Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Volume 75, Issue 3
Displaying 1-37 of 37 articles from this issue
Award Reviews
  • Kazumitsu UEDA
    2011 Volume 75 Issue 3 Pages 401-409
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Human MDR1, a multi-drug transporter gene, was isolated as the first of the eukaryote ATP Binding Cassette (ABC) proteins from a multidrug-resistant carcinoma cell line in 1986. To date, over 25 years, many ABC proteins have been found to play important physiological roles by transporting hydrophobic compounds. Defects in their functions cause various diseases, indicating that endogenous hydrophobic compounds, as well as water-soluble compounds, are properly transported by transmembrane proteins. MDR1 transports a large number of structurally unrelated drugs and is involved in their pharmacokinetics, and thus is a key factor in drug interaction. ABCA1, an ABC protein, eliminates excess cholesterol in peripheral cells by generating HDL. Because ABCA1 is a key molecule in cholesterol homeostasis, its function and expression are highly regulated. Eukaryote ABC proteins function on the body surface facing the outside and in organ pathways to adapt to the extracellular environment and protect the body to maintain optimal health.
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  • Shigeaki KATO, Ryoji FUJIKI
    2011 Volume 75 Issue 3 Pages 410-413
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    The Editorial Board of Bioscience, Biotechnology, and Biochemistry has recently confirmed that the Award Review article above written by Shigeaki KATO and Ryoji FUJIKI, contains a number of references to previous publications by the same authors, several of which contain some problematic images, which have resulted in the conclusions of those papers to be brought into question and those papers retracted. These images include fabrication and falsification of images through inappropriate processing and duplicate use of identical image data for different publications. These instances of misconduct have been confirmed by the University of Tokyo Scientific Research Code of Conduct Committee which raised questions over the validity of the Award Review article, published in Bioscience, Biotechnology, and Biochemistry. Following COPE guidelines and in consultation with the author, the Japan Society for Bioscience, Biotechnology, and Agrochemistry has decided to rescind the award given to the authors. After discussion with the authors, the Editorial Board and the authors have jointly decided to retract this Award Review article from Bioscience, Biotechnology, and Biochemistry.

    As Editor-in-Chief, I regret the time that peer reviewers and others spent on reviewing this paper.
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Analytical Chemistry Regular Papers
  • Zhong-Lan SHEN, Dong YUAN, Qing-De SU, Hui ZHANG, Jun WANG, Jian-Hua Z ...
    2011 Volume 75 Issue 3 Pages 473-479
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    An analytical methodology for the analysis of methamidophos in water and soil samples incoporating a molecularly imprinted solid-phase extraction process using methamidophos-imprinted polymer was developed. Binding study demonstrated that the polymer exhibited excellent affinity and high selectivity to the methamidophos. Evidence was also found by FT-IR analysis that hydrogen bonding between the CO2H in the polymer cavities and the NH2 and P=O of the template was the origin of methamidophos recognition. The use of molecularly imprinted solid-phase extraction improved the accuracy and precision of the GC method and lowered the limit of detection. The recovery of methamidophos extracted from a 10.0 g soil sample at the 100 ng/g spike level was 95.4%. The limit of detection was 3.8 ng/g. The recovery of methamidophos extracted from 100 mL tap and river water at 1 ng/mL spike level was 96.1% and 95.8%, and the limits of detection were 10 and 13 ng/L respectively. These molecularly imprinted solid-phase extraction procedures enabled selective extraction of polar methamidophos successfully from water and soil samples, demonstrating the potential of molecularly imprinted solid-phase extraction for rapid, selective, and cost-effective sample pretreatment.
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  • Satoshi INOUYE, Jun-ichi SATO, Satoko SASAKI, Yuiko SAHARA
    2011 Volume 75 Issue 3 Pages 568-571
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    The fusion protein of streptavidin to aequorin (STA-AQ) was highly purified from inclusion bodies in Escherichia coli cells and applied to a bioluminescent sandwich immunoassay. α-Fetoprotein (AFP), which is a serological marker of liver cancer, was used as a model analyte to test STA-AQ in an immunoassay. The measurable range of AFP by the sandwich immunoassay, using the complex of STA-AQ and the biotinylated anti-AFP antibody, was 0.02–200 ng/mL with an average coefficient of variation of 4.9%. The detection sensitivity with the complex of STA-AQ and the biotinylated anti-AFP antibody was similar to that with the complex of biotinylated aequorin, streptavidin and the biotinylated anti-AFP antibody. STA-AQ would be a useful reporter protein for immunoassays.
