Mechanism of complexing of chlorin <Emphasis Type="Italic">e</Emphasis><Subscript>6</Subscript> and tetra(<Emphasis Type="Italic">p</Emphasis>-carboxyphenyl)porphyrin with malate dehydrogenase

The mechanism of complexing of the tetrapyrrole-nature photodynamic sensitizers chlorin e₆ and tetra(p-carboxyphenyl)porphyrin (TCPP) with the Krebs-cycle enzyme malate dehydrogenase (MDG) has been investigated by spectral-luminescence methods. It is shown that each subunit entering into the composition of the MDG dimer molecule forms an equilibrium complex with one dye molecule. However, if the sites of bonding of TCPP on a protein molecule are independent, the MDG-chlorin e₆ complex has a negative cooperativity since after the dye is incorporated into the first subunit of the macromolecule, its penetration to the second subunit becomes difficult. This is explained by the fact that conformation transformations, arising as a result of the incorporation of chlorin into one of the MDG subunits, are transferred to the site of its bonding on other MDG subunits through the intersubunit contacts of the enzyme. It has been established that photosensitizers compete with the hydrophobic fluorescent probe 8-aniline-1-naphthalenesulfonate (ANS) (whose position in the MDG macromolecule was described in detail in the literature) for the sites of bonding on the protein molecule, which allows the conclusion that TCPP and chlorin e₆ are localized in the catalytic domain of MDG. In this case, the sites of bonding of these dyes and the sites of bonding of nicotinamide-adenine dinucleotide, occupying the MDG domain that bonds coenzymes, do not interact with each other.__________Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 72, No. 1, pp. 111–118, January–February, 2005.     

电聚合修饰碳纤维电极及乳酸脱氢酶活性的测定

用电聚合方法将亚甲基绿 (MG)修饰在碳纤维电极 (直径 7μm)上 ,并用该修饰电极测定了乳酸脱氢酶 (LDH)活性。在烟酰胺腺嘌呤二核苷酸 (NAD+ )浓度为 6 .0× 10 - 4mol L ,乳酸浓度为 5 .0× 10 - 3mol/L的pH7.0NaOH KH2 PO4 缓冲介质中 ,电位恒定在 +0 .10V下 ,用电流法测定乳酸脱氢酶活性 ,线性范围为 15～ 2 4 0U/mL ,检测限为 10U/mL ,响应时间为 15s。该修饰微电极稳定性好、灵敏度高、测定干扰小     

纳米ZnO修饰电极上低电位检测烟酰胺腺嘌呤二核苷酸和乙醇

纳米ZnO修饰玻碳电极在0.23 V对烟酰胺腺嘌呤二核苷酸(NADH)的氧化具有很好的催化活性, 与裸电极上NADH的氧化电位0.70 V相比, 该电位降低了0.47 V, 同时增强了抗干扰能力, 并在很大程度上减小了电极污染. 以乙醇脱氢酶(ADH)为例, 制备了ADH/ZnO修饰电极, 可用于脱氢酶底物乙醇的快速、灵敏检测, 并具有良好的重现性和稳定性. 研究表明纳米ZnO为构建脱氢酶底物的电化学传感器提供了一种新的生物兼容性材料.     

Preliminary kinetic characterization of xylose reductase and xylitol dehydrogenase extracted from Candida guilliermondii FTI 20037 cultivated in sugarcane bagasse hydrolysate for xylitol production

Candida guilliermondii FTI 20037 was cultured in sugarcane bagasse hydrolysate supplemented with 2.0 g/L of (NH₄)₂SO₄, 0.1 g/L of CaCl₂·2H₂O, and 20.0 g/L of rice bran at 35°C; pH 4.0; agitation of 300 rpm; and aeration of 0.4, 0.6, or 0.8 vvm. The high xylitol production (20.0 g/L) and xylose reductase (XR) activity (658.8 U/mg of protein) occurred at an aeration of 0.4 vvm. Under this condition, the xylitol dehydrogenase (XD) activity was low. The apparent K _M for XR and XD against substrates and cofactors were as follows: for XR, 6.4×10⁻² M (xylose) and 9.5×10⁻³ mM (NADPH); for XD, 1.6×10⁻¹ M (xylitol) and 9.9×10⁻² mM (NAD+). Because XR requires about 10-fold less xylose and cofactor than XD for the condition in which the reaction rate is half of the V _max, some interference on the overall xylitol production by the yeast could be expected.     

