Microbial oxygenases and dehalogenases are key enzymes in the degradation of highly chlorinated compounds, which often become significant environmental pollutants. Oxygenases engineered by the methods of directed evolution exhibit enhanced degradation of PCBs and other chlorinated solvents such as trichloroethene and pentachloroethane. Dehalorespiration is an efficient dechlorination mechanism that is coupled with energy-yielding phosphorylation. Recently, a variety of chloroethene-dehalorespiring anaerobes have been isolated, and their reductive dehalogenases have been characterized in biochemical and genetic bases. This review describes our recent studies on dioxygenases and reductive dehalogenases.

抄録全体を表示

The active ingredients of ‘golden root’ of Scutellaria baicalensis GEORGI (Huang-Qin), a valuable traditional Chinese medicine, are polyhydroxyflavonoids, namely baicalein, oroxylin A and wogonin. With the objective of overcoming their poor solubility and to investigate their structure and activity relationships, baicaleinyl 7-O-sulfate was prepared, and extensive comparative antioxidative and anti-inflammatory tests were conducted. All the polyhydroxyflavonoids exhibited significant antioxidative and free-radical scavenging activities. In respect of their nitric oxide (NO) inhibition, wogonin was superior to all the other flavonoids, while oroxylin A was most potent in the inhibition of lipid peroxidation. Wogonin proved to be the most potent (82.9% inhibition, p<0.05) in its anti-inflammatory activity against carrageenan-induced rat hind paw edema. There was a correlation between the in-vivo anti-inflammatory activity and the in-vitro antioxidative activities.

抄録全体を表示

A MUC1 type of glycopeptide was synthesized by the 9-fluorenylmethoxycarbonyl (Fmoc) solid-phase peptide synthesis (SPPS) protocol using benzyl and benzylidene-protected β-D-Gal-(1→3)-β-D-GalNAc-Ser/Thr (TF-β: a stereoisomer of the Thomsen-Friedenreich antigen). The synthetic glycopeptide was released from the resin with reagent K, and the resulting benzylated glycopeptide was deprotected under conditions of low-acidity trifluoromethanesulfonic acid (TfOH). The glycopeptide carrying duplicate non-natural O-glycans was dominant in the products, but was accompanied by a considerable amount of the glycopeptide missing one of the O-glycans. In contrast, the native α-glycoside was relatively stable under the acidic debenzylation conditions as shown by a parallel experiment with the glycopeptide involving α-D-GalNAc-Ser/Thr linkage. Enzymatic glycosylation with CMP-NeuAc was successful with both natural and non-natural O-glycans of the synthetic glycopeptide.

抄録全体を表示

Gerfelin, an inhibitor of human geranylgeranyl diphosphate (GGPP) synthase that has been isolated from a culture broth of Beauveria felina QN22047, was synthesized in 4 and 3 steps starting from 2,4-dihydroxy-6-methylbenzoic acid and 3,4,5-trihydroxytoluene, respectively. An effective ligand, 2-(di-tert-butylphosphino)biphenyl, was used in the palladium-catalyzed diaryl ether-forming reaction. Five analogous compounds of gerfelin were also synthesized for a study of the structure-activity relationship.

抄録全体を表示

A pair of enantiomers of trans-p-menth-2-en-1-ol, an aggregation pheromone of Platypus quercivorus, was synthesized from (S)- and (R)-limonene. The retention time of the aggregation pheromone from the insect coincided with that of (1S,4R)-p-menth-2-en-1-ol synthesized from (S)-limonene from GC analyses with a chiral column, enabling the absolute configuration of the aggregation pheromone to be determined as (1S,4R).

抄録全体を表示

An enantioselective synthesis of the (1S,5R)-enantiomer of litseaverticillols A and B was accomplished in line with our previously reported synthetic pathway for their (1R,5S)-enantiomer. The use of “EtSCeCl₂” prepared from EtSLi and CeCl₃, instead of previously employed EtSLi itself, for the formation of thiol ester intermediates prevented any undesirable epimerization occurring in the process.

