the molecular actions of flavonoids that underly their anti-atherosclerotic activity in vivo.
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  • Takashi INAOKA, Kozo OCHI
    2011 Volume 75 Issue 4 Pages 618-623
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Microorganisms possess the ability to produce a variety of commercially important secondary metabolites such as antibiotics. Although it becomes harder and harder to discover useful new compounds, microorganisms still have the potential to produce unknown compounds. One of the reasons for the difficulty in finding new compounds is that the expression level of many secondary metabolite genes is insufficient in wild-type strains. Therefore, a new method of activating gene expression might be a powerful tool for the screening of novel compounds and for strain improvement to overproduce useful compounds. We found that the rifampicin-resistant RNA polymerase mutations stimulate the expression of antibiotic synthetic gene clusters in several microorganisms. In the case of the Gram-positive model organism Bacillus subtilis, one of the rifampicin-resistance mutations resulted in the activation of a dormant secondary metabolism, neotrehalosadiamine synthesis. To clarify this activation mechanism, we first identified the neotrehalosadiamine biosynthetic operon and investigated its transcriptional regulation. Here we summarize our findings on the transcriptional regulation of the neotrehalosadiamine biosynthetic operon and discuss a crucial effect of the rifampicin-resistance mutation on the expression of dormant genes.
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Analytical Chemistry Regular Paper
  • Satoshi SEKIGUCHI, Hideki KOHNO, Kiyoshi YASUKAWA, Kuniyo INOUYE
    2011 Volume 75 Issue 4 Pages 752-756
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Leptin is one of the representative adipocyte-derived protein hormones. Measuring the serum leptin concentration gives an important index for preventing and treating diabetes mellitus and other diseases. We constructed in this study a chemiluminescent enzyme immunoassay (CLEIA) for measuring leptin by using the anti-leptin polyclonal antibody and alkaline phosphatase (ALP). The method applies the IgG-conjugated ferrite particle to capture leptin in a sample and the ALP-conjugated Fab fragment to detect the captured leptin. We tested Block ace, CE510, and bovine serum albumin (BSA) for their abilities to block non-specific binding of ALP-conjugated anti-leptin Fab to the ferrite particle and found BSA to be the most effective. The measurable range with this ELISA for leptin was 0.1–1.0 pg/mL of leptin and the detection limit (blank+2SD) was 0.1 pg/mL of leptin. These results demonstrate sufficient sensitivity with our system to measure the serum leptin concentration and its clinical usefulness. The results also suggest that a sensitive enzyme immunoassay can be constructed by using only one polyclonal antibody.
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Analytical Chemistry Note
  • Yasuhiro KAKIGI, Takashi HAKAMATSUKA, Takeshi ICHO, Yukihiro GODA, Nao ...
    2011 Volume 75 Issue 4 Pages 777-779
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    The rapid and highly separative ultra high-performance liquid chromatography (UHPLC)-UV method was adopted and validated to investigate the flavonol glycoside compositions in ginkgo leaf products on the Japanese market. The result indicates that certain products contained amounts of flavonol glycosides approximately equivalent to the medicinal product. Additionally, we examined the correlations between the total amount of flavonol glycosides and of terpene lactones in various ginkgo leaf products.
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Organic Chemistry Regular Paper
  • Kengo SHIGETOMI, Shoko OMOTO, Yasuo KATO, Makoto UBUKATA
    2011 Volume 75 Issue 4 Pages 718-722
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    The structure-activity relationship was investigated to evaluate the antifungal activities of tuliposides and tulipalins against tulip pathogenic fungi. 6-Tuliposide B was effectively synthesized via the asymmetric Baylis-Hillman reaction. Tuliposides and tulipalins showed antifungal activities against most of the strains tested at high concentrations (2.5 mM), while Botrytis tulipae was resistant to tuliposides. Tulipalin formation was involved in the antifungal activity, tulipalin A showed higher inhibitory activity than 6-tuliposide B and tulipalin B. Both the tuliposides and tulipalins showed pigment-inducing activity against Gibberella zeae and inhibitory activity against Fusarium oxysporum f. sp tulipae. These activities were induced at a much lower concentration (0.05 mM) than the antifungal MIC values.
