传感器在“生物对环境的影响”一课中的应用

在"生物对环境的影响"一课中设计了2个以传感器为实验工具的学生实验,为构建"生物的活动能够影响环境"的概念提供事实依据,并激发了学生的探究兴趣。     

“酶的活性”实验教学设计与实施

在"酶的活性"的主题教学中,为落实"酶的空间结构影响酶的活性"这一核心概念,教师通过创设情境,让学生提出问题、作出假设、设计实验、实施实验、收集数据、讨论分析,最终解决科学问题,亲身经历"科学大实验",在反复琢磨中深刻理解核心概念,在科学探究中落实生物学核心素养。     

“研究影响光合作用的环境因素”的实验教学改进

在"研究影响光合作用的环境因素"一课中,对实验装置进行了改进设计,组织学生同时探究二氧化碳浓度和光照强度对光合作用的影响,完成实验数据的定量分析,有效提高课堂教学效率,培养学生的科学探究能力。     

“探究pH对酶活性的影响”的创新设计

在进行"探究p H对酶活性的影响"的实验教学中不断质疑,从实验材料、实验装置、实验步骤等方面进行改进,使实验效果更为直观、更有趣,不断促使学生发现问题,也有助于提高学生的实验能力、探究能力及创新设计能力,能在教学中启发学生积极思考,使教学效果更有效。     

探究温度对酶活性影响的实验设计与实施

"新陈代谢与酶"是高中生物学"新陈代谢"一章的一个课题.早在1986年,我校就组织学生通过实验探究酶及其作用特性.1996年7月,周湘老师曾在<生物学通报>发表一篇文章--"酶与代谢"一节的实验探究法教学,文章中提到对酶活性的影响因素的实验探究,本组教师曾先后尝试用唾液淀粉酶催化淀粉水解、过氧化氢酶催化H2O2分解来研究温度和pH对酶活性的影响,但实验效果均不是很理想.     

探究“环境因素对光合作用强度的影响”实验改进建议和实验延伸

在探究"环境因素对光合作用强度的影响"的实验教学中,以教材中的参考案例"探究光照强弱对光合作用强度的影响"为基础,对实验方法进行了优化,并探究了温度、CO_2浓度、光质对光合作用强度的影响。本实验采用分组分工合作的方法,使每位学生都成为实验的参与者,充分提高了课堂的时效性和可操作性,并取得了较好的实验效果,同时激发了学生的改革意识,培养了学生的创新精神。     

关于"pH对酶活性的影响"3种实验方案的比较分析

关于"pH对酶活性的影响"的实验教学中,学生设计了3种不同的实验方案,在课堂交流研讨的过程中引起学生激烈的争论.现将这3种实验方案陈述如下,并将学生的争论进行了整理.     

关于非人灵长类的公正行为的研究

人类关注公正, 非人灵长类也表现出公正行为。本文先以现有研究资料为基础, 以理毛为例分析后认为, 非人灵长类关注投入—收益的对称性, 说明它们可能具备不公正规避这一心理特质; 关于非人灵长类公正行为的实验也表明, 它们不仅比较自身的投入—收益对称性, 而且能在社会比较过程中与其它个体相比。有实验得出期望假设和挫折效应能更好地解释被试的行为, 本文认为, 这些实验结论不一致的主要原因, 是研究者未充分考虑"投入"对被试行为的影响。文章在最后进行了总结并提出了三点研究展望。     

探究水污染对生物的影响及生物的净化作用

本实验是初中生物学"探究环境污染对生物的影响"关于水污染的拓展创新实验,学生积极参与共创最优实验方案,深刻理解生物与环境的关系。从发现问题、选取实验材料,设计实验思路,再到数据统计和作图分析等,在科学探究过程中运用理性思维,实现在教材基础上的深度学习。     

“用比色法探究pH对酶活性的影响”教学设计

在"探究p H对酶活性的影响"实验教学过程中,尝试运用目测比色法简便、快速获取直观的实验结果,达到预期实验目的;同时注重培养学生运用数学方法分析实验现象,养成数学与生物学科相结合的思维方式。     

垃圾填埋场土壤酶活性与化学性质和微生物数量的关系研究

通过气相色谱／质谱定性分析表明,垃圾填埋场土壤的有机物污染成分复杂,共检出有机物50种。研究表明,随着填埋深度的加深,土壤理化性质,微生物数量和土壤酶活性有很大不同。运用典型相关分析对垃圾填埋场的土壤酶活性与土壤化学性质和微生物数量的关系进行了研究。结果表明。土壤酶活性与化学性质总体上密切相关;土壤酶活性与微生物数量总体上密切相关;但在土壤酶中过氧化氢酶活性与化学性质和微生物数量不相关。     

