Effect of substrate concentration on enzyme activity experiment pdf


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Effect of substrate concentration on enzyme activity experiment pdf

1. TITLE Experiment 3: Enzyme Kinetics Experiment OBJECTIVES 1. (William D. Increasing Substrate Concentration increases the rate of reaction. The Lab In this lab, students investigate the activity of the enzyme diastase, and examine the effects of enzyme concentration, temperature, and pH on the ability of diastase to digest starch. Hypothesis I believe that as the concentration of the hydrogen peroxide (substrate) decreases, the rate of reaction will decrease as well. Students will perform dilutions to produce the various enzyme concentrations. The effect of substrate and enzyme concentration . Validity and Reliability As I’ve just discussed, various factors affect enzyme activity. INTRODUCTION. In this part of the activity, you will use the Available Enzyme slider to simulate how doubling the amount of enzyme affects the enzyme catalysis rate, amount of substrate, and amounts of reaction products. pdf Bookmark it to view later. The six factors are: (1) Concentration of Enzyme (2) Concentration of Substrate (3) Effect of Temperature (4) Effect of pH (5) Effect of Product Concentration and (6) Effect of Activators. CATALASE ACTIVITY: EFFECT OF ENZYME CONCENTRATION . intentionally by the cell to regulate enzyme activity. 1 1. 5 and 2. The rate of an enzyme catalyzed reaction depends directly on the concentration of an enzyme. Students will study the effect of various factors on the rate of reaction between an enzyme and its substrate. The relative reaction rate increased with pH - up to the optimum value (around 6. , U. Enzymes are influence by several factors such as pH, temperature, enzyme concentration and substrate concentration. Effect ofsubstrate concentration. , Z. The effect of substrate concentration on enzyme activity. Substrate concentration is the amount of substrate present that can be turned into product and is most commonly measured in molarity (moles per liter). The rate at which the enzyme substrate complex is formed is dramatically increased or decreased in changes in substrate concentration, temperature, pH and presence of an competitive inhibitor the effect of these 4 factors on the enzyme activity is The Effect of pH on Catalase g The title mentions pH and enzyme activity, but these are not such as temperature and substrate concentration. For humans, the optimum temperature is 37oC, but for bacteria living in hot springs, the optimal temperature would be about 85oC. The enzyme activity in the presence of leucomalachite green, an acceptor, and hydrogen peroxide, a substrate, has been studied in the usual manner and the Michaelis constant determined. 2015 Biology 110 Laboratory Manual – page 75 D. Label Enzyme activity can be which is the substrate concentration required for an site of the usual substrate and exert an allosteric effect to change PART C: EFFECT OF TEMPERATURE SUBSTRATE CONCENTRATION ON REACTION RATE. Introduction Effect Of Substrate Concentration On The Activity Of Catalase Concentration of enzyme: The higher the concentration, the higher the rate of reaction will be. Two lines are shown distinguishing the rate of the reaction and the line of best fit; correlating closely. Repeat practical 1 using different concentrations of hydrogen peroxide solution – 5%, 10%, 20%, 30%, 40%, and 50%. It can spontaneouslyThis paper describes a detailed study of the compound of horseradish peroxidase and hydrogen peroxide, an enzyme-substrate compound. 4 3 4. Choose the purified enzyme preparation from part C that yielded the highest activity while still being linear over the entire 20 min. activity at 2X enzyme concentration was approximately 2 times that at 1X enzyme concentration. Starch is the substrate used and amylase is the enzyme. Substrate concentration has many different effects Thus, the effect of alcalase on the DH of tuna viscera protein extract was initially done on enzyme concentrations of 1. This lab deals with how an enzyme is affected by different concentrations of substrate--specifically what effect will different concentrations of a substrate (H 2 O 2) have on an enzyme (catalase in potato juice). 2. Students also learn to determine the optimal conditions under which the enzyme activity is maximized. The enzyme used in this experiment will be catalase from liver. Aim. To analyse the effect of substrate concentration on the activity of enzyme. 5 mg lysozyme m1-1 distilled water. Keep the other factors constant. Determination of the effect of enzyme concentration on catalysis using starch an amylase. Explain how enzymes work. Part D. The catalytic activity of an enzyme is the number of substrate molecules converted to product per second per 1 molecule of enzyme. As the Effect of pH on Enzyme Activity . The effect of temperature, pH, time course, substrate concentration, enzyme concentration and metal ions was studied and results were noted as follows. Catalase Activity: Effect of Enzyme Concentration Westminster College SIM Page 2 GUIDING QUESTIONSThe Effect Of Substrate Concentration On The Activity Of The Enzyme Catalase A Level Biology Project Aims This is an experiment to examine how the concentration of the substrate hydrogen peroxide affects the rate of reaction of the enzyme catalase. Thus need to be constant to keep experiment fair. In this experiment, the effects of pH, temperature, enzyme concentration, and substrate concentration will be tested on the enzyme catecholase. pH is the measurement of the concentration of the Hydrogen ions (H+) 3. Compare the two rates. The substrate-enzyme complex then reacts (Artioli, 2008). 4 In this experiment amylase is use to break down the starch molecules. With a fixed concentration of an enzyme and with increasing substrate concentration , a rapid increase in the rate of the reaction is observed at first . This is an experiment to examine how the concentration of the substrate hydrogen peroxide affects the rate of reaction of the enzyme catalase. The more enzymes available, the quicker the reaction will occur until the substrate is all used up. This reaction will result into two products namely water and oxygen. Activity is usually highest at pH 10 and lowest at pH 4. This experiment aimed to produced glucose syrup from red sorghum starch by hydrolysis Enzyme activity. Curves showing effect of peptone on activity of diluted pepsin solutions. Enzymes such as Catalase are protein molecules which are found in living cells. 041 %/s. Practical Assessment 2 – The effect of substrate concentration on the rate of enzyme activity of Catalase Aim To investigate the effect of substrate concentration (manipulated by increasing concentration of hydrogen peroxide) on the rate of enzyme activity of catalase, produced by liver cells, on the decomposition of hydrogen peroxide. Dilute the 0. To understand the effect of enzyme and substrate concentration on reaction rate To maintain a constant temperature and pH during the experiment, reaction. EXPERIMENT OBJECTIVE: The objective of this experiment is to introduce students to the concept of enzyme catalysis. This is done because increasing the sum of enzymes for different conditions will accordingly increase the figure of active sites that the substrates can adhere to 3, therefore increasing enzyme activity until the enzyme concentration surpasses the substrate concentration, where the enzyme activity will level out. 2O2 and 5 mL of water. SPECTROPHOTOMETRIC DETERMINATION OF ENZYME ACTIVITY Today you will examine the enzyme activity of tyrosinase by assaying its activity with the substrate DOPA using the spectrophotometer. We have already discovered the optimum pH level is pH7, this experiment looks at varying concentrations of the substrate, and its effect on the rate of activity. There are four factors that affect the enzyme activity that is temperature, pH, substrate concentration and enzyme concentration. 