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Organic Chemistry Notes
Biochemistry & Molecular Biology Regular Papers
  • Sung June BYUN, Sung Woo KIM, Kyung-Woon KIM, Jeom Sun KIM, In-Sul HWA ...
    2011 Volume 75 Issue 4 Pages 646-649
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    In this study, we confirmed the ability of the 2-kb promoter fragment of the chicken ovalbumin gene to drive tissue-specific expression of a foreign EGFP gene in chickens. Recombinant lentiviruses containing the EGFP gene were injected into the subgerminal cavity of 539 freshly laid embryos (stage X). Subsequently the embryos were incubated to hatch using phases II and III of the surrogate shell ex vivo culture system. Twenty-four chicks (G0) were hatched and screened for EGFP with PCR. Two chicks were identified as transgenic birds (G1), and these founders were mated with wild-type chickens to generate transgenic progeny. In the generated transgenic hens (G2), EGFP was expressed specifically in the tubular gland of the oviduct. These results show the potential of the chicken ovalbumin promoter for the production of biologically active proteins in egg white.
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  • Shun Zong SONG, Yun Ho CHOI, Guang Yu JIN, Guang Zhao LI, Guang Hai YA ...
    2011 Volume 75 Issue 4 Pages 656-661
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    This study elucidated the effects of cornuside on carbon tetrachloride (CCl4)-induced hepatotoxicity. Rats were treated intraperitoneally with 0.5 mL/kg of CCl4. Sixteen h after CCl4 treatment, the levels of serum aminotransferases, tumor necrosis factor-α (TNF-α), and lipid peroxidation were significantly elevated, whereas the hepatic antioxidative enzyme activities were decreased. These changes were attenuated by cornuside. Histological studies also indicated that cornuside inhibited CCl4-induced liver damage. Furthermore, the contents of hepatic nitrite, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were elevated after CCl4 treatment, while cytochrome P450 2E1 (CYP2E1) expression was suppressed. Cornuside treatment inhibited the formation of liver nitrite, and reduced the overexpression of iNOS and COX-2 proteins, but restored the liver CYP2E1 content as compared with the CCl4-treated rats. Our data indicate that cornuside protects the liver from CCl4-induced acute hepatotoxicity, perhaps due to its ability to restore the CYP2E1 function and suppress inflammatory responses, in combination with its capacity to reduce oxidative stress.
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  • Yan ZENG, Jing YU, Jing CANG, Lijie LIU, Yongchao MU, Junhong WANG, Da ...
    2011 Volume 75 Issue 4 Pages 681-687
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Supplementary material
    Carbohydrate accumulation is common in frost-resistant plants, and many enzymes participate in this process. The sugar content and expression levels of metabolic enzymes related to sugar biosynthesis in response to drops in temperature were measured in two cultivars of winter wheat (Triticum aestivum) with different cold tolerances. The results indicate that the two cultivars examined, Dongnongdongmai 1 and Jimai 22, accumulated high levels of carbohydrate before November 4 (above 0 °C), and that accumulation decreased as temperatures fell. However, this decrease was more modest in Dongnongdongmai 1, which had a higher sugar content. Sucrose and fructose were the main soluble sugars, indicating an important role in freezing tolerance. Gene expression studies revealed that expression of the genes encoding chloroplastic enzymes was significantly upregulated in the tillering nodes. Expression upregulation of TaSS and TaTPT may be helpful for sugar accumulation before November 4.