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Organic Chemistry Regular Paper
  • Kazutoshi SHINDO, Ayako TACHIBANA, Ayumi TANAKA, Shizuka TOBA, Emi YUK ...
    2011 Volume 75 Issue 3 Pages 505-510
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    We performed combinational bioconversion of substituted naphthalenes with PhnA1A2A3A4 (an aromatic dihydroxylating dioxygenase from marine bacterium Cycloclasticus sp. strain A5) and prenyltransferase NphB (geranyltransferase from Streptomyces sp. strain CL190) or SCO7190 (dimethylallyltransferase from Streptomyces coelicolor A3(2)) to produce prenyl naphthalen-ols. Using 2-methylnaphthalene, 1-methoxynaphthalene, and 1-ethoxynaphthalene as the starting substrates, 10 novel prenyl naphthalen-ols were produced by combinational bioconversion. These novel prenyl naphthalen-ols each showed potent antioxidative activity against a rat brain homogenate model. 2-(2,3-Dihydroxyphenyl)-5,7-dihydroxy-chromen-4-one (2′,3′-dihydroxychrysin) generated with another aromatic dihydroxylating dioxygenase and subsequent dehydrogenase was also geranylated at the C-5′-carbon by the action of NphB.
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Biochemistry & Molecular Biology Regular Papers
  • Michael Angus KREZEL, Linda Adriana REZMANN, Naghmeh VARGHAYEE, Josefa ...
    2011 Volume 75 Issue 3 Pages 414-418
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Supplementary material
    To investigate the expression of the unknown angiotensin II type 2 receptor interacting protein (ATIP) isoforms in the rat we used the known sequences of human and mouse ATIP to design sequencing primers to enable us to sequence rat ATIP3 and ATIP4. Exon 4, which is present in human but not mouse ATIP, was not identified in the coding region of rat ATIP. The expression levels of these genes in a range of rat tissues were examined, and we concluded that there is little similarity in the relative tissue distribution of the various ATIP isoforms in rat and human.
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  • Li LI, Wei LI, Sang-Won JUNG, Yong-Woo LEE, Yong-Ho KIM
    2011 Volume 75 Issue 3 Pages 434-442
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    The protective effects of decursin (D) and decursinol angelate (DA) purified from Angelica gigas Nakai on amyloid β-protein (Aβ)-induced neurotoxicity and the underlying mechanisms were investigated. Aβ plays a major role in the pathogenesis of Alzheimer’s disease (AD) by eliciting oxidative stress. It significantly increased cytotoxicity and lipid peroxidation, but decreased glutathione contents and antioxidant enzyme activities. All of these results were markedly reversed by pretreatment with D or DA. Nuclear transcription factor Nrf2, which regulates the expression of antioxidant enzymes, was significantly increased by D or DA pretreatment. Furthermore, D and DA suppressed Aβ aggregation. These results suggest that D and DA increase cellular resistance to Aβ-induced oxidative injury in the rat pheochromocytoma (PC12) cells, presumably through not only the induction of Nrf2 and related antioxidant enzymes, but also the anti-aggregation of Aβ. Thus D and DA have therapeutic potential in treating AD and other oxidative stress-related diseases.
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  • Xu LIU, Lan HONG, Xiao-Yun LI, Yao YAO, Bo HU, Ling LI
    2011 Volume 75 Issue 3 Pages 443-450
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Supplementary material
    The NAC (NAM, ATAF, and CUC) proteins share a highly conserved NAC domain and constitute a large family of plant-specific transcriptional factors. We have isolated a drought-induced NAC gene from Arachis hypogaea, named AhNAC2 (Arachis hypogaea NAC2) but its specific role remains unknown. In this study, we found that transgenic Arabidopsis overexpressing AhNAC2 lines were hypersensitive to ABA in root growth, seed germination, and stomatal closure compared to wild type Arabidopsis. The transgenic lines exhibited enhanced tolerance to drought and salinity stress, and the expression levels of 12 stress-related genes in the AhNAC2 transformed plants were higher than in wild type Arabidopsis. These results indicate that AhNAC2 is a major player in the NAC gene family involved in ABA signaling. Its role as a candidate gene for engineering drought and salt tolerance in cultivated plants is discussed.