抄録全体を表示

Cochliophilin A (5-hydroxy-6,7-methylenedioxyflavone, 1), known as a host-specific attractant towards the zoospores of Aphanomyces cochlioides, a cause of root rot and damping-off diseases of Chenopodiaceae, was found in the Amaranthaceae plant, Celosia cristata, that is susceptible to the pathogen. The content of 1 in Celosia seedlings was quantified as 1.4 μg/g fresh weight. A new isoflavone, cristatein (5-hydroxy-6-hydroxymethyl-7,2′-dimethoxyisoflavone, 2), and five known flavonoids were also identified.

抄録全体を表示

Bioassay-guided fractionation of an ethanol extract of the brown alga, Dictyota dichotoma, led to the isolation of a novel chlorine-containing perhydroazulene diterpene together with two known diterpenes, dictyolactone and sanadaol. The structure of the novel compound, named dictyol J, was elucidated on the basis of spectroscopic information. Dictyolactone showed the highest algicidal activity against red-tide phytoplanktons among the three isolated compounds.

抄録全体を表示

Aspergillus nidulans possessed 16 putative amylolytic genes consisting of 7 α-glucosidase (agdA-F), 7 α-amylase (amyA-F), and 2 glucoamylase (glaA and B) genes on the genome. Among them, the agdA, agdB, agdE, agdF, amyA, amyB, amyF, and glaB genes were induced by isomaltose. AmyR, a Zn(II)₂Cys₆ transcription factor, was required for the induction. The isomaltose-inducible genes possessed at least one consensus sequence for AmyR binding, 5′-CGGN₈CGG, on each promoter region. None of the amylolytic genes was induced by maltose.
The mRNA levels of the amylolytic genes except for agdC, amyD, and amyG increased under carbon-starved conditions. Release from CreA-dependent carbon catabolite repression was the main cause of the increase, but, the mRNA levels of agdB, agdF, amyB, amyF, and glaB increased to some extent even in a creA mutant. Therefore, both CreA-dependent and -independent mechanisms are involved in the up-regulation of the amylolytic genes under carbon-starved conditions.

抄録全体を表示

The effects of cadmium stress on growth, morphology, and protein expression were investigated in Rhodobacter capsulatus B10 using two-dimensional polyacrylamide gel electrophoresis and a scanning electron microscope with an energy dispersive X-ray spectrometer. The bacterium grew in the presence of 150 μM CdCl₂ and highly induced heat-shock proteins (GroEL and Dnak), S-adenosylmethionine synthetase, ribosomal protein S1, aspartate aminotransferase, and phosphoglycerate kinase. Interestingly, the ribosomal protein S1 was proportionally expressed as the amount of cadmium in the medium, suggesting that S1 may be required for the repair of cadmium-mediated cellular damage. On the other hand, we identified five cadmium-binding proteins: 2-methylcitrate dehydratase, phosphate peripalsmic binding protein, inosine-5′-monophosphate dehydrogenase/guanosine-5′-monophosphate reductase, inositol monophosphatase, and lytic murein transglycosylase. The cadmium-treated cells had a filamentous structure and contained less phosphorous than the untreated cells. We propose that these characteristics of the cadmium-treated cells may be due to the inactivation of the phosphate peripalsmic binding protein and lytic murein transglycosylase by cadmium.

抄録全体を表示

Transcripts of thionin genes encoding antimicrobial peptides were present at a high level in rice coleoptiles just after germination, and decreased to an undetectable level after about 3 d, but this decline was suppressed by co-treatment with gibberellic acid (GA₃) and brassinolide (BL). The temporal expression patterns of key enzyme genes for the biosyntheses of gibberellins (GAs) and brassinosteroids (BRs) were correlated with the fluctuation of thionin mRNAs. Jasmonic acid (JA) replaced the effect of GA₃ and BL, and its change in endogenous level was parallel to that of the thionin genes. These results strongly suggest that thionin gene expression was positively regulated by JA, whose endogenous level was synergistically regulated by GAs and BRs. In contrast, thionin gene expression in etiolated seedlings remained high while the endogenous level of JA was low, suggesting the presence of another signaling pathway in the dark to maintain the thionin level.