1 1. Table 2 – Effect of Substrate Concentration on Enzyme Activity Substrate Concentration However, since the enzyme concentration is unchanged, the substrate concentration can be increased to a point where all active sites of the present enzymes are already occupied with substrates, ensuing in impregnation of the enzymes, such that increasing the substrate concentration will no longer increase the enzyme activity. 4. To investigate the effect of surface area on the rate of reaction using the enzyme Catalase, found in potato, and the substrate Hydrogen Peroxide. When the concentration of substrate increase, the rate of reaction is also increase. However, after a certain concentration, any increase will have no effect on the rate of reaction,Catalyst is not consumed in the reaction itself. 2O2 and 5 …SPECTROPHOTOMETRIC DETERMINATION OF ENZYME ACTIVITY Today you will examine the enzyme activity of tyrosinase by assaying its activity with the substrate DOPA using the spectrophotometer. In this experiment we will evaluate the effects on hydrogen peroxidase. Factors affecting enzyme function substrate concentration reaction rate Substrate concentration as substrate = reaction rate more substrate = more frequently collide with enzyme reaction rate levels off all enzymes have active site engaged enzyme is saturated maximum rate of reaction At this form, the substrate is converted into product. The substrate used in this experiment is hydrogen peroxide. e. Stock enzyme solution concentration is 0. com/introbiochem/Enzymes. An Investigation of the Effect Enzyme Concentration has on the Rate of Catalase Activity Essay Sample. 25% H2O2 3. Thus an addition in substrate amylum causes an addition in enzyme contact action. Figure 3 is a line graph representing the effect of Substrate Concentration on enzyme activity. Because of this high rate of activity, only a very small amount of enzyme is needed to act on a relatively large amount of substrate, the substance the enzyme acts upon. . Lab #4: . In this given experiment, it was to be tested what impact the concentration level of substrate will have on the reaction rate. Stoltze 2010) There are four variables that affect enzyme activity: Temperature, pH, and Substrate/enzyme concentration. Therefore, enzymes are also affected by modifications of pH. The effects of cofactors, reversible inhibitors (competitive and non-competitive) and irreversible inhibitors on the rate of enzymatic reactions. In this experiment the catalase enzyme in the liver acting as a catalyst will break down the substrate, hydrogen peroxide. Amylase is an enzyme present in saliva and pancreatic juice. Figure 1 shows that DH of tuna viscera increased as the alcalase concentration increases. Introduction: Enzyme speed up chemical reaction without being changed itself. The binding provides better chemical conditions to activate the reaction and, in turn, lowers the activation energy (Artioli, 2008). This is because more substrate molecules will be colliding with enzyme molecules, so more product will be formed. between enzyme concentration and enzyme activity Illustration of the effect of substrate concentration on reaction rate. Enzymes, which in the experiment was peroxidase extracted from turnip, are protein catalysts that break up the substrate, which in this experiment was hydrogen peroxide. When the substrate concentration is kept high, the reaction rate is proportional to enzyme concentration. Most enzymes function efficiently over a narrow pH range. This is an equilibrium reaction, and will be favored by a high concentration of enzyme and/or substrate. • 3. The research outlined in this paper investigated this property of effect the stated change, X1, in the substrate, then, as in Equation 3, Kl’ 1 =-- nh x E where K1’ is a constant representing the proportionality of reaction velocity and enzyme concentration in the second, non-reference preparation. Mamot, Mohamad Yusof Maskat, S. In this experiment, you will measure the rate of enzyme activity under various conditions, Westminster College. Specifically, it was found that a 1 % starch solution resulted in the highest rate of enzyme activity. Define catalyst, enzyme, active site, substrates, enzyme-substrate complex, products, synthesis, degradation, oxidation/reduction, isomerization, and enzyme specificity. 75% H2O2 1. The catalase you used for your preliminary trials was inside yeast cells. S. 05 M sodium phosphate buffer pH 6. What Effects of Concentrations of a Substrate on Enzyme Lab Answers. The reactants are called substrate. Part II Testing the Effect of Enzyme Concentration. Enzyme Activity Measuring the Effect of Enzyme Concentration. Substrate concentration has many different effects on enzyme activity, depending on the context …Substrate Concentration. Any changes in these may affect the ability of enzymes to carry out their function, as they may work at a slower rate, become denatured, or not work at all. You are designing an experiment to examine the effect of substrate concentration on enzyme activity. Because of this high rate of activity, only a very small amount of enzyme is needed to act on a relatively large amount of substrate, the substance the enzyme acts upon. Enzyme Review. If the substrate concentration is increased then the rate of the reaction will increase. Calculate the rate of enzyme action in toothpicks per second. With a larger number of catalase molecules, the chance of successful collisions between enzyme and substrate will be increased. BY BRITTON CHANCE (From the Johnson Research Foundation, University of Pennsylvania, Philadelphia, Pennsylvania) (Received for publication, June 1, 1951) The Effect Of Substrate Concentration On The Activity Of The Enzyme Catalase A Level Biology Project Aims This is an experiment to examine how the concentration of the substrate hydrogen peroxide affects the rate of reaction of the enzyme catalase. 2–7. enzymes-1 Laboratory Inquiry 1: Enzymes and Enzymatic Reactions Figure 1. This lab contains the use of a catalase which is an enzyme found in liver. Indeed, above a certain point, virtually all enzyme molecules will have the cofactor they need, and thus increasing cofactor concentration will have no further influence on reaction rate. Feb 20, 2014 Effects of substrate concentration on enzyme activity of catalase from For the results of the experiment to be highly accurate there were a The effect of substrate and enzyme concentration . Describe’the’relationship’between’substrate’concentration’and’the’initial’reaction’rate’of’an’enzyme‘ To understand the role of guaiacol in this experiment. In our assay, substrate is ethanol (alcohol) and enzyme is ADH. The x-axis describes the concentration and the y-axis describes the rate of the enzymes reaction. If the enzyme cannot keep up, unprocessed alcohol will accumulate in the body. Amylase works at a pH optimum of around 7. Hypothesis: 1. Effect of Enzyme Concentration During catalysis, the first step is the substrate (S) binding to the enzyme (E), giving an enzyme-substrate complex (ES). Part 1: Effect of Enzyme Concentration. CATALASE ACTIVITY: EFFECT OF ENZYME CONCENTRATION . In the presence of a given amount of enzyme, the rate of an enzymatic reaction increases as the substrate concentration increases until a limiting rate is reached, after which further increase in the substrate concentration produces no significant change in the reaction rate (part 3. 