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1.
Enzymatic desulfurization of dibenzothiophene by a cell-free system of Rhodococcus erythropolis D-1 总被引:1,自引:0,他引:1
Abstract Pseudomonas syringae cells were exposed to Cu2+ alone or in the precence of acetate, proline or cysteine, at concentrations that reduced free Cu2+ to 1/10 of the total copper. Ligand concentrations (designated as isoeffective) were determined experimentally using a Cu2+ -selective electrode and confirmed by computer calculations using published stability constants. Exposure of P. syringae cells to Cu2+ alone resulted in rapid and pronounced cell death, and binding of most of the copper in solution. The addition of acetate, proline or cysteine, a few minutes after Cu2+ treatment, resulted in a significant reduction in cell death, and in the amount of copper bound to the cells. For short exposures to Cu2+ , cysteine was more effective than acetate or proline, but after 60 min of treatment, similar results were observed with these ligands. The addition of ligands before Cu2+ resulted in even more reduced copper toxicity. The results showed that, at isoeffective concentrations, weak and moderate copper-ligands can effectively antagonize copper toxicity, and that this protective effect does not require previously equilibrated copper-ligand solutions and is not very dependent of the nature of the ligand. 相似文献
2.
A respiration-deficient (RD) mutant was isolated from the petite-negative, salt-tolerant yeast Zygosaccharomyces rouxii. One strain among sixteen glycerol-non-utilizing mutants exhibited vigorous liberation of CO2 but no uptake of O2. Furthermore, this strain lacked cytochrome aa3 and had a reduced level of cytochrome b. The few mitochondria found in cells of this strain contained few or no cristae. Salt tolerance and intracellular accumulation of glycerol by the RD strain were almost equal to that of the wild-type strain in media containing NaCl up to 2.5 M. In media with more than 3 M NaCl, the growth of the RD mutant was retarded and the intracellular accumulation of glycerol was depressed in spite of ample production. 相似文献
3.
Yoshikazu Izumi Pijush Kanti Nath Hiroshi Yamamoto Hideaki Yamada 《Applied microbiology and biotechnology》1989,30(4):337-342
Summary A convenient and efficient method of NADPH production from NADP+ has been established using a glucose dehydrogenase system involving whole cells and immobilized cells of Gluconobacter suboxydans IFO 3172. Using airdried cells of the bacterium, the optimum conditions for NADPH production were examined, including the cell and glucose concentrations, NADP+ concentration, pH, buffer and reaction temperature. Under suitable conditions, the conversion ratio and the amount of NADPH accumulated reached about 100% and 73 mg/ml of the reaction mixture, respectively, after 1-h reaction. Intact cells of the bacterium also showed high NADPH production even in the reaction mixture without a surfactant. The addition of Triton X-100 to the reaction mixture and freeze-thawing treatment of intact cells enhanced the production. The NADPH production method was further improved by using cells of the bacterium immobilized by entrapment in a -carrageenan gel lattice. The immobilized cells had almost the same enzymatic properties as the air-dried cells. The conditions for the continuous production of NADPH with an immobilized cell column were also investigated. NADPH was produced in a good yield (about 95%) with this continuous process. 相似文献
4.
Toshikuni Sasaoka Norio Kaneda Yoshikazu Kurosawa Keisuke Fujita Toshiharu Nagatsu 《Neurochemistry international》1989,15(4):555-565
The chromosomal gene for human phenylethanolamine N-methyltransferase (PNMT; EC 2.1.1.28) was isolated from a human genomic library using a cloned human PNMT cDNA as a probe, and the nucleotide sequence was determined. PNMT is encoded in a single gene which consists of three exons. We observed newly the presence of minor PNMT mRNA (type B) besides the major mRNA (type A) as reported previously (Kaneda et al., J. Biol. Chem. 263, 7672–7677, 1988) by Northern hybridization. Type B mRNA carries an approximately 700 nucleotide-long untranslated region in the 5′ terminus. This suggests that two types of mRNA are produced from a single gene through the use of two alternative promoters. A TATA-like sequence locates 30 base pair upstream from the cap site of type A mRNA. Upstream of the cap site, there are several sequences resembling Spl binding sites and glucocorticoid responsive elements, with the latter also found in the first intron. 相似文献
5.
Mutagenesis of human granulocyte colony stimulating factor 总被引:9,自引:0,他引:9
T Kuga Y Komatsu M Yamasaki S Sekine H Miyaji T Nishi M Sato Y Yokoo M Asano M Okabe 《Biochemical and biophysical research communications》1989,159(1):103-111
To define the structure-function relationship, we have made a number of mutants of human granulocyte colony-stimulating factor (hG-CSF) by in vitro mutagenesis. The results indicate that most of the mutations located in the internal and C-terminal regions of the molecule abolished the activity, whereas the mutants without N-terminal 4, 5, 7, or 11 amino acids retained the activity. N-terminal amino acids were also altered by cassette mutagenesis using a synthetic oligonucleotide mixture. Among them, KW2228, in which Thr-1, Leu-3, Gly-4, Pro-5 and Cys-17 were respectively substituted with Ala, Thr, Tyr, Arg and Ser, showed more potent granulopoietic activity than that of intact hG-CSF both in vitro and in vivo. 相似文献
6.