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  • Seung-Soo KIM, So-Ra KIM, Jung-Rok KIM, Jin-Kyoo MOON, Bong-Hwan CHOI, ...
    2011 Volume 75 Issue 3 Pages 451-458
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Supplementary material
    There are phenotypic differences between Korean native pig (KNP) and Yorkshire (YS) breeds due to different interests in selection. YS has been selected for industrial interests such as a growth rate and lean meat production, while KNP has been maintained as a regional breed with local interests such as disease resistance and fat content in and between muscle. A comparison of gene expression profiles from liver tissue reflected overall long-term effects of artificial selection for these two pig breeds. Based on minimum positive false discovery rate (less than 10%) and fold change (|FC|>1.5), 73 differentially expressed genes (DEGs) were identified. Functional analysis of these DEGs indicated clear distinctions in signaling capacity related to epidermal growth factor (EGF), extracellular structure, protein metabolism, and detoxification. Hepatic DEGs demonstrated the importance of hormonal and metabolic capabilities to differences between these two pig breeds.
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  • Shuhei SO, Tatsuo MURAKAMI, Naoki IKEDA, Takahito CHIJIWA, Naoko ODA-U ...
    2011 Volume 75 Issue 3 Pages 480-488
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    The cDNAs encoding venom phospholipase A2 (PLA2) inhibitors (PLIs), named Protobothrops elegans (Pe)γPLI-A, PeγPLI-B, PeαPLI-A, and PeαPLI-B, were cloned from the P. elegans liver cDNA library. They were further divided into several constituents due to nucleotide substitutions in their open reading frames. For PeαPLI-A, two constituents, PeαPLI-Aa and PeαPLI-Ab, were identified due to three nonsynonymous substitutions in exon 3. Far-western blot and mass-spectrometry analysis of the P. elegans serum proteins showed the presence of γPLIs, and αPLIs, which can bind venom PLA2s. In αPLIs from Protobothrops sera, A or B subtype-specific amino acid substitutions are concentrated only in exon 3. A comparison of γPLIs showed that γPLI-As are conserved and γPLI-Bs diversified. Mathematical analysis of the nucleotide sequences of Protobothrops γPLI-B cDNAs revealed that the particular loops in the three-finger motifs diversified by accelerated evolution. Such evolutionary features should have made serum PLIs acquire their respective inhibitory activities to adapt to venom PLA2 isozymes.
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  • Mitsuhiko KUWAHARA, Takashi TAMURA, Kentaro KAWAMURA, Kenji INAGAKI
    2011 Volume 75 Issue 3 Pages 516-521
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Mammalian thioredoxin reductases (TrxRs) contain selenium as selenocysteine (Sec) in the C-terminal redox center –Gly-Cys-Sec-Gly-OH to reduce Trx and other substrates; a Sec-to-Cys substitution in mammalian TrxR yields an almost inactive enzyme. The corresponding tetrapeptide sequence in Drosophila melanogaster TrxR (Dm-TrxR), –Ser-Cys-Cys-Ser-OH, endows the orthologous enzyme with a catalytic competence similar to mammalian selenoenzymes, but implementation of the Ser-containing tetrapeptide sequence SCCS into the mammalian enzyme does not restore the activity of the Sec-to-Cys mutant form (turnover number <2/min). MOPAC calculation suggested that the C-terminal hexapeptide Pro-Ala-Ser-Cys-Cys-Ser-OH functions as a redox center that alleviates the necessity for selenium in Dm-TrxR, and a mutant form of human lung TrxR that mimics this hexapeptide sequence showed improved catalytic turnover (17.4/min for DTNB and 13.2/min for E. coli trx) compared to the Sec-to-Cys mutant. MOPAC calculation also suggested that the dominant form of the Pro-containing hexapeptide is a C+ conformation, which perhaps has a catalytic advantage in facile reduction of the intramolecular disulfide bond between Cys497 and Cys498 by the N-terminal redox center in the neighboring subunit.
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  • Takayuki ISHIMARU, Kazunari ITO, Miho TANAKA, Syunpei TANAKA, Naotoshi ...
