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Kyuichi Matsubayashi Hideo Hakusui Mitsuji Sano 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1977,143(6):571-580
A sensitive and specific method for the determination of lofepramine and its metabolites, desipramine and 2-hydroxydesipramine, in human plasma and urine is described. Lofepramine, desipramine and 2-hydroxydesipramine were derivatized to ethyl p-chlorobenzoate, the bis(heptafluorobutyryl) derivative and the N,O-bis(trifluoroacetyl) derivative, respectively, and then analysed by gas chromatography—mass fragmentography. Corresponding deuterated compounds were used as internal standards. Determination was possible at levels as low as 2 ng/ml for lofepramine and desipramine and 20 ng/ml for 2-hydroxydesipramine. 相似文献
3.
Saccharification of Complex Cellulosic Substrates by the Cellulase System from Clostridium thermocellum 总被引:37,自引:15,他引:22
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Eric A. Johnson Mitsuji Sakajoh Geoffrey Halliwell Ashwin Madia Arnold L. Demain 《Applied microbiology》1982,43(5):1125-1132
True cellulase activity has been demonstrated in cell-free preparations from the thermophilic anaerobe Clostridium thermocellum. Such activity depends upon the presence of Ca2+ and a thiol-reducing agent of which dithiothreitol is the most promising. Under these conditions, native (cotton) and derived forms of cellulose (Avicel and filter paper) were all extensively solubilized at rates comparable with cellulase from Trichoderma reesei. Maximum activity of the Clostridium cellulase was displayed at 70°C and at pH 5.7 and 6.1 on Avicel and carboxymethylcellulose, respectively. In the absence of substrate at temperatures up to 70°C, carboxymethylcellulase was much more unstable than the Avicel-hydrolyzing activity. 相似文献
4.
C Kamibayashi R Estes C Slaughter M C Mumby 《The Journal of biological chemistry》1991,266(20):13251-13260
Protein phosphatase 2A consists of a heterotrimeric complex composed of a catalytic subunit (C) and two associated subunits (A and B). Limited tryptic digestion of the heterotrimeric ABC form resulted in the selective degradation of the Mr = 55,000 B subunit to a 48-kDa polypeptide. The cleavage sites were determined to be within a 3-7-kDa region of the COOH terminus. Proteolysis led to dissociation of the B subunit from the enzyme complex and correlated with an increase in cardiac myosin light chain, smooth muscle myosin light chain peptide, and Leu-Arg-Arg-Ala-Ser-Leu-Gly (Kemptide) phosphatase activity. Purification of the digestion products and native gel electrophoresis indicated that dissociation of the B subunit was responsible for the increase in phosphatase activity. Kinetic analyses with several substrates revealed that dissociation of the B subunit resulted in a 2-7-fold increase in Vmax and a 1.6-5 fold increase in Km. Proteolytic dissociation of the B subunit increased the sensitivity of protein phosphatase 2A to inhibition by okadaic acid. Inhibition of the trypsinized enzyme was very similar to that observed for the purified AC form of protein phosphatase 2A. Incubation of the ABC complex with N-ethylmaleimide resulted in dissociation of the C subunit and generation of an AB complex. Selective release of the C subunit indicated that the B subunit interacts directly with the A subunit and that one or more free sulfhydryls are required to maintain the heterotrimeric structure of protein phosphatase 2A. Treatment of the enzyme with heparin resulted in an increase in specific activity that was due to the release of the B subunit from the complex. These results provide evidence that the B subunit binds directly to the A subunit to modulate enzyme activity and substrate specificity and that the COOH-terminal region of this protein is important for interaction with the AC complex. Dissociation of the B subunit by polyanionic substances related to heparin may represent a mechanism for regulating the activity of this enzyme. 相似文献
5.
Effect of Chlorine Substitution on the Biodegradability of Polychlorinated Biphenyls 总被引:29,自引:22,他引:7
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Thirty-one isomers of polychlorinated biphenyl (PCB) were examined for biodegradability by two species of Alcaligenes and Acinetobacter. The following relationships between chlorine substitution and biodegradability of PCBs were observed. (i) Degradation decreased as chlorine substitution increased. PCB isomers containing more than four chlorines were less susceptible to degradation. (ii) PCBs containing two chlorines on either the ortho position of a single ring (i.e., 2,6-) or on both rings (i.e., 2,2′-) showed very poor degradability. (iii) PCBs containing all chlorine atoms on only a single ring were generally degraded faster than when the same number of chlorines were substituted on both rings. (iv) Preferential ring fission of the molecules occurred with nonchlorinated or lesser chlorinated rings. (v) The formation and accumulation of a yellow intermediate was always observed in 4′-chloro-substituted PCBs. (vi) Significant differences between the two organisms with respect to degradability were not observed except for 2,4,6-trichlorobiphenyl. 相似文献
6.
Keita Saito Miho Takahashi Masato Kamibayashi Toshihiko Ozawa 《Free radical research》2013,47(7):668-676
This study compared the superoxide detection abilities of four spin traps, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide (DEPMPO), 5-(diphenylphosphinoyl)-5-methyl-1pyrroline N-oxide (DPPMPO) and 5-(2,2-dimethyl-1,3-propoxy cyclophosphoryl)-5-methyl-1-pyrroline N-oxide (CYPMPO) in living cells. Electron spin resonance (ESR) signals of the superoxide adducts were observed when spin traps were added to a suspension of human oral polymorphonuclear leukocytes (OPMNs) stimulated by phorbol 12-myristate 13-acetate. The ESR signal of the CYPMPO-superoxide adduct (CYPMPO-OOH) increased for 24 min after the initiation of the reaction, whereas the signals from DMPO-OOH and DPPMPO-OOH peaked at 6 and 10 min, respectively. The maximum concentrations of DMPO-OOH, DPPMPO-OOH and CYPMPO-OOH in OPMNs were 1.9, 6.0 and 10.7 µM, respectively. Furthermore, CYPMPO could more efficiently trap superoxide in blood PMNs compared with DEPMPO. From these results, it was concluded that CYPMPO performs better than DMPO, DPPMPO and DEPMPO for superoxide measurements in living cell systems because it has lower cytotoxicity and its superoxide adduct has a longer lifetime. 相似文献
7.
