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CHAPTER 9: AGITATION AND AERATION

NEW PROBLEM
A 10,000 liter (of liquid) bioreactor contains 5g/L of growing cells, QO2 = 20 mmol O2 / (g
cells.hr),DT = 2 m, Di = 1 m, (6 - blade turbine agitator) x 3 blades. For 1 liquid volume per minute
aeration rate (air), can the OTR(oxygen transfer rate) = OUR(oxygen uptake rate) for N = 100 rpm?
GIVEN:
VL=10,000L
QO2 = 20 mmol O2 / (g cells.hr)
DT = 2 m
Di = 1 m
(6 - blade turbine agitator) x 3 blades
N = 100 rpm
REQD: Is OTR = OUR?
SOLN:
Re = Reynold's Number= LNDi2 / L
L= 1000 kg/m3

L =10-3 N.s/m2

Re = (1000 kg/m3)(100/60rps)(1m)2(1N/kg.m/s2) / 10-3 N.s/m2

Re = 1.67x10-6

*Np = 4(from Blanch and Clark Pmo Correlation)

Pmo = 4 (LN3Di5) for 1 impeller

Pmo = 4(1000 kg/m3) (100/60rps)3(1m)5

Pmo = 1.852x104kg.m2.s2/s (watts) x 3(impellers)

Pmo = 5.62x104 watts = 74.5hP

Pm: NA (aeration no.) = Qa / NDi3

NA = (10,000Lpm)(10-3m3/L) / (100min-1)(1m)3

NA = 0.10

*Pm/Pmo = 0.42(from Blanch and Clark NA Correlation)

Pm = (0.42)(5.62x104 Watts)
Pm = 2.335x104 Watts = 31.3hP

kLa (mmol O2 / ( hr.atm) = 0.60(Pm/VL(hP/103 liters))0.4 (Vs)0.5 (N,rpm)0.5

Pm/VL =31.3hP/(10)(103liters) = 3.13hP/103liters



Vs =104 Lpm(103 cm3/L)/ 4 (2m)2(10cm/m)2 = 318.3 cm/min

kLa = 0.60(3.13)0.4 (318.3)0.5 (200)0.5

kLa = 169 (mmol O2 / (hr .atm)

OUR = X qO2 = (5g cells/ L)(20 mmol O2/g cells.hr)

OUR = 100 mmol O2/L.hr

OTR = kLa(PO2 - P*)

P * for CL = 1 mg O2/liter = HO2 CL

= (0.21 atm/(8mg O2/liter))(1 mg O2/liter)

=0.0263 atm

OTR = 169 mmoles O2/liter.hr.atm (0.21 0.0263) atm

OTR = 31.05 mmol O2/liter.hr

*Since OUR > OTR, we must modify the bioreactor operation in order to bring them into balance

increase N

use pure O2 rather than air.

http://www.chem.mtu.edu/~drshonna/cm4710f07/lectures/chapter10.pdf
CHAPTER 2: ENZYME KINETICS
NEW PROBLEM

The following data were obtained for the reaction A B, catalyzed by the enzyme Aase. The
reaction volume was 1mL and the stock concentration of A was 5.0mM. Seven separate reactions were
examined, each containing a different amount of A. The reactions were initiated by adding 2.0L of a
10M solution of Aase. After 5 minutes, the amount of B was measured.

Reaction Volume of A Amount of B present


added(L) at 5 minutes (nmoles)
1 8 26
2 10 29
3 15 39
4 20 43
5 40 56
6 60 62
7 100 71

(a) Calculate the initial velocity of each reaction (in units of M.min-1)
(b) Determine the KM and Vmax of Aase from a Lineweaver-Burk plot.
(c) Calculate kcat.

SOLN:

(a) o = (26nmol/5min) / (1.0mL) x (103 mL/L) x (.001 mol/1nmol) = 5.2 M.min-1

Reaction o(M.min-1)
1 5.2
2 5.8
3 7.8
4 8.6
5 11.2
6 12.4
7 14.2

(b) Calculate [S] for each reaction


[A] = (.008mL)(5mM)(1mL) x (1000 M/1 mM) = 40 M

Reaction [S] M (x) 1/[S] M-1 (M.min-1) (y) 1/ (min-1/ M)


1 40 0.025 5.2 0.192
2 50 0.02 5.8 0.172
3 75 0.0133 7.8 0.128
4 100 0.010 8.6 0.0116
5 200 0.005 11.2 0.089
6 300 0.0033 12.4 0.081
7 500 0.002 14.2 0.070

y-int = 1/vmax = 0.06

Vmax = 16 M/min

x-int = -1/KM = 1/0.012

KM = 83 M

(c) Calculate [E]T = (0.002mL)(10 M Aase) / 1mL = 0.02 M

Kcat = Vmax / [E]T = (16 M/min) / 0.02 M

Kcat= 800 /min

https://ww2.chemistry.gatech.edu/~lw26/bCourse_Information/3511/stud_comp/chap12_17.pdf