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11.
Shinji Tokuyama Hiroyuki Miya Kazunori Hatano Takeshi Takahashi 《Applied microbiology and biotechnology》1994,40(6):835-840
A novel enzyme, N-acylamino acid racemase, was purified to homogeneity from Streptomyces atratus Y-53 and characterized. This enzyme catalyzes the interconversion of optically active N-acylamino acids. The relative molecular mass (Mr) of the enzyme was estimated to be about 41 000 and 244 000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, respectively, indicating that the enzyme is composed of six subunits with an equal Mr. The enzyme showed a broad substrate specificity toward N-acylamino acids, such as N-acetylmethionine, N-chloroacetylphenylalanine and N-chloroacetylvaline. The apparent Michaelis constant (Km) values for N-acetyl-l-methionine and N-acetyl-d-methionine were calculated to be 15.2 and 5.6 mm, respectively. Enzyme activity was markedly enhanced by divalent metal ions, such as Co2+, Mg2+ and Mn2+, and was inhibited by metal-chelating reagent, indicating that the enzyme is a metalloenzyme. We propose to name the enzyme N-acylamino acid racemase (acylamino acid racemase).
Correspondence to: S. Tokuyama 相似文献
12.
Taishin Nomura Shunsuke Sato Shinji Doi Jose P. Segundo Michael D. Stiber 《Biological cybernetics》1994,72(1):55-67
The Bonhoeffer-van der Pol (BVP) oscillator is a valuable dynamical system model of pacemaker neurons. Isochrons, phase transition curves (PTC), and two dimensional bifurcation diagrams served to analyze the neuron's response to periodic pulse stimuli. Responses are described and explained in terms of the nonlinear dynamical system theory. An important issue in the generation of spikes by pacemaker neurons is the existence of both slow and fast dynamics in the state point's trajectory in the phase plane. It is this feature in particular that makes the BVP oscillator a faithful model of living pacemaker neurons. Comparison of the model's responses with those of a living pacemaker was based also on return maps of interspike intervals. Analyzed in detail were the complex discharges called stammering which involve interspike intervals that arise unpredictably and exhibit histograms with several modes separated by the equal intervals.Supported by Trent H. Wells Jr. Inc. 相似文献
13.
Chunyu Cao Hisao Kurazono Shinji Yamasaki Keiko Kashiwagi Kazuei Igarashi Yoshifumi Takeda 《Microbiology and immunology》1994,38(6):441-447
The gene encoding a Verotoxin 2 variant, VTvp1, was mutated by oligonucleotide-directed site-specific mutagenesis. Among 6 mutant toxins encoded by the mutated genes, E167Q-R170L (glutamic acid at position 167 and arginine at position 170 from N-terminus of the A subunit were replaced by glutamine and leucine, respectively) was found to have markedly decreased activities; inhibition of protein synthesis, Vero cell cytotoxicity and mouse lethality of the purified E167Q-R170L were 1/1,900, 1/125,000 and 1/2,000, respectively, of those of the purified wild-type VT2vp1. Since the antigenic property of the E167Q-R170L was demonstrated to be similar to that of the wild-type VT2vp1 by Ouchterlony double gel diffusion test and by neutralization test of Vero cell cytotoxicity of the VT2vp1, a possibility to use the mutant VT2vp1, E167Q-R170L, as a toxoid is discussed. 相似文献
14.
Shunya Sogon Shinji Imakawa Makoto Masutani Nobuyoshi Takeuchi Katsuhiko Kubo 《Journal of Ethology》1993,11(1):1-10
The purposes of this study were two-fold. First, it compared Japanese infants' (N=129) abilities to regulate emotions at 13
months and 18 months of age, using the Strange Situation procedure. Second, it examined the relationship between the development
of emotion regulation and the mother's emotion expression style as assessed by the Emotion Expression Style Questionnaire
(EESQ). The total number of subjects who successfully completed all 8 episodes of the Strange Situation procedure increased
significantly during the aging between 13 and 18 months of age, indicating that as a group these infants increased their ability
to cope with stressful situations. However, infants who had mothers with negative emotion expression styles did not show greater
capacity for emotion regulation at 18 months. These findings suggest that the development of emotion regulation is mediated
by the mother's emotion expression style. 相似文献
15.
Sakaba Tomoka Soejima Akiko Fujii Shinji Ikeda Hajime Iwasaki Takaya Saito Hiroaki Suyama Yoshihisa Matsuo Ayumi Kozhevnikov Andrey E. Kozhevnikova Zoya V. Wang Hongfeng Wang Siqi Pak Jae-Hong Fujii Noriyuki 《Journal of plant research》2023,136(4):437-452
Journal of Plant Research - A group of temperate grassland plant species termed the “Mansen elements” occurs in Japan and is widely distributed in the grasslands of continental East... 相似文献
16.
17.
Kazumasa Tsuji Masaharu Iijima Hiroshi Matsuzawa Shinji Sakamoto 《Biotechnology Techniques》1997,11(6):395-398
Escherichia coli JM109(DE3) harboring expression plasmid pkAQNÆC30, which carries the Thermus protease aqualysin I (AQI) gene, was cultivated with glucose as a sole carbon source. The final cell concentration was over 15 g dry weight/l and the amount of AQI produced reached approximately 130 kU/ml broth. Moreover, by using two carbon sources, glucose and glycerol, the production yield was increased to over 200 kU AQI/ml, while suppressing the formation of inhibitory acetic acid. 相似文献
18.