抄録全体を表示

An enzyme that has both β-1,4-glucanase and chitosanase activities was found in the culture medium of the soil bacterium Lysobacter sp. IB-9374, a high lysyl endopeptidase-producing strain. The enzyme was purified to homogeneity from the culture filtrate using five purification steps and designated Cel8A. The purified Cel8A had a molecular mass of 41 kDa, as estimated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. A pH optimum of 5.0 was found for the β-1,4-glucanase activity, and pH optima of 5.0 and 7.0 were found for the chitosanase activity. Nucleotide sequencing of the Cel8A gene yielded a deduced amino acid sequence that comprises a 33-amino acid, N-terminal signal peptide and a mature enzyme consisting of a 381-residue polypeptide with a predicted molecular mass of 41,241 Da. The amino acid sequence of the Cel8A, which contains the catalytic module of glycosyl hydrolase family 8, is homologous to β-1,3-1,4-D-glucanase from Bacillus circulans WL-12 and endoglucanase N-257 from B. circulans KSM-N257.

抄録全体を表示

Small heat shock protein (sHSP) is a family of ubiquitous polypeptides involved in a variety of physiological phenomena. From the silkworm, Bombyx mori, we isolated and sequenced the following cDNAs encoding sHSPs: shsp19.9, shsp20.1, shsp20.4, shsp20.8, shsp21.4, and shsp23.7. shsp21.4 was nearly twice as large in size as other shsps. The deduced amino acid sequence of sHSP21.4 was similar to that of Drosophila melanogaster CG14207-PA. Other sHSPs were highly similar to each other and, in a phylogenetic tree, formed a cluster including Plodia interpunctella αCP25. It was speculated that shsp21.4 has at least one intron in genome while other shsps do not. The transcripts of all shsps were subtle, but were constitutively detected in various tissues. Heat shock triggered a substantial increase in the transcripts other than shsp21.4. The B. mori sHSPs are perhaps classified into at least two groups: sHSP21.4 and others.

抄録全体を表示

The cellular target of curvularol, a G₁-specific cell-cycle inhibitor of mammalian cells, was identified by a genetic approach in Saccharomyces cerevisiae. Since the wild-type W303 strain was highly resistant to curvularol, a drug hypersensitive parental strain was constructed in which various genes implicated in general drug resistance had been disrupted. Curvularol resistant mutants were isolated, and strains that exhibited a semi-dominant, curvularol-specific resistance phenotype were selected. All five strains examined were classified into a single genetic complementation group designated YCR1. A mutant gene responsible for curvularol resistance was identified as an allele of the RPL3 gene encoding the ribosomal protein L3. Sequence analysis of the mutant genes revealed that Trp255Cys and Trp255Leu substitutions of Rpl3p are responsible for curvularol resistance. Rpl3p mutants in which Trp255 residue was replaced by other amino acids were constructed. All of these replacements led to varying degrees of increased resistance to curvularol and growth defects.

抄録全体を表示

In the detergent industry, fungal endoglucanases are used to release microfibrils from the surfaces of dyed cellulosic fabrics to enhance color brightness. Family 45 endoglucanase (glycoside hydrolase family 45, GH45) EGL3 from Humicola grisea is more resistant to anionic surfactants and oxidizing agents than family 45 endoglucanase RCE1 from Rhizopus oryzae, while in the present study, a catalytic domain of RCE1 had higher defibrillation activity on dyed cotton fabrics than did that of EGL3. To identify the amino acid regions involved in these properties, we compared the characteristics of RCE1, EGL3, and three chimeric endoglucanases, in which each of the three regions of the catalytic domain of EGL3 was replaced by the corresponding region of the catalytic domain of RCE1. Amino acids in the N-terminal region were involved in resistance to anionic surfactants and oxidizing agents. Furthermore, amino acids in the region adjacent to the N-terminal region were involved in releasing microfibrils and in binding to dyed cotton fabrics, indicating that the binding of the amino acids in this region might be important in the release of microfibrils from dyed cotton fabrics.