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  • Jun IWASHITA, Kyohei OSE, Hiroki ITO, Jun MURATA, Tatsuya ABE
    2011 Volume 75 Issue 4 Pages 688-693
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Mucin production by epithelial cells is modulated by many soluble factors, including epidermal growth factor (EGF). E-Cadherin promotes EGF receptor (EGFR)-mediated MUC5AC mucin production in airway epithelial cells in dense cultures, suggesting the involvement of E-cadherin in activating EGFRs and mucin production. However, the role of E-cadherin in modulating mucin production is not completely understood. We examined its role in MUC5AC production in a human lung epithelial cell line, NCI-H292. Treatment of low density NCI-H292 cells with an anti-E-cadherin monoclonal antibody (SHE78-7) inhibited cell-cell contact in the dispersed colonies, but promoted MUC5AC production. Furthermore, treatment of the NCI-H292 cells with anti-E-cadherin antibody stimulated phosphorylation of extracellular signal-regulated kinase (ERK). The enhanced production of MUC5AC was inhibited with an EGFR inhibitor and with a MEK inhibitor, but not with a Src family kinase inhibitor. These results suggest that inhibition of E-cadherin activates EGFRs independently of Src and promotes MUC5AC production through the ERK signaling pathway in sparsely cultured NCI-H292 cells.
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  • Tomoki NAKAGAWA, Ryoko SATAKE, Masaru SATO, Kuniki KINO
    2011 Volume 75 Issue 4 Pages 700-704
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Depsipeptides are peptide-like polymers consisting of amino acids and hydroxy acids, and are expected to be new functional materials for drug-delivery systems and polymer science. In our previous study, D-alanyl-D-lactate, a type of depsipeptide, was enzymatically synthesized using D-alanine-D-alanine ligase from Thermotoga maritima ATCC 43589 (TmDdl) by Y207F substitution. Thereafter, in this study, further mutagenesis was introduced, based on structural comparison between TmDdl and a well-characterized D-alanine-D-alanine ligase from Escherichia coli. The S137A/Y207F mutant showed higher D-alanyl-D-lactate and lower D-alanyl-D-alanine synthesizing activity than the Y207F mutant. This suggests that substitution at the S137 residue contributes to product selectivity. Saturated mutagenesis on S137 revealed that the S137G/Y207F mutant showed the highest D-alanyl-D-lactate synthesizing activity. Moreover, the mutant showed broad substrate specificity toward D-amino acid and recognized D-lactate and D,L-isoserine as substrates. On the basis of these characteristics, various depsipeptides can be produced using S137G/Y207F-replaced TmDdl.
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  • Sew-Fen LEU, Song Ling POON, Hsiang-Yin PAO, Bu-Miin HUANG
    2011 Volume 75 Issue 4 Pages 723-731
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Cordycepin, a pure compound of Cordyceps sinensis (CS), is known as an adenosine analog. We have found that CS stimulated Leydig cell steroidogenesis. Here we investigated the in vivo and in vitro effects of cordycepin in primary mouse Leydig cell steroidogenesis. The results indicate that cordycepin increased the plasma testosterone concentration. Cordycepin also stimulated in vitro mouse Leydig cell testosterone production in dose- and time-dependent manners. We further observed that cordycepin regulated the mRNA expression of the A1, A2a, A2b, and A3 adenosine receptors in the mouse Leydig cells, and that antagonists of A1, A2a, and A3 suppressed testosterone production 20–50% testosterone production. Furthermore, Rp-cAMPS (cAMP antagonist) and Protein Kinase A (PKA) inhibitors (H89 and PKI) significantly decreased cordycepin-induced testosterone production, indicating that the PKA-cAMP signal pathway was activated by cordycepin through adenosine receptors. Moreover, cordycepin induced StAR protein expression, and H89 suppressed cordycepin-induced steroidogenic acute regulatory (StAR) protein expression. Conclusively, cordycepin associated with adenosine receptors to activate cAMP-PKA-StAR pathway and steroidogenesis in the mouse Leydig cells.