5mM) of Substrate Solution to test the effects of pH on enzyme activity. Each enzyme has an optimum temperature at which its reaction rate is fastest. Describe the experiment using sucrase and interpret the resulting data. Meanwhile, in chemical reaction s, one or more reactants are converted into one or more products. Roughton and V. The enzyme concentration also speed up the rate of enzyme activity, but if the enzyme reach on a certain level than it stay the same. 02%, 0. mobilis was investigated. Fresh tissue contains blood. effect of substrate concentration on enzyme activity experiment pdfLab #4: Enzymes p. But as substrate concentration continues to increase , the increase in rate of The object of this experiment is to demonstrate the effect of performing the Oct 23, 2013 The purpose of this lab is to investigate the impact of temperature, substrate concentration, the concentration of the enzyme or substrate is increased, the rate of the . 5 units/mL and transfer to clean, labeled test tube. Lab #4: Enzymes p. Cow liver was homogenized to provide the supply of hydrogen peroxidase. The effect of substrate concentration, pH and other factors upon the activity of carbonic anhydrase F. This means that the more substrate there is, the more enzyme activity can be observed. As the amount of enzyme present increases, the rate of reaction increases. The conformation of a protein is influenced by pH and as enzyme activity is crucially dependent on its conformation, its activity is likewise affected. As enzyme concentration increases, reaction rate increases As the reaction rate levels off, the substrate becomes a limiting factor-Not all enzyme molecules can find the substrate Whoops! There was a problem previewing Enzyme Inquiry Investigation F15. pdfenzymatic reactions proceed - temperature, pH, enzyme concentration, substrate concentration, and the presence of any inhibitors or activators. Chemical activities that happen within the cell produces hydrogen peroxide, which is poisonous and can kill the organism. The effect of substrate concentration Having successfully completed the practical work to determine the effect of enzyme concentration, modify your experimental procedure to show how you would investigate the effect of substrate concentration on initial rate of enzyme reaction. enzyme and substrate concentration, temperature, and substrate specificity, as well as calculate the concentration of enzymes and substrates, V o, V max, K M and reaction rate. " Enzymes: What’s in your spit? substrate (though the enzyme can act with many substrates at the same time) Part 3 – How does concentration affect enzyme activity? (Hydrogen Peroxide and Catalase) Hydrogen peroxide is a molecule made up of hydrogen and oxygen. pdf from BIO 1107 at Georgia Gwinnett College. This is the optimum pH found inside most of the living cells. The Effect of Substrate Concentration on the Activity of the Enzyme Catalase. Experiments. 1 0. These catalysts are made by the cells in very small amounts which are not consumed during a chemical reaction. A change in pH above or below this range reduces the rate of …The hydrolysis time, hydrolysis temperature, pH, E/S ratio and substrate concentration had a significant influence on degree of hydrolysis, metal-chelating activity, DPPH and superoxide radical scavenging activity on casein hydrolysate of goat milk by Alcalase, the results were beneficial for further provide theoretical basis for production of antioxidant peptides. Enzymes are globular proteins which act as catalysts of chemical reaction. The activity of an enzyme is affected by temperature. (cf. The effects of temperature, pH, enzyme concentration and substrate concentration on the rate of . The enzyme catalase, found in potato juice, was used for the catalyst along with a substrate known as hydrogen peroxide (H2O2). Gen Nelson, Catalase Lab, http://www. • Predict the effects of varying environmental conditions such as pH and temperature on enzyme structure and activity. Evaluate metabolic activities using experimental knowledge of enzymes. The amount of enzyme is not always sufficient for the amount of alcohol to be broken down. We will assume that each filter disk is coated with the same amount of catalase (except in the investigation of the effect of enzyme concentration of enzyme activity). 2 The activity of an enzyme is affected by many factors including temperature and pH. worthington-biochem. • Design and conduct an experiment to explore the effect of another variable on the rate of the enzymatic reaction. pdf . Background Information. When we plot a graph with substrate concentration on the X axis and corresponding velocity on Y axis. 3. , 2011). accessexcellence. 5. • Define enzyme, catalyst, active site, substrate, activation energy, product, denaturation and explain how enzymes function. - The Concentration of Substrate's Effect on the Rate of an Enzyme Controlled Reaction Plan ==== In this investigation I aim to find out whether the concentration of hydrogen peroxide affects the rate at which catalyse breaks it down. The level of Measuring DHFR Catalytic Activity – Effect of enzyme concentration of reaction rate PDF. The presence of the enzyme catalase in the cell helps to quickly convert this toxic substrate into safer products of water and oxygen. Enzymes are biocatalysts that speed up chemical reactions. Dependent Variable Effect of Substrate Concentration on Enzyme Activity In this experiment, if the sucrose concentration The relative reaction rate (volume of oxygen evolved during five minutes) was plotted against pH. Concentration Enzyme Concentration Time to Float Disc (in seconds) % (units/mL) Trial 1 Trial 2 Average Rate Class Average 100 100 75 75 50 50 25 25 0 0 Table 2 – Effect of Substrate Concentration on Enzyme Activity Substrate Concentration Time to Float Disc (in seconds) (from serial The effect of substrate concentration on the rate of an enzyme catalyzed reaction. Table I is a summary of an experiment illustrating this point. They are used to speed up specific reactions in the cells. Finally a point is reached, beyond which the increase in V0 will not depend on the [S]. Incubation Time. Enzyme concentration Substrate concentration decrease collisions between enzyme & substrate. Effect of substrate concentration on the enzyme activity of GFOR/GL in calcium alginate-immobilized Z. Enzyme activity was greater when using the chicken liver rather than the cow liver. The Effect of pH and Enzyme Concentration on Enzyme Activity; Abstract; Enzymes help many chemical reactions to occur in living organisms. Most studies examining the effect of substrate composition on enzyme activity have used changes in plant community composition as a proxy for changes in substrate composition. One way to demonstrate this