    2011 Volume 75 Issue 3 Pages 544-549
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    To provide a molecular explanation of the role of the disulfide (SS) bridge in the thermostability and structural integrity of ovalbumin (OVA), we prepared SS-mutated OVAs in which SS-forming residues were replaced by Ala or Ser (C73A, C73S, C120A, and C73/120A), and compared the conformation, thermostability, susceptibility to elastase, and formation of heat-stable OVA (S-OVA) with those of the wild-type. The circular dichroism (CD) and tryptophan fluorescence spectra revealed that the SS-mutated OVAs assumed a native-like conformation similar to the wild-type. The thermal denaturation temperature for the SS-mutated OVAs was significantly lower than that for the wild-type. C73S, C120A, and C73/120A mutants converted to S-OVA on alkaline treatment. Analyses for elastase digestion fragments showed that a non-native SS bridge was generated in all SS-mutated OVAs, but non-native SS-pairing did not contribute to thermostability. Hence, we concluded that the presence of the original SS bridge in OVA contributes to conformational stability but is not directly related to the conversion to S-OVA.
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  • Hajime NAKATANI, Naohito AOKI, Daita NADANO, Tsukasa MATSUDA
    2011 Volume 75 Issue 3 Pages 550-555
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    The interaction between mammary epithelial and stromal tissue is considered to be important in breast tissue development. In this study, we developed a transplantation procedure for the mammary stromal fibroblastic cell line (MSF) to examine its life in vivo. First we established MSF cells which stably expressed lacZ (lacZ/MSF) and had characteristics of mammary stromal cells. The lacZ/MSF cells were then transplanted into a cleared mammary fat pad of syngenic mice with and without mammary primary epithelial organoids. Whole mount X-gal and carmine staining of the transplants revealed that a number of undifferentiated lacZ/MSF cells survived around the mammary epithelial tissue when transplanted with organoids. These results indicate that transplantation of MSF cells into mammary fat pad was accomplished by co-transplantation with primary mammary organoids. Finally, we discuss the application of transplantation procedure for in vivo studies of the mammary stromal tissue development and stromal-epithelial interactions.
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  • Takeshi HAYASHI, Nobue TAKAMATSU, Takashi NAKASHIMA, Takashi ARITA
    2011 Volume 75 Issue 3 Pages 556-560
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    We encountered a fourth case of honey allergy in Japan. We characterized and identified the IgE-binding proteins in honey using the serum of a honey-allergenic patient. Immunoblot analysis revealed that IgE in the patient serum specifically bound to four proteins in each honey sample. At least three of these IgE-binding proteins were N-linked glycoproteins. To identify the 60-kDa IgE-binding protein in dandelion honey, the N-terminal sequences of the fragmented protein were analyzed, revealing the protein to be major royal jelly protein 1 (MRJP 1). Three IgE-binding proteins removed of N-linked oligosaccharide showed a large reduction in IgE-binding activity as compared with the intact protein. This suggests that the carbohydrates in the IgE-binding proteins are a major epitope for patient IgE.
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  • Takuya SAKAMOTO, Takehiro KAMIYA, Kaori SAKO, Junji YAMAGUCHI, Mutsumi ...
    2011 Volume 75 Issue 3 Pages 561-567
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Supplementary material
    RPTs (regulatory particle triple-A-ATPase) are components of 26S proteasome. We found novel roles of RPT2a and RPT5a in Zn deficiency-tolerance. Arabidopsis thaliana mutants carrying T-DNA in RPT2a and RPT5a were more sensitive to Zn deficiency than the wild-type. In the rpt mutants, the shoot Zn contents were similar to those of the wild-type. Transcripts of Zn deficiency-inducible genes were highly accumulated in the rpt mutants, suggesting that the rpt mutants suffer from various Zn deficiency symptoms, although the Zn levels are not reduced. Lipid peroxidation levels, known to be increased under Zn deficiency, were higher in the rpt mutants than in the wild-type. Poly-ubiquitinated proteins were accumulated upon exposure to Zn deficiency, especially in the rpt mutants. Overall, this study indicates that RPT2a and RPT5a are involved in Zn deficiency-tolerance, possibly through alleviation of oxidative stresses and/or processing of poly-ubiquitinated proteins.
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Biochemistry & Molecular Biology Note
Food & Nutrition Science Regular Papers
  • Yu ISHIDA, Yusuke SHIBATA, Ikuo FUKUHARA, Yuki YANO, Isao TAKEHARA, Ky ...