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  • Mitsuaki SANADA, Kouichi KURODA, Mitsuyoshi UEDA
    2011 Volume 75 Issue 4 Pages 740-747
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Supplementary material
    GTS1 in yeast Saccharomyces cerevisiae is a pleiotropic gene, the expression of which affects a wide range of biological phenomena. A genome-wide transcriptional analysis identified genes upregulated in response to GTS1 induction, most of which were found to be regulated by the Tup1-Cyc8 co-repressor. Among the upregulated genes, FLO1 was indispensable for cell aggregation, one of the pleiotropic effects of GTS1. Deletion of genes encoding the chromatin remodeling complex SWI/SNF abolished FLO1 upregulation and decreased cell aggregation, although the nuclear localization of Gts1p required for cell aggregation was not affected. GTS1 induction caused chromatin remodeling at the FLO1 promoter in a SWI/SNF-dependent manner on micrococcal nuclease accessibility assay. Cyc8p was detected in Gts1p-mediated protein aggregates by agarose gel electrophoresis, indicating that Gts1p inhibited Cyc8p function. These results suggest that inhibition of the Tup1-Cyc8 complex and subsequent SWI/SNF-dependent chromatin remodeling result in Gts1p-mediated gene derepression.
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  • Itaru URIKURA, Tatsuya SUGAWARA, Takashi HIRATA
    2011 Volume 75 Issue 4 Pages 757-760
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Fucoxanthin, a major carotenoid in brown algae, has various beneficial effects. In this study, we evaluated the effect of topical fucoxanthin on UVB-induced skin photoaging in hairless mice. The dorsal skins were treated topically with a 0.001% fucoxanthin solution 2 h each time before UVB irradiation (5 times a week) for 10 weeks. The formation of wrinkles in UVB-irradiated skin treated with vehicle alone significantly increased, as compared with the non-irradiated control. Treatment with fucoxanthin tended to suppress UVB-induced wrinkle formation, but there was no significant difference between wrinkle formation in the control group and the fucoxanthin treatment group. However, topical treatment with fucoxanthin significantly lessened UVB-induced epidermal hypertrophy, VEGF, and MMP-13 expression in the epidermis and thiobarbituric acid reactive substances (TBARS) in the skin. These results indicate that topical treatment with fucoxanthin prevents skin photoaging in UVB-irradiated hairless mice, possibly via antioxidant and antiangiogenic effects.
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Biochemistry & Molecular Biology Notes
Biochemistry & Molecular Biology Communications
  • Yuji TANAKA, Shinya NAKAMURA, Makoto KAWAMUKAI, Nozomu KOIZUMI, Tsuyos ...
    2011 Volume 75 Issue 4 Pages 804-807
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    We made two series of Gateway binary vectors, pGWBs and R4pGWBs, possessing a UDP-N-acetylglucosamine: dolichol phosphate N-acetylglucosamine-1-P transferase (GPT) gene driven by the nopaline synthase promoter (Pnos) as a tunicamycin resistance marker for the transformation of Arabidopsis thaliana. The reporters and tags employed in this system are sGFP, GUS, LUC, EYFP, ECFP, G3GFP, mRFP, TagRFP, 6xHis, FLAG, 3xHA, 4xMyc, 10xMyc, GST, T7, and TAP. Selection of transformants was successful on plates containing 0.15 mg/L of tunicamycin. These vectors were compatible with existing pGWB and R4pGWB vectors for kanamycin, hygromycin B, and BASTA® selection, and are useful new tools for making transgenic Arabidopsis.
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  • Kenji NAKAJIMA, Yoshiki NAKAJIMA
    2011 Volume 75 Issue 4 Pages 808-811
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    The effects of intraperitoneal administration of dimethylsulfonioacetate (DMSA), dimethlsulfoniopropionate (DMSP), and methylmethionine (MeMet) solutions (10 mM each) on the body weights and the hematological parameters (red and white blood cells) of Ehrlich ascites carcinoma (EAC)-bearing mice were examined for up to 10 d. Body weights significantly increased in the EAC-bearing mice treated with and without MeMet in contrast to those with DMSA and DMSP. This increase was attributed to the increased amounts of ascitic fluid. EAC-bearing mice with and without MeMet both showed abnormal values of hematological parameters, while those with DMSA and DMSP exhibited almost normal levels on the 10th day.
