Lineweaver - Burk plot.pptx
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The document discusses Lineweaver-Burk plots, which are double reciprocal plots used to transform the Michaelis-Menten equation for enzyme kinetics into a linear form. This allows for a more accurate determination of the maximum velocity (Vmax) and Michaelis constant (Km) of an enzymatic reaction. The document provides examples of how to construct Lineweaver-Burk plots and extract Km and Vmax values from them. It also discusses how the plots can be used to determine the type of inhibition occurring in the presence of an inhibitor.
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![Need of MM equation transformation
Fig-1 : Michaelis – Menten plot
Fig-1 : A typical curve of Michaelis – Menten
plot of enzyme kinetics is hyperbole curve.
The curve is not perfectly horizontal means
maximum velocity (Vmax) can be achieved only at
infinite substrate concentration.
Because of this there are errors in Vmax and Km
estimation.
Hence in 1934 Hans Lineweaver and Dean Burk
transformed Michaelis – Menten equation into
Lineweaver – Burk equation.
Lineweaver – Burk plot has the great
advantage of allowing a more accurate
determination of Vmax and Km.
V or V0 - Reaction velocity
[s] - Substrate concentration
Vmax - Maximum velocity
Km - Michaelis – Menten constant](data:image/gif;base64,R0lGODlhAQABAIAAAAAAAP///yH5BAEAAAAALAAAAAABAAEAAAIBRAA7)
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This document presents a dynamical model of a vibrocompactor used in railroad construction. The model represents the vibrocompactor as a two-mass mechanical system with two degrees of freedom. Differential equations are derived to describe the forced vibrations of the system excited by an inertial disturbance. Numerical analysis is performed to examine the system's behavior under different forcing frequencies, including its natural frequencies and frequency response. The results indicate the machine can operate steadily over a wide range of frequencies except near its first natural frequency, where non-stationary beating vibrations occur.
Suspension_report
The document summarizes a Matlab simulation and optimization of a 1/4 suspension model for a Tesla vehicle over various road types. The simulation models the road profile and suspension system using differential equations. Initial analysis found maximum velocities and damping forces for each road type using standard suspension parameters. Optimization focused on minimizing damping force for a given maximum velocity by testing different spring and damping coefficients. The optimal values found were the lowest tested, suggesting lower values should be tested. The merit function was found to overvalue damping force and undervalue maximum velocity.
17.pmsm speed sensor less direct torque control based on ekf
This document presents a speed sensorless direct torque control method for permanent magnet synchronous motors (PMSM) using an Extended Kalman Filter (EKF). The EKF is used to estimate the stator flux linkage and rotor speed without needing a mechanical speed sensor. This overcomes issues with traditional direct torque control methods like large current and flux ripples. Simulation results show the EKF method maintains the fast torque response of direct torque control while improving dynamic and static performance and robustness to parameter and load variations compared to traditional methods.
Virtual post rig
This document describes the development and implementation of a virtual suspension analysis system called the Virtual Post Rig. The system aims to analyze and characterize suspension parameters and response through computer simulation, providing advantages over physical testing such as lower cost and faster experimentation. The system models the vehicle suspension using different levels of simplification. It obtains transfer functions relating sprung and unsprung mass response to road inputs. These are used to calculate performance metrics and compare suspension designs, balancing performance and comfort factors.
33208 10 modeling_biochemical_networks
This document discusses modeling of chemical reactions using differential equations based on the law of mass action. It provides the differential equation model for a basic chemical reaction A + B -> C. It then discusses a more complex enzymatic reaction involving the formation of an enzyme-substrate complex and derives the classic Michaelis-Menten differential equation model. It introduces techniques such as non-dimensionalization, the quasi-steady state approximation, and the Lineweaver-Burk transformation to simplify and analyze the model.