    2011 Volume 75 Issue 3 Pages 427-433
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    A double-blind, placebo-controlled, randomized clinical trial was conducted to evaluate the effects of ingesting an excess of tablets containing casein hydrolysate, incorporating angiotensin I-converting enzyme (ACE) inhibitory peptides such as Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP), in subjects with blood pressure ranging from normal to mild hypertension. A total of 48 subjects were given either 5 times more than the effective amount of casein hydrolysate or a placebo in tablet form for 4 weeks. In the active group, systolic blood pressure (SBP) decreased significantly as compared with the placebo group. In stratified analysis, however, this antihypertensive effect was not found in normotensive subjects. In addition, neither an acute or nor an excessive reduction in blood pressure nor clinically important adverse events were observed in this study. These findings suggest that intake of a 5-fold excess of tablets containing casein hydrolysate can lead to a mild improvement in hypertension without side effects.
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  • Jung-Bum LEE, Chihiro YAMAGISHI, Kyoko HAYASHI, Toshimitsu HAYASHI
    2011 Volume 75 Issue 3 Pages 459-465
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Three antiviral and immunostimulating substances (LC1, LC2 and LC3) were isolated from a hot water extract of seeds of Pimpinella anisum by combination of anion-exchange, gel filtration and hydrophobic interaction column chromatographies. Chemical and spectroscopic analyses revealed them to be lignin-carbohydrate-protein complexes. These lignin-carbohydrate complexes (LCs) showed antiviral activities against herpes simplex virus types 1 and 2 (HSV-1 and -2), human cytomegalovirus (HCMV) and measles virus. LCs were also found to interfere with virus adsorption to the host cell surface and directly inactivate viruses. Furthermore, they enhanced nitric oxide (NO) production by inducing iNOS mRNA and protein expression in RAW 264.7 murine macrophage cells. The induced mRNA expression of cytokines including IL-1β and IL-10 was also apparent. These results suggest that the lignin-carbohydrate-protein complexes from P. anisum possessed potency as functional food ingredients against infectious diseases.
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  • Takashi FUJII, Katsumi IKEDA, Morio SAITO
    2011 Volume 75 Issue 3 Pages 489-495
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    The compounds present in rose hips exerting an inhibitory action against melanogenesis in B16 mouse melanoma cells were investigated by dividing an aqueous extract of rose hips (RE) into four fractions. The 50% ethanol eluate from a DIAION HP-20 column significantly reduced the production of melanin and was mainly composed of procyanidin glycosides. We also found that this 50% ethanol eluate reduced the intracellular tyrosinase activity and also had a direct inhibitory effect on tyrosinase obtained as a protein mixture from the melanoma cell lysate. We also investigated the effect of orally administering RE on skin pigmentation in brown guinea pigs, and found that the pigmentation was inhibited together with the tyrosinase activity in the skin. These data collectively suggest that proanthocyanidins from RE inhibited melanogenesis in mouse melanoma cells and guinea pig skin, and could be useful as a skin-whitening agent when taken orally.
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  • Masashi HIGUCHI, Junichi OSHIDA, Koichi ORINO, Kiyotaka WATANABE
    2011 Volume 75 Issue 3 Pages 496-499
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Wheat bran had a protective effect against diquat toxicity in rats fed a purified diet (PD). We studied the effects of wheat bran on the antioxidant system in the liver of rats treated with saline and diquat. Although feeding wheat bran did not affect the concentration of hepatic non-protein sulfhydryl or the activity of glucose 6-phosphate dehydrogenase in the saline-injected rats, these values were significantly higher in the rats fed PD containing wheat bran (W-PD) than in rats fed only PD after administering diquat. The glutathione peroxidase and reductase activities were significantly elevated by wheat bran in the saline-injected rats. Although the glutathione peroxidase activity was unchanged in both the PD-fed rats and W-PD-fed rats after the diquat treatment, the glutathione reductase activity was significantly decreased in both the PD-fed and W-PD-fed rats. Feeding the rats with PD containing 0.15 ppm selenium as well as with W-PD elevated the activity of hepatic glutathione peroxidase and attenuated the diquat toxicity. These results indicate that wheat bran protected against diquat toxicity by activating the hepatic antioxidant system, and that selenium was the key antioxidant in wheat bran.
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  • Robert Antoni OLEK, Wieslaw ZIOLKOWSKI, Jan Jacek KACZOR, Tomasz Henry ...
