新疆甘草化学成分研究

从新疆甘草(GlycyrrhizauralensisFisch.)根中分离得到5个化合物,分别鉴定为:槲皮素(1)、甘草次酸(2)、阿魏酸(3)、5,7,3′,4′-四羟基-7-葡萄糖基二氢黄酮醇(4)、8,5′-二羟基3′α-鼠李糖黄酮(5)。其中化合物4,5为首次从该植物中分离得到。     

冬葵果多糖的抗氧化作用研究

研究冬葵果多糖(Fructus Malvae polysaccharides,FMP)对氧自由基的清除作用及对脂质过氧化的抑制作用。采用分光光度法测定FMP清除Fenton体系产生的羟自由基(.OH)、邻苯三酚自氧化体系产生的超氧阴离子自由基(O2-.)的能力及评价对Fe(Ⅱ)-H2O2体系(.OH)诱导的脂质过氧化终产物丙二醛(MDA)的抑制作用。结果显示,在化学模拟体系中,冬葵果多糖对.OH具有很强的清除作用,与VC比较达到显著水平(P<0.01),对O2-.的清除能力与VC相当。在体外,多糖浓度达到1.72 mg/mL可明显降低MDA的含量,与空白液比较达到显著水平(P<0.01)。以上结果表明,FMP具有明显的抗氧化活性。     

Directed evolution and the creation of enantioselective biocatalysts

Directed evolution has emerged as a key technology to generate enzymes with new or improved properties that are of major importance to the biotechnology industry. A directed evolution approach starts with the identification of a target enzyme to be optimized and the cloning of the corresponding gene. An efficient expression system is needed before the target gene is subjected to random mutagenesis and/or in vitro recombination, thereby creating molecular diversity. Subsequently, improved enzyme variants are identified, preferably after being secreted into culture medium, by screening or selection for the desired property. The genes encoding the improved enzymes are then used to parent the next round of directed evolution. Enantioselectivity is a biocatalyst property of major biotechnological importance that is, however, difficult to deal with. We discuss recent examples of creating enantioselective biocatalysts by directed evolution.     

血管抑素的临床应用研究进展

血管抑素是第一个被发现的肿瘤源性血管生成抑制因子,能抑制血管内细胞的增殖和迁移,从而抑制肿瘤的生长。通过一些实验方法能够检测肿瘤患者体内的血管抑素水平。血管抑素在未来的肿瘤治疗、诊断、预后判断和新生血管疾病的治疗中有着广阔的应用前景。     

大肠杆菌细胞的软X射线显微成像研究

软X射线显微术是研究含水甚至活性生物样品的有力工具。相对于光学显微镜 ,它具有更高的成象分辨率 ;相对于电子显微镜 ,它的样品制备简单—无须对样品进行脱水、染色和超薄切片等。报道的是利用合肥同步辐射X射线源和接触显微成象技术 ,对自然状态下含水的完整XL1 blueMRF′细菌细胞进行显微成象研究。从获得的显微图象中可以看出一些新的现象。含有DNA、蛋白质的拟核以及中体对波长 2 .4nmX射线具有较弱的吸收能力 ;不少细菌细胞的两端对 2 .4nm波长的X射线的吸收也具有很大的差异。这些有趣现象产生的根本原因和生物学意义有待进一步研究。     

初始底物浓度对序批式培养光合细菌产氢动力学影响

实验研究了初始底物浓度对序批式培养光合细菌生长、降解及产氢过程的影响,根据最大比生长速率实验数据拟合得到其关于初始底物浓度影响的关联式,并在建立的修正Monod模型基础上建立了光合细菌比生长速率、基质比消耗速率和比产氢速率关于底物初始浓度影响的数学模型,模型预测值与实验结果在光合细菌生长期和稳定期内得到较好吻合,反映了光合细菌生长、降解和产氢过程中受底物初始浓度限制性和抑制性影响的基本规律。分析发现光合细菌生长、降解基质和产氢过程中最适底物浓度为50 mmol/L,初始底物浓度低于或高于该浓度时,光合细菌生长、降解及产氢过程都受到限制性或抑制性影响,且抑制性影响较限制性影响效果更明显;底物比消耗速率受初始底物浓度影响较小。     