is by determining the effect of enzyme concentration on the rate of activity, while using a substrate concentration which is in excess. Skip the theory and go straight to: For practical purposes, Km is the concentration of substrate which permits the enzyme to achieve half Vmax. While these studies have demonstrated the importance of Introduction. Tube Buffer (mL) Guaiacol (mL) Enzyme (mL) Hydrogen peroxide (mL) 1 (blank) 4. Based on the consequences it can said that if the enzyme concentration is kept changeless, as it was in this experiment, so the rate of reaction is straight relative to the sum of substrate nowadays. Enzyme kinetics is the study of catalytic reactions, or reaction rate, which occurs in the presence ofThe objective of this experiment is to introduce students to the concept of enzyme catalysis. View Essay - The Effect of Substrate Concentration and pH on Enzyme. concentration of its substrate, which is constantly being replenished by the previous enzyme, and of its product, which is steadily being removed by the next enzyme, all enzymeshavingthesameflux,orsteadystatenetforward rate. State null and alternative hypotheses for this experiment: Null: Alternative: Fill in the blanks below to complete the design. This experiment uses one concentration (0. Fill each test tube with 5 mL of 3. J. The study of the rate at which an enzyme works is called enzyme kinetics. To test the effect of enzyme concentration, dilutions have been changed so that a variable amount of enzyme is added to a fixed amount of substrate. Adsorb microscale quantities of plant extract onto filter paper discs and assess catalase activity by comparing times taken for the discs to surface in hydrogen peroxide solution. 1% amylase solution was pipetted into one test tube and 5cm³ starch into another. Enzyme Activity 6 Laying the Foundation in Biology 316 Enzyme Activity Measuring the Effect of Enzyme Concentration Enzymes are proteins that serve as biological catalysts in a wide variety of life sustaining chemical reactions that take place in cells. AP Biology Enzymes and temperature molecules that reduce enzyme activity- The Effect of pH on Enzyme Activity pH is a measure of the concentration of hydrogen ions in a solution. e0is the enzyme concentration at time zero (initial concentration). Enzyme Activity Measuring the Effect of Enzyme Concentration Objective Students will measure the length of time it takes for various concentrations of catalase-soaked filter paper disks to float to the top of a cup filled with hydrogen peroxide. O-nitrophe nol, the y ellow produc t of the bre akdown of ONPG, absorbs light maximally at 420nm. Expectations: Overall: B2. enzyme activity with implications for ecosystem functioning (Hooper et al. When temperature The effect of temperature and pH on the activity of GFOR was assessed for the purified enzyme [1], intact Z. them with water for the following experiments. Place your hands in the pail of iced water for 10 minutes. An enzyme with a high Km relative to the physiological concentration of substrate, as shown above, is not normally saturated with substrate, and its activity will vary as the concentration of substrate varies, so that the rate of formation of product will depend on the availability of substrate. In a typical experiment, you will measure the change (decrease) in substrate concentration or th e increase in product concentration over time. • Determine the relationship between other environmental variables (temperature) and the rate of the enzymatic reaction. Feb 20, 2014 Effects of substrate concentration on enzyme activity of catalase from For the results of the experiment to be highly accurate there were a The effect of substrate and enzyme concentration . Substrate determination. Reaction rate depends on many different factors, including concentration of enzyme, concentration of substrate, and the pH, osmolarity, and temperature of the reaction environment. 3) and then decreased at higher pH values. Effect of enzyme concentration. In these works, pH values in the ranges of 5. period. Introduction: This enzyme helps to convert hydrogen peroxide into oxygen and water. Finally, the product will be released. 0%. The Effect Of Substrate Concentration On The Activity Of The Enzyme Catalase A Level Biology Project. 10B - Explain cell functions and processes in terms of chemical reactions and energy changes. Test the effect of temperature on enzyme activity. The inhibitor is the substance that decreases or abolishes the rate of enzyme action. Effect of enzyme concentration. SPECTROPHOTOMETRIC DETERMINATION OF ENZYME ACTIVITY that one can know how much substrate Effects of Enzyme Concentration experiment with enzyme kinetics in a “modern” way, controlling the pH of the solution etc. was highest at neutral pH (7) and showed only 1/3 enzyme activity at pH 11while pH of 1 showed no. 340 nm and the X-AXIS will be time. 0 2 Explain your answers: Vocabulary list Enzyme During the experiment, factors such as: amount of enzyme, substrate concentration, and pH were tested to find a correlation with enzyme activity. activity increases steeply as substrate concentration increases. lation between the rate and the concentration of substrate or enzyme. *Note: This is an abbreviated Project Idea, without notes to start your background research, a specific list of materials, or a procedure for how to do the experiment. 7–6. Effects of Enzyme ConcentrationEffect Of Substrate Concentration On The Activity Of Catalase Concentration of enzyme: The higher the concentration, the higher the rate of reaction will be. Test what effect the chemical reaction has on the enzyme itself. To investigate the effect of amylase concentration on its activity. The control showed the same enzyme activity each time the experiment was run. Retrying. 5 ml, 5 ml, 7. Introduction. Upon the completion of this experiment it was found that when temperature and pH were aid constant the activity of an enzyme is determined by the comparative concentration of the enzyme and its substrate, amylum in this instance. Lab Report (Effect of concentration on enzyme activity) Biology Noor Alawadhi 11-KC Introduction: An Enzyme is a protein, which is capable of starting a chemical reaction, which involves the formation or breakage of chemical bonds. 2%. Table 1 gives the relative proportions of buffer, enzyme and cell-wall suspension which have been utilized in testing the effect of substrate concentration of enzyme activity. Figure 3 is a line graph representing the effect of Substrate Concentration on enzyme activity. Enzymes are said to be catalytic proteins which increases the rate of a chemical reaction without being altered in the process of that reaction. Salwanee, Wan Aida Wan Mustapha, S. 0, 1. Factors Affecting Enzyme Activity: 6 Factors. To understand why peroxidase is a necessary enzyme for all aerobic cells. To determine the effect of enzyme concentration the effect of substrate concentration on enzyme activity. Links to PubMed are also available for Selected References. Use appropriate keywords to use in your searchEnzymes work at their optimum temperature and pH. The concentration of enzyme, E,‘, of this preparation necessary to effect the of enzyme, the enzyme activity is 1. After this point, increases in substrate concentration will not increase the velocity enzyme activity in terms of reaction speed from measurable laboratory data. Catalase is an enzyme found in food such as potato and liver. The cell-wall stock solution is 0. STANDARDS more peroxide substrate is left, O2 is no longer produced. To understand the effect of enzyme and substrate concentration on reaction rate To maintain a constant temperature and pH during the experiment, reaction. Then more the concentration of the enzyme, the higher the rate of the reaction. It can be noted that both enzyme solution used had different results. How is the rate of enzyme activity affected by increasing the concentration of the substrate? 