Kensuke Furukawa Kenzo Tonomura Akira Kamibayashi 《Bioscience, biotechnology, and biochemistry》2013,77(7):1577-1583
Metabolism of 2,4,4′-trichlorobiphenyl by Acinetobacter sp. strain P6 has been studied. When the incubation was carried out without shaking at 15°C, two isomeric monohydroxy compounds, a dihydrodiol compound, a dihydroxy compound, a meta-cleaved yellow compound and a dichlorobenzoic acid were detected by combined gas liquid chromatograph-mass spectrometry. As an additional metabolite, dichlorodihydroxy biphenyl, a dechlorinationhydroxylation product, was also detected. When the incubation mixture was shaken at 30°C, a meta-cleaved yellow compound was readily produced and predominantly accumulated in the reaction mixture upon further incubation. The major pathway of 2,4,4′-trichlorobiphenyl by Acinetobacter sp. P6 was considered to proceed oxidatively via 2.′3′-dihydro-2′,3′-diol compound, concomitant dehydrogenated 2′,3′-dihydroxy compound and then the 1′,2′-meta-cleaved yellow compound, i.e., 3-chloro-2-hydroxy-6-oxo-6(2,4-dichlorophenyl)hexa-2,4-dienoic acid. 相似文献
8.
Hideoki Tanaka Hidekatsu Maeda Hideo Suzuki Akira Kamibayashi Kenzo Tonomura 《Bioscience, biotechnology, and biochemistry》2013,77(6):1429-1438
A photosynthetic bacterium, which can grow photosynthetically on benzoate, was isolated from sewage mud. Various kinds of aromatic compounds including heterocyclic aromatic compounds were photometabolized by the washed cells grown photosynthetically on benzoate with no lag period. Among these, thiophene-2-carboxylate was metabolized most rapidly to its (+)-tetrahydro derivative. The same strain could also grow on succinate under photosynthetic conditions. However, thiophene-2-carboxylate was only photometabolized after a long lag period by the washed cells grown photosynthetically on succinate, and the metabolite was not its (+)-tetrahydro derivative but (+)-3-hydroxytetrahydrothiophene-2-carboxylate. In the presence of chloramphenicol, an inhibitor of protein synthesis, the photometabolism of thiophene-2-carboxylate by the washed cells grown photosynthetically on benzoate was not affected at all, but the photometabolism of the same substrate by the washed cells grown photosynthetically on succinate was completely inhibited. These results indicate that a reduction system of broad substrate specificity for aromatic rings is already present in the benzoate-grown cells but absent in the succinate-grown cells. It seems that such a reduction system for aromatic rings is induced by an aromatic substrate. 相似文献
9.
Masami Uebayasi Sugio Kawamura Noboru Tomizuka Akira Kamibayashi 《Bioscience, biotechnology, and biochemistry》2013,77(6):1799-1807
For Hyphomicrobium 53-49 capable of growing under various conditions, aerobic methanol, anaerobic methanol (with denitrification), autotrophic (H2-O2-CO2), aerobic ethanol and aerobic acetate, investigation and comparison of the specific activities of the following enzymes were performed: alcohol dehydrogenase (NAD-ethanol linked and NAD-methanol linked), primary alcohol dehydrogenase, formaldehyde dehydrogenase (NAD-GSH linked and DCPIP linked), formate dehydrogenase, serine hydroxymethyl transferase, hydroxypyruvate reductase, isocitrate lyase (icl), malate lyase, malate dehydrogenase, ribulosebisphosphate (RuBP) carboxylase, phos-phoenolpyruvate (PEP) carboxykinase (ADP linked), PEP carboxylase (phosphorylating), pyruvate carboxylase (NADH linked and NADPH linked) and α-ketoglutarate carboxylase (NADH linked and NADPH linked). On the basis of the data obtained, it was concluded that during growth on methanol, aerobically and anaerobically, the icl+ serine pathway operated, while during autotrophic growth on H2-O2-CO2, CO2 was incorporated through the RuBP pathway and others, and during growth on ethanol or acetate, neither the serine pathway nor the RuBP pathway operated. The organism changed its metabolism through the regulation of the metabolic enzymes according to the growth conditions. 相似文献
10.
Physical and Functional Interactions between Type I Transforming Growth Factor β Receptors and Bα, a WD-40 Repeat Subunit of Phosphatase 2A
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Irene Griswold-Prenner Craig Kamibayashi E. Miko Maruoka Marc C. Mumby Rik Derynck 《Molecular and cellular biology》1998,18(11):6595-6604
We have previously shown that a WD-40 repeat protein, TRIP-1, associates with the type II transforming growth factor β (TGF-β) receptor. In this report, we show that another WD-40 repeat protein, the Bα subunit of protein phosphatase 2A, associates with the cytoplasmic domain of type I TGF-β receptors. This association depends on the kinase activity of the type I receptor, is increased by coexpression of the type II receptor, which is known to phosphorylate and activate the type I receptor, and allows the type I receptor to phosphorylate Bα. Furthermore, Bα enhances the growth inhibition activity of TGF-β in a receptor-dependent manner. Because Bα has been characterized as a regulator of phosphatase 2A activity, our observations suggest possible functional interactions between the TGF-β receptor complex and the regulation of protein phosphatase 2A. 相似文献