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21.
24S-hydroxycholesterol (HC) is most abundant oxysterols in the brain, passes through blood brain barrier, and is therefore regarded as an intermediary for brain cholesterol elimination. We reported that large-conductance Ca2+- and voltage-activated K+ (slo1 BK) channels are suppressed by this oxysterol, which is presumably intercalated into cell membrane to access the outer surface of the channel. Such an outer approach would make it difficult to interact with the inner, ion-conducting part of the channel. The present findings showed that 24R-HC, the racemic counterpart of 24S-HC, also suppressed slo1 BK channel but in a different voltage-dependent manner. There was a difference between the effects of the two enantiomers on activation kinetics but not on deactivation kinetics. It is suggested that the chirality contributes to the efficacy of channel blockers that act from outer lipophilic parts of channels, as with those which act on the inner, ion-permeable surface. 相似文献
22.
Kaliappan Gopal Perumal Nagarajan Jose Jedy Avinash T. Raj S. Kalai Gnanaselvi Parveen Jahan Yogendra Sharma Esaki M. Shankar Jerald M. Kumar 《PloS one》2013,8(6)
Abdominal aortic aneurysm (AAA) is a common chronic degenerative disease characterized by progressive aortic dilation and rupture. The mechanisms underlying the role of α-tocopherol and β-carotene on AAA have not been comprehensively assessed. We investigated if α-tocopherol and β-carotene supplementation could attenuate AAA, and studied the underlying mechanisms utilized by the antioxidants to alleviate AAA. Four-months-old Apoe−/− mice were used in the induction of aneurysm by infusion of angiotensin II (Ang II), and were orally administered with α-tocopherol and β-carotene enriched diet for 60 days. Significant increase of LDL, cholesterol, triglycerides and circulating inflammatory cells was observed in the Ang II-treated animals, and gene expression studies showed that ICAM-1, VCAM-1, MCP-1, M-CSF, MMP-2, MMP-9 and MMP-12 were upregulated in the aorta of aneurysm-induced mice. Extensive plaques, aneurysm and diffusion of inflammatory cells into the tunica intima were also noticed. The size of aorta was significantly (P = 0.0002) increased (2.24±0.20 mm) in the aneurysm-induced animals as compared to control mice (1.17±0.06 mm). Interestingly, β-carotene dramatically controlled the diffusion of macrophages into the aortic tunica intima, and circulation. It also dissolved the formation of atheromatous plaque. Further, β-carotene significantly decreased the aortic diameter (1.33±0.12 mm) in the aneurysm-induced mice (β-carotene, P = 0.0002). It also downregulated ICAM-1, VCAM-1, MCP-1, M-CSF, MMP-2, MMP-9, MMP-12, PPAR-α and PPAR-γ following treatment. Hence, dietary supplementation of β-carotene may have a protective function against Ang II-induced AAA by ameliorating macrophage recruitment in Apoe−/− mice. 相似文献
23.
Toru Nakayama Nobuyoshi Esaki Woon-Joo Lee Izumi Tanaka Hidehiko Tanaka Kenji Soda 《Bioscience, biotechnology, and biochemistry》2013,77(9):2367-2369
Four types of β-xylosidases from a concentrated culture filtrate of Pénicillium wortmanni IFO 7237, designated as xylosidase-1, -2, -3, and -4 were purified to homogeneity on SDS polyacrylamide gel electrophoresis by an alcohol precipitation, DEAE-Sephadex A-25 ion exchange chromatography, and isoelectric focusing. The molecular weights of xylosidase-1, -2, -3, and -4 were estimated to be 110,000, 195,000, 210,000, and 180,000 respectively and their isoelectric points to be 3.7, 4.28, 4.6, and 4.8. The pH optima of β-xylosidase activities were from 3 to 4.5. The optimum temperature for enzyme activities was from 55°C to 65°C. On the enzymic hydrolysis of phenyl ß-d- xyloside, the reaction product of each enzyme was found to be β-d-xylose with retention of configuration. All the four ß-xylosidases were free of α-xylosidase and ß-glucosidase activities. All the enzyme activities of four β-xylosidases were strongly inhibited by Hg2+ and N- bromosuccinimide. With respect to the hydrolysis patterns and HPLC analysis of hydrolyzates from xylooligosaccharides, xylosidase-2 was totally different from other three as a distinct enzyme. Xylosidase-1 was also in a separate group although xylosidase-3 and -4 showed closely related action patterns as a different group. 相似文献
24.
