0 2 40 60 80 100 120 140 160 s (mg/t) @ Plot of VS indicates thal this is subshot 025 “2 eguatios a fs lice 0.15 4-0-49332 =n +1535£ 0.05 ° Vi 0.05 e. Bey fon 004-002 0 002 004 006 008 01 0.12 ine, He effect : us w is af T20 Ne = 0.049333 Ven = 220.3 mg) t-h aff eae f72 —* 0+ 0.019833 + 65355 C/km) = 2.09 4, =--/ im 2 @ Tass pe, Keb dtl gfeMoblern 213 Determine Ksx. from... Makinnin ree etron rate § which is detecained by sething GSI =O LS Tmae >A bm’ Kor fom plot, CSTnee 2 mgt Le, Kate af reaction at_S:70 vag| £2 a Stace’, Mn bibs bon effect rs observed at fis Suostrafe, we mest use /U-rt ejuclon U=_Vm fs] = Go3Yw) Ko’ + £5] (31.4 (0) Y= HHO maf thr aProblem ! Ale @ Pht Ns /s for ail cases te dekermiat the type 9 hi han . w . © 160 0 1126mM 4 [6195mM 0.15782 + 0.19404« = 9.15917 + 0.3098. y= 0.15343 + 0.40383% Nn a L . vs SS ee x 8 the Sonne, regardless off inhibbr, but aS XL inemasesy Thm clecreases Capparend), that iS an increased! vo lus &f Koyapp , Wsul ting in a Nelechon ia reaction rake. This (3 Chorackristic of Competitive Tnhibibo, @Dj Pablem — Bb6 Cogn). © Determine. Vm, Km, Ke Vin, Bm tan. be oktermined fom the T=.0. Case. | eto CW) +. 0.15782 + 0.19904 C '/s) Wm at Cls)2Q. + Wms 0.15782 i eee Vor 34 inh | i ‘| | —Vkm at (Wn): 07 01S7EAtOURIOICY, : 0.18 R= 0.19404 Km | Ke must. be clteranned fom the T>.O case ' Il z= paemy peers i ee i Ds Smt Bs 597+ 0.30% C15) Te haemt : (n= 18917 40,8098 (hema) no} i a. BORE = Tee: Fivegat Le GF om ! 0.3098 Koc | (teh %m4: CW) 0153434 0.40383C/5) 0 = 0/5343 10.4038 307 /kngge) ij Ou SR Abeer le ree. ~ 2.6 0.40383 Knegpj Hj i, —__— ain H | DetaGmt : AGS att hadmM eS + tabatt) Ar an are . a, Witte tat Ci. \ a Be ii bs 4 Az = _47/mM We would expect Mat. Kets the same at different - | acea trations. of in hibiter. The aliffecenas in the hwo | ialues.shocon here we dire._fp_epecimentel enor.alg ee eae aver 2s k (s,- oe ki=2.16 ilies: om /sec. Sb= lecong/L = my we 9? -a35,40,2=0 = 9.62 9y/fnF= 625 roll b. Ve MuSs =k (S,-Ss) Kets = er (0-42) we SOD (4-0.62) =0.076, - (o.2+4+6.62) wy} {orProblem 3.9 By inspection, Vmax = “a5. pmellmin , Km = 20 pel] Plot Vv -vereus Yits] (Eadie - Hofstee) where y-int. = Vn; X-int, = Vm] Km and Slope = -Kme 140 120 100 80 60 40 20 0 0 1 3 4 2 vi{S] Plotting the deta and using the data points far high eRe te coReeN ALON i a Vin = 130 parnol/min 5 Kin = 8.24 pmol i From the plot it is obvious that the data do not Fit into Michaelis-Menten Kinetics at low substrate Concentrations. Seme of the error may be atributed +o the method Since both axes Contain v. However, there are +wo possible explanations for the deviations from Michaelis - Menten Kinetics: () the enzyme is immobilized, thus diffusional effects became important 3 (i) there might be unspecific binding of substrate to the Enzyme thus requiring @ eritical Substrate. concentration for the reaction to follow Michaelis- Menten Kinetics.