EM_and_Dynamic_Modeling_of_an_Maglev
This document summarizes a PhD dissertation on dynamic modeling of an eddy-current device called an electrodynamic wheel, which is an application of magnetic levitation technology. It presents the steady-state and transient analytic modeling of forces on a conductive guideway from a rotating magnet configuration, validates the models against finite element analysis and experimental testing, and applies the modeling to an electromechanical dynamic simulation. The modeling and validation lay the groundwork for further experimental work on a 4-wheeled electrodynamic wheel maglev system.
talk @ KEKPH 201.03.05
The document discusses using reactor neutrino experiments to determine the neutrino mass hierarchy. It finds that a baseline length of around 50km is optimal. An energy resolution with statistical error less than 3% and systematic error less than 1% is required to determine the mass hierarchy with reactor neutrinos. Specifically, a 20GW reactor with a 5 kiloton detector running for 5 years could achieve this resolution and determine the mass hierarchy.
Lecture 10
This document provides an overview of enzyme kinetics concepts including:
1) Rate constants like k1 and k-1 describe the rates of individual reaction steps. The overall rate v depends on reactant concentrations according to rate laws.
2) Michaelis-Menten kinetics describe enzyme-catalyzed reactions using parameters like Km, Vmax, and kcat/Km. Km represents substrate binding affinity, Vmax is the maximum reaction rate, and kcat/Km is the catalytic efficiency.
3) Reversible inhibitors are classified as competitive, non-competitive, or uncompetitive depending on whether they bind the enzyme (E), enzyme-substrate complex (ES), or both.
Acceleration Modelling of EV1_FR8695
This document models and simulates the acceleration of a GM EV1 electric vehicle from 0 to 60 mph using MATLAB. It considers three mass scenarios: the base mass of 1560kg, a reduced mass of 1326kg, and an increased mass of 1794kg. The acceleration is modeled in two phases using differential equations. Simulation results show that acceleration decreases as mass increases, with times to reach 60 mph being 9.1, 7.8, and 10.5 seconds respectively. A mathematical solution to the differential equations is also derived and verified against MATLAB simulations, showing close matches.
Similar to Lineweaver - Burk plot.pptx (20)
Improving EV Lateral Dynamics Control Using Infinity Norm Approach with Close...
Improving EV Lateral Dynamics Control Using Infinity Norm Approach with Close...
Vibrations of a mechanical system with inertial and forced disturbance
Vibrations of a mechanical system with inertial and forced disturbance
17.pmsm speed sensor less direct torque control based on ekf
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Recently uploaded
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...
Context. With a mass exceeding several 104 M⊙ and a rich and dense population of massive stars, supermassive young star clusters
represent the most massive star-forming environment that is dominated by the feedback from massive stars and gravitational interactions
among stars.
Aims. In this paper we present the Extended Westerlund 1 and 2 Open Clusters Survey (EWOCS) project, which aims to investigate
the influence of the starburst environment on the formation of stars and planets, and on the evolution of both low and high mass stars.
The primary targets of this project are Westerlund 1 and 2, the closest supermassive star clusters to the Sun.
Methods. The project is based primarily on recent observations conducted with the Chandra and JWST observatories. Specifically,
the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
−1
. The X-ray sources exhibit a highly concentrated spatial distribution,
with 1075 sources located within the central 1 arcmin. We have successfully detected X-ray emissions from 126 out of the 166 known
massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
Compexometric titration/Chelatorphy titration/chelating titration
Classification
Metal ion ion indicators
Masking and demasking reagents
Estimation of Magnisium sulphate
Calcium gluconate
Complexometric Titration/ chelatometry titration/chelating titration, introduction, Types-
1.Direct Titration
2.Back Titration
3.Replacement Titration
4.Indirect Titration
Masking agent, Demasking agents
formation of complex
comparition between masking and demasking agents,
Indicators/Metal ion indicators/ Metallochromic indicators/pM indicators,
Visual Technique,PM indicators (metallochromic), Indicators of pH, Redox Indicators
Instrumental Techniques-Photometry
Potentiometry
Miscellaneous methods.
Complex titration with EDTA.
Thornton ESPP slides UK WW Network 4_6_24.pdf
ESPP presentation to EU Waste Water Network, 4th June 2024 “EU policies driving nutrient removal and recycling
and the revised UWWTD (Urban Waste Water Treatment Directive)”
Authoring a personal GPT for your research and practice: How we created the Q...