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  • Stefano ZANCONATO, Giovanni MINERVINI, Irene POLI, Davide De LUCREZIA
    2011 Volume 75 Issue 4 Pages 812-815
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Phage display relies on an iterative cycle of selection and amplification of random combinatorial libraries to enrich the initial population of those peptides that satisfy a priori chosen criteria. The effectiveness of any phage display protocol depends directly on library amino acid sequence diversity and the strength of the selection procedure. In this study we monitored the dynamics of the selective pressure exerted by the host organism on a random peptide library in the absence of any additional selection pressure. The results indicate that sequence censorship exerted by Escherichia coli dramatically reduces library diversity and can significantly impair phage display effectiveness.
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  • Tadashi HARA, Atsushi TERADA, Hiroki YAMAGUCHI, Takashi NAKASHIMA, Yos ...
    2011 Volume 75 Issue 4 Pages 816-819
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    We investigated the contribution of peripheral stem-loops to the catalytic activity of an archaeal RNase P RNA, PhopRNA, from Pyrococcus horikoshii OT3. PhopRNA mutants, in which the stem-loops were individually deleted, were prepared and characterized with respect to precursor tRNA (pre-tRNA) cleavage activity in the presence of five RNase P proteins. All the mutants retained the activity to some extent, indicating that they are moderately implicated in catalysis. Further characterization suggested that the stem-loops serve largely as binding sites for the proteins, and that their interactions are predominantly involved in stabilization of the active conformation of PhopRNA.
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Food & Nutrition Science Regular Papers
  • Chia-Yu CHANG, Tain-Junn CHENG, Fang-Rong CHANG, Hsien-Yi WANG, Wei-Ch ...
    2011 Volume 75 Issue 4 Pages 624-632
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    It has been reported that medicinal mushrooms might induce different types of immune responses. Anthodia camphorata (A. camphorata) has attracted much attention for its therapeutic effects in treating hepatoma. We tested this anti-tumor effects using immunomodulation of macrophages and extracts of A. camphorata. We evaluated the anti-proliferation effects of various extracts of A. camphorata from fruiting bodies (AC-FB), mycelium of solid-state cultures (AC-SS), liquid-state cultures (AC-LS) and polyaccharide extracts from liquid-state cultures (AC-PS), and extracts of A. camphorata stimulated RAW 264.7 macrophage cell-conditioned mediums (MC-CMs). We measured cell proliferation and, did migration assays by cell cycle analysis and by observing apoptosis-related proteins (AKT, PARP-1, and NF-κB) and the mRNA expression of cytokines (TNF-α and IL-1β) of macrophages in human hepatoma cell lines. Our results revealed that two of the extracts (AC-FB and AC-SS) had better anti-proliferation effects, implying an immunomodulatory role the macrophages might play. This outcome is consistent with findings that AC-FB and AC-SS increase mRNA expression of TNF-α and the corresponding expression of apoptosis-related proteins on activation of MC-CMs, while A. camphorata polysaccharides induce macrophage-derived anti-tumor activities in human hepatoma cells via IL-1β and Akt activation. These results indicate that anti-tumor effects exerted by modulation of macrophage activation of A. camphorate may be influenced by the other constituents which (contained little or no polysaccharide) of A. camphorata.
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  • Hideki TAKAHASHI, Yuri BUNGO, Katsuhiko MIKUNI
    2011 Volume 75 Issue 4 Pages 633-637
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    We investigated the effects of various cyclodextrins (CDs) on the aqueous solubility and thermal stability of α-lipoic acid, a compound with low water solubility. α-CD, β-CD, and γ-CD had little effect on the aqueous solubility of α-lipoic acid. In contrast, 6-O-α-maltosyl-CDs increased it in a concentration-dependent manner, 6-O-α-maltosyl-β-CD enhancing solubility the most. The thermal stability of α-lipoic acid in the solid state was improved by the addition of G2-β-CD®, a commercial product of 6-O-α-maltosyl-β-CD. The thermal stability of α-lipoic acid was also improved by the addition of β-CD. Analysis by differential scanning calorimetry showed that G2-β-CD®, a mixture of maltosyl-β-CDs, and β-CD efficiently formed complexes with α-lipoic acid. These results suggest that the sizes and shapes of these β-CD compounds are compatible with complexation with α-lipoic acid. Moreover, both the formation of an aqueous complex with G2-β-CD® and an insoluble complex with β-CD increased the thermal stability of α-lipoic acid.
