Enzyme kinteics of amylase using l2 in Kl was used to determine starch conc at t = 0 min andt = 1 min. Tubes were set absorbance measured after 1 minute of incubation. The reactionwas halted with HCl and 5 mL of I, in KI indicator was added. 1 ml of each was used assample in the spectrophotometer.Using this table,Concentration ofVolume ofVolume ofVolume of l2Test tube stock starch,starchdistilledin KI solution, Absorbancemg/mLsolution, mL water, mLmL1100.00092100.44.650.33063100.84.20.589110|1.23.80.840910|1.63.41.0885102.03.01.2572What is the standard curve for the conc. of the starch?

The standard curve allows estimating the concentration of unknown samples by comparing them to known…

Answered: Enzyme kinteics of amylase using l2 in…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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Similar questions

Enzyme X has a molecular weight of 48,000. It converts substrate Z into product Y. Z absorbs at 340 nm, and Y absorbs at 480 nm. A.) At what wavelength would you measure the change in absorbance to assay for enzyme X? Would the absorbance increase or decrease over time? B.)If Vmax = 60 μmol/min and you used 400 μL of a 0.1 mg/mL solution of enzyme, what is the turnover number?
please fill out the table I need it for lab tomorrow At the start of the lab, you will be given stock preparations of alkaline phosphatase and PNPP onice—keep them that way. You will also be instructed on the volume of enzyme solution to beadded to reaction mixtures. Typically, this lab uses 5 μl of enzyme dilution. Before the start of lab, calculate thevolumes in μl of each component you need to mix together for each of these samples.For all of the following assays, use the amount of enzyme noted above and vary the substrateconcentration. Use the following assay format and PNPP concentrations of 0.0, 0.20, 0.40, 0.60,0.80, and 1.0 mM. Calculate the volumes of the 100 mM PNPP stock solution and buffer (finalvolume of 1.00 ml) needed to get each of the substrate concentrations. A single blank tube, withalkaline buffer only, is sufficient, because PNPP has no absorbance at 405 nm.
Two experiments were performed with the enzymeribonuclease. In experiment 1 the effect of increasingsubstrate concentration on reaction velocity was measured.In experiment 2 the reaction mixtures were identical tothose in experiment 1 except that 0.1 mg of an unknowncompound was added to each tube. Plot the data accordingto the Lineweaver-Burk method. Determine the effect of theunknown compound on the enzyme’s activity. (Substrateconcentration is measured in millimoles per liter. Velocity ismeasured in the change in optical density per hour.)
An enzyme “happyase” catalyzes the reaction: sad to happy. For total enzyme concentration of 4 nM, the measured Vmax was 3.2 μM/S. What is the kcat for happyase ______ and what is its unit _____ (shown in fraction format, eg. M/S)
Mike has determined that enzyme he is attempting to purify has an isoelectric point of 4.5 (pI = 4.5). He has decided to examine anion exchange chromatography as a potential purification step. He tested out using 2 different buffer and linear NaCl gradient on HPLC (like what you did). His results are shown below. Which buffer should Mike uses for his purification (both buffer has pH higher than enzyme’s pI)? Why?
During enzyme analysis to determine glucose concentration (sample used is corn syrup stock sol. and enzymes are glucose oxidase and peroxidase), what conditions must be in place to ensure accurate results for such coupled reaction?
Utilising the provided class data generate the following graphs: I) Michaelis Menten; II) Lineweaver-Burk; and III) Hanes-Woolf. Ensure that you clearly label each graph,and add the relevant trendlines with equations. Table 1: Class data demonstrating the Absorbance at 700nm obtained for the alkaline phosphatase enzyme reaction Table 1 tube Abs700mm 1 0.000 2 0.060 2 0.090 4 0.140 5 0.190 6 0.250 7 0.290 The equipment we used are • 20mM Tris Buffer pH 8.5 • 33mM MgCl2 • Alkaline Phosphatase (2mg/ml) in 20mM Tris Buffer pH 8.5 • 4mM Glucose-1-phosphate • Acid Molybdate pH 5.0 • Reducing Agent • Distilled Water • Glass Test tubes • Tube Rack • Cuvette • Pipettes and Tips • Water bath set to 37oC The method we used is Method/Protocol: 1. Read the protocol in its entirety before starting. Take note of any additional information that appears in subsequent steps that may influence how previous steps are performed. 2. Using glass tubes, generate the reactions mixtures…
Explain how acetaminopen gives such spectroscopic and chromatographic graph by interpreting the peaks that appeared in the graphs. Fig1. Mass Spectrometric Chromatogram of Acetaminophen (the two graphs) Fig2. IR Spectrograph of Acetaminophen
Lab: Isolation of beta-amylase from sweet potato Objective: To isolate the enzyme β -amylase using sweet potato as a source Materials Required: Sweet potato.Knife/peeler.Mortar and Pestle.A Blender.Blue capped tubes.20mM sodium Phosphate buffer at pH 7.Vortexer. Procedure: 1.Take a clean sweet potato and peel the skin off.2.Weigh the peeled sweet potato and note the weight.3.The sweet potato is cut into small pieces and transferred into a mortar and pestle.4.The pieces are crushed and then transferred into a blender.5.Add 40 ml of cold 20mM sodium phosphate buffer saline. Blend it until it forms a paste.6.Gently transfer the potato slurry into a blue capped tube.7.Allow the enzyme to extract over a 1 hour period at room temperature, with frequent vigorous stirring on a vortex mixer.8.Then the extract is filtered using a GF A glass fibre filter and the filtrate is collected in a new blue capped tube.9.Centrifuge the filtrate at 12000rpm for 20 minutes at 4 degree Celsius.10. After…
An enzymatic reaction follows M-M kinetics with Vmax= 2.5 mol m-3s-1and Km = 5 mM.Calculate the time required for 50% conversion of the substrate in a batch reactor if theinitial substrate concentration is0.2 M.Show your calculation steps.
So we did an experiment and I am curious on the following because the prof did not elaborate too much. She just gave the postive result and reagent. 1. What type of reaction occurred when the samples (enumerated) reacted with the Bial’s reagent? Give the chemical equation for each.a. Glucoseb. Sucrosec. Fructosed. Lactosee. Galactosef. Ribose 2. Explain the mechanism behind the test in simple terms. 3. What are the important products of the reaction of each sample?
A certain plant storage protein, after being isolated, was digested using both 8N H2SO4 and conc. Ba(OH)2. The resulting hydrolysates were then analyzed for amino acid composition using paper chromatography with EtOH-NH3-H2O (80:10:10) as the solvent system and ninhydrin as the visualizing agent. Name the amino acids found in the acid hydrolysate labelled 1-5.

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