Example 6.1. A strain of mold was grown in a batch culture on glucose and the following data were ob- tained. Cell Glucose Time concentration concentration wy (et) C2) 0 1 100 9 245 7 16 5 904 B 105 769 30 2 48.1 34 33 206 36 315) 938 40 4l 063 a Calculate the maximum net specific growth rate, bb. Calculate the apparent growth yield What maximum cell concentration could one expect if 150 g of glucose were used the same size inoculum? Solution A plot of In X versus f yields a slope of 0.1 h. LAX 41-125 ‘AS 0625-100 ©) Xue = Xo + PS, = 1.25+ 0.4150) = 60.25 g cells Example 6.2. Logistie Equation Ethanol formation from glucose is accomplished in a batch culture of Saccharomyces cere- visiae, and the following data were obtained. Time (h) Glucose (S), gL. Biomass (X), g/L. Ethanol (P), g/l. by y= 0.4 g cells/g substrate ° 100 0s 00 2 95 10 25 5 85 2 713 0 38 48 200 15 30 2) uo 20 2 96 43.0 25 5 lo4 47s 0 2 107 490 By fitting the biomass data to the logistic equation, determine the carrying-capucity coefficient k b. Determine yield coefficients ¥ps and YsSolution - 1) Equation 6.95 can be rewritten as: or ptf; xX at where X is average biomass concentration during Af, and X, is about 10.8 g/L, since growth is almost complete at 30 h. Thus’ ari) Kig/l) 1 axsardh) (ie kor) 2 075 om 0931 036 3 138 0236 886 028 5 Mas 0186 0681 023 5 os 03 ote 022 5 ses oo 0200 022 5 1000 core om 022 5105s omns7 i) 02s A value of k= 0.24 hr! would describe most of the data, although it would slightly under- estimate the initial growth rate. Another approach would be to take the log of the above ‘equation to gi Lax LB tog k+tog{ 1-2 oa eg x. ‘and (o fit the data to this equation and estimate & from the intercept. In this case k would be about 0.25 h, b) The yields are estimated directly from the data as: The above estimate of Yys is only approximate, as maintenance effects and endogenous, metabolism have been neglected.Example 64. A new strain of yeast is being considered for biomass production. The following data were oblained using a chemostat. An influent substrate concentration of 800 mg/l and an excess of oxygen were used at a pH of 5.5 and T= 35°C. Using the following data, calculate 41, K,, Yl, ky, and m,, assuming gg, Ue SI(K, +S) — ky Carbon substrats Coll Dilution rate!) concentration (mg/D)__ concentration (mg/D) o1 167 366 02 335 407 03 504 408 4 101 404 os 169, a7 06 298 299 07 702 30 Solution The first step is to plot UY ¥% versus 1/D. ¥ is calculated from X/(Sp— S$). The ine tercept is I/Y'¥, = 1.58 or ¥¥, = 0.633 g X/z S. The slope is 0.06 g Sig X-h, which is the value form, Recall m, = k,/¥ i; then ky =m, ¥s = 0.06 g Sig X-h- 0.633 2 X/g 8 =0.038 h-! For the second step, recall that @ Dik, tin ie S o ‘We now plot LAD + 4,) versus US. The intercept is 1.25 b or p= 0.8 br! The slope is 100 ‘hi(mgM). Thus, Kt, 100 or X, = 80 mg/l Example 65. ‘The specific growth rte for inhibited growth ina chemostat i given by the following equation: Mm May Ke S+IK IR, @ where Sy- 10g KI gh 1-005en Yi Xy=0 K=001g1 y= OSh1 hy a Determine X and 5 asa function of D when 1=0. b. With inhibitor added to a chemostat, determine the effluent substrate concentration and X as a function of D. «. Determine the cell productivity, DX, as a function of dilution rate,Solution a) s-KD___D i,-D 035-D 5 _s-ol D_) X= ¥eis(S 9) 0.1 10 — 15-p) b) In the presence of inhibitor X= MS, -9)= 0410 - 2 ) 05-D 6D ©) Pr, = DX = DYys(Sp 5)=0.10{ 10 5) Example 9.1 Ina chemostat with cell recycle, as shown in Fig. 9.1, the feed flow rate and culture volumes are F= 100 mivh and V= 1000 ml, respectively. The system is operated under glucose limita tion, and the yield coefficient, Pi, is 0.5 gdw cells/g substrate. Glucose concentration in the Figure 9.2. Comparison of biomass con- S329 centrations and ouput rates in steady satos se cf cherostat cles wih and witout ey- E815 ‘Sk. Symbols X= homnsscomentation i Es ‘hemostat withou recycle; Xp = biomass § B10 encentation in chemostat clare wth = 83 Cycle; Ry = biomass output rate per unit vol- 8 fos {ne without eyele; biomass carat & § ‘rate of chemostat with recycle; fijg= 1.00 EB fr: $,=20 gh Ky =0.010 gf Fys= BS GT OF OS OS TO TE TS TS TS ZO OS pis concen air, C~ 20; and as 05. Ditution rate (Ho!) recycle rat,feed is Sp = 10 g glucose/l. The kinetic constants of the organisms are t1,,= 0.2 h!, K,= 1g ¢glucose/l. The value of C is 1.5, and the recycle ratio is 0: = 0.7. The system is at steady state a Find the substrate concentration in the recycle stream (S). 