Tadanobu Nakadai Seiichi Nasuno Nobuyoshi Iguchi 《Bioscience, biotechnology, and biochemistry》2013,77(8):1343-1352
To elucidate the constitution of peptidases from Aspergillus oryzae, systematic separation of the enzymes was carried out by batchwise treatment with Amberlite IRC-50 and precipitation with rivanol. Proteases were separated to two fractions. They were Amberlite IRC-50 adsorbed and the non-adsorbed fractions and the latter fraction was further separated to two fractions, rivanol precipitable and non-precipitable fractions.Acid carboxypeptidase I was purified from the rivanol non-precipitable fraction by column chromatography on DEAE-cellulose, DEAE-Sephadex A-50 and SE-cellulose. The purified enzyme was not homogeneous on disc electrophoresis, although symmetric peaks were obtained for enzyme protein and activity in Sephadex gel filtration. The optimum pH is at pH 4.0 for carbobenzoxy-l-alanyl-l-glutamic acid. The enzyme activity was inhibited by SH reagents, but not inhibited by metal chelating agents. The molecular weight of the enzyme was estimated to be about 120,000 by gel filtration. 相似文献
25.
Tadanobu Nakadai Seiichi Nasuno Nobuyoshi Iguchi 《Bioscience, biotechnology, and biochemistry》2013,77(9):1481-1488
Acid carboxypeptidase III from Aspergillus oryzae was purified from the rivanol non-precipitated fraction. The optimum activity of the enzyme occurred at pH 3.0 for carbobenzoxy-l-glutamyl-l-tyrosine. The enzyme was inhibited by diisopropylphosphorofluoridate and SH reagents such as p-chloromercuribenzoate and monoiodoacetate, but not by such metal chelating agents as ethylenediaminetetraacetate, αα′-dipyridyl and o-phenanthroline. The molecular weight of the enzyme was estimated to be about 61,000. The enzyme hydrolyzed the peptides that possess masked or bulky N-terminal. 相似文献
26.
Nobuyoshi Esaki Hidehiko Tanaka Edith Wilson Miles Kenji Soda 《Bioscience, biotechnology, and biochemistry》2013,77(12):2861-2864
The α2β2 complex of tryptophan synthase from Escherichia coli catalyzes β-replacement reactions of l-serine and its derivatives (e.g., β-chloro-l-alanine and O-methyl-Dl-serine) with various alkanethiols. The products from thiobenzyl alcohol and ethanethiol were isolated to demonstrate the enzymatic synthesis of the corresponding S-substituted l-cysteines. Reactivities of various S-substituent donors were examined, and thiols such as thiobenzyl alcohol, 1-propanethiol and 1-butanethiol were found to be much more efficient substituent donors than the physiological substrate, indole. In addition, tryptophan synthase catalyzes β-replacement reactions of l-threonine with thiols to form the corresponding S-substituted β-methylcysteines, which are also produced by β-addition reactions of l-vinylglycine with thiols. These enzymatic reactions facilitate the synthesis of various sulfur-containing amino acids. 相似文献
27.
Tornio Kimura Nobuyoshi Esaki Hidehiko Tanaka Kenji Soda 《Bioscience, biotechnology, and biochemistry》2013,77(12):3157-3159
Methods are investigated for evaluating the kinetic parameters in a modified Monod’s equation which give the best fit to the growth thermograms for bacterial cultures observed in batch calorimeters. Four mathematical methods were employed as parameter fitting techniques. The growth thermograms observed for soil microbes cultured with glucose as a limiting substrate were used as the objects of the analysis. For the calculation of the heat evolution rate, the Runge-Kutta method, which is commonly used for the numerical analysis, was employed. A comparison of the results obtained by the four methods in terms of closeness of fit to the actual thermograms showed that optimization by direct searching with the Simplex method is the most effective procedure for obtaining the best values of the parameters to reproduce the observed thermograms. 相似文献
28.
