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101.
以牛血球为材料,经溶血等处理和丙酮沉淀,获得牛血球超氧化物歧化酶粗品。此粗酶可以通过DEAE-Sepharose和CM-Sepharose快速柱层析,获得超氧化物歧化酶纯品。纯化的酶比活可达13500u/mg,经PAGE、SDS-PAGE和快速蛋白液相色谱(FPLC)检测,结果表明,纯化酶是均一的Sephadex G-100凝胶过滤测得该酶分子量为31,800,SDS-PAGE测得亚基分子量为15 相似文献
102.
利福霉素生产菌产生钝化RifSV物质的分离及性质研究 总被引:1,自引:0,他引:1
利福霉素SV(简称RifSV)生产菌──地中海拟无枝菌酸菌在生物合成RifSV过程中,产生一种能钝化自身产物(RifSV)的物质.实验证实,该物质是由谷氨酸、天冬氨酸、赖氨酸及缬氨酸等14种常见氨基酸组成的蛋白质.分子量(MW)约2.5×104D,等电点(PI)为5.7—6.1.在100℃下加热10min,其活性丧失.作用于RifSV的最适pH值范围为7.4—8.6,最适温度为29℃,初步证实该物质是一种酶(暂称利福霉素钝化酶) 相似文献
103.
104.
大瓶螺碱性磷酸酶的分离纯化及部分性质研究李清漪,曾和期(西南师范大学生物系,重庆630715)碱性磷酸酶(EC3.1.3.1,简称AKP)是广泛存在于动物组织中的水解酶。对软体动物中AKP的研究仅有少数报道[1,2],对属于单壳贝类的水生食用螺──大... 相似文献
105.
Mitogen- and isoproterenol-induced changes of [Ca2+]i in T cells attached to a glass substrate were examined. Murine (C57BL/6) splenic T cells were attached to coverslips or 35-mm dishes (MatTek) precoated with Cell Tak® (3.5 µg/cm2). The cells were then loaded with fluorescent dye (2 µg/ml of fura2-AM or fluo3-AM) and changes in [Ca2+]i in a population of cells (using a spectrofluorometer) or in single cells (using a confocal microscope) were measured during continuous superfusion. Population measurements of [Ca2+]i demonstrated that concanavalin A (Con A, 2 or 5 µg/ml) caused an increase in [Ca2+]i that rose to a peak and then declined to a steady state. The concentration-response relationship (0.05–5 µg/ml) had an EC50 of 0.3 µg/ml. Isoproterenol decreased the Con A-induced elevation of steady state [Ca2+]i. In single cell studies, the increase in [Ca2+]i in response to Con A typically occurred in about 50% of the cells in a microscope field, and the delay before activation varied among cells. Taken together, these data demonstrate that Cell Tak® can be used to attach T cells to glass coverslips and will be useful for the study of signaling mechanisms in T cells. 相似文献
106.
B. O. Solomon A. -P. Zeng H. Biebl A. O. Ejiofor C. Posten W. D. Deckwer 《Applied microbiology and biotechnology》1994,42(2-3):222-226
Product formation during anaerobic degradation of glycerol byKlebsiella pneumoniae DSM 2026, under glycerol limitation and glycerol excess in continugius cultures, has been investigated. Major and minor products and by-products as well as gaseous products were measured. The results indicated a positive correlation between specific glycerol uptake and most product formation rates under glycerol limitation. The production of 1,3-propanediol, lactate, formate, acetate, succinate and the by-products of anaerobic glycerol degradation byK. pneumoniae, acetoin and 2,3-butanediol, was favoured by glycerol excess, while hydrogen generation and ethanol formation were best under glycerol limitation. It was also found that under glycerol limitation the rate of hydrogen evolution was generally higher than the CO2 production rate while under excess glycerol the reverse was true. Hence, on the basis of the ratio of the specific rates of evolution of H2 and CO2 (q
H
2/q
CO
2), it is possible to infer the existence of glycerol limitation. On the basis of the carbon and available electron balances, which are independent of metabolic pathways, the data are consistent. The NADH2 balance, which took into consideration the pathways of product formation, was also tested to check the validity of the assumed pathways and to check critically the consistency of the data. Good balances were also obtained.[ 相似文献
107.
108.
Intramolecular excimer formation of 1,3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to evaluate the effect of ethanol on the rate and range of the lateral mobility and the range of the rotational mobility of bulk bilayer structures of the plasma membrane vesicles (ATCC-PMV) isolated from cultured hybridoma cells (ATCC TIB 216). In a concentration-dependent manner, ethanol increased the excimer to monomer fluorescence intensity ratio (I/I) of Py-3-Py in the ATCC-PMV and decreased the anisotropy (r), limiting anisotropy (r) and order parameter (S) of DPH in the ATCC-PMV. This indicates that ethanol increased both the lateral and rotational mobility of the probes in the ATCC-PMV. Selective quenching of DPH by trinitrophenyl groups was utilized to examine the range of transbilayer asymmetric rotational diffusion of the ATCC-PMV. The anisotropy (r), limiting anisotropy (r
) and order parameter (S) of DPH in the inner monolayer were 0.024, 0.032, and 0.069, respectively, greater than calculated for the outer monolayer of the ATCC-PMV. Selective quenching of DPH by trinitrophenyl groups was also used to examine the transbilayer asymmetric effects of ethanol on the range of the rotational mobility of the ATCC-PMV. Ethanol had a greater increasing effect on the range of the rotational mobility of the outer monolayer as compared to the inner monolayer of the ATCC-PMV. It has been proven that ethanol exhibits a selective rather than nonselective fluidizing effect within the transbilayer domains of the ATCC-PMV.This paper was supported in part by a research grant from the Korea Science and Engineering Foundation (KOSEF 88-1013-01) and from the Korea Research Foundation (1991–1993). 相似文献
109.
Soo Wan Nam Byung Moon Kim Bong Hyun Chung Dae Ook Kang Jong Seog Ahn 《Biotechnology letters》1994,16(9):897-902
Summary For the secretion of human lipocortin-1 (LC-1) in yeast, a expression and secretion vector was constructed by using the promoter and signal sequence of glucoamylase gene (STA1) of Saccharomyces diastaticus. After the cDNA of human LC-1 was ligated with the secretion vector, the resulting hybrid plasmid was transformed into S. diastaticus. When the recombinant S. diastaticus was cultivated in YPD medium, LC-1 was expressed and secreted into the extracellular medium, yielding LC-1 protein at a concentration of 2.5 g/mL. 相似文献
110.
D Chen C Q Zeng M J Wentz M Gorziglia M K Estes R F Ramig 《Journal of virology》1994,68(11):7030-7039