1b Find the specific growth rate (\1qq)of the organisms. c. Find the cell (biomass) concentration in the recycle stream Find the cell concentration in the centrifuge effluent (X)). Solution Using ¢q. 9.9, we determine ftp Hae = [+ Cl ~ C)D = [1 + (1 = 1.5)0.710.1) = 0.065 ht Then 5 Kina 10069) _ 9.48 9/1 HaHa 0.2— 0.065 DS Ss O.G0- 0.48) Me 73g/ A biomass balance around the concentrator yields (1+ 0)K, = 00K, +X, X= (1+ aX, = (1.7)(7.3) — (0.7)(1.5)(7.3) =48 2 Example 9.3 In a fed-batch culture operating with intermittent addition of glucose solution, values of the following parameters are given at time f= 2 h, when the system is at quasi-steady state. V=1000 ml = 300 mith S, = 100 g glucose/t =03h"4 K,-Olgglucose/t Yr, 05 gdw cells! glucose X30 g a. Find Vp (the initial volume of the culture) b, Determine the concentration of growth-limiting substrate in the vessel at quasi-steady state. ¢.Determine the concentration and total amount of biomass in the vessel at ¢ = 2h (at quasi-steady state). 4. I gp = 0.2 g product/g cells, Py = 0, determine the concentration of product in the vessel at 2h.Solution a VeVy+ Ft Vg = 1000 ~ 200(2) = 600 ml b. D=FPV=02h' KD__ (0.102) KD _ =02 g glucose/I i,-D 03-02 S2u0o" ce. X= XS + PYM Sy = 30+ (0.2)(0.5(100)(2) = 50 g¢ Mog gx (Sig Dt HPht aXe Be Str =04102y20{ 2, 222 > =16 g/l Example 9.4 Glucose is converted to ethanol by immobilized S. cerevisiae cells entrapped in Ca-alginate beads in a packed column. The specific rate of ethanol production is gp = 0.2 g ethanol’ cell - h, and the average dry-weight cell concentration in the bed is X= 25 g/l bed. Assume that growth is negligible (i.c., almost all glucose is converted to ethanol) and the bead size is sufficiently small that 7 = 1. The feed flow rate is F = 400 Wh, and glucose concentration in the feed is Sp; = 100 g glucose/l. The diameter of the column is I m, and the product yield co- efficient is ¥p5 ~ 0.49 g ethanol/g glucose. a. Write a material balance on the glucose concentration over a differential height of the col- umn and integrate it to determine S = S(z) at steady state. b. Determine the column height for 98% glucose conversion at the exit of the column. ¢. Determine the ethanol concentration in the effluent.Solution, a, A material balance on the glucose concentration over a differential height of the column vyields =r ds, = 4X gy = 10% 4 ae Y, Tos Integration yields S. zo" “Ff as, Raf dz Ns, Yas This equation differs from the form of eq. 9.72 because Sy, is high and the reaction rate is effectively zero order. b. Sy= 0.02(100) = 2 g glucose/1. Substituting the given values into the above equation yields (0.225) (nA) LOY 10)" 049— 400 H=49dm=49 m & P= Ypys (Soy — So) = 0.4998) = 48 gil. Example 9.2 Derive the relationship between the overall mass-transfer coefficient for liquid phase A; and the individual mass-transfer coefficients, kr and ko. How can this relationship be simplified for sparingly soluble gases? Solution: According tolEqs. (9.7) and (9.10),RC, -C)=K,(C, -C;) @.L) ‘Therefore, by rearranging Eq. (9.11) ALi G-G Ky GO 12) Since FY(Cr, — C1) = hg (Ce — Co.) (9.13) By substituting Eq. 9.13) to Eq. (9.12), we obtain Pili gq-q 1. (0.14) Kp hy ke Cg—Co, hy gM which is the relationship between K;, ky, and kg. M is the slope of the line connecting (Cr, Cz) and (Cz. Ca) as shown in Figure 9.4. For sparingly soluble gases, the slope of the equilibrium curve is very steep: therefore, Mis much greater than 1 and from Ea. (9.14) Kk, @.15) Similarly, for the gas-phase mass-transfer coefficient, K,=ke 9.16) Example 9.4 Estimate the mass-transfer coefficient