3. 5cm³ of undiluted 0. Adsorb microscale quantities of plant extract onto filter paper discs and assess catalase activity by comparing times taken …Effect of pH Each enzyme has an optimum pH. an experiment was set up to measure the activity of an enzyme using a substrate that produced a coloured product. *The reaction time for tube 1 was the fastest due to the high substrate concentration and lower dH20 concentration. W. SAFTEY 1. 2. Finally, we tested the effect of pH on the enzyme with pH 7, pH 1 and pH 11. However, studying the effects of substrate concentration is elaborated by the fact that during the course of an in vitro reaction, [S] changes due to the conversion of substrate …3 The effect of substrate concentration on catalase activity Rate of reaction can be increased when there are more substrate molecules available to bind with the active site of the catalase enzyme. Enzymes work at their optimum temperature and pH. Week 2 – Effect of Enzyme & Substrate Concentration on Enzyme Activity Week 2 Learning Goals: To understand the role of enzymes in catalyzing chemical reactions To understand the effect of enzyme and substrate concentration on reaction rate To learn how to perform enzyme assays During enzyme substrate reaction, the initial velocity V0 gradually increases with increasing concentration of the substrate. Other factors such as concentration of enzymes and substrates affect the rate of reaction. An enzyme unit is the amount of enzyme that will catalyse the transformation of 1 μmol of substrate/min under specified conditions of pH and temperature. 05 M sodium phosphate buffer pH 6. Solution concentrations, volumes and observations for Experiment 2: The effect of substrate concentration on enzyme activity. Test normal catalase activity. Effect of Concentration on Enzyme Activity Lab Report. Since any deviation from the pH optimum reduces obligatorily the enzyme activity, such an effect can easily be misinterpreted as enzyme inhibition: the more of the particular component is added, the lower the enzyme activity. We can describe the reaction Mar 21, 2018 Effect of Substrate Concentration on the Rate of Activity of Catalase Because the enzyme is the limiting factor, the reaction will stop completely This will minimise the effect of temperature on the results of the experiment. 0 % H2O2 0. An Investigation of Catalase/Hydrogen Peroxide Enzymatic Kinetics Lauren Perryman With Racquel Robinson, Kimberly Roberts, Halie Choate Abstract Catalase is an enzyme that breaks down hydrogen peroxide into water and gaseous oxygen. 25 mg m1-1 cell-wall material suspended in a 0. Figure 1. 05%, 0. This molecule is known as the substrate. However, the effect of substrate on enzyme activity is not simply to increase it. Aims. The substance that an example: jones_lab5. Initiate the first enzyme reaction. What were the independent and dependent variables in this substrate-enzyme complex (Artioli, 2008). Upon the completion of this experiment it was found that when temperature and pH were aid constant the activity of an enzyme is determined by the comparative concentration of the enzyme and its substrate, amylum in this instance. An enzyme with a high Km has a low affinity for its substrate, and requires a greater concentration of substrate to achieve Vmax. . mobilis cells Initially, the effect of substrate concentrations on the catalytic activity of the immobilized GFOR/GL from Z. Next: Effects of Inhibitors PDF version of Introduction to Enzymes This means that the more substrate there is, the more enzyme activity can be observed. Objective: To investigate the influence of substrate concentration over enzyme activity. Enzyme activity can be affected by the change of pH, temperature and the concentration of substrate or enzyme in the solution. Enzymes work at their optimum temperature and pH. Define the term optimum with respect to peroxidase activity. of the enzyme much like a change in temperature does, with similar effects on enzyme activity. A water bath was heated to 35°C. When the rate of reaction Substrate Concentration – The binding of an enzyme to its substrate is an essential part of the enzyme catalyzed reaction. As for temperature, it will affect the enzyme activity when there is a rise in temperature. However, at very high substrate concentrations, the enzymes become saturated with substrate and a higher concentration of substrate does not increase the reaction rate. pH Subtrate concentration Effects Of temperature, pH and substrate concentration on enzyme activity Temperature: Optimum pH at which enzyme activity is fastest (pH 7 is optimum for most enzymes) At low substrate concentrations, enzyme. This experiment aimed to produced glucose syrup from red sorghum starch by hydrolysis Enzyme activity . 005%, 0. Enzyme Concentration In order to study the effect of increasing the enzyme concentration upon the reaction rate, the substrate must be present in an excess amount; i. 0% H. Let us examine enzyme kinetics as a function of the concentration of substrate available to the enzyme. Thus, in experiments that involve enzyme activity in fresh tissue, it is helpful to wash off the blood in a beaker of cold water before grinding the tissue. The time it takes for the filter paper disk to rise (from the bottom of the cup) is an indication of the rate of enzyme activity. Enzymes the rate of a chemical reaction by decreasing the activation energy, the energy required to initiate a reaction. 2 0. Concentration of Substrate. the enzyme concentration and/or the substrate concentration. Enzyme experiments often provide real ‘messy’ data, because their activity can change dramatically from one lesson to the next. Investigate the chemical structures, functions, and chemical properties of biological molecules involved in some common cellular processes and biochemical reactions; B3. 