竞争性抑制的非稳态酶动力学布尔函数图论研究

以非稳戊酶动力学的布尔函数图形方法,来研究一类竞争性抑制的非稳态酶动力学问题,推导出此类反应的百稳态酶动力学方程,并对此动力学方程进行了讨论,分析了此类竞争性抑制酶反应体系的非稳态酶动力学问题。     

反竞争性抑制的非稳态酶动力学布尔函数图解研究

以非稳态酶动力学的布尔函数图形方法,来研究一类反竞争性抑制的非稳态酶动力学问题,推导出此类反应的非稳态酶动力学方程,并对此动力学方程进行了讨论,分析了此类反竞争性制酶反应体系的非稳态酶动力学问题。     

Ping Pong Bi Bi机制的非稳态酶动力学分布函数图论研究

以非稳态酶动力学的布尔函数图形方法,来研究一类PingPongBiBi机制的非记酶动力学问题,推导出此类反应的非稳态酶动力学方程,并对此动力学方程进行了讨论,分析了此类PingPongBiBi机制酶反应体系的非稳态酶动力学方程。     

Random Bi Bi机制的非稳态酶动力学布尔函数图论研究

本文以非稳态酶动力学的布尔函数图形方法^[1],来研究一类Random Bi Bi机制的非稳态酶动力学问题,推导同此类反应的非稳态酶动力学方程,并对此动力学方程进行了讨论,分析了此类Random Bi Bi机制酶反应体系的非稳态酶动力学方程。     

非稳态酶活化动力学的布尔函数图论分析

以非稳态酶动力学的布尔函数图形方法研究非稳态酶活化动力学问题,推导出此类反应的非稳态酶动力学方程,并对此动力学方程进行了讨论,分析了酶活化反应体系的非稳态酶动力学过程．     

3-硝基-N-甲基水杨酰胺抑制琥珀酸细胞色素c还原酶的动力学特征

 用可逆非竞争性抑制的动力学方程分析3-硝基-N-甲基水杨酰胺对琥珀酸细胞色素c还原酶的抑制作用,发现在高浓度抑制剂条件下实验数据偏离动力学方程。以大豆磷脂取代酶中的天然磷脂后,不再出现这种偏离。这一现象可能与呼吸链酶的双体特征有关。     

酶动力学的极值原理假说

平衡态假说和稳态假说是酶动力学的基础。本文提出极值原理假说,用于完善平衡态假说和稳态假说的不足之处,扩大了酶动力学的应用范围,并以Michaelis-Menten方程为例进行了说明。     

胆碱脱氢酶的动力学性质

本文对增溶胆碱脱氢酶的稳态初速度及产物抑制动力学做了。底物胆碱和ＰＭＳ的相互影响：变化一个底物的浓度,另一个底物的Ｋｍ及Ｖｍａｘ均变化。该酶的产物三甲胺 地 制表现为对底物胆碱非竞争性而地ＰＭＳ竞争性,在胆碱饱和的情况下,三甲胺乙醛对酶的抑制仍表现为对ＰＭＳ竞争性。这些结果表明增溶胆碱脱氢酶的催化机制搂双底物双产物乒乓机制。１－ＰＣ与９－ＡＣ对增溶胆碱脱氢酶均有抑制作用,且均为混和型抑制,Ｋ１分别     

乙醇对F_1-ATP酶和H~+-ATP酶的抑制与其核苷酸结合位点状态的关系

 本文利用动力学方法研究了乙醇对F_1-ATP酶和H~(+)-ATP酶复合体的抑制与其结合核苷酸位点状态的关系,结果表明天然情况下乙醇对F_1呈现反竞争性抑制类型,对H~(+)-ATP酶呈现非竞争性抑制类型,且乙醇对F_1和H~(+)-ATP酶的抑制与核苷酸结合位点的构象密切相关。游离状态下和膜结合状态下的F_1在部分结合的核苷酸被洗脱前后动力学行为的不同,反映了二种状态下的F_1具有不同的构象,且F_0和膜脂对F_1起着一定的调控作用。     

固定化细胞的混合糖连续发酵动力学模型

利用固定化啤酒酵母和固定化毕赤酵母在两个串联的固定床内连续发酵由葡萄糖和木糖组成的混合糖制取酒精的过程,建立了连续发酵的非结构动力学模型。该模型以带抑制项的米氏动力学方程为酶动力学基础,考虑了抑制物抑制、底物抑制、轴向弥散及膜传质等因素。成功地引入了一个综合考虑颗粒相内外传质的总有效因子简化模型的计算,并取得了较为满意的仿真结果。     

Graphic rules in steady and non-steady state enzyme kinetics

Graphic methods, when applied to enzyme kinetics, can provide a visually intuitive relation between calculations and reaction graphs. This will not only greatly raise the efficiency of calculations but also significantly help the analysis of enzyme kinetic mechanisms. In this paper, four graphic rules are presented. Rules 1-3 are established for steady state enzyme-catalyzed reaction systems and Rule 4 is for non-steady state ones. In comparison with conventional graphic methods which can only be applied to steady state systems, the present rules have the following merits. 1) Complicated and tedious calculations can be greatly simplified; for example, in calculating the concentrations of enzyme species for the bi-bi random mechanism, the calculation work can be reduced 8-fold compared with the King-Altman's method. 2) A great deal of wasted labor can be avoided; for example, in calculating the rate of product formation for the same mechanism, the operation of finding and removing the 96 reciprocally canceled terms is no longer needed because they automatically disappear during the derivation. 3) Final results can be easily and safely checked by a formula provided in each of the graphic rules. 4) Non-steady state systems can also be treated by the present graphic method; for example, applying Rule 4, one can directly write out the solution for a non-steady state enzyme-catalyzed system, without the need to follow more difficult and complicated operations to solve differential equations. The mathematical proofs of Rules 1-4 are given in Appendices A-D (in the Miniprint), respectively.     

