Professional Documents
Culture Documents
Ass. 5
Ass. 5
BY
YAKINDRA PRASAD TIMILSENA (ID 111332, Asian Institute of Technology, Thailand)
Q.N. 4.7 Klebsiella aerogenes is produced from glycerol in aerobic culture with
ammonia as nitrogen source. The biomass contains 8% ash, 0.40g biomass is
produced for each g of glycerol consumed and no major metabolic products are
formed. What is the oxygen requirement for this culture in mass terms?
Solution:
MW of glycerol = 92
MW of Biomass (Klebsiella aerogenes), CH1.73O0.43N0.24 = 23.97/0.92 = 26.1
Degree of reduction for substrate i.e. glycerol (ᵞs) = 4.7
Degree of reduction for biomass (ᵞB) = 4x1+1x1.73-2x0.43-3x0.24 = 4.15
No. of carbon atoms in glycerol, C3H8O3 (w) = 3
Yield of biomass (Yxs) = 0.40 gg-1
MW substrate
Now, c = Yxs = 0.4x92/26.1 = 1.41 g mol biomass/g mol substrate
MWcells
And oxygen requirement (a) = ¼(𝑤 ∗ ᵞs − c ∗ᵞB) = ¼(3 ∗ 4.7 − 1.41 ∗ 4.15)= 2.1
MW of acetic acid = 60
MW of Biomass (methane bacteria), CH1.4O0.40N0.20 = 22.6
Degree of reduction for substrate i.e. acetic acid (ᵞs) = 4.0
Degree of reduction for biomass (ᵞB) = 4x1+1x1.4-2x0.40-3x0.20 = 4.0
No. of carbon atoms in acetic acid, CH3COOH (w) = 2
Degree of freedom of product (ᵞp) = 8
No. of carbon atoms in the product i.e. methane (j) = 1
N balance: z + bi = cᵟ + fm
Or, 0 + b*1 = c*0.20 + f*0
Or, c = b/0.2
Or, c = 5b .................... (i)
O balance: y + 2a + bh = cβ + 2d +e + fl
Or, 2 + 0 + b*0 = c*0.40 + 2*0.91 + e + f*0
Or, 2 = 0.40c + 1.82 + e
Or, 0.40c + e = 0.18
Or, 0.4*5b + e = 0.18 (from eq (i))
Or, 2b + e = 0.18 ........................ (iii)
H balance: x + bg = cα + 2e + fk
Or, 4 + b*3 = c*1.4 + 2e + f*4
Or, 4 + 3b = 1.4c + 2e + 4f
Solving (ii) and (iv) we get: 16b –2e = 0.36 ......... (v)
Solving eq (iii) and eq (v) we get: b = 0.036
From (i): c = 5* 0.036 = 0.18
From (iii): e = 0.18 – 2* 0.036
Or, e = 0.108
From (ii): f = 1.09 – 5*0.036
Or, f = 0.91
Question 4.9
a) Cellulomonus bacteria used as a single cell protein for human or animal food
are produced from glucose under anaerobic condition. All carbon in the substrate is
converted into biomass; ammonia is used as nitrogen source. The molecular
formula for biomass is CH1.56O0.54N0.16; the cells also contain 5% ash. How does
the yield of biomass from substrate in mass and molar terms compare with the
maximum possible biomass yield?
b) Another system for manufacture of SCP is Methylophilus methylotrophus. This
organism is produced aerobically from methanol with ammonia as nitrogen source.
The molecular formula for the biomass is CH1.68O0.36N0.22; the cells contain 6% ash.
1) How does the maximum yield of biomass compare with the above? What is the
main reason for the difference?
2) If the actual yield of biomass from methanol is 42% the thermodynamic
maximum, what is the oxygen demand?
