5-氟尿苷的微生物转化

Conditions for biotransformation and purification of FUR were investigated.The result showed that when the cell concentration of E.aerogenes was 10%(w/v),the temperature and pH were 7.8 and 60℃ respectively,59.7% UR was converted to FUR.It also demonstrated that the ideal procedure for the purification of FUR is silica gel column chromatography by two elution systems (S 1∶CHCl 3∶CH 3OH∶H 2O 61∶13∶1,and S 2;CH 3COOCH 2CH 3∶CH 3OH∶CH 3COOH∶H 2O 12∶3∶3∶2)with the purity and yield of 98.0% and 86.0% …     

大叶藤黄果的大叶藤黄醇

大叶藤黄(Garcinia xanthochymus Hook.f.ex J.Anders.)又名人面果,岭南倒捻子,最近我们从果分泌的树脂中分离、鉴定了大叶藤黄醇(xanthochmol)。 大叶藤黄醇为黄色针状晶,mp 135℃,IRν_(max)~(KBr).(cm~(-1)):3200—3400(OH),1640—1660,1715(>C=O),3079(C=CH基的νc-H振动),890(末端甲烯基)。 ~1H NMR(WH-90MHz.TMS为内标,CDC_3为溶剂)δ(ppm):1.02,1.17(2s,各3H,>CMe_2),且1.54×2,1.70×2(2s,各6H,—CH=CMe_2×2)。     

外源H_2O_2和·OH对大麦幼苗根系线粒体膜脂和流动性的伤害

以大麦 (H ordeum vulgare L.)为材料 ,研究了外源 H2 O2 和· OH对大麦根系呼吸速率、线粒体膜流动性和膜脂脂肪酸组成的影响。结果表明 ,1 0 mmol/L H2 O2 或· OH处理 4d,大麦幼苗根系呼吸速率和线粒体膜脂不饱和脂肪酸含量及脂肪酸不饱和指数下降 ,线粒体膜脂荧光强度增加 ,膜流动性下降 ,且 H2 O2 或· OH处理浓度 (在 0 .1～ 1 0 mmol/L范围内 )越高 ,膜脂流动性下降越明显。 H2 O2 和· OH处理的同时加入同浓度的抗坏血酸 (As A)和甘露醇 ,膜流动性明显增强或恢复     

鼎湖山针阔叶混交林地表温室气体排放的日变化

利用静态箱 -气相色谱法对鼎湖山针阔叶混交林地表 3种温室气体 CO2 、CH4 、N2 O通量进行了原位观测 .结果表明 ,鼎湖山针阔叶混交林地表为 CO2 、N2 O的排放源 ,为 CH4 的弱汇 ,通量日变幅分别是 4 88.99～ 70 0 .5 7,0 .0 4 9～ 0 .10 8mg/ (m2· h)和- 0 .0 2 5～ - 0 .0 5 3mg/ (m2 · h) ;地表凋落物分解释放 CO2 约占总排放的 1/ 3;凋落物层和林下灌木对 CH4 和 N2 O的通量无明显影响 ;CO2 、N2 O的 9∶ 0 0观测值与其日平均值有明显差异     

水杨酸提高香蕉幼苗的抗冷性与H2O2代谢有关

探讨了水杨酸(salicylic acid,SA)提高香蕉幼苗抗冷性的可能机理。在常温下(30／22℃)用不同浓度(0—3．5mmol／L)的SA水溶液喷洒叶片1d,置于7℃低温下冷胁迫3d,随后于常温下恢复2d后测定电解质泄漏率,结果表明：SA0．3～0．9mmol／L能显著提高香蕉幼苗的抗冷性,以0．5mmol／L效果最佳。若把冷胁迫温度降到5℃,SA0．5mmol／L预处理可显著减少幼苗叶片的萎蔫面积。但当SA浓度高于1．5mmol／L时,恢复期间的电解质泄漏甚至高于对照(蒸馏水处理),表明它们加剧了冷害。SA提高香蕉幼苗的抗冷性可能需要H2O2的参与：1)SA0．5mmol／L常温处理诱导了H2O2的积累和活性氧造成的膜脂过氧化——三氯乙酸反应物质(TBARS)的增加,这可能与H2O2的清除酶——过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性的受抑和H2O2的产生酶—超氧化物歧化酶(SOD)活性几乎不受影响有关;2)外源H2O2(1．5—2．5mmoL／L)也能显著降低低温胁迫期间的电解质泄漏,表明也能提高抗冷性;3)而用H2O2的捕捉剂——二甲基硫脲(DMTU)可明显抑制SA诱导的抗冷性;4)在低温胁迫与恢复期间,SA预处理明显提高了CAT和APX的活性,抑制了H2O2与TBARS的快速上升。     

