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951.
The synthesis of four β-lactam antibiotics (penicillin G, pivaloyloxymethyl ester of penicillin G, ampicillin and pivampicillin) catalyzed byEscherichia coli penicillin acylase has been investigated in water-methanol mixtures. The enzyme reactions were either thermodynamically or kinetically controlled at the same conditions using phenylacetic acid andd--phenylglycine methyl ester as acyl donors and 6-aminopenicillanic acid and pivaloyloxymethyl 6-aminopenicillanic acid as acyl acceptors. It has been found that the influences of the cosolvent content on the reaction rates and synthetic yields are significantly different depending on the substrates used in the experiments. On the other hand, within certain ranges of the methanol content (up to ca. 40% (v/v) the residual activities of the enzymes in water-methanol mixtures were only slightly lower than those in aqueous media. To analyze the factors that determine the reaction rate in water-cosolvent mixtures, the effect of methanol on the apparent pK values of the substrates has been investigated, and a mathematical model has been developed on the basis of the assumption that the enzyme binds non-ionized substrates. Model simulation results indicate that the solvent effect on reaction rates is mainly attributed to the kinetic effects of changes in apparent pK values. 相似文献
952.
Douglas L. Mann 《Cytokine & growth factor reviews》1996,7(4):341-354
The ability of myocardium to successfully compensate for, and adapt to, stress ultimately determines whether the heart will decompensate and fail, or whether it will instead maintain preserved function. Despite the importance of the myocardial response to environmental stress, very little is known with respect to the biochemical mechanisms that are responsible for mediating and integrating the stress response in the heart. In the present review we will summarize recent experimental material which suggests that cytokines that are expressed within the myocardium in response to a environment injury, namely tumor necrosis factor-alpha (TNF-α), interleukin-1 (IL-1) and interleukin-6 (IL-6), may play an important role in initiating and integrating homeostatic responses within the heart. However, these ‘stress-activated’ cytokines all have the potential to produce cardiac decompensation when expressed at sufficiently high concentrations. Accordingly, the theme that will emerge from this discussion is that the short-term expression of stress-activated cytokines within the heart may provide the heart with an adaptive response to stress, whereas long-term expression of these molecules may be frankly maladaptive by producing cardiac decompensation. 相似文献
953.
A bacterium which has phospholipase A1 activity was isolated from soil. It is aerobic, motile, oxidase-negative and has flagella. The G+C content of the DNA was 58.1 mol%. The major isoprenoids of cell was were Q-8 and MK-8. The main cellular fatty acids were saturated straight chain (n-16) and cyclic (17:) fatty acids. Based on its morphological, physiological and chemotaxonomic characteristics, this organism was placed in the genusSerratia. Nutritional factors affecting enzyme production were explored. Xylose and ammonium sulfate were the best carbon and nitrogen sources, respectively. Ferrous ions exerted a considerable positive effect on enzyme production. The optimal pH and temperature for phospholipase A1 production were 7.0 and 30°C, respectively. 相似文献
954.
The scale up of earlier work from these laboratories using the enzyme serinehydroxymethyltransferase has resulted in the use of this enzyme for the synthesis of 2-amino-3-hydroxy-1,6-hexanedicar☐ylic acid (7) on a preparative scale. This compound, which has been barely described in the literature, is potentially useful for the synthesis of carbocyclic β-lactams and carbocyclic nucleosides. 相似文献
955.
Ben M. De Rooij Jan N.M. Commandeur Jane R. Ramcharan Hilda C.P. Schuilenburg Ben L.M. Van Baar Nico P.E. Vermeulen 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1996,685(2):1402
Epichlorohydrin (ECH) is used in many industrial processes. Different toxic effects of ECH were found in rodents. The metabolism of ECH was investigated before in rats using [14C]ECH. The aim of this investigation was the development of non-radioactive quantitative analytical methods for measuring two urinary metabolites of ECH, namely 3-chloro-2-hydroxypropylmercapturic acid (CHPMA) and α-chlorohydrin (α-CH). The identity of CHPMA and α-CH excreted in urine of rats treated with 5 to 35 mg/kg ECH was confirmed by GC-MS. The quantitative analysis of CHPMA, involving ethyl acetate extraction from acidified urine and subsequent methylation and analysis by gas chromatography-flame photometric detection (GC-FPD), showed a method limit of detection of 2 μg/ml. The analysis of α-CH, based on ethyl acetate extraction and subsequent analysis by GC-ECD, showed a method limit of detection of 2 μg/ml. CHPMA and α-CH derivatives could be determined quantitatively down to concentrations of 0.5 and 0.4 μg/ml urine, respectively, by selected-ion monitoring GC-MS under EI conditions. Cumulative urinary excretion of CHPMA and α-CH by rats treated with ECH were found to be 31 ± 10 and 1.4 ± 0.6% (n = 13) of the ECH dose, respectively. For CHPMA, the dose-excretion relationship suggested partially saturated ECH metabolism. For α-CH, the dose-excretion relationship was linear. With fractionated urine collection it was found that approximately 74 and 84% of the total cumulative excretion of CHPMA and α-CH, respectively, took place within the first 6 h after administration of ECH. From these investigations it is concluded that the GC-FPD and GC-ECD based methods developed are sufficiently sensitive to measure urinary excretion of CHPMA and α-CH in urine from rats administered 5 to 35 mg/kg ECH. It is anticipated that the analysis of CHPMA and α-CH based on GC-MS may be sufficiently sensitive to investigate urinary excretion from humans occupationally exposed to ECH. 相似文献
956.
