游离和溶胶-凝胶固定化甲醛脱氢酶酶促反应动力学的研究

采用溶胶-凝胶法对甲醛脱氢酶进行固定化，酶的包埋率超过了98％．在pH7附近、37℃下，以游离酶和固定化酶作催化剂，NADH为电子供体，进行了甲酸转化为甲醛的酶促反应．游离和固定化甲醛脱氢酶酶促反应都遵循Michaelis-Menten反应机理，用Dalziel提出的双底物酶促反应动力学方程进行拟合．固定化酶酶促反应速率为游离酶酶促反应速率的50％左右．固定化酶的动力学常数φ和米氏常数K高于游离酶，估计是凝胶基质孔中存在扩散效应所致．     

Mutation scanning-coupled analysis of haplotypic variability in mitochondrial DNA regions reveals low gene flow between human and porcine Ascaris in endemic regions of China

Haplotypic variation within and among the Ascaris populations representing six provinces in China was investigated. Mitochondrial DNA regions in the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes were amplified by PCR from total genomic DNA samples (n > 720) from Ascaris individuals from humans and pigs, and subjected to mutation scanning and subsequent selective sequencing. For the cox1, ten different electrophoretic profiles were recorded for human Ascaris, and the same number for pig Ascaris, one of them being common to both host species. For the nad1, 11 different profiles were detected for human Ascaris, and 15 for pig Ascaris. Having defined all haplotypes (20 for pcox1 and 26 for pnad1) by sequencing, their frequencies were estimated in each of the two host species and each of the six provinces. For each mitochondrial region, the frequency of the different haplotypes varied considerably, depending on host species and geographical origin. Analysis of the sequence data (representing all haplotypes for each mitochondrial locus) by F-statistics indicated restricted gene flow between human Ascaris and pig Ascaris, and supported the conclusions from previous molecular epidemiological investigations that pigs are not a significant source of Ascaris infection in humans in endemic regions.     

基于药效团模型的DHODH抑制剂构效关系研究

利用药效团模型研究二氢乳清酸脱氢酶(Dihydroorotate dehydrogenase,DHODH)抑制剂的构效关系,为DHODH抑制剂的虚拟筛选提供新的方法.以31个具有DHODH抑制活性的化合物为训练集化合物,半数抑制浓度(IC50)范围为7~63000 nmol/L,利用Catalyst/HypoGen算法构建DHODH抑制剂药效团模型,通过对训练集化合物多个构象进行叠合,提取药效团特征及三维空间限制构建药效团模型.利用基于CatScramble的交叉验证方法及评价模型对已知活性化合物的活性预测能力,确定较优药效团模型.模型包含1个氢键受体、3个疏水中心,表征了受体配体相互作用时可能发生的氢键相互作用、疏水相互作用和π-π相互作用,4个药效特征在三维空间的排列概括了DHODH抑制剂产生活性的结构特点.所得较优模型对训练集化合物及测试集化合物的计算活性值与实验活性值的相关系数分别为0.8405和0.8788.利用药效团模型对来源于微生物的系列化合物进行虚拟筛选,筛选出59个预测活性较好的化合物,可作为进一步药物研发的候选化合物.     

HPLC法测定鸡精中谷氨酸钠的含量

以邻苯二甲醛与谷氨酸钠中的氨基进行柱前在线衍生化反应,采用C18色谱柱分离、荧光检测器（激发340nm,发射450nm）进行测定,建立了柱前衍生反相高效液相色谱测定鸡精中谷氨酸钠含量的方法。该方法相对标准偏差为0．69％,加标回收率为99．1％～101％,在0．10~50．0mg／L范围内,谷氨酸钠的峰面积和浓度之间的相关系数为0．9999,保留时间和峰面积的相对标准偏差分别为1．22％和0．71％,鸡精中谷氨酸钠定量下限为0．2μg／g。     