高中生物学新教材"pH对酶活性的影响"实验中用斐林试剂的道理

高中生物学新教材 [实验七 ]探索影响淀粉酶活性的条件 (第 4 9页 ) ,其中“p H对酶活性的影响”实验中 ,若用碘液检验淀粉是否被淀粉酶水解 ,产生的现象将是 1、2号试管中的液体不变蓝色 ,3号试管溶液变蓝色。原因是 :1号试管内既没加碱 ,也没加酸 ,溶液近似中性 ,适于淀粉酶发挥催化作用 ,所以淀粉被分解成麦芽糖 ,而麦芽糖遇碘不变蓝色 ;2号试管内加入了Na OH溶液 ,使得 p H过高 ,淀粉酶失活 ,淀粉未分解 ,但加入碘液检验仍不变蓝色 ,这是因为 I2 与 Na OH发生了反应 ,即 :3I2 +6 Na OH 冷 5 Na I+Na IO3 +3H2 O(歧化反应 ) ,消…     

稻田CH4和N2O排放关系及其微生物学机理和一些影响因子

用静态箱法原位观测和分析了我国北方稻田 3～ 1 2月CH4和N2 O的排放及其关系 ,并研究了这一关系发生的微生物学机理 .同时 ,监测了土壤湿度、pH、水分及Eh的变化 .结果表明 ,稻田CH4和N2 O排放之间存在着互为消长的关系 (R2 =0 0 4 94) .土壤湿度、pH及Eh变化范围分别在 0～ 2 4℃、6 87～ 7 0 2和 41 5～ 30 0mv之间 ,水分从非淹水期的 38～ 72 ?至 5～ 1 0cm浅水淹灌 .土壤Eh对CH4和N2 O的释放起重要的调控作用 .在整个观测期内 ,与CH4和N2 O释放密切相关的 6种菌群 (发酵细菌、产氢产乙酸细菌、产甲烷细菌、甲烷氧化菌、硝化细菌、反硝化细菌 )各有其数量消长及酶活性变化规律 ,稻田CH4和N2 O排放之间互为消长的关系受这些相关微生物数量及酶活性变化的共同调控 .     

外源H2O2对水稻幼苗抗寒性的调节作用

在常温下用不同浓度的外源H2O2(0～20 mmol·L-1)预处理水稻幼苗,再进行12 h 6℃低温胁迫,根据幼苗相对含水量和质膜相对透性筛选最佳外源H2O2处理浓度,并分析最佳外源H2O2浓度下幼苗的渗透调节物质和活性氧相关指标的变化.结果表明:(1)0～8 mmol·L-1 H2O2预处理可以增加水稻幼苗的相对含水量,降低其质膜相对透性,并以4 mmol·L-1 H2O2的效果最佳.(2)低温胁迫后,与对照组相比,4 mmol·L-1外源H2O2预处理降低了水稻幼苗萎蔫程度,并使其总呼吸速率、交替途径容量都有增加,同时还抑制了丙二醛的含量,增加了可溶性糖、可溶性蛋白质和脯氨酸的含量.(3)外源H2O2预处理对水稻幼苗的内源H2O2含量以及O(-)/(·)2产生速率没有显著影响.研究发现,外源H2O2可以通过提高呼吸速率、降低脂质过氧化程度、增加碳氮代谢来有效增强水稻幼苗的抗寒性,它可能以一种独立于内源活性氧系统之外的方式发挥作用.     

沙棘果皮多糖清除氧自由基的活性研究

沙棘(Hippophae rhamnosides)果皮经80℃恒温水浴提取,乙醇沉淀得粗多糖.Sevag法去蛋白,经50%和70%乙醇分级,得3种级分沙棘多糖H1、H2和H3;以Fenton反应,即H2O2/Fe2 /水杨酸为·OH产生和检测体系;以邻苯三酚/EDTA/Tris-HCl为O2-.产生体系,对沙棘多糖H1、H2和H3进行抗氧自由基活性研究.结果表明,沙棘多糖对·OH和O2-.有较显著的清除能力.不同级分多糖H1、H2和H3浓度达200μg·mL-1时,对·OH的清除率分别为44.9%、49.0%和26.4%,抗O2-.活性分别为36.9%,15.4%和23.1%.多糖质量浓度增大时,两种自由基清除率增加,且呈量效关系.     