抄録全体を表示

Enzymatic removal of p-alkylphenols from aqueous solutions was investigated through the two-step approach, the quinone conversion of p-alkylphenols with mushroom tyrosinase (EC 1.14.18.1) and the subsequent adsorption of quinone derivatives enzymatically generated on chitosan beads at pH 7.0 and 45 °C as the optimum conditions. This technique is quite effective for removal of various p-alkylphenols from an aqueous solution. The % removal values of 97–100% were obtained for p-n-alkylphenols with carbon chain lengths of 5 to 9. In addition, removal of other p-alkylphenols was enhanced by increasing either the tyrosinase concentration or the amount of added chitosan beads, and their % removal values reached >93 except for 4-tert-pentylphenol. This technique was also applicable to remove 4-n-octylphenol (4NOP) and 4-n-nonylphenol (4NNP) as suspected endocrine disrupting chemicals. The reaction of quinone derivatives enzymatically generated with the chitosan’s amino groups was confirmed by the appearance of peaks for UV–visible spectrum measurements of the chitosan films incubated in the p-alkylphenol and tyrosinase mixture solutions. In addition, 4-tert-pentylphenol underwent tyrosinase-catalyzed oxidation in the presence of hydrogen peroxide.

抄録全体を表示

A unique N-linked glycosylation motif (Asn⁷⁹-Tyr-Thr) was found in the sequence of type-A feruloyl esterases from Aspergillus spp. To clarify the function of the flap, the role of N-linked oligosaccharides located in the flap region on the biochemical properties of feruloyl esterase (AwFAEA) from Aspergillus awamori expressed in Pichia pastoris was analyzed by removing the N-linked glycosylation recognition site by site-directed mutagenesis. N79 was replaced with A or Q. N-glycosylation-free N79A and N79Q mutant enzymes had lower activity than that of the glycosylated recombinant AwFAEA wild-type enzyme toward α-naphthylbutyrate (C4), α-naphthylcaprylate (C8), and phenolic acid methyl esters. Kinetic analysis of the mutant enzymes indicated that the lower catalytic efficiency was due to a combination of increased K_m and decreased k_cat for N79A, and to a considerably decreased k_cat for N79Q. N79A and N79Q mutant enzymes also exhibited considerably reduced thermostability relative to the wild-type.

抄録全体を表示

The occurrence of positional isomers of minor C₂₄ unsaturated fatty acids in female gonad lipids of the limpet Cellana grata was clarified by gas chromatography–mass spectrometry of the combination of their 4,4-dimethyloxazoline and picolinyl ester derivatives. In this study, in addition to 5,9-24:2, 9,15-24:2, 5,9,15-24:3, and 5,9,17-24:3, previously identified, 24:4n-6, 24:5n-3, and four novel nonmethylene-interrupted fatty acids, 9,17-24:2, 9,15,18-24:3, 5,9,15,18-24:4, and 5,9,15,18,21-24:5, were newly recognized. All C₂₄ unsaturated fatty acids detected were present only in triacylglycerols.

抄録全体を表示

In our previous paper (Kimura, Y., et al., Biosci. Biotechnol. Biochem., 67, 1852–1856, 2003), we found that a complex type N-glycans containing β1-3 galactose residue occurs on royal jelly glycoproteins. During structural analysis of minor components of royal jelly N-glycans, we found complex type N-glycans bearing both galactose and N-acetylgalactosamine residues. Detailed structural analysis of pyridylaminated oligosaccharide revealed that the newly found N-glycan had a complex type structure harboring a tumor marker (T-antigen) unit: Galβ1-3GalNAcβ1-4GlcNAcβ1-2Manα1-6 (Galβ1-3GalNAcβ1-4GlcNAcβ1-2Manα1-3) Manβ1-4GlcNAcβ1-4GlcNAc. To our knowledge, this may be the first report of the presence of the T-antigen unit in the N-glycan moiety of eucaryotic glycoproteins.

