酶促核苷类化合物酯类衍生物的合成

核苷类化合物酯类衍生物在功能高分子材料、寡聚核苷酸和医药化学等领域具有重要的应用价值。本文就近年来核苷类化合物酯类衍生物酶法合成的研究进行综述。     

水稻组蛋白脱乙酰化酶HD2 HDACs蛋白质的生物信息学分析

HD2 HDACs家族成员是植物特异性的组蛋白脱乙酰化酶并参与基因的转录调控.本研究利用生物信息学方法对水稻组蛋白脱乙酰化酶HD2 HDACs家族成员（HDT701、HDT702、HDT2201和HDT2202）的生物学功能进行研究.结果表明,HD2 HDACs家族成员位于不同的染色体上,分布于细胞的不同部位;它们编码的蛋白均含有依赖于cAMP-和cGMP的蛋白激酶、蛋白激酶C、酪蛋白激酶II与酰胺化4个相同的位点;它们均无跨膜区,都为不稳定的亲水性蛋白质,都无信号肽.多序列比对发现粳稻与籼稻HD2 HDACs的同源性非常高;水稻HD2 HDACs蛋白质的结构域比较特殊;这些蛋白可能参与植物的基因复制、信号传导、转录过程和免疫应答等过程.     

壳聚糖固定化酶载体小球的研究及制备方法的改进

比较了不同粘均相对分子质量壳聚糖(50×104,100×104,200×104)的成球情况及壳聚糖相对分子质量对固定化酶活力、酶活性回收率的影响,确定了50×104的壳聚糖为最适固定化β-半乳糖苷酶载体. 自设计方法制备了毫米级壳聚糖小球,较传统方法更加简单、快速. 湿润壳聚糖小球直接干燥和采用体积分数为30%的甘油处理后干燥对比实验表明,后者能够保持小球湿态时的结构,既保证了酶的固定化效果,又有利于载体的工业储存.     

不同碳源条件下里氏木霉纤维降解酶的蛋白组解析

用双向电泳研究里氏木霉在4种 碳源条件下分泌组中纤维降解酶的分布情况。结果表明:在以甘油或葡萄糖为碳源时仅CBHⅡ、AxeI和BXL 3种纤维降解酶有微量表达。在以纤 维二糖为碳源时检测到较高表达水平的CBHⅠ、CBHⅡ、EGⅠ、EGⅢ、BXL、AxeⅠ和AglⅠ。纤维素纸浆可诱导CBHⅠ、CBHⅡ、EGⅠ、EGⅡ、EGⅢ 、EGⅣ、ManⅠ、ManⅡ、AxeⅠ、BXL和AglⅠ等11种纤维降解酶的大量表达,表达量总和占总分泌组的61％。纤维素诱导的纤维降解酶组分情况 和纤维二糖条件相似,但表达量和酶活性都有显著上升。     

产气气杆菌产普鲁兰酶发酵培养基的优化

对一株产气气杆菌产普鲁兰酶的发酵培养基进行了优化,确定了该菌株的最适发酵培养基配方为(g/L):玉米淀粉 15,豆饼粉 10,CH3COONH4 8,K2HPO4·3H2O 0.5,MgSO4·7H2O 0.5,KCl 0.5,FeSO4·7H2O 0.05.采用该培养基在5 L 发酵罐水平发酵 55h,普鲁兰酶活力达到 54.64 U/mL.研究了不同铵盐形式对该菌株产普鲁兰酶的影响,结果发现:以CH3COONH4为无机氮源,该菌株产普鲁兰酶活力高,且该酶绝大部分能分泌至胞外.而以NH4Cl,NH4NO3及(NH4)2SO4为无机氮源,该菌产普鲁兰酶活力较低,且为胞内酶.     