    2011 Volume 75 Issue 3 Pages 500-504
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Although a number of studies have focused on the higher ethyl pyruvate antioxidative activity than its sodium salt under various stress conditions, and the greater protective properties of the ester form have been suggested as the effect of better cell membrane penetration, the molecular mechanism has remained unclear. The aim of the present study was therefore to compare the antioxidative activities of sodium and ethyl pyruvate under in vitro conditions by using a liver homogenate as the model for cell membrane transport deletion. The potential effect of ethanol was also evaluated, and hypochlorous acid was used as an oxidant. Our data indicate the concentration-dependent scavenging potency of both sodium and ethyl pyruvate, with the ester having higher activity. This effect was not related to the presence of ethanol. Better protection of the liver homogenate by ethyl pyruvate was also apparent, despite the fact that cell membrane transport was omitted.
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Food & Nutrition Science Notes
Microbiology & Fermentation Technology Regular Papers
  • Pattaraporn YUKPHAN, Taweesak MALIMAS, Yuki MURAMATSU, Wanchern POTACH ...
    2011 Volume 75 Issue 3 Pages 419-426
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Isolates AH11T and AH13T were isolated from flowers of lantana and candle bush respectively collected in Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates formed an independent cluster, which was then connected to the type strain of Saccharibacter floricola. The calculated pair-wise 16S rRNA gene sequence similarities of isolate AH11T were 95.7–92.3% to the type strains of the type species of the 12 genera of acetic acid bacteria. The DNA base composition was from 51.2 to 56.8 mol % G+C, with a range of 5.6 mol %. When isolate AH11T was labeled, DNA-DNA similarities were 100, 12, 4, 5, and 4% respectively to isolates AH11T and AH13T and the type strains of Saccharibacter floricola, Gluconobacter oxydans, and Acetobacter aceti. The two isolates were non-motile and did not oxidize either acetate or lactate. No growth was found in the presence of 0.35% acetic acid w/v. The two isolates were not osmophilic but osmotolerant, produced 2,5-diketo-D-gluconate from D-glucose, and did not oxidize lactate, thus differing from strains of Saccharibacter floricola, which showed weak lactate oxidation. The two isolates contained unsaturated C18:1ω7c fatty acid as the major fatty acid, and were unique in the presence of a considerable amount of straight-chain C18:12OH fatty acid. Q-10 was present as the major isoprenoid quinone. Neokomagataea gen. nov. was proposed with the two species, Neokomagataea thailandica sp. nov. for isolate AH11T (=BCC 25710T=NBRC 106555T), which has 56.8 mol % G+C, and Neokomagataea tanensis sp. nov. for isolate AH13T (=BCC 25711T=NBRC 106556T), which has 51.2 mol % G+C.
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  • Michiro TABATA, Ryo ENDO, Michihiro ITO, Yoshiyuki OHTSUBO, Ashwani KU ...
    2011 Volume 75 Issue 3 Pages 466-472
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    A γ-hexachlorocyclohexane (HCH)-degrading bacterium, Sphingomonas sp. MM-1, was isolated from soil contaminated with HCH isomers. Cultivation of MM-1 in the presence of γ-HCH led to the detection of five γ-HCH metabolites, γ-pentachlorocyclohexene, 2,5-dichloro-2,5-cyclohexadiene-1,4-diol, 2,5-dichlorohydroquinone, 1,2,4-trichlorobenzene, and 2,5-dichlorophenol, strongly suggesting that MM-1 has the lin genes for γ-HCH degradation originally identified in the well-studied γ-HCH-degrading strain Sphingobium japonicum UT26. Southern blot, PCR amplification, and sequencing analyses indicated that MM-1 has seven lin genes for the conversion of γ-HCH to β-ketoadipate (six structural genes, linA to linF, and one regulatory gene, linR). MM-1 carried four plasmids, of 200, 50, 40, and 30 kb. Southern blot analysis revealed that all seven lin genes were dispersed across three of the four plasmids, and that IS6100, often found close to the lin genes, was present on all four plasmids.