沙棘果皮多糖清除氧自由基的活性研究

沙棘(Hippophae rhamnosides)果皮经80℃恒温水浴提取, 乙醇沉淀得粗多糖。Sevag法去蛋白,经50%和70%乙醇分级, 得3种级分沙棘多糖H1、H2和H3; 以Fenton 反应, 即H2O2/Fe2+/水杨酸为.OH产生和检测体系; 以邻苯三酚/EDTA/Tris-HCl为O2 -. 产生体系, 对沙棘多糖H1、H2和H3进行抗氧自由基活 性研究。结果表明, 沙棘多糖对.OH和O2 -. 有较显著的清除能力。不同级分多糖H1、H2和H3浓度达200mg.mL-1时, 对.OH的清除率分别为44.9%、49.0%和26.4%, 抗O2-. 活性分别为36.9%,15.4%和23.1%。多糖质量浓度增大时,两种自由基清除率增加, 且呈量效关系。     

Metal-independent production of hydroxyl radicals by halogenated quinones and hydrogen peroxide: an ESR spin trapping study

The metal-independent production of hydroxyl radicals (*OH) from H(2)O(2) and tetrachloro-1,4-benzoquinone (TCBQ), a carcinogenic metabolite of the widely used wood-preservative pentachlorophenol, was studied by electron spin resonance methods. When incubated with the spin trapping agent 5,5-dimethyl-1-pyrroline N-oxide (DMPO), TCBQ and H(2)O(2) produced the DMPO/*OH adduct. The formation of DMPO/*OH was markedly inhibited by the *OH scavenging agents dimethyl sulfoxide (DMSO), ethanol, formate, and azide, with the concomitant formation of the characteristic DMPO spin trapping adducts with *CH(3), *CH(CH(3))OH, *COO(-), and *N(3), respectively. The formation of DMPO/*OH and DMPO/*CH(3) from TCBQ and H(2)O(2) in the absence and presence, respectively, of DMSO was inhibited by the trihydroxamate compound desferrioxamine, accompanied by the formation of the desferrioxamine-nitroxide radical. In contrast, DMPO/*OH and DMPO/*CH(3) formation from TCBQ and H(2)O(2) was not affected by the nonhydroxamate iron chelators bathophenanthroline disulfonate, ferrozine, and ferene, as well as the copper-specific chelator bathocuproine disulfonate. A comparative study with ferrous iron and H(2)O(2), the classic Fenton system, strongly supports our conclusion that *OH is produced by TCBQ and H(2)O(2) through a metal-independent mechanism. Metal-independent production of *OH from H(2)O(2) was also observed with several other halogenated quinones.     

The interaction of native DNA with dimethyltin(IV) species

The reaction of aqueous native DNA (calf thymus) with the solvated organotin(IV) species [(CH3)2SnCl2(C2H5OH)n], as well as with [(CH3)2Sn(OH)(H2O)n]+ and (CH3)2Sn(OH)2 (i.e., the hydrolysis products of aqueous (CH3)2SnCl2 at pH approximately 5 and pH approximately 7.4 respectively), was investigated by 119Sn M?ssbauer spectroscopy. The addition of [(CH3)2SnCl2(C2H5OH)n] to DNA yielded a solid product, possibly (CH3)2Sn(DNA phosphodiester)2, where the environment of the tin atom is trans-octahedral with linear CSnC skeleton, and the equatorial atoms may consist of oxygen or nitrogen from water as well as from the nucleic acid constituents. No interaction with DNA apparently takes place due to hydrolyzed dimethyltin(IV) species, which occur in aqueous phases at approximate physiological pH values. The reaction pathway is then assumed to require weakly solvated, easily dissociable species such as [(CH3)2SnCl2(C2H5OH)n], which would imply in vivo reactivity of cellular DNA with organotins from hydrophobic sites.     