实验诊断学PBL教学案例的设计原则

由临床检验专业人员承担的《实验诊断学》教学应以临床检验学为教学重点,案例的教学目标及学习重点应围绕临床检验基本原理,检验结果质量保证和检验结果影响因素三方面进行。案例选择应遵循以下原则：①选择质量保证的过程中容易出现问题的环节;②选择物质代谢过程清楚,相关疾病的病理生理改变明确的检验项目,如血糖;③选择在,临床检验中应用较多的方法,比如酶的连续监测法。     

案例教学法在《酶工程》教学中的应用

结合几年来的教学实践,从酶工程教学中实施案例教学法的优势、必要性、过程条件及意义等方面介绍了实施案例教学法在生物工程专业《酶工程》课程教学中的重要地位。通过该教学法的实践,取得了良好教学效果,增强了学生的应用能力。     

Specific arginine modification at the phosphatase site of muscle carbonic anhydrase

Mammalian carbonic anhydrase III has previously been shown to catalyze the hydrolysis of p-nitrophenyl phosphate in addition to possessing the conventional CO2 hydratase and p-nitrophenylacetate esterase activities. Modification of pig muscle carbonic anhydrase III with the arginine reagent phenylglyoxal yielded two clearly distinctive results. Reaction of the enzyme with phenylglyoxal at concentrations equivalent to those of the enzyme yielded stoichiometric inactivation titration of the enzyme's phosphatase activity, approaching 100% loss of activity with the simultaneous modification of one arginine residue, the latter based on a 1:1 reaction of phenylglyoxal with arginine. At this low ratio of phenylglyoxal to enzyme, neither the CO2 hydratase activity nor the acetate esterase activity was affected. When the modification was performed with a significant excess of phenylglyoxal, CO2 hydratase and acetate esterase activities were diminished as well. That loss of activity was accompanied by the incorporation of an additional half dozen phenylglyoxals and, presumably, the modification of an equal number of arginine residues. The data in their entirety are interpreted to show that the p-nitrophenylphosphatase activity is a unique property of carbonic anhydrase III and that excessive amounts of the arginine-modifying reagent lead to unspecific structural changes of the enzyme as a result of which all of its enzymatic activities are inactivated.     

Aspartokinase Activity and the Developmental Cycle of Myxococcus xanthus

The relationship between aspartokinase activity and fruiting body formation in Myxococcus xanthus was investigated. Two required amino acids, methionine and isoleucine, which stimulated the enzyme in vitro also inhibited fruiting body formation when added to 0.1% Casitone agar. Threonine, a potent feedback inhibitor of the aspartokinase, completely reversed the effects of methionine and isoleucine both on enzyme activity and fruiting body formation. A mutant, M. xanthus FB-S, which had the unusual property of forming fruiting bodies on 1.0% Casitone agar, also exhibited an altered regulation of aspartokinase activity. Spermidine, which is a strong stimulator of the enzyme in vitro, interfered with the developmental cycle of both M. xanthus FB and FS-S. During glycerol induction of myxospores the level of aspartokinase dropped more than 75% during the first hour. These data indicate a strong correlation between aspartokinase activity and the induction of the developmental cycle in M. xanthus. It is suggested that the decrease in aspartokinase activity results in diaminopimelic acid starvation, blockage of cell wall growth, and subsequent induction of the developmental cycle.     

The buried diversity of bovine seminal ribonuclease: shape and cytotoxicity of the swapped non-covalent form of the enzyme

Bovine seminal ribonuclease exists in the native state as an equilibrium mixture of a swapped and an unswapped dimer. The molecular envelope and the exposed surface of the two isomers are practically indistinguishable and their diversity is almost completely buried in the interior of the protein. Surprisingly, the cytotoxic and antitumor activity of the enzyme is a peculiar property of the swapped dimer. This buried diversity comes into light in the reducing environment of the cytosol, where the unswapped dimer dissociates into monomers, whereas the swapped one generates a metastable dimeric form (NCD-BS) with a quaternary assembly that allows the molecule to escape the protein inhibitor of ribonucleases. The stability of this quaternary shape was mainly attributed to the combined presence of Pro19 and Leu28. We have prepared and fully characterized by X-ray diffraction the double mutant P19A/L28Q (PALQ) of the seminal enzyme. While the swapped and unswapped forms of the mutant have structures very similar to that of the corresponding wild-type forms, the non-covalent form (NCD-PALQ) adopts an opened quaternary structure, different from that of NCD-BS. Moreover, model building clearly indicates that NCD-PALQ can be easily sequestered by the protein inhibitor. In agreement with these results, cytotoxic assays have revealed that PALQ has limited activity, whereas the single mutants P19A and L28Q display cytotoxic activity against malignant cells almost as large as the wild-type enzyme. The significant increase in the antitumor activity, brought about by the substitution of just two residues in going from the double mutant to the wild-type enzyme, suggests a new strategy to improve this important biological property by strengthening the interface that stabilizes the quaternary structure of NCD-BS.     