Thematic analysis in qualitative research is a time-consuming and systematic task, typically done using teams. Team members must ground their activities on common understandings of the major concepts underlying the thematic analysis, and define criteria for its development. However, conceptual misunderstandings, equivocations, and lack of adherence to criteria are challenges to the quality and speed of this process. Given the distributed and uncertain nature of this process, we wondered if the tasks in thematic analysis could be supported by readily available artificial intelligence chatbots. Our early efforts point to potential benefits: not just saving time in the coding process but better adherence to criteria and grounding, by increasing triangulation between humans and artificial intelligence. This tutorial will provide a description and demonstration of the process we followed, as two academic researchers, to develop a custom ChatGPT to assist with qualitative coding in the thematic data analysis process of immersive learning accounts in a survey of the academic literature: QUAL-E Immersive Learning Thematic Analysis Helper. In the hands-on time, participants will try out QUAL-E and develop their ideas for their own qualitative coding ChatGPT. Participants that have the paid ChatGPT Plus subscription can create a draft of their assistants. The organizers will provide course materials and slide deck that participants will be able to utilize to continue development of their custom GPT. The paid subscription to ChatGPT Plus is not required to participate in this workshop, just for trying out personal GPTs during it.
如何办理(uvic毕业证书)维多利亚大学毕业证本科学位证书原版一模一样
原版纸张【微信：741003700 】【(uvic毕业证书)维多利亚大学毕业证】【微信：741003700 】学位证，留信认证（真实可查，永久存档）offer、雅思、外壳等材料/诚信可靠,可直接看成品样本，帮您解决无法毕业带来的各种难题！外壳，原版制作，诚信可靠，可直接看成品样本。行业标杆！精益求精，诚心合作，真诚制作！多年品质 ,按需精细制作，24小时接单,全套进口原装设备。十五年致力于帮助留学生解决难题，包您满意。
本公司拥有海外各大学样板无数，能完美还原海外各大学 Bachelor Diploma degree, Master Degree Diploma
1:1完美还原海外各大学毕业材料上的工艺：水印，阴影底纹，钢印LOGO烫金烫银，LOGO烫金烫银复合重叠。文字图案浮雕、激光镭射、紫外荧光、温感、复印防伪等防伪工艺。材料咨询办理、认证咨询办理请加学历顾问Q/微741003700
留信网认证的作用:
1:该专业认证可证明留学生真实身份
2:同时对留学生所学专业登记给予评定
3:国家专业人才认证中心颁发入库证书
4:这个认证书并且可以归档倒地方
5:凡事获得留信网入网的信息将会逐步更新到个人身份内，将在公安局网内查询个人身份证信息后，同步读取人才网入库信息
6:个人职称评审加20分
7:个人信誉贷款加10分
8:在国家人才网主办的国家网络招聘大会中纳入资料，供国家高端企业选择人才
3D Hybrid PIC simulation of the plasma expansion (ISSS-14)
3D Particle-In-Cell (PIC) algorithm,
Plasma expansion in the dipole magnetic field.
原版制作(carleton毕业证书)卡尔顿大学毕业证硕士文凭原版一模一样
原版纸张【微信：741003700 】【(carleton毕业证书)卡尔顿大学毕业证】【微信：741003700 】学位证，留信认证（真实可查，永久存档）offer、雅思、外壳等材料/诚信可靠,可直接看成品样本，帮您解决无法毕业带来的各种难题！外壳，原版制作，诚信可靠，可直接看成品样本。行业标杆！精益求精，诚心合作，真诚制作！多年品质 ,按需精细制作，24小时接单,全套进口原装设备。十五年致力于帮助留学生解决难题，包您满意。
本公司拥有海外各大学样板无数，能完美还原海外各大学 Bachelor Diploma degree, Master Degree Diploma
1:1完美还原海外各大学毕业材料上的工艺：水印，阴影底纹，钢印LOGO烫金烫银，LOGO烫金烫银复合重叠。文字图案浮雕、激光镭射、紫外荧光、温感、复印防伪等防伪工艺。材料咨询办理、认证咨询办理请加学历顾问Q/微741003700
留信网认证的作用:
1:该专业认证可证明留学生真实身份
2:同时对留学生所学专业登记给予评定
3:国家专业人才认证中心颁发入库证书
4:这个认证书并且可以归档倒地方
5:凡事获得留信网入网的信息将会逐步更新到个人身份内，将在公安局网内查询个人身份证信息后，同步读取人才网入库信息
6:个人职称评审加20分
7:个人信誉贷款加10分
8:在国家人才网主办的国家网络招聘大会中纳入资料，供国家高端企业选择人才
Deep Software Variability and Frictionless Reproducibility
Deep Software Variability and Frictionless ReproducibilityUniversity of Rennes, INSA Rennes, Inria/IRISA, CNRS
The ability to recreate computational results with minimal effort and actionable metrics provides a solid foundation for scientific research and software development. When people can replicate an analysis at the touch of a button using open-source software, open data, and methods to assess and compare proposals, it significantly eases verification of results, engagement with a diverse range of contributors, and progress. However, we have yet to fully achieve this; there are still many sociotechnical frictions.