Graphical rules for non-steady state enzyme kinetic

The exiting graphical methods in enzyme kinetics can be used only within the scope of steady state reactions. In this paper, two graphical rules are presented to deal with the non-steady state enzyme catalysed reaction systems. According to Rule 1 we can immediately write out the phase concentration of enzyme species. The calculation work such as setting up differential equations, making Laplace transformation, expanding determinants, which are both tedious and liable to error, are completely saved. By means of Rule 2 the secular equations for the consecutive first-order reactions can be written out directly without need of setting up differential equations, expanding determinants, etc., that would otherwise be laborious and prone to errors. In addition, two check formulae are also presented for these two graphical methods, respectively. They are useful in order for avoiding the omission of terms during calculations, especially, for complicated mechanisms.     

Asymmetric Effect of Mechanical Stress on the Forward and Reverse Reaction Catalyzed by an Enzyme

The concept of modulating enzymatic activity by exerting a mechanical stress on the enzyme has been established in previous work. Mechanical perturbation is also a tool for probing conformational motion accompanying the enzymatic cycle. Here we report measurements of the forward and reverse kinetics of the enzyme Guanylate Kinase from yeast (Saccharomyces cerevisiae). The enzyme is held in a state of stress using the DNA spring method. The observation that mechanical stress has different effects on the forward and reverse reaction kinetics suggests that forward and reverse reactions follow different paths, on average, in the enzyme''s conformational space. Comparing the kinetics of the stressed and unstressed enzyme we also show that the maximum speed of the enzyme is comparable to the predictions of the relaxation model of enzyme action, where we use the independently determined dissipation coefficient for the enzyme''s conformational motion. The present experiments provide a mean to explore enzyme kinetics beyond the static energy landscape picture of transition state theory.     

Dynamic modeling of an enzymatic membrane reactor for the treatment of xenobiotic compounds

A membrane enzymatic reactor, consisting of a stirred tank coupled to an ultrafiltration membrane was set up for the enzymatic oxidation of xenobiotic compounds. The azo dye Orange II was selected for the model compound and manganese peroxidase for the oxidative enzyme. The ligninolytic cycle was initiated and maintained by the controlled addition of all factors (reactants, mediators, and stabilizers) at suitable rates. Considering the distinctiveness of this process, in which the substrate to be oxidized is not the primary substrate for the enzyme, a kinetic model was developed. The azo dye concentration and hydrogen peroxide addition rate were found to be the main factors affecting the process. The reaction kinetics was defined using a Michaelis-Menten model with respect to the Orange II concentration and a first-order linear dependence relative to the H(2)O(2) addition rate. The dynamic model, which takes into account both the kinetics and the hydraulics of the system, was validated by comparing the experimental results in continuous operation under steady and non-steady state to model predictions. In particular, the model predicted the behavior of the system when unexpected alterations in steady-state operation occurred. Furthermore, the model allowed us to obtain the most appropriate H(2)O(2)/Orange II ratio in the feed to maximize the process efficiency.     

Transient model of thermal deactivation of enzymes

The kinetics of enzyme deactivation provide useful insights on processes that determine the level of biological function of any enzyme. Photinus pyralis (firefly) luciferase is a convenient enzyme system for studying mechanisms and kinetics of enzyme deactivation, refolding, and denaturation caused by various external factors, physical or chemical by nature. In this report we present a study of luciferase deactivation caused by increased temperature (i.e., thermal deactivation). We found that deactivation occurs through a reversible intermediate state and can be described by a Transient model that includes active and reversibly inactive states. The model can be used as a general framework for analysis of complex, multiexponential transient kinetics that can be observed for some enzymes by reaction progression assays. In this study the Transient model has been used to develop an analytical model for studying a time course of luciferase deactivation. The model might be applicable toward enzymes in general and can be used to determine if the enzyme exposed to external factors, physical or chemical by nature, undergoes structural transformation consistent with thermal mechanisms of deactivation.     

Microscopic diffusion-reaction coupling in steady-state enzyme kinetics

The theory of diffusion-controlled processes is applied to describe the steady state of a reversible enzymatic reaction with special emphasis on the effects of enzyme saturation. A standard macroscopic steady-state treatment requires only that the average diffusional influx of substrate equals the net reaction flux as well as the average diffusional efflux of product. In contrast, the microscopic diffusion-reaction coupling used here takes properly into account the conditional concentration distributions of substrate and product: Only when the enzyme is unoccupied will there be a diffusional association flux; when the enzyme is occupied, the concentration distributions will relax towards their homogeneous bulk values. In this way the relaxation effects of the non-steady state will be constantly reoccurring as the enzyme shifts between unoccupied and occupied states. Thus, one is forced to describe the steady state as the weighted sum of properly time-averaged non-stationary conditional distributions. The consequences of the theory for an appropriate assessment of the parameters obtained in Lineweaver-Burk plots are discussed. In general, our results serve to justify the simpler macroscopic coupling scheme. However, considerable deviations between the standard treatment and our analysis can occur for fast enzymes with an essentially irreversible product release.