一株产纤溶酶菌株的分离鉴定及其纤溶组分分析

【目的】筛选性能良好的产纤溶酶菌株,对菌株进行多项分类鉴定,分析其纤溶酶系的组成特征及纤溶能力。【方法】通过酪蛋白培养基初筛,琼脂-纤维蛋白双层平板复筛,从海泥、土壤等环境中筛选纤维蛋白降解菌,以尿激酶为标准测定纤溶酶活性。通过形态学、生理生化特征研究,结合16S rDNA基因序列分析菌株种类及系统分类地位。通过SDS-PAGE和纤维蛋白酶谱法分析胞外纤溶酶系的组成特征。【结果】筛选到一株能降解纤维蛋白的细菌CNY16,鉴定其为沙福芽孢杆菌(Bacillus safensis)。该酶为胞外酶,SDS-PAGE和纤维蛋白酶谱结果表明该纤溶酶系有至少两种分子量大小不同的纤溶酶,分别约33 kD和23 kD。能有效溶解血块中纤维蛋白,并且对红细胞无降解作用。【结论】细菌CNY16是一株新的纤溶酶产生菌,纤溶酶活性及稳定性较好,具有潜在开发价值。为获取新型纤溶酶提供了一种新的菌源。     

The inflow of the substrate can regulate the persistence of memory of enzymes having the properties of hysteresis: theoretical analysis

A simple kinetic model of hysteretic enzymes with the influx of substrate or ion (transported by the enzyme) is considered. Two alternative steady activity levels are shown to arise in the system with a hysteretic enzyme. The transition between these levels can proceed in an oscillatory manner. The duration of the initial steady activity level is shown to be determined by the initial substrate (or ion) level, and the oscillatory transition between the activity levels is the property of hysteretic enzymes. It was shown for plasma membrane Ca2+-ATPase as an example that the level of the signal can be encoded into the time interval in which the enzyme retains the memory about this signal.     

Conversion of the noncooperative Bacillus subtilis aspartate transcarbamoylase into a cooperative enzyme by a single amino acid substitution.

Allosteric enzymes are part of a unique class of enzymes which regulate metabolic pathways. On the molecular level, allosteric regulation is the result of interactions between discrete binding sites on the enzyme. In order to accommodate these multiple binding sites, allosteric enzymes have evolved with oligomeric quaternary structures. However, only a few oligomeric enzymes are known to have regulatory interactions between binding sites. Is regulatory activity an inherent property of oligomeric enzymes? The trimeric Bacillus subtilis aspartate transcarbamoylase catalyzes the first committed step of the pyrimidine biosynthetic pathway and is not known to be a regulatory enzyme. When an alanine residue is substituted for the active-site residue Arg-99 by site-specific mutagenesis, the regulatory activity of homotropic substrate cooperativity (Hill coefficient of 1.5) is observed in the resulting mutant enzyme. These results suggest that homotropic regulation may have evolved by a relatively small number of mutations to an oligomeric enzyme.     

Spin-trapping agent alpha-phenyl N-tert-butylnitrone binds to trypsin and enhances heparin-induced inhibition of amidolytic activity and structural degradation of the enzyme.

The effects of heparin on trypsin have recently been demonstrated to involve inhibition of catalytic activity and degradation of the enzyme by means of an oxidative mechanism. The possibility that alpha-phenyl N-tert-butylnitrone protects heparin-induced radical formation on trypsin was investigated by measuring amidolytic activity and changes in the structure of trypsin in the presence of heparin with and without alpha-phenyl N-tert-butylnitrone. The results show that alpha-phenyl N-tert-butylnitrone does not only prevent, but it even significantly enhances effects of heparin on the enzyme. This is due to the unique property of alpha-phenyl N-tert-butylnitrone, independently of spin-trapping capacity, to modify the trypsin structure by binding irreversibly to the catalytic triad, at sites distinct from those to which heparin binds.