Synthetic analogs for oxovanadium(IV/V)-glutathione interaction: an NMR, EPR, synthetic and structural study of oxovanadium(IV/V) compounds with sulfhydryl-containing pseudopeptides and dipeptides

The reaction of [VO(CH3COO)2(phen)] (phen = 1,10-phenanthroline) with the sulfhydryl-containing pseudopeptides (scp), N-(2-mercaptopropionyl)glycine (H3mpg), N-(2-mercaptopropionyl)cysteine (H4m2pc), N-(3-mercaptopropionyl)cysteine (H4m3pc) and the dipeptides glycylglycine (H2glygly) and glycyl-L-alanine (H2glyala), in the presence of triethylamine, results in the formation of the compounds Et3NH[VO(mpg)(phen)] (1), (Et3NH)2[VO(m2pc)] (4), [(Et3NH)2[VO(m3pc) (5), [VO(glygly)(phen)] x 2CH3OH (2 x 2CH3OH) and [VO(glyala)(phen)] x CH3OH (3 x CH3OH). Evidence for the molecular connectivity in 2 x CH3OH was established by X-ray crystallography, showing the vanadium(IV) atom ligated to a tridentate glygly2- ligand at the N(amine), N(peptide) and O(carboxylato) atoms. Combination of the correlation plot of the EPR parameters gz versus Az, together with the additivity relationship supported the prediction of the equatorial donor atom sets of the V(IV)O2+ center at various pH values for the V(IV)O2+-glutathione system considered in this study. Model NMR studies (interaction of vanadium(V) with the scp H3mpg) showed that there is a possibility of vanadium(V) ligation to glutathione.     

5—氟尿苷的微生物转化

5 氟尿苷 (简称ＦＵＲ)是抗肿瘤核苷药物脱氧氟尿苷 (Ｆｌｏｘｕｒｉｄｉｎｅ ,简称ＤＦＵＲ)的合成中间体。脱氧氟尿苷是一种抗代谢类抗肿瘤药 ,在体内可以部分转化为氟尿嘧啶 (简称ＦＵ) ,二者具有相似的作用途径和抗肿瘤谱。与ＦＵ相比 ,由于ＤＦＵＲ的抗肿瘤活性高且毒副反应小 ,主要用于治疗晚期结直肠癌和各种类型肝癌。在国内 ,采用化学法合成的ＤＦＵＲ业已进入临床研究阶段[1]采用化学合成法生产ＤＦＵＲ时 ,由于反应过程中需将碱基或核糖残基的部分基团进行保护 ,而且产物为多种核苷异构体和其它副产品的混合物 ,需要进一步分离 ,…     

Metabolic mechanisms of methanol/formaldehyde in isolated rat hepatocytes: carbonyl-metabolizing enzymes versus oxidative stress

Methanol (CH(3)OH), a common industrial solvent, is metabolized to toxic compounds by several enzymatic as well as free radical pathways. Identifying which process best enhances or prevents CH(3)OH-induced cytotoxicity could provide insight into the molecular basis for acute CH(3)OH-induced hepatoxicity. Metabolic pathways studied include those found in 1) an isolated hepatocyte system and 2) cell-free systems. Accelerated Cytotoxicity Mechanism Screening (ACMS) techniques demonstrated that CH(3)OH had little toxicity towards rat hepatocytes in 95% O(2), even at 2M concentration, whereas 50 mM was the estimated LC(50) (2h) in 1% O(2), estimated to be the physiological concentration in the centrilobular region of the liver and also the target region for ethanol toxicity. Cytotoxicity was attributed to increased NADH levels caused by CH(3)OH metabolism, catalyzed by ADH1, resulting in reductive stress, which reduced and released ferrous iron from Ferritin causing oxygen activation. A similar cytotoxic mechanism at 1% O(2) was previous found for ethanol. With 95% O(2), the addition of Fe(II)/H(2)O(2), at non-toxic concentrations were the most effective agents for increasing hepatocyte toxicity induced by 1M CH(3)OH, with a 3-fold increase in cytotoxicity and ROS formation. Iron chelators, desferoxamine, and NADH oxidizers and ATP generators, e.g. fructose, also protected hepatocytes and decreased ROS formation and cytotoxicity. Hepatocyte protein carbonylation induced by formaldehyde (HCHO) formation was also increased about 4-fold, when CH(3)OH was oxidized by the Fenton-like system, Fe(II)/H(2)O(2), and correlated with increased cytotoxicity. In a cell-free bovine serum albumin system, Fe(II)/H(2)O(2) also increased CH(3)OH oxidation as well as HCHO protein carbonylation. Nontoxic ferrous iron and a H(2)O(2) generating system increased HCHO-induced cytotoxicity and hepatocyte protein carbonylation. In addition, HCHO cytotoxicity was markedly increased by ADH1 and ALDH2 inhibitors or GSH-depleted hepatocytes. Increased HCHO concentration levels correlated with increased HCHO-induced protein carbonylation in hepatocytes. These results suggest that CH(3)OH at 1% O(2) involves activation of the Fenton system to form HCHO. However, at higher O(2) levels, radicals generated through Fe(II)/H(2)O(2) can oxidize CH(3)OH/HCHO to form pro-oxidant radicals and lead to increased oxidative stress through protein carbonylation and ROS formation which ultimately causes cell death.     