Inspired by David Donoho's vision, this talk aims to revisit the three crucial pillars of frictionless reproducibility (data sharing, code sharing, and competitive challenges) with the perspective of deep software variability.
Our observation is that multiple layers — hardware, operating systems, third-party libraries, software versions, input data, compile-time options, and parameters — are subject to variability that exacerbates frictions but is also essential for achieving robust, generalizable results and fostering innovation. I will first review the literature, providing evidence of how the complex variability interactions across these layers affect qualitative and quantitative software properties, thereby complicating the reproduction and replication of scientific studies in various fields.
I will then present some software engineering and AI techniques that can support the strategic exploration of variability spaces. These include the use of abstractions and models (e.g., feature models), sampling strategies (e.g., uniform, random), cost-effective measurements (e.g., incremental build of software configurations), and dimensionality reduction methods (e.g., transfer learning, feature selection, software debloating).
I will finally argue that deep variability is both the problem and solution of frictionless reproducibility, calling the software science community to develop new methods and tools to manage variability and foster reproducibility in software systems.
Exposé invité Journées Nationales du GDR GPL 2024
Sharlene Leurig - Enabling Onsite Water Use with Net Zero Water
Sharlene Leurig - Enabling Onsite Water Use with Net Zero WaterTexas Alliance of Groundwater Districts
Presented at June 6-7 Texas Alliance of Groundwater Districts Business MeetingTHEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...
THEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...Abdul Wali Khan University Mardan,kP,Pakistan
hematic appreciation test is a psychological assessment tool used to measure an individual's appreciation and understanding of specific themes or topics. This test helps to evaluate an individual's ability to connect different ideas and concepts within a given theme, as well as their overall comprehension and interpretation skills. The results of the test can provide valuable insights into an individual's cognitive abilities, creativity, and critical thinking skillsApplied Science: Thermodynamics, Laws & Methodology.pdf
When I was asked to give a companion lecture in support of ‘The Philosophy of Science’ (https://shorturl.at/4pUXz) I decided not to walk through the detail of the many methodologies in order of use. Instead, I chose to employ a long standing, and ongoing, scientific development as an exemplar. And so, I chose the ever evolving story of Thermodynamics as a scientific investigation at its best.
Conducted over a period of >200 years, Thermodynamics R&D, and application, benefitted from the highest levels of professionalism, collaboration, and technical thoroughness. New layers of application, methodology, and practice were made possible by the progressive advance of technology. In turn, this has seen measurement and modelling accuracy continually improved at a micro and macro level.
Perhaps most importantly, Thermodynamics rapidly became a primary tool in the advance of applied science/engineering/technology, spanning micro-tech, to aerospace and cosmology. I can think of no better a story to illustrate the breadth of scientific methodologies and applications at their best.
Describing and Interpreting an Immersive Learning Case with the Immersion Cub...