A thionine-based reversible redox sensor in a sequential injection system

According to the current demands of Green Analytical Chemistry and regarding the need for lower reagent consumption with improved analytical performance, an automatic methodology with a flow-through optosensor incorporating solid-phase spectrophotometric detection was developed. The sensor used in this work was based on the redox state of thionine whose oxidized form is blue and reduced form is colorless with monitoring carried out at 621 nm. This redox indicator was immobilized on gel beads and subsequently packed into a flow-through cell. It was then assembled into a sequential injection system and was shown to be an excellent alternative to monitor enzymatic redox reactions automatically as the redox catalysis is performed by glucose dehydrogenase. This enzyme is a representative dehydrogenase enzyme and uses NAD⁺ as cofactor, promoting the oxidation of glucose to glucono-lactone and reduction of NAD⁺ to NADH. The produced NADH promotes color depletion on the surface of the sensor. The calibration graph for glucose was linear between 5.74 × 10⁻⁴ and 2.00 × 10⁻³ mol L⁻¹ and detection limit was 1.72 × 10⁻⁴ mol L⁻¹. Glucose concentration in different samples including sera, salines, perfusion solutions, powder for preparing oral solutions and solutions for hemodialysis was determined. The method proved to be reproducible with a RSD < 5% for glucose determinations.     

五氯酚对好氧活性污泥中不同酶活性的影响

以五氯酚为研究对象,采用分光光度法测定其对城市污水处理厂好氧活性污泥中的脲酶、转化酶、脱氢酶活性的影响。结果表明,随着五氯酚暴露浓度的增大,脲酶活性总体呈上升趋势;转化酶活性先升高后降低,暴露时间为0.5 h,转化酶升高率达到最大值49.3%;暴露15 h后转化酶活性开始呈现降低态势,实验最大暴露时间25 h时降低率达到76.8%。脱氢酶活性则大多呈现降低趋势,在暴露0.5 h时其酶活性随五氯酚浓度变化呈明显负相关,酶活性降低率为52%,之后脱氢酶活性被抑制情况呈现平缓趋势。综合生理生化反应指标的敏感度和分析方法的精密度进行评价,五氯酚对转化酶影响幅度大且分析精密度高,而脱氢酶响应快,可作为急性毒性应激敏感指标。     

用静息细胞培养法研究L－谷氨酸氧化酶的生物合成

建立了静息细胞培养系统，并以此研究了Ｌ－谷氨酸氧化酶生物合成的影响因素。结果表明，发酵１６ｈ的菌体在有诱导物情况下有酶的合成，无诱导物则无酶的合成，发酵４０ｈ的菌体在有、无诱导物的条件下均有酶的合成。对发酵１６ｈ的菌体研究表明：β－丙氨酸（Ⅰ）、α－酮戊二酸（Ⅱ）、丙酮酸钠（Ⅲ）是酶合成的诱导物，谷氨酸（Ⅳ）则不是。而对发酵４０ｈ菌体而言：ＭｇＣｌ＿２，ＣａＣｌ＿２能促进酶的合成；Ⅰ不能作氮源利用；Ⅰ～Ⅳ均具有高浓度抑制产酶、低浓度促进产酶的双重作用。     

三元全酶系统的共固定化和辅酶再生

本文研究了用对-氨基苯砜乙基(ABSE)-交联琼脂糖做载体,通过戊二醛共价交联,对醇脱氨酶、乳酸脱氢酶、辅酶Ⅰ进行了位置对位置的共固定化。按本实验方法得到的固定化三元全酶系统的辅酶再生能力为相应游离酶系统的5倍。本文还对该三元全酶系统的固定化机理和辅酶再生性能做了一定的讨论。     

离子交换层析法分离大鲵乳酸脱氢酶同工酶

采用DEAE—纤维素阴离子交换层析,以0.02M tris-HCl+0.06M NaClpH7.8和0.02M tris-HCl0.15M NaCl pH7.4缓冲液分部洗脱,可以分离大鲵乳酸氢酶同工酶LDH_1和LDH_5。     

甲醛对人红细胞的影响

生产、科研、生活中,有大量人群长期接触甲醛。甲醛可引起肝脏病变,致癌、致突变,已日益为人们所关注。本文采用人红细胞为材料,研究甲醛的毒害作用。人红细胞悬液,经0.1～1.0×10~(-6)mol/l甲醛处理后,通过生化分析,实验结果表明,在本实试条件下甲醛增强红细胞溶血作用,抑制乙酰胆碱酯酶活力和引起可滴定巯基含量降低。同时使磷酸甘油醛脱氢酶、乳酸脱氢酶活力提高和促进细胞内Ca~(3+)的泄漏。