7-chlortetracycline is produced by high cell density cultivation of an engineered strain of

Streptomyces aureofaciens in fed-batch culture with the intermittent addition of fructose

solution to the culture medium.

The initial culture volume at quasi-steady state is V0 = 300 L, and fructose-containing

nutrient solution is added with a flow rate of F = 100 L/h.

Fructose concentration in the feed solution and initial cell concentration is S0 = 400 g/L
and X0 = 15 g DW /L, respectively. Assume that the organism follows Monod kinetics and
the kinetic and yield coefficients of the organism are given as µ m = 0.2 h-1, KS = 0.5 g/L,
and YX/S = 0.3 g DW/ g fructose

1. Determine the culture volume at quasi-steady state when t = 5 h (in L)

Culture volume is given as V = V0 + Ft

At t = 5 h, given V0 = 300 L and F = 100 L/h

V = V0 + Ft = 300 L + (100 L/h x 5 h) = 800 L

2. The dilution rate after 5 h of fed-batch operation is (in h -1)

D = F/V = 100/800 = 0.125 h-1
3. Determine the concentration of fructose at t = 5 h at a quasi-steady state (in g/L)
Under quasi-steady state conditions,

𝐷≈𝜇
According to Monod Kinetics,
𝜇 𝑆
𝜇=
𝐾 +𝑆
At quasi-steady state

𝜇= =D

. . /
𝑆= = = 0.833 g/L
. .
 4. Determine the total amount of cells at quasi-steady state when t = 5 h (in kg)

At quasi steady state and at high cell density, s<<si and

Also applying 𝐷 ≈ 𝜇 in the mass balance equation for biomass gives,

X = X + (Y S F)t

Xt0 = X0*V0

X = (15 g DW/L ∗ 300L) + 0.3 ∗ 400 ∗ 100 ∗5h

= 64.5 kg

5. Determine the concentration of cells at quasi-steady state when t = 5 h (in g/L)

Concentration of cells : X = Xt/V = 64.5 kg / 800 L = 80.625 g/L

6. If qP = 0.05 g product/g cells h and P0 = 0.1 g/L, determine the product

concentration in the vessel at t = 5 h (in g/L)

Based on product balance ,

𝑃𝑉 𝑉 𝐷𝑡
𝑃= + 𝑞 𝑋 ( + )𝑡
𝑉 𝑉 2
𝑃𝑉 𝑉 𝐷𝑡
𝑃= + 𝑞 Y S ( + )𝑡
𝑉 𝑉 2
𝑔 𝑝𝑟𝑜𝑑𝑢𝑐𝑡
0.1 𝑥 300𝐿
= 𝐿
800𝐿
𝑔 𝑝𝑟𝑜𝑑𝑢𝑐𝑡 g cells g fructose 300 𝐿 (0.125 h 𝑥 5 ℎ)
+ 0.05 𝑥 0.3 𝑥 400 + 5ℎ
𝑔𝑐𝑒𝑙𝑙𝑠 ℎ g fructose L 800 𝐿 2

= 20.66 g/L