0 µmole per min per ml enzyme. The hydrolysis of viscera protein extract (VPE) and enzyme concentration influence enzymatic activity …Cited by: 6Publish Year: 2013Author: S. In an enzyme action lab, a student wanted to know the effect of salt concentration on the action of an enzyme. active site, also has a specific shape. Enzymes are proteins that serve as biological catalysts in a wide variety of life-sustaining chemical reactions that take place in cells. Materials and Methods. - The Effect of pH on Enzyme Activity pH is a measure of the concentration of hydrogen ions in a solution. Hydrogen ion will affect the hydrogen and ionic bonds within the This means that the more substrate there is, the more enzyme activity can be observed. The focal aim of this experiment is to see what pH our protease works at best to determine where it is made in the body. It is said that the substrate is complimentary to their substrate. The purpose of this experiment was to investigate the effects of pH on catalase activity. A. Schraer Hert J. • Predict the effects of varying enzyme and substrate concentrations on enzyme activity. Enzyme inhibition means decreasing or cessation in the enzyme activity. Kinetic energy, in turn is the energy in motion. This one is Erol Altug's the negative effect of denaturing more and more enzyme molecules. II. To analyse the effect of substrate concentration on the activity of enzyme. Substrate concentration is the amount of substrate present that can be turned into product and is most commonly measured in molarity (moles per liter). enzymes-1 Laboratory Inquiry 1: Enzymes and Enzymatic Reactions is equal to the substrate concentration at which the reaction the effect of pH on enzyme The time it takes for the filter paper disk to rise (from the bottom of the cup) is an indication of the rate of enzyme activity. Effect of Catalase on Hydrogen Peroxide Under the influence of an enzyme called catalase, the hydrogen Repeat the experiment using uncooked pieces of muscle G. An enzyme can be denatured (unfolded) by extreme heat, pH, or ionic concentration. 25 mg m1-1 cell-wall material suspended in a 0. Based on the consequences it can said that if the enzyme concentration is kept changeless, as it was in this experiment, so the rate of reaction is straight relative to the sum of substrate nowadays. For many enzymes, this corresponds to pH values of around 7. Table 2. The purpose of this lab was to observe the effect of enzyme concentration on the reaction The binding of an enzyme with an specific substrate produces an enzyme substrate complex (ES). 0 0. The contact between the enzyme and substrate is the most essential pre-requisite for enzyme activity. The activity of enzymes is affected by both the concentrations of enzymes present and the concentration of substrate present. The relative activity is determined by noting the time taken for the starch substrate to break down. Enzyme Lab Activity Questions With Answers dumas,kawasaki kle500 2004 2007 workshop service manual pdf,pro motocross reacted the substrate binds to the active site of enzyme reaction questions and answers pdf free download in The higher the substrate concentration was, the more substrate the enzyme could convert to products. poisons, radiation, the concentration of enzymes, and the concentration of the substrate. 7 0. At higher substrate concentration the rate of reaction increase smaller and smaller amount in response to increase in substrate concentration. Enzymes are very sensitive organic molecules. Students also learn to determine the At a concentration of 5 drops, the rate in the above experiment should be about 0. What do you think would happen if you increased the substrate concentration to 40. 147965772-James-Lee-Enzyme-Kinetics-Solution. Thus, the effect of alcalase on the DH of tuna viscera protein extract was initially done on enzyme concentrations of 1. The effect of substrate concentration can be investigated using the following investigation. Oct 23, 2013 Abstract: The purpose of this lab is to investigate the impact of temperature, substrate concentration, enzyme concentration, and the presence of an inhibitor on the effectiveness and rate of an enzyme. To determine the effect of substrate concentration on the rate of enzyme action. The single most important property of enzymes is the ability to increase the rates of reactions occurring in living organisms, a property known as catalytic activity . Substrate Concentration. Enzyme Practical - Biology bibliographies - in Harvard style The Effect Of Substrate Concentration On The Activity Of The Enzyme Effect of Substrate intensity increases we can calculate the activity of the enzyme. The specific activity of an enzyme is expressed as the number of units per milligram of protein. Experiment 3: The Effect of pH on Catalase Activity Lab Results Page 02 section 1 - 25 marks attempt all questions 1. After the substrate is bound, the reaction takes place, and then the product is released. 0 cm3 starch solution, measured using measuring cylinder (± 0. 0% H2O2 Analysis(Questions:(’ 1. pdf) Consider the Concepts. Usually, the enzyme activity increases from pH 4 to 10. 2% starch to …Part II Testing the Effect of Enzyme Concentration. g. PURPOSE To determine the effect of different concentrations of a substrate have on an enzyme. Enzyme assays are used also to determine the concentration of substrates in samples Enzyme experiments often provide real ‘messy’ data, because their activity can change dramatically from one lesson to the next. : Effect of substrate concentration over enzyme activity. Each enzyme has its own optimal pH which it works under,To investigate the effect of amylase concentration on its activity. , the reaction must be independent of the substrate concentration. Enzyme kinetics is the study of catalytic reactions, or reaction rate, which occurs in the presence of However, since the enzyme concentration is unchanged, the substrate concentration can be increased to a point where all active sites of the present enzymes are already occupied with substrates, ensuing in impregnation of the enzymes, such that increasing the substrate concentration will no longer increase the enzyme activity. Effect of substrate concentration: In part A, the concentration of starch used in the assay was 0. Thus OD 420 in this experiment is a measure of the Volume of substrate Increased volume of substrate will increase product (oxygen) produced. At relatively low concentration of substrate, the rate of reaction increase linearly with an increase in substrate concentration. The induced-fit model of enzyme action. PDF alternative if you need it. However, since the enzyme concentration is unchanged, the substrate concentration can be increased to a point where all active sites of the present enzymes are already occupied with substrates, ensuing in impregnation of the enzymes, such that increasing the substrate concentration will no longer increase the enzyme activity. Starch is a common nutrient used Part 2: Effect of Substrate Concentration on Reaction Rate. 1 Euler, H. It really should be evident that the substrate and enzyme concentration levels would effect the rate of reaction the was they did as it was noted in the intro of the As the substrate concentration increases, the enzyme reaction rate increases. The job of catalase in this experiment was to accelerate the …The substrate concentration, [S] is one of the important parameter that affecting the rate of a reaction that catalyzed by an enzyme. As time increases there will less abundance of starch and more of the sugar present. If you do not remember how to use the III. Place three test tubes in a rack and label them 1, 2, and 3. 7 0. More substrates will also mean a faster reaction rate until the enzyme is fully saturated so that it cannot continue increasing its activity. Changing the enzyme concentration The amount of enzyme available to process the substrate can control an enzyme catalytic reaction. But as substrate concentration continues to increase , the Full text is available as a scanned copy of the original print version. Then the spectrophotometer was turned on and allowed to warm up and Find Study Resources The Effect Of Substrate Concentration On The Activity Of The Enzyme Catalase A Level Biology Project Aims This is an experiment to examine how the concentration of the substrate hydrogen peroxide affects the rate of reaction of the enzyme catalase. The second experiment showed the dependence of the effective work of the enzymes of their concentration. Get a printable copy (PDF file) of the complete article (657K), or click on a page image below to browse page by page. Then dip a construction paper disk into different solutions that consist of either 2. Student answers may vary.  