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11.
基于可见吸收信号的乳酸脱氢酶光纤传感器 总被引:1,自引:0,他引:1
报道一种测定乳酸脱氢酶活力的基于可见吸收信号的光纤生物传感器,在该传感体系中,通过辅酶I的氧化还原对(NAD^+/NADH)将乳酸脱氢酶和心肌黄酶催化的两个脱氢过阳以耦合,第一个脱氢过程对分析对象进行了化学识别,第二个脱氢过程引起可见吸收信号的变化,该传感器对0~400U/L的乳酸脱氢酶有线性响应关系,检测下限为48UL,该传感器已用于人体血清中乳酸脱氢酶活力的测定。 相似文献
12.
介体型乳酸脱氢酶生物传感器的研制及应用 总被引:3,自引:0,他引:3
本报道了能对丙酮酸产生良好响应的电流型乳酸脱氢酶生物传感器。该传感器以耐尔兰A修饰浸石墨电极为基体电极,将酶直接固化在蚕丝蛋白膜上。在PH7.4的NaH2PO4-NaOH介质中,当2.4×10^-4mol/L的辅酶I(NADH)存在下,该传感器的响应电流与丙酮酸浓度在3.2×10^-5mol/L范围内有良好的线性关系。响应时间为60s.本讨论了影响传感器响应的各种因素,用此传感器测定了人血清中 相似文献
13.
Nils Siegbahn Mats-Olle Maånsson Klaus Mosbach 《Applied biochemistry and biotechnology》1987,12(2):91-105
A soluble, bifunctional enzyme complex has been prepared by crosslinking lactate dehydrogenase and alcohol dehydrogenase with
glutaraldehyde. The crosslinking was performed on a solid phase while the active sites of alcohol dehydrogenase and lactate
dehydrogenase were held adjacent to one another with the aid of a bis-NAD analog. Subsequently, the enzyme complex was released
from the solid phase. The soluble enzyme complex was then purified by using NAD-Sepharose as an affinity adsorbent. Based
on gel filtration experiments, the complex was estimated to consist of one of each dehydrogenase.
By using a third enzyme, lipoamide dehydrogenase, which competes with lactate dehydrogenase for NADH produced by alcohol dehydrogenase,
the effect of site-to-site orientation was studied. It was found that about 83% of the NADH produced by alcohol dehydrogenase
was oxidized by site-to-site oriented lactate dehydrogenase compared to a figure of only about 61% obtained in an identical
system of separate enzymes. This indicates that given two alternative routes, the preference for the one to lactate dehydrogenase
over the one to lipoamide dehydrogenase is enhanced when lactate dehydrogenase and alcohol dehydrogenase are site-to-site
oriented. 相似文献
14.
Fumihiko Hasumi Yasumitsu Miyamoto Ichiro Okura 《Applied biochemistry and biotechnology》1995,55(1):1-4
Glutamate synthesis by reductive amination of 2-oxoglutarate was performed by the combination of NADH regeneration system
and glutamate dehydrogenase (GluDH). The conversion of 2-oxoglutamate to glutamate was 98% after 3 h, and the turnover number
of NAD+was 17. 相似文献
15.
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17.
介体型乙醇生物传感器的研制及应用 总被引:1,自引:0,他引:1
本文报道了电流型乙醇生物传感器的研制与应用。该传感器以健合型耐尔蓝修饰浸蜡石墨电极为基体电极,将乙醇脱氢酶及NAD^+固定在人造丝网上,成为一种无试剂的乙醇生物传感器。在pH8.8的Tris/HCl介质中,该传感器的响应电流与乙醇浓度在0.10-1.0mmol/l范围内有良好的线性关系。 相似文献
18.
Fumihiko Hasuni Katsunori Fukuoka Shuichi Adachi Yasumitsu Miyamoto Ichiro Okura 《Applied biochemistry and biotechnology》1996,56(3):341-344
Alanine synthesis by reductive amination of pyruvate was performed by the combination of NADH regeneration system and alanine
dehydrogenase (AlaDH). The conversion of pyruvate to alanine was 99% after 1 h. Leucine synthesis was also carried out by
the combination of NADH regeneration system and leucine dehydrogenase (LeuDH). The conversion of 4-methyl-2-oxovalerate to
leucine was 60% after 1.5 h. 相似文献
19.
含离子液体[bmim]Cl的反应介质中马肝醇脱氢酶的催化特性 总被引:2,自引:0,他引:2
研究了马肝醇脱氢酶(HLADH)在含离子液体1-丁基-3-甲基咪唑氯酸盐([bmim]Cl)的反应介质中的催化特性. 以乙醇为底物时,该酶在[bmim]Cl含量≤0.15 g/ml的体系中的活力高于在不含离子液体的体系中的活力; 离子液体浓度过高(>0.15 g/ml)对酶活性有明显的抑制作用. 反应温度和pH对含离子液体的反应介质中酶活力的影响规律与不含离子液体时的规律相似. 与不含离子液体的反应介质相比, HLADH在含0.05 g/ml [bmim]Cl的体系中催化乙醇氧化的活化能下降,酶反应的Vmax和Km均升高. 反应体系中低浓度(≤0.1 g/ml)的离子液体能提高酶的热稳定性,但高浓度(>0.1 g/ml)的离子液体可降低酶的热稳定性. 紫外二阶导数光谱显示,在含不同浓度离子液体的反应介质中酶分子构象的变化有较大的差异. 相似文献
20.
We have evaluated the potential of near-infrared spectroscopy (NIRS) as a technique for rapid analysis of lactate in whole blood. To test the NIRS technique, a comparison was made with a standard clinical method using whole blood samples taken from five exercising human subjects at three different stage of exercise. To expand lactate concentration within the physiological range, standard additions method was used to generate 45 unique data points. Spectra were collected over the 2050-2400 nm spectral range with a 1 mm optical path length quartz cell. Reference lactate concentrations in the samples were determined by enzymatic measurements. Estimates and calibration of the lactate concentration with NIRS was made using partial least squares (PLS) regression analysis and leave-N-out cross validation on second derivative spectra. Separate calibrations were determined from each of the subject samples and cumulative PRESS was used to determine the number of PLS factors in the final model. The results from the PLS model presented are generated from the five individual calibration coefficient vectors and provided a correlation coefficient of 0.978 and a standard error of cross validation of 0.65 mmol l−1 between the enzymatic assay and the NIRS technique. To study the parameters that impact the spectra baseline and the correlation between the calculated model and the data, referenced measurements of lactate against baseline spectrum were made for each individual. A correlation coefficient of 0.992 and a standard error of cross validation of 0.21 mmol l−1 were found. The results suggest that NIRS may provide a valuable tool to assess physiological status for both research and clinical needs. 相似文献