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  • Miho DAIFUKU, Yusuke YAGUCHI, Kosuke NISHI, Takeaki OKAMOTO, Hiroyuki ...
    2011 Volume 75 Issue 4 Pages 638-645
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    The immunostimulation effects of yellowtail heart extracts were examined. Screening various parts of the yellowtail viscera, we found that extracts from the yellowtail heart enhanced IgM production by human hybridoma HB4C5 cells. Yellowtail heart extracts heated at 121 °C for 20 min and dialyzed showed the highest IgM production-stimulating activity toward HB4C5 cells. Also, immunoglobulin production by mouse spleen lymphocytes was stimulated by yellowtail heart extracts in vitro, and lymphocytes derived from mice administered the extract for 20 d were activated in vivo. Yellowtail heart extracts were partially purified by anion-exchange chromatography, and fractions containing a 33 kDa-protein exhibited immunostimulating activity. LC-MS/MS analysis revealed that the 33 kDa-protein was most similar to tropomyosin-4 from various fishes. Purified tropomyosin from porcine muscle enhanced IgM production by HB4C5 cells. This means that tropomyosin-4 is one of the immunostimulating substances in the yellowtail heart.
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  • Kayoko KAWAKAMI, Hiroshi NISHIDA, Naoto TATEWAKI, Yuki NAKAJIMA, Tetsu ...
    2011 Volume 75 Issue 4 Pages 650-655
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Persimmon leaf (PL) has been commonly recognized for its wide variety of health benefits. A previous study has reported that persimmon leaf extract (PLE) contained flavonols with the 2″-galloly moiety (PLEg). Galloylated homologues generically show stronger activity in their biological function, so enhanced functions can be expected for PLEg. We investigated in this present study the effect of PLEg on the cellular DNA damage checkpoint signaling to sensitize cancer chemotherapy. Treatment with PLE and PLEg significantly increased the cytotoxicity of doxorubicin (DOX) in A549 adenocarcinoma cells. PLE and PLEg reduced the phosphorylation of checkpoint proteins such as structural maintenance of chromosomes 1 (SMC1), checkpoint kinase 1 (Chk1), and p53 in DOX-treated cells. Moreover, PLE decreased the phosphorylation of ATM (ataxia telangiectasia mutated) in a dose-dependent manner. PLE, and especially PLEg, abrogated the G2/M checkpoint during DOX-induced DNA damage. These results suggest that PLEg specifically inhibited ATM-dependent checkpoint activation by DOX, and that PLEg might be a useful sensitizer in cancer chemotherapy.
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  • Jiyoung KIM, Jung Nam CHOI, Kang Mo KU, Daejung KANG, Jong Sang KIM, J ...
    2011 Volume 75 Issue 4 Pages 674-680
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Supplementary material
    Liquid chromatography tandem mass spectrometry (LCMS/MS)-based metabolite profiling was applied to elucidate the correlation between metabolite release and antioxidant activity during water blanching of Chrysanthemum coronarium L. (CC). Some major metabolites showing differences between fresh CC and blanched CC (BCC) were selected by principal component analysis (PCA) and partial least-square discriminate analysis (PLS-DA) loading plots, and were identified as dicaffeoylquinic acid (DCQA), succinoyl-DCQA, and acetylmycosinol. By PLS regression analysis of the correlation between antioxidant components and effects, candidate antioxidative metabolites were predicted due to strong positive correlations with DCQA and succinoyl-DCQA, and by a relatively weak positive correlation with acetylmycosinol.
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  • Jiyoung KIM, Jung Nam CHOI, Daejung KANG, Gun Hee SON, Young-Suk KIM, ...