抄録全体を表示

Bovine β-lactoglobulin (β-LG) was conjugated with chitosan (CHS) by means of a water-soluble carbodiimide to reduce the immunogenicity of β-LG. Each β-LG-CHS conjugate was purified by ion-exchange chromatography and hydrophobic chromatography. The conjugation between β-LG and CHS was confirmed by SDS–PAGE, the isoelectric point of the conjugate being higher than that of β-LG. Two types of the β-LG-CHS conjugate were obtained with molar ratios of β-LG to CHS of 1:1 (F1) and 1:2 (F2). Structural analyses by fluorescence measurement, ELISA with monoclonal antibodies and retinol-binding activity indicated that the conjugates had almost maintained the native structure of β-LG. The antigenicity of the β-LG-CHS conjugates was similar to that of β-LG in C3H/He mice. Reduction of the immunogemicity of β-LG was achieved by conjugation with CHS. In particular, F2 showed very low immunogenicity. B cell epitopes of β-LG and the conjugates recognized in C3H/He mice were determined with 15-mer multi-pin peptide; the linear epitope profiles of the conjugates were found to be similar to those of β-LG, while the antibody response to each epitope was dramatically reduced. Conjugation of β-LG with chitosan was effective for reducing the immunogenicity of β-LG.

抄録全体を表示

Encapsulated calcium in liposome (L-Ca) produced by using egg phosphatidyl choline in the laboratory was injected into rabbit to evaluate the effect of calcium injection on the ageing of meat. After injecting L-Ca into the blood vessels of rabbit to increase the Ca²⁺ concentration in the body for 24 h, the fragmentation rate of myofibrils was observed. The fragmentation rates in the loin from the control group and L-Ca injected group were 2.56% and 3.10% in 2 days, 12.27% and 16.18% in 6 days, and 33.56% and 49.60% in 10 days, respectively (p<0.05). SDS–PAGE patterns of connectin and nebulin show that the total degradation of connectin by the control group took longer than 2–3 days, while it was within 1 day for the L-Ca-injected group. The control group took 8–10 days for nebulin, while the L-Ca-injected group took 2–3 days for total degradation. These results indicate that, injecting L-Ca into rabbit was effective for reducing the ageing period of meat without resulting in any physical shock or contamination.

抄録全体を表示

The hyperhomocysteinemia induced by a dietary addition of 1% methionine was significantly suppressed by the concurrent addition of 1% glycine or 1.4% serine to the same degree. The methionine-induced increase in the hepatic concentration of methionine metabolites was significantly suppressed by glycine and serine, but the hepatic cystathionine β-synthase activity was not enhanced by these amino acids. When the methionine-supplemented diet was changed to the methionine plus glycine or serine diet, the plasma homocysteine concentration rapidly decreased during and after the first day. The hyperhomocysteinemia induced by an intraperitoneal injection with methionine was also suppressed by concurrent injection with glycine or serine, although the effect of serine was significantly greater than that of glycine. These results indicate that glycine and serine were effective for suppressing methionine-induced hyperhomocysteinemia: serine and its precursor glycine are considered to have elicited their effects mainly by stimulating cystathionine synthesis by supplying serine, another substrate for cystathionine synthesis.

抄録全体を表示

We examined in rats whether or not the prolonged ingestion of recrystallized amylose (RCA) would prevent the development of insulin resistance. Rats were fed on a diet containing waxy cornstarch (WCS) as carbohydrate or a diet containing 30% RCA in place of WCS for 18 wk. Glucose tolerance test (GTT) was conducted at every four weeks. On wk 16, the plasma insulin response as assessed by the area under the curve was lower in the RCA diet group than in the WCS diet group. The fasting plasma insulin level tended to increase over time in both groups, but was lower in the RCA diet group on wk 16. An autopsy revealed that the adipose tissue mass and serum free fatty acid concentrations were significantly higher in the WCS diet group. The results suggest that prolonged ingestion of RCA had the effect of slowing the development of insulin resistance through a lower concentration of serum free fatty acids, presumably due to the prevention of adipocyte hypertrophy.