αsl-Casein can be made either soluble or insoluble by adjusting the concentration of coexisting calcium ions. In this study, we tried to make a soluble-insoluble interconvertible enzyme through the formation of a conjugate of an enzyme and αsl-casein using a heterobifunctional crosslinking reagent, N-succinimidyl 3-(2-pyridyldithio)propionate. The conjugate of phosphoglyceromutase and native αs1-casein did not exhibit sufficient calcium-dependent precipitation. However, conjugates of enzymes (phosphoglyceromutase, enolase or peroxidase) and αsl-casein polymerized by transglutaminase precipitated almost completely in the presence of more than 50 mM CaCl2. Most of the enzyme conjugates precipitated as calcium caseinates could be solubilized reversibly with EDTA, without a significant loss of activity. A mixture of the enzyme ? polymerized αs1-casein conjugates prepared with phosphoglyceromutase, enolase and pyruvate kinase could catalyze sequential reactions which convert d-3-phosphoglycerate into pyruvate with the same efficiency as a mixture of free enzymes. These results indicate that conjugates of enzymes and polymerized αs1-casein can be useful as soluble-insoluble interconvertible enzymes. 相似文献
29.
Hiroko Shimoi Shinji Nagata Nobuyoshi Esaki Hidehiko Tanaka Kenji Soda 《Bioscience, biotechnology, and biochemistry》2013,77(12):3375-3381
The leucine dehydrogenase (l-leucine: NAD+ oxidoreductase, deaminating, EC 1.4.1.9) gene of Clostridium thermoaceticum was cloned and expressed in Escherichia coli C600 with a vector plasmid, pICD242, which was constructed from pBR322 and the leucine dehydrogenase gene derived from C. thermoaceticum. The enzyme overproduced in the clone was purified about 12 fold to homogeneity by heat treatment and another two steps with a yield of 46%. The enzyme of E. coli- pICD242 was immunochemically identical with that of C. thermoaceticum. The enzyme has a molecular weight of about 350,000 and consists of six subunits identical in molecular weight (56,000). The enzyme is not inactivated by heat treatment: at pH 7.2 and 75°C for 15 min; at 55°C and various pH’s between 6.0 and 10.0 for 10 min. The enzyme catalyzes the oxidative deamination of branched-chain l-amino acids and the reductive amination of their 2-oxo analogues in the presence of NAD+ and NADH, respectively. The pro-S hydrogen at C-4 of the dihydronicotin- amide ring of NADH is exclusively transferred to the substrate; the enzyme is B stereospecific. The enzymological properties are very similar to those of the Bacillus stearothermophilus enzyme [T. Ohshima, S. Nagata and K. Soda, Arch. Microbiol., 141, 407 (1985)]. 相似文献
30.
Seiichi Nasuno Tadahiko Ohara Nobuyoshi Iguchi 《Bioscience, biotechnology, and biochemistry》2013,77(2):291-293
Hydrochloric acid treatment of methyl 3-(4-isobutylphenyl)-3-methylglycidate and methyl 2-hydroxy-3-(4-isobutylphenyl)-3-butenoate, a rearrangement product of the former, in acetic acid gave 3-(4-isobutylphenyl)-3-methylpyruvic acid and 2-(4-isobutylphenyl)-pro-panal. The same treatment of 2-hydroxy-3-(4-isobutylphenyl)-3-butenoic acid gave 2-(4-isobutylphenyl)-propanal. Both 3-(4-isobutylphenyl)-3-methylpyruvic acid and 2-(4-iso-butylphenyl)-propanal were oxidized to 2-(4-isobutylphenyl)-propionic acid. 相似文献