Current descriptions of immersive learning cases are often difficult or impossible to compare. This is due to a myriad of different options on what details to include, which aspects are relevant, and on the descriptive approaches employed. Also, these aspects often combine very specific details with more general guidelines or indicate intents and rationales without clarifying their implementation. In this paper we provide a method to describe immersive learning cases that is structured to enable comparisons, yet flexible enough to allow researchers and practitioners to decide which aspects to include. This method leverages a taxonomy that classifies educational aspects at three levels (uses, practices, and strategies) and then utilizes two frameworks, the Immersive Learning Brain and the Immersion Cube, to enable a structured description and interpretation of immersive learning cases. The method is then demonstrated on a published immersive learning case on training for wind turbine maintenance using virtual reality. Applying the method results in a structured artifact, the Immersive Learning Case Sheet, that tags the case with its proximal uses, practices, and strategies, and refines the free text case description to ensure that matching details are included. This contribution is thus a case description method in support of future comparative research of immersive learning cases. We then discuss how the resulting description and interpretation can be leveraged to change immersion learning cases, by enriching them (considering low-effort changes or additions) or innovating (exploring more challenging avenues of transformation). The method holds significant promise to support better-grounded research in immersive learning.
Immersive Learning That Works: Research Grounding and Paths Forward
We will metaverse into the essence of immersive learning, into its three dimensions and conceptual models. This approach encompasses elements from teaching methodologies to social involvement, through organizational concerns and technologies. Challenging the perception of learning as knowledge transfer, we introduce a 'Uses, Practices & Strategies' model operationalized by the 'Immersive Learning Brain' and ‘Immersion Cube’ frameworks. This approach offers a comprehensive guide through the intricacies of immersive educational experiences and spotlighting research frontiers, along the immersion dimensions of system, narrative, and agency. Our discourse extends to stakeholders beyond the academic sphere, addressing the interests of technologists, instructional designers, and policymakers. We span various contexts, from formal education to organizational transformation to the new horizon of an AI-pervasive society. This keynote aims to unite the iLRN community in a collaborative journey towards a future where immersive learning research and practice coalesce, paving the way for innovative educational research and practice landscapes.
waterlessdyeingtechnolgyusing carbon dioxide chemicalspdf
The technology uses reclaimed CO₂ as the dyeing medium in a closed loop process. When pressurized, CO₂ becomes supercritical (SC-CO₂). In this state CO₂ has a very high solvent power, allowing the dye to dissolve easily.
在线办理(salfor毕业证书)索尔福德大学毕业证毕业完成信一模一样
学校原件一模一样【微信：741003700 】《(salfor毕业证书)索尔福德大学毕业证》【微信：741003700 】学位证，留信认证（真实可查，永久存档）原件一模一样纸张工艺/offer、雅思、外壳等材料/诚信可靠,可直接看成品样本，帮您解决无法毕业带来的各种难题！外壳，原版制作，诚信可靠，可直接看成品样本。行业标杆！精益求精，诚心合作，真诚制作！多年品质 ,按需精细制作，24小时接单,全套进口原装设备。十五年致力于帮助留学生解决难题，包您满意。
本公司拥有海外各大学样板无数，能完美还原。
1:1完美还原海外各大学毕业材料上的工艺：水印，阴影底纹，钢印LOGO烫金烫银，LOGO烫金烫银复合重叠。文字图案浮雕、激光镭射、紫外荧光、温感、复印防伪等防伪工艺。材料咨询办理、认证咨询办理请加学历顾问Q/微741003700
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三.真实教育部学历学位认证（教育部存档！教育部留服网站永久可查）
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◇在校期间，因各种原因未能顺利毕业……拿不到官方毕业证【q/微741003700】
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◇回国时间很长，忘记办理；
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◇企事业单位必须要求办理的
◇需要报考公务员、购买免税车、落转户口
◇申请留学生创业基金
留信网认证的作用:
1:该专业认证可证明留学生真实身份
2:同时对留学生所学专业登记给予评定
3:国家专业人才认证中心颁发入库证书
4:这个认证书并且可以归档倒地方
5:凡事获得留信网入网的信息将会逐步更新到个人身份内，将在公安局网内查询个人身份证信息后，同步读取人才网入库信息
6:个人职称评审加20分
7:个人信誉贷款加10分
8:在国家人才网主办的国家网络招聘大会中纳入资料，供国家高端企业选择人才
Equivariant neural networks and representation theory
Or: Beyond linear.
