共查询到20条相似文献,搜索用时 8 毫秒
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An enzymatic route to L-alpha-glycerylphosphorylcholine 总被引:5,自引:0,他引:5
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C A Marsh 《Analytical biochemistry》1985,145(2):266-272
A method was developed for determination of D-glucaric acid by treatment with a bacterial extract containing glucarate dehydratase and ketodeoxyglucarate aldolase. This led to the quantitative formation of pyruvate, which was then assayed by use of lactate dehydrogenase. Measurements of D-glucarate in individual samples of human urine by this technique were compared with those by the commonly used method of beta-glucuronidase inhibition, and gave values for D-glucarate content which were about 25% higher, but with otherwise good correlation. 相似文献
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Over-production of lactate dehydrogenase from Plasmodium falciparum opens a route to new antimalarials 总被引:2,自引:0,他引:2
Dilek Turgut-Balik Debbie K. Shoemark Kathleen M. Moreton Richard B. Sessions J. John Holbrook 《Biotechnology letters》2001,23(11):917-921
Over-production of lactate dehydrogenase (PfLDH) from Plasmodium falciparum from E. coli TG2 cells transformed with a pKK223-3 plasmid containing the wild type gene isolated by Bzik DJ, Fox BA, and Gonyer K (1993) Mol. Biochem. Parasit.
59, 155–166, gave mostly an inactive protein after isolation. Sequencing the N-terminus of the over-produced protein showed that the major product commenced at an internal methionine. Truncation of the protein occurred due to the inappropriate priming from a Shine–Dalgarno (SD) sequence upstream of Met 35. Silent mutations of this SD sequence to remove the purine-rich region allowed over-production of the full length PfLDH up to 15 mg protein l–1 broth. The purified protein exhibited biochemical properties of an authentic LDH enzyme. However, high activity with 3-acetylpyridine adenine dinucleotide as well as with the natural cofactor, NAD, was also observed. The high-resolution X-ray structure obtained from the recombinant enzyme has provided the opportunity for the development of inhibitors specific to PfLDH. 相似文献
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One-cell mouse embryos were cultured in several concentrations of pyruvate and lactate. Maximum development to blastocysts occurred when one-cell ova were cultured in media containing 0.25 mM pyruvate during the first cleavage division and 30.00 mM lactate plus 0.25 mM pyruvate after the first cleavage division. The unusual sensitivity of one-cell ova to both the kind and quantity of energy source was not evident on day 2 of development; normal appearing two-cell ova were formed under extreme conditions of up to 100.00 mM pyruvate and 90.00 mM lactate. The data demonstrate that the successful development of one-cell ova in vitro depends on satisfying separate requirements for the first cleavage division versus development after the first cleavage division. The formation of morphologically normal two-cell ova cannot be used as the sole criterion for satisfying the requirements of the first cleavage divison. 相似文献
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Gas exchange following lactate and pyruvate injections 总被引:1,自引:0,他引:1
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The kinetics of pyruvate reduction by lactate dehydrogenase from Phycomyces blakesleeanus NRRL 1555 (-) have been determined at pH 6.0. Initial rate studies performed in the pyruvate reduction direction suggest that a sequential mechanism is operating. Product inhibition studies with NAD+ and L(+)-lactate are consistent with an ordered sequential mechanism if we considered that NAD+ mimics the NADH that binds cooperatively on the enzyme and also the existence of dead-end complex responsible for substrate inhibition by pyruvate at this pH value. 相似文献
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Summary A basic procedure was developed to produce high-protein amaranth flour (HPAF) using a commercial preparation of heat-stable alpha-amylase. Slurries (20%, w/v) of gelatinized whole flour were liquefied at 70 and 90°C, pH 6.5, 0.1% (w/v) enzyme concentration and 30 min hydrolysis time. Protein content of raw flour was increased from 15 to 29.6 or 39.3% at liquefaction temperatures of 70 or 90°C, respectively. Some physicochemical and functional properties of HPAF were assessed. HPAF might be used as a dry milk extender. 相似文献