    2011 Volume 75 Issue 4 Pages 732-739
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Supplementary material
    Liquid chromatography mass spectrometry and multivariate analysis were employed to investigate the correlation between fermentation time-dependent metabolite changes in cheonggukjang, a traditional fermented soybean product, and changes in its antioxidant activity over 72 h. The metabolite patterns were clearly distinguished not by strains but by fermentation time, into patterns I (0–12 h), II (12–24 h), and III (24–72 h), which appeared as distinct clusters on principal component analysis. The compounds that significantly contributed to patterns I, II, and III were soyasaponins, isoflavonoid derivatives, and isoflavonoid aglycons respectively. Partial least square analysis for metabolite to antioxidant effects showed correlations between the ferric reducing/antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay during 24–36 h, and 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) test and total phenol content (TPC) during 36–72 h. Compared with the strong negative correlations of glucosylated-isoflavonoids with DPPH, ABTS and TPC during fermentation, the isoflavonoid aglycon displayed strong positive correlations with these compounds during fermentation.
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Food & Nutrition Science Note
Microbiology & Fermentation Technology Regular Papers
  • Hiroto MORITA, Haruka ABO, Ayako OKAMOTO, Hiroshi MAEDA, Youhei YAMAGA ...
    2011 Volume 75 Issue 4 Pages 662-668
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Supplementary material
    Gene AO090103000153 is unique to Aspergillus oryzae RIB40 and A. flavus NRRL3357, and is speculated to encode a serine-type carboxypeptidase. In this study, we purified and characterized a heterologously expressed gene product of AO090103000153. 5′-Rapid amplification of cDNA ends indicated that the translation start site of the gene is located 1,586 bp downstream of the translation start site predicted by the genome sequencing project. The gene, starting from the revised translation start codon, termed ocpC, was transcribed constantly in A. oryzae RIB40. Purified recombinant OcpC exhibited the enzymatic properties of a serine-type carboxypeptidase. This protease was stable at temperatures below 45 °C and a low pH, and had broad substrate specificity for N-acylpeptides, but it exhibited significantly lower specific activity and a lower kcat value for substrates than previously reported serine-type carboxypeptidases from A. oryzae.
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  • Tohru KOBAYASHI, Osamu KOIDE, Shigeru DEGUCHI, Koki HORIKOSHI
    2011 Volume 75 Issue 4 Pages 669-673
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    A chitosanase of deep-biosphere Bacillus thuringiensis strain JAM-GG01 was purified. The optimal pH and temperature for the purified enzyme (Cho-GG) were about pH 6 and 60 °C, but Cho-GG was unexpectedly unstable under incubation at over 40 °C. This discrepancy between higher activity and lower stability in the same range of temperature was abolished by the addition of reaction products, chitotriose and chitotetraose. The Cho-GG gene was amplified by PCR and sequenced. The deduced amino acid sequence of Cho-GG showed more than 98% identity to those of other Bacillus enzymes belonging to GH family 8. Although Cho-GG did not show the definite characteristics of a sub-seafloor ectoenzyme, the thermal stability of many chitosanases of B. turingienesis and other related strains can be improved by adding chitotriose or chitotetraose.
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  • Wei ZHAO, Jia ZHENG, Hong-bo ZHOU
    2011 Volume 75 Issue 4 Pages 694-699
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    The combined effect of macronutrients in the extraction medium on α-amylase produced by Bacillus subtilis were studied by using response surface methodology in shaken flask cultures. The production of amylase was significantly affected by the interaction between wheat bran and the cotton seed extract in the extraction medium and by the interaction between the cotton seed extract and starch. The optimal combination in the extraction medium for maximum α-amylase production was determined as 10.80 g·L−1 of wheat bran, 9.90 g·L−1 of cotton seed extract, 0.5 g·L−1 of starch, 2.0 g·L−1 of yeast extract, 5.00 g·L−1 of NaCl and 2.00 g·L−1 of CaCl2. A 12.55-fold increase of enzyme activity was recorded in the optimized medium compared to the result acquired in a minimum essential medium. The optimized medium was used to compare different cultivation strategies in fermenters. The pH-stat strategy for reducing cellular stress response and the substrate concentration-stat strategy for reducing substrate inhibition were independently investigated. The temperature-limited strategy has been proposed to solve the proteolytic digestion problem, although the high-pressure strategy resulted in high productivity. A hybrid strategy simultaneously controlling pH, temperature, substrate concentration and pO2 was finally investigated to enhance the efficiency of the process. This hybrid strategy resulted in high activity of α-amylase, increasing the productivity almost three-fold as compared to an ordinary fed-batch culture.