Studies on Neotriptetraolide Isolated from Tripterygium wilfordii Hook. f.

A new diterpenoid bisepoxide, neotriptetraolide, was isolated from the leaves and root of Tripterygium wilfordii Hook. f. Neotriptetraolide was crystallized from CH3OH as colorless needle-like crystal, with amp of 237～238 ℃. Its molecular formula was C20H2608. Its structure was elucidated by means of spectral analysis (UV, IR, MS, 1H-NMR, 13C- NMR, 2D-COSY, 2D-NOESY, 13C-NOE, and selective long-range DEPT spectroscopy) and the computation of molecular mechanics and molecular graph.     

Structural Study of Dichlorotriptetraolide Isolated from Tripterygium wilfordii

A new chlorinated diterpene epoxide, dichlorotriptetraolide (L5) was isolated from the leaves of Tripterygium wilfordii Hook. f. Dichlorotriptetraolide was crystallized from CH3OH as colorless needle-like crystal, with amp of 252.0～253.5 ℃. Its molecular formula is C2oH26O7Cl2. The structure was identified on the basis of spectral analyses (UV, 1H-NMR, 13C-NMR. 13C-NOE. 2D-cosy. LD/FTMS. EIMS, selective long-range DEPT spectrum and X-ray fluorescence spectrum) and computerization of molecular mechanics and molecular graph.     

Study on the Structure of a New Triterpene Saponin B from Polygala japoniea Houtt

A new triterpene saponin B has been isolated from the earial parts of Polygala japonica Houtt in folk-lore medicine. Its molecular: C48H78O20, m.p. 199–202℃, [α]D23+30.0 (C, 0.5, CH3OH). Acidic hydrolysis of this saponin gave a sapogenin (2α, 3α, 24-tri-hydroxyolean-12-ene-28-oic acid) and D-glucose. The structure of saponin B was elucidated as 28-O- [β-D-glucopyranosyl (1→2) -β-D-glucopyranosyl (1→2) -β-D-glucopy- ranosyl] 2α, 3α, 24-trihydroxyolean-12-ene-28-oic acid mainly by 13C-NMR, MS and some chemical transfomations.     

New, partially hydrolyzable synthetic analogues of lecithin, phosphatidyl ethanolamine, and phosphatidic acid

The synthesis of two new synthetic analogues of lecithin, two of phosphatidyl ethanolamine ("cephalin"), and one new phosphatidic acid analogue is described. They comprise one of each of the following types: the "isosteric" diether lecithin and cephalin analogues ROCH(2)CH(OR)- CH(2)CH(2)P(O) (O(-))OCH(2)CH(2)N(+)R'(3) (R = C(18)H(37); R' = H or CH(3)); and the "hydrocarbon" analogues of phosphatidic acid, lecithin, and cephalin, C(17)H(35)CH(2)CH(C(18)H(37))CH(2)P(O)(R) = (R'); [R = R' = OH; R = O(-), R' = OCH(2)CH(2)N(+)(CH(3))(3); and R = O(-), R' = OCH(2)CH(2)N(+)H(3)]. Infrared spectra and other properties of these compounds are described.     

Oxidation of hydrogen and reduction of methanol to methane is the sole energy source for a methanogen isolated from human feces.

A methanogenic coccus isolated from human feces requires H2 and CH3OH for growth and uses H2 to reduce CH3OH to CH4. Growth does not occur with CH3OH alone. The organism does not grow or produce CH4 from acetate or methylamines without or with H2 or from H2 and CO2 or formate. In a complex medium. CO2 is required for formation of approximately 50% of cell carbon, whereas the methyl carbon from methanol is not incorporated into cell carbon.