Enzymesare proteins …2 The activity of an enzyme is affected by many factors including temperature and pH. effect of substrate concentration on enzyme activity experiment pdf As catalysts, enzymes lower the amount of energy required to make a reaction occur. Furthermore, as the amount of substrate increases, the rate of reaction will initially increases. Initial velocity experiments in which the substrate concentration is varied generally yield information about substrate binding. Select 10 toothpicks. How does changing the substrate concentration compare to changing the enzyme concentration in this experiment? 5. pH is a measure of hydrogen ion and sodium hydroxide ion concentration. Substrate concentration has many different effects on enzyme activity, depending on the context of the reaction and the nature of the enzyme. Effects of Enzyme ConcentrationFound that the enzyme catecholase is the most efficient in pH 6, though the increasing of acidity or alkalinity of solution reduces enzyme activity. § Your cells are constantly exposed to poisonous - Investigating the Effect of Substrate Concentration on Catalase Reaction Planning -Aim: The aim of the experiment is to examine how the concentration of the substrate (Hydrogen Peroxide, H2O2) affects the rate of reaction of the enzyme (Catalase). 4 ice. 5% H2O2 2. The concentration of substrates is often used to measure enzyme activity, which is based on the rate of a reaction (product formed over time). Catalyst is not consumed in the reaction itself. The optimum pH of a particular enzyme corresponds to the pH of its natural environment. Enzyme Concentration. From the previous lab exercise on succinate dehydrogenase, you know the effect of enzyme concentration. There is a change when amylase reacts with starch. Precautions should be taken while handling enzymes, since parameters like pH, temperature, and presence of ions affect the enzyme activity. Introduction This is an A-level biology project. 11. enzyme activity (i. 2000; Waldrop and Firestone 2006). EFFECT OF pH UPON THE REACTION KINETICS OF THE ENZYME-SUBSTRATE COMPOUNDS OF CATALASE*. The effects of temperature, pH, enzyme concentration and substrate concentration on the rate of enzyme reactions. that would improve the experiment. CONCLUSION It was confirmed in this experiment that changes in the environment like temperature, pH levels, substrate and enzyme concentrations did effect the rate of reaction. Related experiments Investigating an enzyme-controlled reaction: catalase and hydrogen peroxide concentration 1. Ibrahim[PDF]Introduction to Enzymes - Worthington Biochemicalwww. Generally the enzyme activity increases as the drops of the enzyme (concentration) increases. Hypothesize about the effect of changing enzyme concentration on the time of starch disappearance. Substrate concentration also affects the enzyme activity. As substrate concentration increases, reaction rate increases until the example, if you wish to determine the effect of pH on enzyme activity, you might ask “Does lower pH change the rate of reaction of an enzyme?” 2. E is an enzyme molecule and italics lowercasefor the concentration: e. Volume of liquid detergent: 2drops of liquid detergent were added to each test tube throughout the experiment. Booth Physiology Laboratory, University of Cambridge, England and the Fatigue Laboratory, Harvard University, Boston, Mass. , temperature, acidity, hormones). EDVO-Kit: AP13 Enzyme Activity See Page 3 for storage instructions. The higher the hydrogen ion concentration, the lower the pH. A substrate is the surface or material on or from which an The time it takes for the filter paper disk to rise (from the bottom of the cup) is an indication of the rate of enzyme activity. They are all very specific as each enzyme just performs one particular reaction. Enzyme concentration and substrate concentration also play a role in catalysis (Campbell 2005). Assemble and label clean test tubes to prevent contamination or premature reaction from occurring. 0 0. A small Km indicates that the enzyme requires only a small amount of substrate to become saturated. Effect of temperature and pH on enzyme activity Enzymes are basically the proteins which speed up the rate of reactions in living cells. 3 0. First, extract the catalase enzyme from the potato. Describe how temperature, pH, enzyme concentration, and/or substrate concentration can affect the reaction rate. Enzymes are specific to their substrate, because the shape of their active site will only fit the shape of their substrate. How Substrate Concentration Affects Enzyme Activity. Above or below an enzyme’s optimum pH, its activity is lower. Effect of pH on alkaline phosphatase activity. At this point, temperature, pH and time of incubation were fixed at 40°C, pH8 and 60 min. Effect of Temperature. Test what types of tissues contain the enzyme catalase. Explain in one to two complete sentences why a spontaneous reaction would benefit from an enzyme to catalyze the reaction. When the substrate binds with the enzyme, a product is produced. Measurement of Enzyme Activity Stopped assays To measure the activity of an enzyme one must measure Biology 4A Laboratory: Enzyme Function substrate concentration, enzyme concentration and inhibitors. Optical density (OD) is a measure of the a mount of light absorbe d by a solution. Factors that can alter enzyme activity include concentration of enzyme, substrate, or products; temperature; pH; and protein denaturants. The results are . An enzyme makes a reaction proceed faster, but is not consumed in the reaction. With a fixed concentration of an enzyme and with increasing substrate concentration , a rapid increase in the rate of the reaction is observed at first. [1] A substrate is a substance which an enzyme acts upon. Interested in The Effect of Substrate Concentration and pH on Enzyme. Because the active site is so specific it will only bind with one kind of molecule. Graph the results putting substrate concentration on the x-axis. To determine the effects of substrate concentration, pH, and temperature on enzyme activity. 5 ml, or 10 ml enzyme and enough distilled water to have a final volume of 10 ml. But most of those reactions would not occur fast enough without help from enzymes -- protein molecules that bring substrate molecules together and combine them into new molecules. The enzyme activity. To describe how pH, temperature, and the concentration of an enzyme and its substrate influence enzyme activity. STANDARDS • 3. enzyme activity. 12B - Identify and describe factors affecting metabolic function (e. This means that the more substrate there is, the more enzyme activity can be observed. Title of Lab: The effect of concentration on enzyme activity. org) . Because random. of three environmental factors, temperature, pH, and enzyme concentration level. 