抄録全体を表示

We examined the dietary effects of cyclic nigerosylnigerose (CNN), a dietary indigestible oligosaccharide with four D-glucopyranosyl residues linked by alternating α-(1→3)- and α-(1→6) glucosidic linkages, on the intestinal immune function of mice, and the effects were compared with those of α-(1→3)-linked oligosaccharide (nigerooligosaccharides, NOS) or α-(1→6)-linked oligosaccharide (isomaltooligosaccharides, IMO). BALB/c mice were fed with 1–5% CNN, 5% IMO, or 12.5% NOS for 4 weeks, and the intestinal mucosal immune responses were determined. In the 1–5% CNN fed groups, the amounts of IgA in feces increased significantly. In addition, IgA, transforming growth factor-β1 (TGF-β1), and interleukin-6 (IL-6) secretion by Peyer’s patch (PP) cells were enhanced in CNN fed mice. In the 5% CNN group, pH in the cecum decreased, and the amounts of lactic acid and butyric acid increased. These findings were not observed in the NOS- or IMO-fed group of mice. They suggest that CNN supplementation changes the intestinal environment of microflora and indirectly enhances the immune function in the gut.

抄録全体を表示

Brewer’s and baker’s yeasts appear to have components that protect from liver injury. Whether sake yeast, Saccharomyces cerevisiae Kyokai no. 9, also has a hepatoprotective effect has not been examined. Here we show that sake yeast suppresses acute alcoholic liver injury in mice. Male C57BL/6 mice that had been fed a diet containing 1% sake yeast for two weeks received three doses of ethanol (5 g/kg BW). In the mice fed sake yeast, ethanol-induced increases in triglyceride (TG) and glutamate pyruvate transaminase (GPT) were significantly attenuated and hepatic steatosis was improved. In addition, sake yeast-fed mice showed a smaller decrease in hepatic S-adenosylmethionine (SAM) level and a smaller increase in plasma homocysteine (Hcy) level after ethanol treatment than the control mice, suggesting that a disorder of methonine metabolism in the liver caused by ethanol was relieved by sake yeast. These results indicate that sake yeast protects against alcoholic liver injury through maintenance of methionine metabolism in the liver.

抄録全体を表示

We screened myoga extracts for inhibitors of human platelet aggregation and human 5-lipoxygenase. We identified a novel labdane type of diterpene, together with three known diterpenes (miogadial and galanals A and B) from the flower buds of myoga. Spectroscopic data indicated the structure of the new compound to be 12(E)-labdene-15,16,(8β)17-trial (miogatrial). Miogatrial and miogadial were potent inhibitors of human platelet aggregation and human 5-lipoxygenase (5-LOX). The sesquiterpene, polygodial, also exhibited strong inhibitory activity against human platelet aggregation and 5-LOX. On the other hand, galanals A and B did not have inhibitory activity in either experimental system. It thus appears that a 3-formyl-3-butenal structure was essential for the potent inhibition of human platelet aggregation and human 5-LOX.

抄録全体を表示

Tienchi ginseng tea was prepared from stems and leaves of Tienchi ginseng which is a special product in China. The increase in systolic blood pressure (SBP) was significantly inhibited by the consumption of a 4% this tea solution as drinking water from the prehypertensive stage (6 weeks of age) in male stroke-prone spontaneously hypertensive rats (SHRSP). A similar intake from the hypertensive stage also showed an anti-hypertensive effect. In contrast, a similar intake had no effect on SBP of normotensive Wistar Kyoto rats. We found that the rhizome of Tienchi ginseng contained two types of saponin: 20(s)-protopanaxadiol (PPD) such as ginsenosides Rb₁ and Rd having a hypotensive effect, and 20(s)-protopanaxatriol (PPT) such as ginsenosides Rg₁ and Re having a hypertensive effect. In contrast, the tea sample contained PPD and gamma-aminobutyric acid (GABA), but no PPT. These results suggest that drinking this tea infusion would be useful for controlling hypertension.

抄録全体を表示

Royal jelly (RJ) has diverse physiological and pharmacological functions. We observed its weak estrogenic activity in the previous study. RJ stimulated the proliferation of mouse osteoblast-like MC3T3-E1 cells at 0.1 mg/ml, and the effect was blocked by the specific estrogen receptor antagonist ICI 182,780. The addition of 0.1–1.0 mg/ml RJ enhanced collagen production in culture medium. Oral administration of RJ to normal female mice for 9 weeks increased the ash content of their tibiae. DNA microarray analysis revealed significant changes in gene expression related to extracellular matrix formation when the femurs of mice fed RJ were analyzed. Quantitative reverse transcriptase-PCR (RT-PCR) confirmed up-regulation of procollagen I α1 gene expression. These data suggest that RJ as a whole or some of its individual components stimulates production of type I collagen and other activities for bone formation through action on osteoblasts.

