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71.
72.
Toshihiko Ubuka Masahiro Kinuta Reiko Akagi Shozo Kiguchi Miyabi Azumi 《Analytical biochemistry》1982,126(2):273-277
A procedure for the simultaneous preparation of S-sulfo-l-cysteine and l-alanine 3-sulfinic acid is described. The method is based on the quantitative reaction between sulfite and S-(2-amino-2-carboxyethylsulfonyl)-l-cysteine. The yield was 95% for S-sulfo-l-cysteine and 91% for l-alanine 3-sulfinic acid. The reaction was also applied to the quantitative determination of sulfite in biological materials. In this procedure, sulfite reacts with S-(2-amino-2-carboxyethylsulfonyl)-l-cysteine. Separation of the reaction product, S-sulfo-l-cysteine, is done by ion-exchange fractionation, and it is determined with acid ninhydrin reagent 2 (M. K. Gaitonde, 1967, Biochem. J.104, 627–663). The recovery was 96.8 ± 0.3%. 相似文献
73.
Robert P. Clement Masahiro Kohashi Walter N. Piper 《Archives of biochemistry and biophysics》1982,214(2):657-667
Rat hepatic uroporphyrinogen III cosynthase has been isolated and purified 50-fold with a 36% yield by ammonium sulfate fractionation and sequential chromatography on DEAE-Sephacel and Sephadex G-100SF. Inhibition of uroporphyrinogen III formation with increasing porphobilinogen concentration was observed. Cosynthase was shown to be thermolabile, and a time-dependent loss of enzyme activity during reaction with uroporphyrinogen I synthase and porphobilinogen was observed. The pH optimum for the complete system (synthase and cosynthase) was pH 7.8 in 50 mm Tris-HCl or 50 mm sodium phosphate buffer. Various metals (KCl, NaCl, MgCl2, CaCl2) increased formation of Uroporphyrinogen III. Heavy metals including ZnCl2, CdCl2, and CuCl2 were shown to selectively inhibit cosynthase activity, whereas other metals (HgCl2, PbCl2) were less selective and inhibited both synthase and cosynthase at similar concentrations. 相似文献
74.
Toru Shimizu Masahiro Hatano Seiji Nagao Yoshinori Nozawa 《Biochemical and biophysical research communications》1982,106(4):1112-1118
43Ca NMR spectra of Ca2+-Tetrahymena calmodulin(Tet. CaM.) complexes have been observed under various conditions. Off-rate of Ca2+ from Tet. CaM. is estimated to be approx. 2.7 × 103 s?1 under a certain assumption. Relaxation rates of 43Ca NMR of Ca2+-Tet. CaM. are remarkably increased(by one order in magnitude) by adding trifluoperazine(TFP), a potent calmodulin antagonist. Relaxation parameters estimated suggest that Ca2+ mobility is reduced by the TFP binding. A stoichiometry of TFP is two moles per Tet. CaM. molecule. The relaxation rates of 43Ca NMR signals are increased by adding excessive Mg2+ to the Ca2+-Tet. CaM. solutions. The addition of Mg2+ to the Ca2+-Tet. CaM. complex decreases apparent pKa value of the complex as well. 相似文献
75.
A β-N-acetylhexosaminidase [EC 3.2.1.30] has been purified ~98-fold from an extract of the digestive organs of Saxidomus purpuratus by using ammonium sulfate fractionation, and chromatography on Toyopearl HW-50, CM-cellulose, and Sepharose 4B. The purified enzyme, the molecular weight of which was estimated to be ~66,000 by gel filtration, was composed of two sub-units of molecular weight 30,000 as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The purified enzyme had a pH optimum of 3.8 and an optimum temperature of 55°, and its activity was enhanced ~2-fold in the presence of 0.1m sodium chloride. The Michaelis constants toward p-nitrophenyl 2-acetamido-2-deoxy-β-d-glucoside and -galactoside were 1.2 × 10?4 and 1.3 × 10?4m, respectively. 相似文献
76.
Eisuke Mekada Kenji Kohno Masahiro Ishiura Tsuyoshi Uchida Yoshio Okada 《Biochemical and biophysical research communications》1982,109(3):792-799
We have measured the specific uptake of 125I-labelled diphtheria toxin in the presence of methylamine by a number of cell lines with different sensitivities to diphtheria toxin. The results show a strong correlation between the toxin sensitivities of the cell lines and the amount of specific uptake. The specific association of labelled toxin with cells was clearly demonstrated even with CHO cells, a cell line with relatively low sensitivity. Thus, CHO cell mutants that are resistant to diphtheria toxin could be classified as toxin-binding or non-binding cells by this method. 相似文献
77.
78.
Masashi Fujiwara Keiko Fukushi Mitsuo Takai Jisuke Hayashi Masahiro Fukaya Hajime Okumura Yoshiya Kawamura 《Biotechnology letters》1992,14(7):539-542
Summary Shuttle vector pUF106 was constructed by ligation ofAcetobacter xylinum plasmid pFF6 toEscherichia coli plasmid pUC18. It had unique restriction sites suitable for insertion of a foreign DNA fragment and conferred ampicillin resistance to a host. pUF106 transformed cellulose-producingA. xylinum ATCC10245 as well asE. coli JM109. 相似文献
79.
80.
Koji Sode Masahiro Tatara Haruko Takeyama J. Grant Burgess Tadashi Matsunaga 《Applied microbiology and biotechnology》1992,37(3):369-373
Summary Versatility of gene transfer by transconjugation in marine cyanobacteria was demonstrated. In this study, seven different marine cyanobacteria were used as recipient cells. First, transconjugation was carried out using the mobilizable transposon (Tn5) carrying plasmid pSUP1021. Transconjugants were observed in all marine cyanobacteria tested. Second, the broad-host-range vector pKT230 (IncQ) was tested for transconjugation. pKT230 has been successfully transferred in a marine cyanobacterium Synechococcus sp. NKBG15041C, and replicated as an autonomous replicon without alteration in the restriction enzyme pattern. A maximum transfer efficiency of 5.2 × 10–4 transconjugants/recipient cell was observed, when mating was performed on agar plates containing low salinity (0.015 m NaCl) medium. This is the first study to demonstrate gene transfer in marine cyanobacteria via transconjugation.
Correspondence to: K. Sode 相似文献