小牛类表皮生长因子活性肽(c-EGF)的分离纯化及性质 总被引:1,自引:0,他引:1
本文报道小牛颌下腺通过酸抽提、CM52、DE52柱层析、SephadexG75凝胶过滤及HPLC等步,分离纯化到一个分子量为6kD,等电点为4.6的小肽。该小肽类似人重组表皮生长因子(rh—EGF)的生物活性,具有刺激NRK细胞的增殖及刺激新生小鼠的睁眼、萌牙,并具有增加A431细胞膜蛋白质磷酸化的功能。氨基酸组成缺少苯丙氨酸及苏氨酸残基。我们命名它为小牛类EGF样活性肽(calfEGF-likeactivepeptide,简称c-EGF)。 相似文献
957.
猪精子中与卵透明带糖蛋白ZP3结合的蛋白质 总被引:3,自引:0,他引:3
依次经PSL-Sepharose亲和层析柱和纤维素CM-52离子交换层析柱,从猪精子的CHAPS抽提液分离得4个蛋白质组分。用固相透明带精蛋白结合试验(IZPGBA)检测;表明精子蛋白SP1和SP2具有结合透明带糖蛋白ZP3的活性,SP2并显示凝集血球的活性。精子蛋白SP1与卵预温育明显抑制精卵结合,抑制活性与加入的精子蛋白的浓度呈正相关。用生物素标记的ZP3和蛋白质印迹技术,证明SP1中的68kD精子蛋白与ZP3结合,提示68kD精子蛋白参与精卵结合。 相似文献
958.
Recombinant human pigment epithelium-derived factor (PEDF): characterization of PEDF overexpressed and secreted by eukaryotic cells. 总被引:1,自引:0,他引:1
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E. Stratikos E. Alberdi P. G. Gettins S. P. Becerra 《Protein science : a publication of the Protein Society》1996,5(12):2575-2582
Pigment epithelium-derived factor (PEDF) is a serpin found in the interphotoreceptor matrix of the eye, which, although not a proteinase inhibitor, possesses a number of important biological properties, including promotion of neurite outgrowth and differential expression in quiescent versus senescent states of certain cell types. The low amounts present in the eye, together with the impracticality of using the eye as a source for isolation of the human protein, make it important to establish a system for overexpression of the recombinant protein for biochemical and biological studies. We describe here the expression and secretion of full-length glycosylated human recombinant PEDF at high levels (> 20 micrograms/ mL) into the growth medium of baby hamster kidney cells and characterization of the purified rPEDF by circular dichroism and fluorescence spectroscopies and neurite outgrowth assay. By these assays, the recombinant protein behaves as expected for a correctly folded full-length human PEDF. The availability of milligram amounts of PEDF has permitted quantitation of its heparin binding properties and of the effect of reactive center cleavage on the stability of PEDF towards thermal and guanidine hydrochloride denaturation. 相似文献
959.
960.
Sjaak Peelen Sybren S. Wijmenga Paul J. A. Erbel Robert L. Robson Robert R. Eady Jacques Vervoort 《Journal of biomolecular NMR》1996,7(4):315-330
Summary The 1H, 15N and 13C backbone and 1H and 13C beta resonance assignments of the long-chain flavodoxin from Azotobacter chroococcum (the 20-kDa nifF product, flavodoxin-2) in its oxidized form were made at pH 6.5 and 30°C using heteronuclear multidimensional NMR spectroscopy. Analysis of the NOE connectivities, together with amide exchange rates, 3JHnH coupling constants and secondary chemical shifts, provided extensive solution secondary structure information. The secondary structure consists of a five-stranded parallel -sheet and five -helices. One of the outer regions of the -sheet shows no regular extended conformation, whereas the outer strand 4/6 is interrupted by a loop, which is typically observed in long-chain flavodoxins. Two of the five -helices are nonregular at the N-terminus of the helix. Loop regions close to the FMN are identified. Negatively charged amino acid residues are found to be mainly clustered around the FMN, whereas a cluster of positively charged residues is located in one of the -helices. Titration of the flavodoxin with the Fe protein of the A. chroococcum nitrogenase enzyme complex revealed that residues Asn11, Ser68 and Asn72 are involved in complex formation between the flavodoxin and Fe protein. The interaction between the flavodoxin and the Fe protein is influenced by MgADP and is of electrostatic nature.Abbreviations SQ
semiquinone
- FMN
riboflavin 5-monophosphate; nif, nitrogen fixation
- TSP
3-(trimethylsilyl)propionate sodium salt
- DSS
2,2-dimethyl-2-silapentane-5-sulfonate sodium salt
Supplementary Material is available on request, comprising a Materials and Methods section for the expression and purification of the A. chroococcum flavodoxin, a Table S1 containing the parameters of the titration of A. chroococcum flavodoxin with the Fe protein, and a Table S2 containing the 15N, HN, 13C, 1H, 13C, 1H and 13CO chemical shifts.To whom correspondence should be addressed. 相似文献