Michio Masuda Tsuyoshi Abe Shinji Sato Teruaki Suzuki Minoru Suzuki 《Journal of phycology》1997,33(2):196-208
Many morphologically similar, but chemically distinct, populations have been found in the marine red alga Laurencia nipponica Yamada (Rhodomelaceae, Ceramiales) growing in Japan. Each chemical type is characterized by a specific end-product of halogenated secondaly metabolite synthesis: chamigrane-type sesquiterpenoids such as prepacifenol and halochamigrene epoxide and C15 bromoethers such as laurencin, laureatin, isoprelaurefucin, epilaurallene, and kumausallene. These seven types of secondary metabolite syntheses remained the same in the wild and under various culture conditions. Because bromoethers and terpenoids are probably synthesized by different metabolic pathways, it is virtually certain that different sets of enzymes participate in their synthesis. Prepacifenol- and laureatin-producing populations were selected as representatives of terpenoid and bromoether groups, respectively. F1 tetrasporophytes derived from crosses between reciprocal, female and male gametophytes of prepacifenol- and laureatin-producing strains bore both types of metabolites, suggesting that the genes Producing these enzyme systems are encoded by nuclear genomes. The F1 gametophytes resulting from the reciprocal crosses produced either prepacifenol or laureatin, and the four individuals derived from spore tetrads (a set of tetraspores derived from a single tetrasporangium) produced either prepacifenol or laureatin in a 1:1 ratio, indicating that genes participating in terpenoid synthsis and those participating in bromoether synthesis are on different loci of homologous chromosomes and are segregated at meiosis (tetrasporogenesis). One individual of this interpopulational F1 gamtophyte produced both parental types of metabolite, perhaps indicating the occurrence of a recombination type. Natural hybrid individuals, including such recombination-type gametophytes, were found in a sympatric locality at which these two chemical types occur. F1 tetrasporophytes derived from crosses between respective prepacifenol- and laureatin-producing strains and their F1 gametohytes produced only parental-type metabolite-producing plants. These results indicate that the diverse chemical types can be referred to as races (chemical races). 相似文献
19.
Shinji Hosoi Mitsuo Satoh Hiromasa Miyaji Tatsunari Nishi Tamio Mizukami Mamoru Hasegawa Seiga Itoh Tatsuya tamaoki 《Cytotechnology》1995,19(1):1-10
Pro-UKS1 was designed as a thrombin-resistant derivative of pro-urokinase (pro-UK) by introducing a glycosylation site using site-directed mutagenesis. An expression plasmid for pro-UKS1, pMo1UKS1SEd1-5, was constructed and introduced into Namalwa KJM-1 cells (Hosoiet al., 1988), and cells resistant to G418 and Methotrexate (MTX) were obtained. Amongst them, the highest pro-UKS1 producer (resistant to 500 nM of MTX), clone 41-8, was selected and further characterized. Clone 41-8 was cultured in serum-free ITPSGF medium (Hosoiet al., 1988). Under the conventional conditions, the concentration of pro-UKS1 reached 26 g ml–1. Addition of glucose and tri-iodothyronine (T3) improved productivity, and the maximal productivity of pro-UKS1 was 67 g ml–1 day–1. In this conditioned medium, content of pro-UKS1 was above 80% of total proteins.Abbreviations BSA
bovine serum albumin
- dhfr
dihydrofolate reductase
- HEPES
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- kb
kilobase pairs
- kDa
kilodaltons
- MTX
Methotrexate
- PBS
phosphate buffered saline
- pro-UK
pro-urokinase
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- T3
tri-iodothyronine
- Tween-PBS
phosphate buffered saline containing 0.05% Tween 80 相似文献
20.
Immunochemical study on PHI/PHM with use of synthetic peptides 总被引:2,自引:0,他引:2
N Yanaihara C Yanaihara K Nokihara K Iguchi S Fukata M Tanaka Y Yamamoto T Mochizuki 《Peptides》1984,5(2):247-254
We have synthesized PHI and PHM (human PHI) as well as their fragments, PHI (1-6), PHI (1-15), PHI (14-19), PHI (14-27), PHI (20-27), PHM (1-15) and PHM (13-27), by the solution or solid-phase method for peptide synthesis. Using the highly purified synthetic peptides as immunogens or haptenic immunogens, five kinds of PHI/PHM specific antisera were produced. The major antibody-recognition sites of the five antisera were located respectively in the PHI C-terminal (R8201), in the PHI N-terminal (R8403), in the PHM C-terminal (R8502), and in the PHM whole molecule (R8702 and R8703). Radioimmunoassays (RIAs) with antisera R8201, R8403 and R8502, respectively, showed a wide distribution of immunoreactive (IR) PHI/PHM in porcine and human gastrointestinal and brain tissues. The concentrations of IR-PHI in the porcine gastrointestinal tissues, however, differed between the R8201 and R8403 RIAs employed for measurement. By using these two different PHI RIAs, the IR-PHI in the porcine brain tissue extract was shown to be almost a single component coeluting with synthetic PHI in gel filtration. The IR-PHI in the extract of porcine lower intestine on the other hand, contained, besides a PHI-like component, unidentified component(s) eluting immediately after synthetic PHI in gel filtration; this crossreacted with the PHI C-terminal specific R8201 antiserum but not with the N-terminal specific R8403 antiserum, suggesting the presence of the C-terminal-related fragment(s) of PHI in the tissues. 相似文献