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  • Yuya SUGIMURA, Tatsuro HAGI, Takayuki HOSHINO
    2011 Volume 75 Issue 3 Pages 511-515
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    We measured the adhesion of candidate probiotic lactic acid bacteria (LAB) to carp intestinal mucus. The percentage of adherent bacteria varied among strains. Four strains, two with high adhesion and two with low adhesion in vitro, were tested for in vivo colonization ability. Carp were fed LAB-containing feed for 12 d, and then unsupplemented feed until day 33, and the numbers and compositions of intestinal LAB were analyzed during the entire period. LAB with lower in vitro adhesion disappeared quickly from the intestine after LAB feeding stopped. LAB with higher in vitro adhesion remained in the intestine 3 weeks after LAB feeding stopped, indicating a strong correlation between mucus adhesion in vitro and colonization ability in vivo. Next we isolated nine candidate probiotic LAB with high in vitro mucus-binding ability. Three of them were fed to carp, and all three were stably maintained in the intestine.
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  • Thanongsak CHAIYASO, Ampin KUNTIYA, Charin TECHAPUN, Noppol LEKSAWASDI ...
    2011 Volume 75 Issue 3 Pages 531-537
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
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    Cellulase-free xylanase production by thermophilic Streptomyces thermovulgaris TISTR1948 was cultivated in a basal medium with rice straw as sole source of carbon and as an inducible substrate. Variable medium components were selected in accordance with the Plackett-Burman experimental design. The optimization conditions of physical factors (pH and temperature levels) were then combined in further studies through the response surface methodology approach. Only two significant components, rice straw and yeast extract, were chosen for the optimization studies. A second-order quadratic model was constructed by central composite design (CCD). The model revealed that both pH and temperature levels were significant, and were dependent on xylanase production. Under these experimental designs, the xylanase yield increased from 51.11 to 274.49 U/mL (3,400 to 10,000 U/g of rice straw) or about 537% higher than an unoptimized basal medium. The optimum conditions to achieve maximum yield of xylanase were 27.45 g/L of rice straw and 5.42 g/L of yeast extract under relatively neutral conditions of pH 7.11, 50.03 °C, and a incubation period.
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  • Thitinard NITHERANONT, Akira WATANABE, Yasuhiko ASADA
    2011 Volume 75 Issue 3 Pages 538-543
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
    JOURNAL FREE ACCESS
    A major laccase isozyme (Lac 1) was isolated from the culture fluid of an edible basidiomycetous mushroom, Grifola frondosa. Lac 1 was revealed to be a monomeric protein with a molecular mass of 71 kDa. The N-terminal amino acid sequence of Lac 1 was highly similar to those of laccases of some other white-rot basidiomycetes. Lac 1 showed the typical absorption spectrum of a copper-containing enzyme. The enzyme was stable in a wide pH range (4.0 to 10.0), and lost no activity up to 60 °C for 60 min. The optimal pH of the enzyme activity varied among substrates. The Km values of Lac 1 toward 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), 2,6-dimethoxyphenol, guaiacol, catechol, and 3,4-dihydroxy-L-phenylalanine were 0.0137 mM, 0.608 mM, 0.531 mM, 2.51 mM, and 0.149 mM respectively. Lac 1 activity was remarkably inhibited by the chloride ion, in a reversible manner. Lac 1 activity was also inhibited by thiol compounds.
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Microbiology & Fermentation Technology Notes
Environmental Science Regular Paper
  • Shinichi MIYASHITA, Shoko FUJIWARA, Mikio TSUZUKI, Toshikazu KAISE
    2011 Volume 75 Issue 3 Pages 522-530
    Published: March 23, 2011
    Released on J-STAGE: April 05, 2011
    Advance online publication: March 07, 2011
    JOURNAL FREE ACCESS
    Supplementary material
    We examined the short-term metabolic processes of arsenate for 24 h in a freshwater unicellular green alga, Chlamydomonas reinhardtii wild-type strain CC-125. The arsenic species in the algal extracts were identified by high-performance liquid chromatography/inductively coupled plasma mass spectrometry after water extraction using a sonicator. Speciation analyses of arsenic showed that the levels of arsenite, arsenate, and methylarsonic acid in the cells rapidly increased for 30 min to 1 h, and those of dimethylarsinic acid and oxo-arsenosugar-glycerol also tended to increase continuously for 24 h, while that of oxo-arsenosugar-phosphate was quite low and fluctuated throughout the experiment. These results indicate that this alga can rapidly biotransform arsenate into oxo-arsenosugar-glycerol for at least 10 min and then oxo-arsenosugar-phosphate through both reduction of incorporated arsenate to arsenite and methylation of arsenite and/or arsenate retained in the cells to dimethylarsinic acid via methylarsonic acid as an possible intermediate.
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Environmental Science Note
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