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Nt1310 Unit 5 Lab

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Graph 5 illustrates the relationship between the enzyme parameters of the Michaelis–Menten equation. This equation is represented in equation 3 below. The plot was supposed to be a smooth curve but an error occurred, so the velocity of tube 8 was lower than expected. Tube 8 and tube 9 had the same concentration of substrate, so the velocity of tube 8 should have been the same as the velocity of tube 9. The velocity of tube 9 was 26.296μmoles/liter/min. This error could have been due to the wrong about of enzyme or ONPG added to the tube. The wrong amount of ONPG can be easily added to the tube since too many tubes were set up at the same time. Taking time and carefully adjusting the pipet can prevent this error from occur. Other possible errors …show more content…
Then, we plotted the 1/V versus 1/S to get the Lineweaver-Burk plot for series A. The plot is represented by the graph 7 below.
The same procedure performed for series A was repeated for series B, but 0.4ml of 50mM lactose was added to each tube instead of 0.4 of water. Two absorbance readings of each tube were taken and the average was calculated. The results and what were added to the tube are listed in table 6 below. Table 7 was constructed the same way as table 5.
The information from table 7 was used to produce the Lineweaver-Burk plot for series B with inhibitor. The best-fit line generated an equation of y = 5.287x + 0.0124. This is shown below on graph 8.
Same procedure performed for series C, but 0.4ml of 150mm lactose was added to each tube instead. The results are showed in Table 8 and 9 below. Another Lineweaver-Burk plot was made using the 1/V and 1/S values from table 9.
Finally, the obtained line equations for three plots were used to calculate the Kmapparent and Vmaxapparent of each plot. For each series, the plot produced a y = mx+ b equation, where b is 1/Vmaxapparent and m is Kmapparent / Vmaxapparent . The calculations for the Vmaxapparent and the Kmapparent of each plot are shown

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