for the oxygen dissolution in water 25°C ina mixing vessel equipped with flat-blade disk turbine and sparger by using Calderbank and Moo-Young's correlations.Solution: ‘The diffusivity of the oxygen in water 25°C is 2.5x10* m’/s (Example .1). The viscosity and density of water at 25°C is 8.904x10~“ kg/m s (CRC Handbook of Chemistry and Physics. p. F-38, 1983) and 997.08 ke/m3 (Perry and Chilton, p. 3-71, 1973), respectively. The density of air can be calculated from the ideal gas law. _ PM _1.01325x105(29) Pos "Rp 8314107298) Therefore the Schmidt number, Ny =H = 8.90410" 59579 PD 997.08(25x10") Substituting in Eq. (9.27) for small bubbles, =1.186kg/m* a. 3 y= 0310572)" [roe 186)(8.904<10 dean (997.08) =127x10" mis Substituting in Eq. (9.32) for large bubbles, a 13 042057)" 2 08-1. 186)(8.904x10 9. S| (997.08) =458x10" ms Therefore, for the air-water system, Eqs. (9.27) and (9.32) predict that the mass-transfer coefiicients for small and large bubbles are 1.27x10* and 4.58x107 ms, respectively. which are independent of power consumption and gas-flow rate.Solution: The diffusivity of the oxygen in water 25°C is 2.5x10° m’/s Example 1). The viscosity and density of water at 25°C is 8.904x10™ kg/m s (CRC Handbook of Chemistry and Physics, p. F-38, 1983) and 997.08 kg/m3 Perry and Chilton, p. 3-71, 1973), respectively. The density of air can be calculated from the ideal gas law, = PM _1.01325210°29) Pas RT 8314x1098) ‘Therefore the Schmidt number, w 8.90410 ~ pDyz —997.08(2.5x10°) Substituting in Eq. (9.27) for small bubbles, =1.186kg/m’* N, =3572 ™ 3 i, = 031057 | 08 ~1.186)(8.904 x10 roan (997.08) =1.27%10+ mvs Substituting in Eq. (9.32) for large bubbles, 0.2057 zy [eons 186(8.904 x10) 9.81) |" “_ (997.08)" =458x10" ms Therefore. for the air-water system. Eqs. (9.27) and (9.32) predict that the mass-transfer coefficients for small and large bubbles are 1.27x10~ and 4.58x10* mys, respectively, which are independent of power consumption and gas-flow rate.of)" ose)" (@628 “2 8.931077 981 000416 (29036 P,=687W b. Gas hold-up: The interfacial tension for the air-water interface is 07197 kp’? (CRC Handbook of Chemistry and Puysics . . F-33, 1983). The volume of the the dispersion is v=21.20°0.2)=1.43m" 4 The superficial gas velocity is y= 4Q__ (0.00416) os = = 000356mis Substituting these values into Eq. (9.51) gives oonesei| 4.2:16x10" ‘ou 43)°*997. or] 00356 0.265 0.07197" 0265 The solution of the preceding equation for H gives H=0023 . Sauter-mean diameter: In Eq. (9.45) [__oons7 Pa 413 ar aR) *007 | = 0.00366m=39mm 0.023" +9.0x10*4. Interfacial area a: In Eq. (9.37) SH _6 0.023) _ 37 759-1 Dy 0.00366 . Volumetric mass-transfer coefficient: Since the average size of bubbles is 4 mm, we should use Eq. (9.32). Then, from Example 4 k, =4.58x107 avs Therefore, k,a= 4.58x10*(37.7)=0.017s* The preceding estimated values compare well with those experimental values. The percent errors as defined in Example .1 are —1 4 percent for ‘the power consumption, 15 percent for the gas hold-up, and -21.7 ‘percent for the volumetric mass-transfer coefficient.Example 9.6 To measure ky a, a fermenter was filled with 10 L of 0.5 M sodium sulfite solution containing 0.003 M Cu™ ion and the air sparger was tuned on. After exactly 10 minutes, the air flow was stopped and a 10 mL sample was taken and titrated. The concentration of the sodium sulfite in the sample was found to be 0.21 mol/L. The experiment was carried ont at 25°C and 1 atm Calculate the oxygen uptake and kya. Solution: ‘The amount of sodium sulfite reacted for 10 minutes is 0.5-0.21=0.29molL According to the stoichiometric relation, Eq. (0.59) the amount of oxygen requited to react 0.29 molL is 0.29" =0.145movL ‘Therefore, the oxygen uptake is 32g 0, /mol (0.145mole 0; /L)| =i |=7.7310° g/Ls The solubility of oxygen in equilibrium with air can be estimated by Eq. (9.58) as Po, _ (latm)(0.209mol O, /mot air) Ho) (3atm Limol)(imol/32g) =8.4310* gL. Therefore, the value of kya is, according to the Eq. (9.57). tan lelY _ _173x10% "C=C, ~— @43x10° gL —0) = 09178 9