丙酮酸脱氢酶系：除草剂品种的新靶标

丙酮酸脱氢酶系抑制剂;综述;丙酮酸脱氢酶系：除草剂品种的新靶标     

聚苯胺用作乙醇脱氢反应中的电子传递介质

脱氢酶（ADH）在电化学氧化还原反应中是很重要的一种酶,但它在催化有机分子脱氢反应时需烟酰胺腺嘌呤二核苷酸（NAD＋）参与,后者从底物接收电子生成还原形NADH.脱氢酶电极是根据NADH的电化学氧化产生的阳极电流构成的[1－3].然而NADH与裸体炭电极和铂金电极之间的直接电子传递是非常困难的,往往需一个相当高的过电位[4].另一个问题是生成物易将电极玷污[5,6].克服这些问题的方法是使用均相电子传递介质,例如在底物溶液中加入Meldola蓝、Nile蓝A和NMP＋甲替硫酸盐等[7－9],及复相电子传递介质,例如将镍六氰基高铁酸盐固…     

Acrolein detection based on alcohol dehydrogenase inhibition

This paper presents the effect of acrolein on three dehydrogenases and proposes a fast spectrometric method for acrolein analysis. We have found that alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (AlDH) are inhibited by low acrolein concentrations (0.2?mM) while inhibition of glutamate dehydrogenase (GDH) is not observed even at higher acrolein concentrations (1?mM). Acrolein is a suicide substrate for AlDH and ADH inhibition by acrolein is competitive. Cysteine (L-Cys) and glutathione (GSH) react with acrolein and thus reduce its expected inhibitory effect. ADH was chosen to develop a spectrophotometric method for acrolein analysis based on enzyme inhibition. The calibration curve is linear between 0.2 and 1.0?mM acrolein.     

Some Biochemical Aspects of the Protective Effect of Strontium Chloride on Gamma-Irradiated Rats

The effect of treatment with SrCl₂ (10 mg/100 g rat) on rats 15 minutes prior to whole body γ-irradiation (7,5 Gy) was studied. The hazardeous effect of irradiation were greatly corrected in the treated group. The hyperglycemic effect and liver glycogen accumulation in the untreated group decreased to normal level. The enzymatic activities of serum alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase and lactate dehydrogencase were greatly affected showing insignificant changes in the treated group of animals. Life span calculated on 50% survival was also significantly elongated by 36.3%. These results show the potentiality of SrCl₂ as a radioprotective agent which was not used before.     

Microchip extraction of catecholamines using a boronic acid functional affinity monolith

We report on the development of a rapid enzyme logic gate-based electrochemical assay for the assessment of traumatic brain injury (TBI). The concept harnesses a biocatalytic cascade that emulates the functionality of a Boolean NAND gate in order to process relevant physiological parameters in the biochemical domain. The enzymatic backbone ensures that a high-fidelity diagnosis of traumatic brain injury can be tendered in a rapid fashion when the concentrations of key serum-based biomarkers reach pathological levels. The excitatory neurotransmitter glutamate and the enzyme lactate dehydrogenase were used here as clinically-relevant input TBI biomarkers, in connection to the low-potential detection of the NADH product in the presence of methylene green at a glassy carbon electrode. A systematic optimization of the gate and the entire protocol has resulted in the effective discrimination between the physiological and pathological logic levels. Owing to its robust design, the enzyme-based logic gate mitigates potential interferences from both physiological and electroactive sources and is able to perform direct measurements in human serum samples. Granted further detailed clinical validation, this proof-of-concept study demonstrates the potential of the electrochemical assay to aid in the rapid and decentralized diagnosis of TBI.