Abstract: Equivariant neural networks are neural networks that incorporate symmetries. The nonlinear activation functions in these networks result in interesting nonlinear equivariant maps between simple representations, and motivate the key player of this talk: piecewise linear representation theory.
Disclaimer: No one is perfect, so please mind that there might be mistakes and typos.
dtubbenhauer@gmail.com
Corrected slides: dtubbenhauer.com/talks.html
Recently uploaded (20)
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...
Compexometric titration/Chelatorphy titration/chelating titration
Compexometric titration/Chelatorphy titration/chelating titration
Authoring a personal GPT for your research and practice: How we created the Q...
Authoring a personal GPT for your research and practice: How we created the Q...
3D Hybrid PIC simulation of the plasma expansion (ISSS-14)
3D Hybrid PIC simulation of the plasma expansion (ISSS-14)
mô tả các thí nghiệm về đánh giá tác động dòng khí hóa sau đốt
mô tả các thí nghiệm về đánh giá tác động dòng khí hóa sau đốt
Deep Software Variability and Frictionless Reproducibility
Deep Software Variability and Frictionless Reproducibility
Sharlene Leurig - Enabling Onsite Water Use with Net Zero Water
Sharlene Leurig - Enabling Onsite Water Use with Net Zero Water
THEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...
THEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...
Applied Science: Thermodynamics, Laws & Methodology.pdf
Applied Science: Thermodynamics, Laws & Methodology.pdf
Describing and Interpreting an Immersive Learning Case with the Immersion Cub...
Describing and Interpreting an Immersive Learning Case with the Immersion Cub...
Immersive Learning That Works: Research Grounding and Paths Forward
Immersive Learning That Works: Research Grounding and Paths Forward
waterlessdyeingtechnolgyusing carbon dioxide chemicalspdf
waterlessdyeingtechnolgyusing carbon dioxide chemicalspdf
Equivariant neural networks and representation theory
Equivariant neural networks and representation theory
Lineweaver - Burk plot.pptx
- 1. Lineweaver - Burk plot Double reciprocal plot BTH-3506 : ENZYMOLOGY CE-1 -By 1) Gunali Chaudhari (8111) 2) Nikita Jedhe (218213)
- 2. Need of MM equation transformation Fig-1 : Michaelis – Menten plot Fig-1 : A typical curve of Michaelis – Menten plot of enzyme kinetics is hyperbole curve. The curve is not perfectly horizontal means maximum velocity (Vmax) can be achieved only at infinite substrate concentration. Because of this there are errors in Vmax and Km estimation. Hence in 1934 Hans Lineweaver and Dean Burk transformed Michaelis – Menten equation into Lineweaver – Burk equation. Lineweaver – Burk plot has the great advantage of allowing a more accurate determination of Vmax and Km. V or V0 - Reaction velocity [s] - Substrate concentration Vmax - Maximum velocity Km - Michaelis – Menten constant
- 3. How to calculate x-intercept? Fig-3 : Lineweaver – Burk plot X-intercept at y = 0 y = 1/V0= 0 Put, y = 0 in Lineweaver – Burk equation – 0 = km . 1 + 1 Vmax [s] Vmax - 1 = Km . 1 Vmax Vmax [s] -1 . Vmax = 1 Vmax Km [s] -1 = 1 Km [s]
- 4. 1 = Km+[s] V0 Vmax [s] Reciprocal of MM equation Double reciprocal plot Fig-2 : Lineweaver – Burk plot v0 = Vmax [s] Km + [s] Michaelis – Menten equation Inverting the equation 1 = Km + 1 V0 Vmax [s] Vmax Lineweaver – Burk equation Comparing this equation with straight line equation y = mx + c, we get – y= 1/v0, m(slope)= Km/Vmax, x= 1/[s], c (y-intercept)= 1/Vmax