0, and 6. If a chemical reaction is exergonic, it is considered spontaneous. If changed it would affect the rate of enzyme activity since substrate concentration is one of the factors that affect enzyme activity. Importance: The rate of a chemical reaction is affected by the total number of enzymes as well as the concentration of substrates. Use 1. This is due to increase in frequency of collisions between the substrates molecules and the enzymes increase. org). Substrate Concentration Time to Float Disc (in seconds) (from serial dilutions) Trial 1 Trial 2 Average Rate Class Average 0. It catalyses the hydrolysis of amylose and amylopectin (both starch components) to a mixture of products including maltose and dextrin. The Effect Of Substrate Concentration On The Activity Of The Enzyme Catalase A Level Biology Project Aims This is an experiment to examine how the concentration of the substrate hydrogen peroxide affects the rate of reaction of the enzyme catalase. enzyme and substrate concentration, temperature, and substrate specificity, as well as calculate the concentration of enzymes and substrates, V o, V max, K M and reaction rate. Before considering the factors which affect enzyme reactions, it is important to demonstrate that the enzyme assay shows that the enzyme actually follows accepted chemical principles. This experiment will determine the effect substrate concentration against enzyme. the potential to alter its activity, even abolishing the enzyme's ability to catalyze the reaction altogether, if the effect is great enough. Effect of Substrate Concentration 1. Using a graduated cylinder, measure about 4 mL of the enzyme, glucose oxidase at a concentration of 6. As substrate concentration (alcohol) increases, overall enzyme activity rate increases to keep up. Enzymes are very specific molecules, each enzyme has a certain type of active site necessary for the reaction to happen. 1% and 0. enzyme activity (i. They speed up chemical reactions. Effect of temperature on pectinase activity: The effect of temperature on the activity of crude enzyme was studied by DNS method. The Effect of Temperature on Enzyme Activity 1. 5, 6. Describe how you would set up the positive and negative controls for this experiment. Temperature is the average kinetic energy of a system. Since the enzyme is usually found in a fairly neutral environment, it was hypothesized that the enzyme activity will be slowed or stopped in pH solutions that were either acidic or basic. Hence, the maximum velocity is reached at relatively low substrate concentrations. 1 0. Experiment A: The Influence of Enzyme Concentration on the Rate of Starch Digestion Formulate a question about enzyme concentration and reaction rate. , Effect of temperature, pH, enzyme to substrate ratio, substrate concentration 30 and time on the antioxidative activity of hydrolysates from goat milk casein by Alcalase and enzymatic reactions, and antioxidant enzymes and endogenous antioxidants in the body under synergy of has been removed (Murphy et al. 2) pH level pH level is also a factor which affects the rate of enzyme activity. 5–7. Determine the effect that starch concentration has on activity by performing assays on solutions of 0. The objective of this experiment is to introduce students to the concept of enzyme catalysis. The effects of temperature, pH, enzyme concentration and substrate concentration on the rate of . You can identify abbreviated Project Ideas by the asterisk at the end of the title. Most enzymes have an optimum pH near to 7. Shu et al. 01%, 0. In this experiment you will measure the disappearance of substrate and cofactor by recording the decrease in Absorbance at 340 nm over time. pdf. There is a release of a disaccharide - maltose. According to the similarity between the inhibitor and the substrate, enzyme inhibition is Download the student sheet Investigating effect of temperature on the activity of lipase (72 KB) with questions and answers. A change in pH above or below this range reduces the rate of enzyme reaction considerably. This graph shows the results for the first trial of the substrate experiment, or part 2. After this point, increases in substrate concentration will not increase the velocity enzyme activity in terms of reaction speed from measurable laboratory data. Your breath, the beating of your heart and your thoughts would all stop in the absence of chemical reactions. Repeat step 1. Due to the presence of enzymes in blood that change the carbon dioxide gas that is dissolved in the blood into carbonic acid, the tissue itself may effect pH. Consider the natural environment of yeast, which is ubiquitous. At low substrate concentration, the active site on the enzyme is not saturated by substrate and thus the enzyme is not working at full capacity. Effect of Environmental Variables on Enzyme Reaction Rate in Living Cells A catalyst is a substance that increases the rate of a chemical reaction without being consumed or permanently changed in the reaction. 0% hydrogen peroxide? 4. Summary. A large Km indicates the need for high substrate concentrations to achieve maximum reaction velocity. In order for an enzyme to perform its given job, it needs what is known as a substrate to bind to the active site of the enzyme so that the enzyme can speed up the reaction of the substrate. The effect of substrate concentration on the rate of an enzyme catalyzed reaction. • The convention used for this slides is to use UPPERCASEfor the molecular entity: e. In this experiment, rate of enzyme activity was measured under different pH values. Explain the use of Benedict’s solution in demonstrating sucrase The substrate with the lowest Km upon which the enzyme acts as a catalyst is frequently assumed to be enzyme's natural substrate, though this is not true for all enzymes. The Effect Of Substrate Concentration On The Activity Of The Enzyme Catalase A Level Biology Project Aims This is an experiment to examine how the concentration of the substrate hydrogen peroxide affects the rate of reaction of the enzyme catalase. Table 2 – Effect of Substrate Concentration on Enzyme Activity. The effect of concentration on enzyme activity. PURPOSE To determine the effect of Investigate the effect of substrate concentration on the rate of activity of the enzyme catalase. The most common catalysts in biological systems are special proteins called enzymes. Afterwards, place the paper disk into a solution of hydrogen peroxide. 0, respectively, were referred to as optimal for the action of the enzyme. mobilis[14], and permeabilized and immobilized cells [8]. Safety and COSHH requirements • glasswareEffects of Enzyme Concentration, Temperature, pH and Time on the Degree of degree of hydrolysis (DH) during hydrolysis by using single factor experiment. Enzyme concentration and substrate concentration also play a role in catalysis (Campbell 2005). Experiments with unireactant enzymes results in the estimation of the kinetic parameters, but there is only one possible model for substrate binding. substrate amount remains constant, causes an increase in the rate of the reaction until a maximum point is reached where all enzyme is bound to the substrate present. Test the effect of concentration on enzyme activity. H. Bookmark The Effect of Substrate Concentration and pH on Enzyme. If this happens, the enzyme will no longer be functional because the shape of the active site will be destroyed. Graph 5 Effect of Substrate Concentration Series 2 Time of Reaction in Seconds there was no reaction time. 0% H. 2 0. Time how long it takes to break the 10 toothpicks as fast as you can