抄録全体を表示

Some blue pigments were formed in the D-xylose (1 M)-glycine (0.1 M) reaction system. A novel blue pigment, designated as Blue-M2 (blue Maillard intermediate-2), was identified as 5-[1,4-dicarboxymethyl-5-(2,3-dihydroxypropyl)-1,4-dihydropyrrolo[3,2-b]pyrrole-2-ylmethylene]-1,4-dicarboxymethyl-2-{5-[N-carboxymethyl(2,3,4-trihydroxytetrahydrofuran-2-yl)methylamino]-2-hydroxymethyl-4-(1,2,3-trihydroxypropyl)tetrahydrofuran-3-yl}-4,5-dihydropyrrolo-[3,2-b]pyrrole-1-ium. Blue-M2 is presumed to have been generated by the reaction between Blue-M1, which was identified as the major blue pigment in a previous paper (Hayase et al., Biosci. Biotechnol. Biochem., 63, 1512–1514 (1999)), and di-D-xyluloseglycine. Blue pigments are important Maillard reaction intermediates through the formation of melanoidins.

抄録全体を表示

The performance-enhancing effects of Pseudosasa japonica were investigated in mice using an adjustable-current water pool. Compared to the control group, a 1.5-fold increase in swimming time was observed in the mouse group administered an 80% ethanol extract (PJE) of the leaves of P. japonica. The blood lactate level, an important indicator of fatigue, was significantly lower (28%, P<0.05) in PJE group than in the control group. These results suggest that PJE possesses stimulatory effects that can enhance exercise endurance and reduce fatigue.

抄録全体を表示

Eighty-seven healthy volunteers ingested a purple-fleshed sweet potato beverage with various contents of anthocyanin (beverage A; 22.1 mg/250 ml, B; 107.8, C; 84.9). An acylated anthocyanin, peonidin 3-caffeoylsophoroside-5-glucoside, was detected in the urine 2 h after ingestion. The concentrations were 15.1±2.2 μg/l of urine (mean±SEM), 46.6±5.3, and 53.3±2.2 for beverages A, B, and C respectively.

抄録全体を表示

We investigated the effects of dietary iron deficiency on bone metabolism by measuring markers of bone turnover in rats. Twelve 3-week-old male Wistar-strain rats were fed a control diet or an iron-deficient diet for 4 weeks. Dietary iron deficiency decreased hemoglobin concentration and increased heart weight. Serum osteocalcin concentration, bone mineral content, bone mineral density, and mechanical strength of the femur were significantly lower in the iron-deficient group than in the control group. These results suggested that dietary iron deficiency affected bone, which might have been due to a decrease in bone formation in rats.

抄録全体を表示

The effects of several types of whole fruits and vegetables on human lymphocytic DNA were investigated by using two versions of the comet assay. The total antioxidative capacity, as the FRAP value, and ascorbic acid (AA) content were also measured to explore the relationship between the effect and antioxidant content.

抄録全体を表示

We investigated the effects of herb extracts, Rhus verniciflua, Agrimonia pilosa, Sophora japonica, and Paeonia suffruticosa, on the lowering of blood glucose levels and thiobarbituric acid reactive substances (TBARS) in streptozotocin (STZ)-induced diabetic rats. After 4 weeks, oral administration of Rhus verniciflua extract (50 mg/kg) exhibited a significant decrease in blood glucose levels in diabetic rats (P<0.05). Blood TBARS concentrations, the products of glucose oxidation in blood, were also lowered by Rhus verniciflua extract supplementation. In addition, Sophora japonica and Paeonia suffruticosa extracts significantly reduced TBARS levels versus diabetic controls. Serum concentrations of liver-function marker enzymes, GOT and GPT, were also restored by Rhus verniciflua (50 mg/kg) supplementation in diabetic rats.