- 5. Practice Problem 1 If the X-intercept on a Lineweaver – Burk plot or double reciprocal plot is equal to -2 , what is the Km value ? 1) -2 2) 0.5 3) 1 4) -0.5 5) -1 Solution - According to Lineweaver – Burk plot, x- intercept = -1/Km -1/Km = -2 ½ = Km Km = 2) 0.5
- 6. Practice problem 2 The following experimental data were collected during a study of the catalytic activity of an intestinal peptidase with the substrate glycylglycine: Glycylglycine + H2O peptidase 2 glycine [S] (Mm) V (µm/min) 1.5 0.21 2.0 0.24 3.0 0.28 4.0 0.33 8.0 0.40 16.0 0.45 1/[s] (Mm)-1 1/v (µm/min)-1 0.67 4.76 0.50 4.16 0.33 3.57 0.25 3.03 0.125 2.50 0.0625 2.22 y = 4.240x + 2.004 R² = 0.991 0 1 2 3 4 5 6 0 0.2 0.4 0.6 0.8 Reciprocal of Reaction velocity 1/v or1/ v 0 (1/µm/min) Reciprocal of Glycylglycine concentration 1/[s] (1/Mm) y= mx + c = y= 4.240x + 2.004 C = 1/Vmax = 2.004 Vmax = 1/2.004 = 0.49 µm/min m = Km/Vmax = 4.240, 4.240= Km/0.49 Km= 4.240 × 0.49 = 2.07 Mm
- 7. Lineweaver – Burk plot of bisubstrate reactions Sequential Mechanism Ping pong Mechanism Fig-4 : This enzyme reaction involving a ternery complex formation. For this mechanism, Lineweaver –Burk plot at varying S1 and different fixed values of S2 give a series of intersecting lines. Fig-5 : This enzyme reaction not involving a ternery complex formation. This is double displacement pathway. For this mechanism, Lineweaver – Burk plot at varying S1 and different fixed values of S2 give a series of parallel lines.
- 8. Practice problem 3 Consider the following set of data and answer the following questions – [s] (mM) V (mMol/min) V (+inhibitor) (mMol/min) 0.5 23.5 16.67 1.0 32.2 25.25 1.5 36.9 30.49 2.5 41.8 37.04 3.5 44.0 38.91 A) Plot the data on a Lineweaver – Burk plot and determine Km and Vmax. B) The 2nd set of velocities represents the rate of the reaction when an inhibitor is added. Plot this data on the same graph as above and determine new Km and Vmax and the type of inhibitor (competitive, uncompetitive, non-competitive). C) Can the effects of the inhibitor be over – ridden by adding more substrate? Why? 1/[s] (1/mM) 1/V (mMol/min)-1 1/v (+ inhibitor) (mMol/min)-1 2 0.042 0.059 1 0.031 0.039 0.66 0.027 0.032 0.40 0.023 0.026 0.28 0.022 0.025
- 9. Lineweaver – Burk plot for Enzyme Inhibition Fig-6 : Competitive Inhibition Slope – Change Y-intercept- Same Km – increased Vmax- unaffected Fig-7 : Non competitive Inhibition Slope – Change Y- intercept - Change Km – unaffected Vmax- reduced Fig-8 : Uncompetitive Inhibition Slope – Same Y- intercept - Change Km – reduced Vmax- reduced
- 10. y = 0.011x + 0.018 R² = 0.997 y = 0.020x + 0.018 R² = 0.998 0 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0 0.5 1 1.5 2 2.5 1/[s] (1/mM) 1/V (1/mM/min) without inhibitor with inhibitor B) y= 0.020x + 0.018 After solving for Km and Vmax We get, Vmax= 55.55 mM/min Km= 1.1 mM m = change, c = same, Vmax= unaffected, Km = increased The type of inhibitor is Competitive. A) y= 0.011 + 0.018 After solving for Km and Vmax We get, Vmax= 55.55 mM/min Km=0.61mM
- 11. Limitations Fig-9 : Limitations of Lineweaver – Burk plot Fig-9 : all data found at large substrate concentrations will be clustered near the origin. small experimental errors are found far to the right of the y-axis calling for a large extrapolation back to obtain x- and y- intercepts. To circumvent these problems, experimental data is often plotted on a Eadie-Hofstee plot, another linear form of the Michaelis-Menten plot or by computer programs that fit the data to the Michaelis-Menten equation itself.
- 12. References • Lehninger Principles Of Biochemistry- 4th edition and 7th edition • www.chem.libretexts.org • www.sciencing.com • www.ncbi.nlm.nih.gov