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  • Chiho SUZUKI, Choong-Soo YUN, Takashi UMEDA, Tsuguno TERABAYASHI, Kazu ...
    2011 Volume 75 Issue 4 Pages 711-717
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Pmr, a histone-like protein H1 (H-NS) family protein encoded on plasmid pCAR1, is a key factor in optimizing gene transcription on both pCAR1 and the host chromosome. To clarify the mode of function of Pmr, we performed gel filtration chromatography analysis and protein-protein cross-linking, and found that Pmr forms homo-oligomers, consisting of its homodimers. We also found, by atomic force microscopy, that Pmr has DNA-bridging capacity. From these results, Pmr was deduced to have features common to H-NS family proteins. Additionally, evaluating protein-DNA affinity is important to clarify the mode of function of Pmr, and hence we performed an electrophoretic mobility shift assay. Though Pmr formed high-order protein-DNA complexes and did not show preference for nucleic acid sequences, the C-terminal region of Pmr did, suggesting that the DNA-binding affinity of Pmr can be evaluated by using its C-terminal region.
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  • Tetsuro MATSUNAGA, Ayumi NAKAYUKI, Yuki SAITO, Akio KATO, Yuichiro NOI ...
    2011 Volume 75 Issue 4 Pages 748-751
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    The periodontopathogenic bacterium Eikenella corrodens has an N-acetyl-D-galactosamine (GalNAc)-specific lectin, that contributes significantly to the pathogenicity of the bacterium. Recently, we reported that plasmid-mediated genomic recombination enhances the activity of this lectin. In this study, we investigated the effects of genomic recombination on certain virulence factors. Introduction of the recombinase gene resulted in hemolysis and significantly increased bacterial adhesion to epithelial cells. It was suggested that the enhanced adhesion was attributable to increased lectin activity due to genomic recombination, because it was inhibited by the addition of GalNAc. In contrast, invasion of the epithelial cells was remarkably reduced by genomic recombination. Although we assumed that this decrease in invasion resulted from a loss of type-IV pili, the phase variant did not show any decrease in invasion activity. This suggests that type-IV pili do not contribute to the invasive ability of E. corrodens. Our results suggest that genomic recombination enhances the pathogenicity of E. corrodens.
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Microbiology & Fermentation Technology Notes
Microbiology & Fermentation Technology Communication
Environmental Science Regular Paper
  • Hideyuki INUI, Taketo WAKAI, Keiko GION, Kiyoshi YAMAZAKI, Yun-Seok KI ...
    2011 Volume 75 Issue 4 Pages 705-710
    Published: April 23, 2011
    Released: May 10, 2011
    [Advance publication] Released: April 22, 2011
    JOURNALS FREE ACCESS
    Supplementary material
    Zucchini cultivars Cucurbita pepo subsp. ovifera cv. Patty Green and subsp. pepo cv. Gold Rush were cultivated hydroponically in a nutrient solution supplemented with a mixture of dioxins and dioxin-like compounds. Patty Green and Gold Rush showed low and high accumulation of these compounds in the aerial parts respectively. In both cultivars, the accumulation of each congener negatively depended on its hydrophobicity. This suggests that desorption and solubilization were partly responsible for congener specificity of accumulation, since this was not found in soil experiments. In contrast, no clear difference in accumulation in the roots was observed between the cultivars, whereas the translocation factors, which are indicators of efficient translocation from the roots to the aerial parts, differed among the congeners hydrophobicity-dependently. There were positive correlations between accumulation in the roots and the hydrophobicity of the polychlorinated biphenyl congeners in both cultivars. These results indicate that translocation was also partly responsible for the congener specificity and accumulation concentrations.
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