Peptidylglycine-alpha-hydroxylating monooxygenase generates two hydroxylated products from its mechanism-based suicide substrate, 4-phenyl-3-butenoic acid

The bifunctional enzyme peptidylglycine-alpha-amidating monooxygenase mediates the conversion of C-terminal glycine-extended peptides to their active alpha-amidated products. Peptidylglycine-alpha-hydroxylating monooxygenase (PHM, EC 1.14.17. 3) catalyzes the first reaction in this two-step process. The olefinic compound 4-phenyl-3-butenoic acid (PBA) is the most potent irreversible, mechanism-based PHM inactivator known. While the details of the inhibitory action of PBA on PHM remain undefined, covalent modification of the protein has been proposed as the underlying mechanism. We report here that, in the process of inactivating PHM, PBA itself serves as a substrate without covalently labeling the enzyme. Approximately 100 molecules of PBA are metabolized per molecule of PHM inactivated, under saturating conditions. The metabolism of PBA by PHM generates two hydroxylated products, 2-hydroxy-4-phenyl-3-butenoic acid and its allylic isomer, 4-hydroxy-4-phenyl-2-butenoic acid. While one enantiomer for each product is significantly favored in the reaction, both are produced. From these observations, we conclude that hydroxylated PBA products are formed by a delocalized free radical mechanism and that the lack of absolute stereospecificity indicates significant freedom of movement within the catalytic site. The ability of PHM to metabolize PBA suggests that the physiological functions of PHM may include the hydroxylation of substrates other than those containing terminal glycines.     

植物种群表观竞争机制及其研究进展

植物表观竞争既影响群落结构和组成,也与生态系统功能密切相关。目前,国内尚缺乏对植物表观竞争的实验研究和机制探索。对植物表观竞争概念和研究进展进行了系统梳理和阶段性总结,分析了草食动物为媒介的表观竞争主要通过食物和生境介导植物竞争过程,以及ELP-采食者关系对表观竞争的影响;植物参与的表观竞争通常与资源竞争共同作用;AM真菌为媒介的表观竞争不仅受到宿主植物菌根依赖性的影响,还与AM真菌对资源的需求和环境资源供给状况有密切关系。未来研究既要注重实验设计的合理性,也要考虑测定方法和评价参数的有效性;重视植物群落和生态系统水平上表观竞争的功能研究。这不仅能够为植物群落结构和物种共存提供机制性解释,而且能够丰富经典的植物竞争理论和多营养级相互作用的食物网理论。     

Regulation of peptide amidation in cultured pituitary cells

The intermediate lobe of the pituitary contains the alpha-amidated peptide alpha-melanotropin and high levels of a copper and ascorbate-dependent peptidylglycine alpha-amidating monooxygenase (PAM) capable of converting peptides terminating in -X-Gly into amidated products (-X-NH2). As reported previously, the ability of cultured intermediate pituitary cells to produce alpha-amidated alpha-melanotropin declined rapidly. A decline in PAM activity assayed in vitro under optimized conditions failed to account quantitatively for the lack of production of alpha-amidated product, while a 100-fold decline in cellular levels of ascorbate could account for the lack of production of alpha-amidated product. Incubation of intermediate pituitary cultures with ascorbate partially restored the ability of the cells to produce alpha-amidated product without significantly increasing the level of PAM activity. In intermediate pituitary cultures made competent to produce alpha-melanotropin by addition of ascorbate, the actual extent of amidation occurring was modulated by the presence of specific secretagogues (bromocriptine or corticotropin-releasing factor). Cultured anterior pituitary cells showed a similar rapid 3-fold decline in PAM activity assayed in vitro under optimized conditions. Cellular levels of ascorbate also declined rapidly to levels 100-fold below those in the intact anterior pituitary. The addition of ascorbate to the anterior pituitary cultures rapidly restored the enzyme activity assayed in vitro to the levels in the initial cell suspension. Thus, production of amidated product peptide may be regulated by cellular levels of ascorbate, by cellular levels of PAM activity, and by the concentration of specific secretagogues to which the cells are exposed.