Enzyme Kinetics Examples and Problems 1. An enzyme is produced for producing a sun protection lotion.

Given kinetic data for the enzyme reaction with Vm=2.5 mmol m!.s" #m=$.% m& and 'o=12m&" what would (e the time re)uired for %5* conversion in a (atch reactor+

2.

An enzyme was assayed at an initial su(strate concentration of 1,-5&. .he #m/ for the su(strate is 201,-!&. At the end of 1 min" 2* of the su(strate had (een converted to product. a. 1hat * of the su(strate will (e converted to product at the end of ! min.+ calculate the product and su(strate concentration at that time. 2f the initial concentration of the su(strate were 1,-3&" what percent of the su(strate will (e converted to product after ! min+ 1hat is the ma0imum attaina(le velocity of the reaction with the enzyme concentration used+ At a(out what su(strate concentration will Vm (e o(served+ At this ' concentration" what * of the su(strate will (e converted to product after ! min+

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c. d. e.

3. An enzyme catalyzed reaction of the form ' 4 has a value of G, = !.5 k6 mole. a. (. 7alculate the e)uili(rium constant at 2%$# 2f we start with a solution containing ,.1& su(strate" what is the at e)uili(rium+

4. Given the reaction 8 9 ' - 8.' -- 8 9 4 where k1 = 101,:& s" k-1=101,2 s and k2=!01,2 s a. 5. 7alculate #m/" #m

An enzymatic assay was carried under two different sets of conditions out using a pure su(strate '. .he results are ta(ulated (elow. a. 7alculate the #m and Vm for given su(strate concentrations and rates for A only. (. c. ;se the ! plots and comment on their differences <or inhi(ition e0ample do = then compare with A A V, >mol ?.min@ ,.21 ,.25 ,.2$ ,.!! ,.55 ,.5, B v >mol ?.min@ ,.,$ ,.1 ,.12 ,.1! ,.13 ,.1$

' >mol ?@ 1.501,-5 2.,01,-5 !.,01,-5 5.,01,-5 $.,01,-5 13.,01,-5 3.

Aou are given a 5., ug m? solution of an enzyme >&1 = 5,",,,Ba@. 2n your e0periment" you have determined the initial reaction velocities vo at several su(strate concentrations" C'D.

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Betermine the #m and Vm using lineweaver-=urke 4lot 7alculate the turnover num(er for the reaction

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' >m&@ V, >m& min@ ,.25 ,.,3 ,.5 ,.,$ ,.: ,.1! 1 ,.13 2 ,.25 ! ,.!1 5 ,.!3 $ ,.!% 12 ,.5! 2, ,.53 4esticide inhi(ition on an active enzyme has (een reported which caused enzyme activities to reduce. 7ollected data are presented for (oth without and with inhi(ition. a. (. Betermine the rate model with and without inhi(itor. Betermine the type of inhi(ition. w/o v >mol ?.min@ 5.3,8-,5 :.1,8-,5 $.$,8-,5 1.2%8-,5 1.5%8-,5 with v >mol ?.min@ !.:,8-,5 5.:,8-,5 3.1,8-,5 1.,!8-,5 1.258-,5

' >mol ?@ !.!,8-,5 5.,,8-,5 3.:,8-,5 1.358-,! 2.218-,! $.

Eate measurements were made on a food enzyme with and without presence of an inhi(itor with the following results. 2nhi(itor concentration is C2D = 2,u& w/o v" umol min $.2 12.$3 15.51 1:.%5 2,.,2 22.35 with v" umol min 5.35 $.13 %.3$ 11.,5 1!.53 15.:1

' u& 25 5, :5 1,, 15, 2,, a. (. c.

;se either lineweaver-=urk or 8adie-Foftstee plot to determine type of inhi(ition Betermine #m and Vo for the enzyme without inhi(ition as well as #!" the inhi(itor constant 1hat further tests are needed to improve the accuracy of the estimate of #!+ Fow would you calculate then+

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An e0periment on the thermal degradation of an enzyme was carried out and some of the results are (elow

.ime" s , 1,, 2,, 5,, 1,,, a. (. c.

*Activity G 357 1,, %2 $5 3: 55

*Activity G:57 1,, $5 :2 55 2,

4lot a graph of log activity vs. time and determine the inactivation rate constants at 357 and :57. Betermine the activation energy for the thermal degradation of the enzyme. 1hen eggs are pasteurized" alpha-amylase activity is often used as an indicator that the pasteurization process has (een satisfactorily completed. 'uggest a reason why alpha-amylase activity may (e used in this way.

10. .he e)uili(rium constant for the reaction '- 4 is 5. 'uppose we have a mi0ture of C'D = 201,-5& and C4D = !01,-5&. #m = !01,-5&" Vm=2 umol >?.min@" Vm=5 umol >?.min@ a. (. 1hich direction will the reaction proceed on addition of an appropriate enzyme+ At what initial velocity will the reaction start towards e)uili(rium+

11. 8stimate the k" the first order rate constant" for an enzyme preparation with a Vm = 5.3 umol ?.min under the given e0perimental condition" #m = 201,-3&

12. An enzyme was assayed at an initial su(strate concentration of 201,-5&. 2n 3 min" half of the su(strate had (een used. .he #m for su(strate is 501,-5&. a. 7alculate k" Vm" and concentration of product (y 15 min.

1!. An enzyme has a #m of 5.:01,-5&. 2f the Vm of the preparation is 22 umol ?.min" what velocity would (e o(served in the presence of 201,-5& su(strate and 501,-5& ofH a. (. c. 7ompetitive inhi(itor Ion competitive inhi(itor ;ncompetitive inhi(itor 2f #i in all cases is !01,-5&

15. .he effect of temperature on the hydrolysis of lactose (y a (eta-galactose is shown (elow a. 7alculate the activation energy ." ,7 2, Vm >umol mg protein.min@ 5.5

!, !5 5, 55 15. 'huler and kargi " pro(lem !.!

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