胰激肽原酶片对早期糖尿病肾病疗效观察

目的:评价胰激肽原酶片治疗早期糖尿病肾病患者的疗效和安全性.方法:将82例早期糖尿病肾病患者随机分成2组,采用糖尿病饮食控制、体育锻炼及根据病情给予口服降糖药物或/和胰岛素治疗,治疗组(42例)加用胰激肽原酶片治疗4周.观察治疗前后尿微量白蛋白与24h尿蛋白定量和不良反应.结果:治疗组治疗前后尿微量白蛋白与24h尿蛋白定量明显降低,差异有统计学意义(P<0.01);对照组治疗前后比较差异无统计学意义(P>0.05);组间比较差异有统计学意义(P<0.01);2组患者无明显的药物不良反应.结论:胰激肽原酶片治疗早期糖尿病肾病患者疗效好,用药安全,可广泛应用于临床.     

酶水解麦糟蛋白制备可溶性肽工艺研究

对碱性蛋白酶水解麦糟蛋白制备多肽的工艺条件进行了研究。通过单因素实验和正交实验,确定了较佳工艺条件：酶解pH10,加酶量3500u/g,酶解温度50℃,酶解时间20min。在此条件下,麦糟蛋白的水解度（DH）达22.18%,氮溶指数（NSI）达23.68%。     

巨大芽孢杆菌AP25内切葡聚糖酶基因的克隆及序列测定

从土壤中分离到一株产纤维素酶的巨大芽孢杆菌AP25,经羧甲基纤维素平板检测,该菌可产生葡聚糖内切酶。根据GenBank登录的β-1,4内切葡聚糖酶基因（DQ782954．1,M28332．1,AY859492．1）的同源性序列,利用PCR方法克隆到该酶的基因,并对其进行测序。测序结果显示其全长为1500bp,推测其含499个氨基酸。与GenBank中的已知葡聚糖内切酶相比较,发现该酶的氨基酸序列与Bacillus subtilis的β-1,4内切葡聚糖酶基因同源性达到达94％。     

Conformation-activity studies on the interaction of berberine with acetylcholinesterase:Physical chemistry approach

Berberine has been reported as an acetylcholinesterase (AChE) inhibitor.With significantly low cytotoxicity,berberine will be developed for the clinical treatment of Alzheimer disease (AD) with higher efficacy and fewer side effects.This work investigated the structure change events of AChE that occur during the interaction with berberine by isothermal titration calorimetry (ITC),fluorescence titration,and circular dichroism (CD).The results show that the binding of berberine to AChE is mainly driven by a favorable entropy increase with a less weak affinity.Berberine causes a loss in enzymatic activity at a concentration much below the concentration which gradually exposed the tryptophan residues to a more hydrophilic environment and unfolded the protein,which indicates that the inhibition of AChE with berberine includes the main contributions of interaction and minor conformation change of the protein induced by the alkaloid.     

Comparative proteomic analysis of the response in resistant and susceptible maize inbred lines to infection by Curvularia lunata

Proteins differentially expressed from maize leaves in response to the infection by Curvularia lunata strain CX-3 were identified through a high-resolution two-dimensional gel electrophoresis (2-DE) method. Two inbred lines, 78599-1 and E28, were used, respectively, as resistant and susceptible lines to CX-3 infection. Proteins were extracted from the fourth leaves of six- or seven-leaf stage plants sampled at 24, 36, 48, 60, and 72 h after inoculation with CX-3. Twenty-seven differentially expressed protein spots resolved on the 2-DE gels were identified by MALDI-TOF MS/MS. The results showed that these proteins are associated with photosynthesis, respiration,oxidative and drought stress tolerance as well as signal transduction in maize. Among stress-related proteins, the 22 kDa drought-inducible protein, putative glutathione peroxidase (GPX), and translation initiation factor (eIF-5A) were up-regulated in the resistant inbred line and were implicated in host defense response to C. lunata infection. It suggests that drought-inducible and oxidation stress-related proteins might directly contribute to maize resistance to C. lunata.