Solution:
Part (a)
By elemental balance we have to find c,
C balance: w = c +d
Or, 6 = c + d
Or, c = 6 – d ....................... (i)
N balance: z + bi = cᵟ
Or, 0 + b*1 = c*0.16
Or, b = 0.16c .......................... (ii)
H balance: x + bg = cα + 2e
Or, 12 + b*3 = c*4 + 2*1.56
Or, 12 + 3b = 4c + 3.12 ....................... (v)
MW cells
Now, Biomass Yield (Yxs ) = c
MWsubstrate = 2.52 *25.73/180 = 0.36 g g-1
Now,
cmax= w*(ᵞs)/ (ᵞB) = 6*4/4 = 6 g mol of biomass / g mol of glucose
Part (b)
i) Methanol + O2 + ammonia CH1.68 O0.36 N0.22 + CO2 + H2 O
(Substrate) (nitrogen source) (Biomass)
Here,
Molecular weight of methanol (CH4O) = 32
Degree of reduction (ᵞs) =6
No. of carbon atoms in the substrate (w) = 1 (for methanol)
Molecular weight of biomass including ash = 22.52 / 0.94 = 23.95
Degree of reduction of biomass (ᵞB) = 4.3
By relation, Maximum yield of biomass (cmax) =1.4
ii) Actual yield of biomass from methanol (Yxs) = 0.42* thermodynamic maxm
= 0.42*(cmax/w)
= 0.42* (1.4/1) = 0.588 gg-1
QUESTION: 4.10
no O
C6H12O6 + NH3 2 = CH1.8O0.5N0.2 + CO2 + H2O + C2H6O
(Substrate) (Biomass) (Product)
N balance: z + bi = cᵟ + fm
Or, 0 + b*1 = 0.8*0.20 + f*0
Or, b = 0.16
Carbon balance: w = c + d + fj
Or, 6 = 0.8 + d + f*2
Or, d + 2f = 5.2 .................... (i)
O balance: y + 2a + bh = cβ + 2d +e + fl
Or, 6 = 0.8*0.50 + 2*d + e + f*1
Or, 2d + e + f = 5.6 ................ (ii)
From (ii) and (iii) we have 3f – e = 4.8
Or, e = 3f - 4.8
H balance: x + bg = cα + 2e + fk
Or, 12 + 0.16*3 = 0.8*4 + 2*e + f*6
Now,
Yield of biomass (Yxs) for z. mobilis = 0.05gg-1
Therefore, c = Yxs X Molecular weight of glucose / Molecular weight of cells
= 0.05 X 180 / 24.6
= 0.37 g mol of biomass / g mol of glucose
N balance: z + bi = cᵟ + fm
Or, 0 + b*1 = 0.37*0.20 + f*0
Or, b = 0.074
Carbon balance: w = c + d + fj
Or, 6 = 0.8 + d + f*2
Or, d + 2f = 5.2 .................... (i)
O balance: y + 2a + bh = cβ + 2d +e + fl
Or, 6 = 0.8*0.50 + 2*d + e + f*1
Or, 2d + e + f = 5.6 ................ (ii)
H balance: x + bg = cα + 2e + fk
Or, 12 + 0.074*3 = 0.8*4 + 2*e + f*6
The yield of ethanol from Z. mobilis is greater than that from S. Cerevisae by 0.39-0.38= 0.01gg-1
4.11 Detecting unknown Products
Given:
The biomass yield from the substrate (YXS) = 0.37g biomass/g glucose
Products = ?
Solution:
c( MWofcells )
Y XS
MWofsubstrate
Y ( MWofsubstrate) 0.37 180
c XS
( MWofcells ) 25.13
= 2.65 g mol of biomass/ g mol of substrate
We know
Hence other product is also synthesized beside increase in the biomass when glucose is
consumed.
MW of Substrate =
MW of Biomass (CH1.83O0.55N0.25) = 26.13
Final Concentration of biomass = 25g l-1
4.13 Oxygen demand for production of recombinant protein
Solution:
Given:
The biomass yield from the substrate (YXS) = 0.48g biomass/g glucose
For ammonia, i= 1
For glucose, w= 6
j =1
Molecular weight of glucose = 180
Molecular weight of biomass = 24.6 (for E. coli)
Degree of reduction for biomass (γB) = (4)(1) +(1)(1.5) +(-2)(0.5) +(-3)(0.2) = 3.9
δ=0.2, m= 0.25
c( MWofbiomass)
Y XS
MWofsubstrate
Y ( MWofsubstrate) 0.48 180
c XS 3.51
( MWofbiomass) 24.6
Case II
Product = 0
bi = cδ
b(1) = (3.51)(0.2)
b = 0.702
Hence when wild –type E. coli is used, 0.702g of ammonia is required per g of glucose used.
Oxygen demand is given by
Hence 2.53 g of oxygen is required per g of glucose consumed if wild type E. coli is used.
Acetic acid is produced from ethanol during growth of Acetobacter aceti, which has the
composition CH1.8O0.5N0.2 . Biomass yield from substrate is 0.14g g-1. Ammonia is used as
nitrogen source. How does growth in this culture affect oxygen demand for acetic acid
production?
Solution:
Oxygen demand = ?
Here,
For ethanol, w = 2
We know,
c( MWbiomass )
YXS
MWsubstrat e
c(24.6)
0.14
46
0.14 4.6
c 0.26
46
f ( MWproduct )
YPS
MWsubstrat e
f (60)
0.92
46
0.92 46
f 0.71
60