抄録全体を表示

Chlorogenic acid possessed a weak caffeine-like psychostimulant property when assessed for its effect on spontaneous locomotor activity in mice. In the evaluation of the effects for the major metabolites of chlorogenic acid which were detected upon incubation with rat feces and/or excreted in urine after oral administration to rats, caffeic and m-coumaric acids were found to be the principal active metabolites, while the others contributed little to this caffeine-like psychostimulant activity.

抄録全体を表示

We investigated a temperature adaptation of Bacillus subtilis 168 in which chromosomal groEL was replaced with a psychrophilic groEL. This strain can grow at 50 °C but not at 51 °C, a temperature at which wild-type B. subtilis can grow. Using in vivo random mutagenesis by the B. subtilis mutator strain (mutS, mutM, mutY), two thermo-adaptants were isolated from the groEL substituted strain at 52 °C. They contained novel amino acid alterations in their ATP binding motif (T93I) and the inter-monomer contact (R285H) region of GroEL. These results suggest that GroEL participates in bacterial temperature adaptation.

抄録全体を表示

In order to establish the molecular phylogeny of the genus Rhizopus, three molecules of the ribosomal RNA-encoding DNA (rDNA), complete 18S, internal transcribed spacer (ITS)1-5.8S-ITS2, and 28S D1/D2 regions of all the species of the genus were sequenced. Phylogenetic trees showed three major clusters corresponding to the three groups in the current morphological taxonomy, microsporus-group, stolonifer-group, and R. oryzae. R. stolonifer var. lyococcos was clustered independently from the major clusters. R. schipperae clustered differently in all trees. Strains of R. sexualis had multiple ITS sequences. A. rouxii clustered with R. oryzae. These results indicate the possibility of molecular identification of species groups using rDNA sequencing. Reclassification of the genus might be appropriate.

抄録全体を表示

The chitinolytic bacterium Aeromonas hydrophila strain SUWA-9, which was isolated from freshwater in Lake Suwa (Nagano Prefecture, Japan), produced several kinds of chitin-degrading enzymes. A gene coding for an endo-type chitinase (chiA) was isolated from SUWA-9. The chiA ORF encodes a polypeptide of 865 amino acid residues with a molecular mass of 91.6 kDa. The deduced amino acid sequence showed high similarity to those of bacterial chitinases classified into family 18 of glycosyl hydrolases. chiA was expressed in Escherichia coli and the recombinant chitinase (ChiA) was purified and examined. The enzyme hydrolyzed N-acetylchitooligomers from trimer to pentamer and produced monomer and dimer as a final product. It also reacted toward colloidal chitin and chitosan with a low degree of deacetylation. When cells of SUWA-9 were grown in the presence of colloidal chitin, a 60 kDa-truncated form of ChiA that had lost the C-terminal chitin-binding domain was secreted.

抄録全体を表示

Acetobacter tropicalis SKU1100 produces a pellicle-forming capsular polysaccharide (CPS), consisting of galactose, glucose, and rhamnose. We cloned the galE gene, a UDP-galactose synthesis gene, from A. tropicalis SKU1100 by PCR. A galE-disruptant was prepared and found not to produce CPS and thus not to form a pellicle under the static condition. Instead, the ΔgalE mutant secreted an extracellular polysaccharide (EPS), which was purified and found to have a unique character, different from the original CPS.

抄録全体を表示

3-Dehydroshikimate was formed with a yield of 57–77% from quinate via 3-dehydroquinate by two successive enzyme reactions, quinoprotein quinate dehydrogenase (QDH) and 3-dehydroquinate dehydratase, in the cytoplasmic membranes of acetic acid bacteria. 3-Dehydroshikimate was then reduced to shikimate (SKA) with NADP-dependent SKA dehydrogenase (SKDH) from the same organism. When SKDH was coupled with NADP-dependent D-glucose dehydrogenase (GDH) in the presence of excess D-glucose as an NADPH re-generating system, SKDH continued to produce SKA until 3-dehydroshikimate added initially in the reaction mixture was completely converted to SKA. Based on the data presented, a strategy for high SKA production was proposed.

抄録全体を表示