Mitoshi Yokota Yutaka Tagawa Daikichi Okada Tooru Yasutake Yoshikazu Mine Hiroshi Ishikawa Kousei Miyashita Masao Tomita Sumihiro Tabuchi 《Biotherapy》1990,2(3):207-212
Pre- and postoperative intradermal administration of OK-432 enhanced the SU-PS skin reaction in patients with gastric cancer, but failed to prevent a fall in the NK activity induced by the operation.The change in NK activity was not associated with a change in the proportion of Leu 7-positive cells, but was related to Leu 11a-positive cells. Intradermal injection of OK-432 increased the proportion of Leu 7-positive cells in the patients in whom they accounted for less than 20% of lymphocyte population. The case was the same with Leu 11a-positive cells.Intravenous injection of OK-432 tended to increase suppressor-inducer T cells (CD4+2HA+ cells), B cells and Leu 7-positive cells. Particularly, the proportions of OK-M1-positive cells and MHC class II antigen-positive cells increased in all patients. Immunotherapy with OK-432 given intravenously at a dose of 0.1 KE appeared to be safe because no side effects were essentially observed. 相似文献
7.
Role of Chitinase and Chitin Oligosaccharides in Lignification Response of Cultured Carrot Cells Treated with Mycelial Walls 总被引:3,自引:0,他引:3
Chitinase activity was induced in cultured carrot cells by incubationwith mycelial walls of a fungus, Chaetomium globosum. Both intra-and extracellular chitinases were resolved into four componentsby gel filtration chromatography. The extracellular enzymesliberated soluble oligosaccharides of different sizes from insolublechitin, suggesting that these carrot chitinases are endo-hydrolases.The solubilized chitinase digests obtained from insoluble mycelialwalls of C. globosum and chitin were fractionated by gel filtrationchromatography, and the elicitor activity of each fraction forthe accumulation of phenolic acids in cultured carrot cellswas determined. In both solubilized fragments of fungal wallsand of chitin, elicitor-active oligosaccharides were distributedin many fractions, however, potent activity for inducing phenolicacid synthesis was observed in the high molecular weight fractions. (Received October 5, 1987; Accepted February 12, 1988) 相似文献
8.
Matuo Y Nishi N Muguruma Y Yoshitake Y Masuda Y Nishikawa K Wada F 《Cytotechnology》1988,1(4):309-318
Among several detergents, a zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS), was found to be least cytotoxic for cultured mammalian cells. CHAPS improved the activity recovery and elution profile of crude and purified fibroblast growth factors (FGFs) during chromatographies. Diluted preparations of FGFs were stabilized by CHAPS against the loss during storage. Amino acid sequence analysis was not disturbed by CHAPS. CHAPS was removable by reversed-phase high-performance liquid chromatography. These results indicate that CHAPS is useful as a non-cytotoxic stabilizing agent in purification of various kinds of bioactive polypeptides.Abbreviations -MEM
Alpha Modification of Eagle's Minimal essential medium
- CHAPS
3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate
- CHAPSO
3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propane sulfonate
- CS
Calf Serum
- EGF
Epidermal Growth Factor
- FGF
Fibroblast Growth Factor
- HPLC
High-Performance Liquid Chromatography
- NGF
Nerve Growth Factor
- NOG
1-O-n-octyl--D-glucopyranoside
- NP-40
Nonidet P-40
- PBS
Phosphate-Buffered Saline
- SB 12
3-(dodecylmethylammonio)-1-propane sulfonate
- SDS
Sodium Dodecyl Sulfate
- TGF- and
Transforming Growth Factor type and 相似文献
9.
Preventive effect of MCI-186 on 15-HPETE induced vascular endothelial cell injury in vitro 总被引:8,自引:0,他引:8
T Watanabe I Morita H Nishi S Murota 《Prostaglandins, leukotrienes, and essential fatty acids》1988,33(1):81-87
Using cultured bovine aortic endothelial cells, the effects of MCI-186, a radical scavenger, were studied on arachidonic acid metabolism and on the cell injury caused by 15-HPETE. MCI-186 at 3 X 10(-5) M enhanced prostacyclin production in the intact endothelial cells without affecting phospholipase A2. When endothelial cell homogenates were used as an enzyme source, it was found that MCI-186 stimulated the conversion of arachidonic acid to prostacyclin like phenol, perhaps by trapping OH radicals produced in the process of the conversion of PGG2 to PGH2. On the other hand, MCI-186 was found to inhibit lipoxygenase metabolism of arachidonic acid in cell free homogenates of rat basophilic leukemia cells. The lipoxygenase inhibition caused by 3 X 10(-5) M MCI-186 was almost equivalent to that caused by 3 X 10(-6) M BW 755C. MCI-186 remarkably protected against endothelial cell damage caused by 15-HPETE. 3 X 10(-5) M of 15-HPETE caused endothelial cell death in about 60% of the population: however, pretreatment of the cells with 10(-5) M of MCI-186 or concomitant addition of 10(-5) M of MCI-186 with 15-HPETE to the cultures prevented the cell death completely. These results suggest that MCI-186 may become an unique anti-ischemic drug. 相似文献
10.
Comparative study on the distribution of six species of bitterling in a creek in Fukuoka Prefecture was carried out. The upper part of the creek was fluvial, and the lower part stagnant.Acheilognathus lanceolatus, A. tabira subsp. (b) of Nakamura (1969),A. rhombeus andRhodeus atremius showed wide distribution in the creek throughout their life.A. limbatus seemed to be a fluvial species, andR. ocellatus smithii a lentic species. In the spawning season, however, all the species of bitterling, exceptingR. o. smithii, probably possess a common spawning ground in which mussels occur at a relatively high density. Adult females ofA. lanceolatus, A. tabira subsp. (b) andR. atremius without ripe eggs aggregated in the lower, stagnant parts of the creek. Most juveniles of these species and ofA. rhombeus seemed to grow in the lower part of the creek, and then migrate upward untill their spawning season. No species among the same phylogenetic group has the same set of the life modes (spawning season, spawning sites, aggregation of females without ripe eggs in the lower part of the creek and probability of upstream migration of 0-year-old fish toward spawning sites). 相似文献