Direct evidence of electron flow via the heme c group for the direct electron transfer reaction of fructose dehydrogenase using a silver nanoparticle-modified electrode

The direct electron transfer reaction of fructose dehydrogenase (FDH) from Gluconobacter sp. on alkanethiol-modified silver nanoparticles (AgNPs) was examined using cyclic voltammetry and surface-enhanced resonance Raman scattering (SERRS). Using cyclic voltammetry, catalytic oxidation currents (based on the direct electron transfer reaction of FDH) were observed from a potential of approximately −100 mV (vs. Ag/AgCl, 3 M NaCl) in the presence of d-fructose, without a mediator. A comparison of the SERRS spectra and the resonance Raman spectra of FDH in solution indicated that the heme c site retained its six-coordinated low-spin heme after immobilization. Moreover, SERRS also demonstrated that the heme c of the adsorbed FDH was the electron transfer site within the enzyme.     

One site fits both: A model for the ternary complex of folate + NADPH in R67 dihydrofolate reductase, a D2 symmetric enzyme

R67 dihydrofolate reductase (DHFR) is a novel enzyme that confers resistance to the antibiotic trimethoprim. The crystal structure of R67 DHFR displays a toroidal structure with a central active-site pore. This homotetrameric protein exhibits 222 symmetry, with only a few residues from each chain contributing to the active site, so related sites must be used to bind both substrate (dihydrofolate) and cofactor (NADPH) in the productive R67 DHFR?NADPH?dihydrofolate complex. Whereas the site of folate binding has been partially resolved crystallographically, an interesting question remains: how can the highly symmetrical active site also bind and orient NADPH for catalysis? To model this ternary complex, we employed DOCK and SLIDE, two methods for docking flexible ligands into proteins using quite different algorithms. The bound pteridine ring of folate (Fol I) from the crystal structure of R67 DHFR was used as the basis for docking the nicotinamide-ribose-P_i (NMN) moiety of NADPH. NMN was positioned by both DOCK and SLIDE on the opposite side of the pore from Fol I, where it interacts with Fol I at the pore's center. Numerous residues serve dual roles in binding. For example, Gln 67 from both the B and D subunits has several contacts with the pteridine ring, while the same residue from the A and C subunits has several contacts with the nicotinamide ring. The residues involved in dual roles are generally amphipathic, allowing them to make both hydrophobic and hydrophilic contacts with the ligands. The result is a `hot spot' binding surface allowing the same residues to co-optimize the binding of two ligands, and orient them for catalysis.     

间接原子吸收法测定乳酸环丙沙星

研究了在弱酸性介质中，乳酸环丙沙星与雷氏盐定量生成缔合物的反应条件，以及通过原子吸收法测定沉淀中Cr的含量而间接测定乳酸环丙沙星含量的分析方法。线性范围在5－40mg/L，相对标准偏差为2．5％，回收率在98％－101％之间。     

基于细菌脱氢酶活性法的水质综合毒性快速测定仪

在无氧条件下,生物染料亚甲基蓝可以作为生物新陈代谢过程中体内脱氢酶催化反应的电子受体,亚甲基蓝与细菌悬液共存时,其退色速度标志着细菌脱氢酶活性的大小。脱氢酶的活性大小在一定程度上能够反映出微生物新代谢的能力,微生物所处的水环境中毒物的存在会引起微生物脱氢酶活性变化,毒物的毒性与脱氢酶的活性相关。本文以动力学比色法测定亚甲基蓝的退色速度为基础设计并制造了一台水质综合毒性相关。本文以动力学比色法测定亚     

乳酸正丁酯的合成

报道了合成乳酸丁酯的新方法 ,以乳酸和正丁醇为原料 ,在甲基磺酸催化下以亚硫酸镁为脱水剂 ,用索氏提取器进行回流合成。应用该方法 ,酯化时间缩短到 3h,收率 97.2 % ,同时对影响收率的各个因素进行了研究。     

Norcholanic acids as substrates for recombinant 3β-hydroxysteroid dehydrogenase and progesterone 5β-reductase, enzymes of the 5β-cardenolide biosynthesis

The conversion of 23-nor-5,20(22)E-choladienic acid-3β-ol and other intermediates of the putative norcholanic acid pathway of cardenolide biosynthesis by recombinant 3β-hydroxysteroid dehydrogenase from Digitalis lanata in dehydrogenation and reduction reactions was investigated. 23-nor-4,20(22)E-choladienic acid-3-one was found to be a substrate of recombinant progesterone 5β-reductases from D. lanata and Arabidopsis thaliana. The role of various substrates in cardenolide biosynthesis is discussed.     

Assay of Serum Glutamic-Oxaloacetic Transaminase Using Malate Dehydrogenase Preparation Obtained from Streptomyces aureofaciens

Abstract

A modified spectrophotometric method for serum glutamic-oxaloacetic transaminase (SGOT) assay was developed. A crude cell-free extract from Streptomyces aureofaciens which showed a high level of malate dehydrogenase (MDH) activity (E.C. 1.1.1.37) was used as the enzymatic indicator. The lyophilized microbial preparation was used without previous purification and was quite stable under refrigeration for one year. Serum sample assays using both the method utilizing the crude cell extract and an enzymatic commercial kit showed good correlation.     

Effects of Micronutrient Niacin on Treatment Efficiency of Textile Wastewater

0 IntroductionMioclroogincuatlri pernotsce pslsaeys afo vrer wyais tmepwoarttaernt t rroelaetmine nbti- .A widerange of micronutrientsis required byactiva-ted sludge[1-5]. Adequate trace nutrient concentra-tions ,including metal ions and vitamins , are re-quiredtosupport all the genera presentinthe activa-ted sludgetreatment systemfor a diverse communityto survive. The lack of specific nutrients may resultin an unbalanced mixed culture,leading to poor-quality effluent , reduced treatment effic…     

Effects of He－Ne laser irradiation on enzyme activity in milking Goats＇ seminal plasma

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Polymorphisms in genes involved in folate metabolism as maternal risk factors for Down syndrome in China

Objective： To explore the relationship between genetic polymorphisms in methylenetetrahydrofolate reductase （MTHFR）, methionine synthase reductase （MTRR）, the central enzymes in folate metabolism that affects DNA methylation and synthesis, and the risk of Down syndrome in China. Methods： Genomic DNA was isolated from the peripheral lymphocytes of 64 mothers of children with Down syndrome and 70 age matched control subjects. Polymerase chain reaction and restriction fragment length polymorphism were used to examine the polymorphisms of MTHFR 677C→T, MTRR 66A→G and the relationship between these genotypes and the risk of Down syndrome was analyzed. Results： The results show that the MTHFR 677C→T polymorphism is more prevalent among mothers of children with Down syndrome than among control mothers, with an odds ratio of 3.78 （95% confidence interval （CI）, 1.78-8.47）. In addition, the homozygous MTRR 66A→G polymorphism was independently associated with a 5.2-fold increase in estimated risk （95% CI, 1.90-14.22）. The combined presence of both polymorphisms was associated with a greater risk of Down syndrome than the presence of either alone, with an odds ratio of 6.0 （95% CI, 2.058-17.496）. The two polymorphisms appear to act without a multiplicative interaction. Conclusion： MTHFR and MTRR gene mutation alleles are related to Down syndrome, and CT, TT and GG gene mutation types increase the risk of Down syndrome.