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The lactate dehydrogenase-catalyzed reduction of pyruvate by NADH was studied using a spectroscopic method. The inhibitory effect exhibited by high concentrations of pyruvate was investigated in phosphate and 2,2-diethylmalonate buffers. Kinetic studies were carried out in which the rate of the enzyme-catalyzed reaction was monitored at various stages of pyruvate hydration, H2O + CH3COCO2? ? CH3C(OH)22C02?. Buffered solutions of different initial relative amounts of ketopyruvate and hydrated pyruvate (2,2-dihydroxypropanoic acid) were also preincubated with the enzyme and NAD+. Kinetic runs were initiated in the resultant solutions at various stages of incubation by the introduction of NADH. The results of the present investigation indicate that hydrated pyruvate is a major inhibitor of lactate dehydrogenase and forms an inhibitory complex with the enzyme and oxidized coenzyme. 相似文献
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K Y Hostetler H R Williams W W Shreeve B R Landau 《The Journal of biological chemistry》1969,244(8):2075-2077
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Regulation of gluconeogenesis from pyruvate and lactate in the isolated perfused rat liver 总被引:1,自引:0,他引:1
The effects of glucagon and the alpha-adrenergic agonist, phenylephrine, on the rate of 14CO2 production and gluconeogenesis from [1-14C]lactate and [1-14C]pyruvate were investigated in isolated perfused livers of 24-h-fasted rats. Both glucagon and phenylephrine stimulated the rate of 14CO2 production from [1-14C]lactate but not from [1-14C]pyruvate. Neither glucagon nor phenylephrine affected the activation state of the pyruvate dehydrogenase complex in perfused livers derived from 24-h-fasted rats. 3-Mercaptopicolinate, an inhibitor of the phosphoenolpyruvate carboxykinase reaction, inhibited the rates of 14CO2 production and glucose production from [1-14C]lactate by 50% and 100%, respectively. Furthermore, 3-mercaptopicolinate blocked the glucagon- and phenylephrine-stimulated 14CO2 production from [1-14C]lactate. Additionally, measurements of the specific radioactivity of glucose synthesized from [1-14C]lactate, [1-14C]pyruvate and [2-14C]pyruvate indicated that the 14C-labeled carboxyl groups of oxaloacetate synthesized from 1-14C-labeled precursors were completely randomized and pyruvate----oxaloacetate----pyruvate substrate cycle activity was minimal. The present study also demonstrates that glucagon and phenylephrine stimulation of the rate of 14CO2 production from [1-14C]lactate is a result of increased metabolic flux through the phosphoenolpyruvate carboxykinase reaction, and phenylephrine-stimulated gluconeogenesis from pyruvate is regulated at step(s) between phosphoenolpyruvate and glucose. 相似文献
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Pyruvate, lactate and lactate dehydrogenase appeared linearly in 2 ml 0.9% NaCl recirculated through the rabbit oviduct for 4 h in vivo. In oviducts from rabbits injected 3 days previously with 100 i.u. hCG, the rate of appearance of all three constituents was considerably reduced. It is considered unlikely that the lactate dehydrogenase secreted brings about the interconversion of pyruvate and lactate in the oviduct lumen. 相似文献
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S Yamamoto 《The International journal of biochemistry》1983,15(2):185-190
1. Low enzymatic activities in low pyruvate concentrations and high Km were observed in sodium chloride solutions. 2. The pyruvate inhibition shown by the % activity at 1 mM pyruvate was lower sodium chloride than in 0.1 M sodium phosphate. 3. At 40 degrees C, as compared with results at 20 degrees C, less pyruvate inhibition was observed in phosphate buffer and in sodium chloride solutions. 4. By using the equilibrium constants between dimer and tetramer, a theoretical explanation is proposed for the pyruvate inhibition. In this explanation, it is suggested that the quaternary complex which is composed of tetrameric enzyme, coenzyme and two kinds of pyruvates was the main cause of the pyruvate inhibition. 相似文献