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INTRODUCTION

Cell growth and product formation are complex processes reflecting the overall kinetics

and stoichiometry of the thousands of intracellular reactions that can be observed within a cell.

Also, we may need to know how close to its thermodynamic limit a system is operating.

Thermodynamic limit –the limit for a large number N of particles where the volume is

taken to grow in proportion with the number of particles.

If a system is close to its thermodynamic limit, it would be unwise to improve production

through mutation or genetic engineering. When cells growth occurs, cells are a product of reaction

and must be represented in reaction equation. Although the cell is complex, the stoichiometry of

conversion of substrates into products and cellular materials is often represented by a simple

pseudo-chemical equation. When cell growth occurs, cells are a product of reaction and must be

represented in the reaction equation.

OTHER DEFINITIONS

𝑿⁄𝑺 ) – maximum yield of cell mass per unit mass of

substrate consumed when no maintenance is considered.

ATP Yield Coefficient (𝒀𝑿⁄𝑨𝑻𝑷 ) – represents the amount of biomass synthesized per mole

of ATP generated.

Relationship between the two: (𝑌𝑋⁄𝑆 = 𝑌𝑋⁄𝐴𝑇𝑃 𝑁) as N = moles ATP/mol substrate

Regularities

𝑌𝑋𝑀⁄𝑆 ≥ 10.5 g dry wt/mol ATP

26.95 kcal/g equivalent of available electrons transferred to oxygen (coefficient of variation of

4%)

4.291 g equivalent of available electrons per quantity of biomass containing 1 g atom carbon

0.462 g carbon in biomass per gram of dry biomass

𝑌𝑋⁄𝑒 − = 3.14 ± 0.11 g dry wt/g equivalent of electrons

Growth – orderly increased of all chemical components

Balaced Growth – growth during which a doubling of the biomass is accompanied by a doubling

of all measurable properties of the population such as protein, DNA, RNA and intracellular water.

Cell growth can be determined by:

- Cell number

- Cell mass

- Cell activity

MEASUREMENT OF CELL NUMBER

Microscopic Counts - the number of cells in a population can be measured under a microscope

by counting cells placed in special counting chambers.

Two types of Chambers:

1. HEMOCYTOMETER – blood cell counting chamber for use with microorganisms of 3

µm in diameter or larger

2. PETROFF-HAUSSER COUNTING CHAMBER – for used primarily with bacteria

Direct-Counting Method

ADVANTAGES DISADVANTAGES

1. Minimal equipment is required 1. Dead cells cannot usually be

2. Results are obtained rapidly distinguished from live cells

3. Morphological characteristics of the 2. The method is not suitable for cell

organisms can be observed suspensions of low density

3. Small cells are difficult to see under the

microscope and can be missed when

counting

4. The actual counting procedure is

tiresome and may cause considerable

eyestrain

5. It is not suitable for highly flocculating

cells such as mycelium

Viable cell – one that is able to divide and form a colony

Two ways of performing plate count

1. Spread Plate Method – a volume of no larger than 0.1 ml is spread over the agar surface

2. Pour Plate Method – sample is mixed with melted agar and poured into a sterile plate

Coulter Counter

- To avoid tedium of direct microscopic counting

- Using this, cell size can also be measured

- Cannot ditinguish between cells and any impure particles

- Difficult to use with microorganisms in chains and is useless with mycelial organisms

MEASUREMENT OF CELL MASS

Cell Dry Weight

- can be measured directly by taking an aliquot of cell suspension and centrifuging it

- most direct approach for the quantitative measurement of cell mass and is probably the

most reliable and reproducible

- can only be used with dense cell suspensions

Turbidity

- based on the fact that small particles scatter light proportionally, within certain limits,

to their concentration

- when a beam of light is passed through a suspension of organisms, the reduction in the

amount of light transmitted as a consequence of scattering is thus a measure of cell

density. Such measurements are usually made in spectrophotometer, which reads in

Absorbency (A) units.

Absorbency – logarithm of the ratio of intensity of light striking the suspension (Io) to that

transmitted by the suspension (I)

𝐼𝑜

A = 𝑙𝑜𝑔 𝐼

Indirect Methods

The indirect methods for measuring cell mass are based on the overallstoichiometry for

growth and product formation, which may be written in the general form:

The change of the cell mass can be monitored indirectly by measuring the following:1

1. Nutrient Consumption

2. Product Formation

3. Cell Components

4. Heat Evolution

5. Viscosity

Temperature - important factor affecting the performance of cells

Groups of Organisms

1. Psychrophiles- (Topt < 20°C)

2. Mesophiles - (Topt = from 20° to 50°C)

3. Thermophiles - (Topt > 50° C)

As the temperature is increased toward optimal growth temperature, the growth rate

approximately doubles for every 10°C increase in temperature. Above the optimal temperature

range, the growth rate decreases and thermal death may occur.

Temperature also affects product formation. However, the temperature optimum for growth

and product formation may be different. The yield coefficient is also affected by temperature. In

some cases, such as single-cell protein production, temperature optimization to maximize the yield

coefficient is critical. When temperature is increased above the optimum temperature, the

maintenance requirements of cells increase resulting in a decrease in the yield coefficient.

Temperature also may affect the rate-limiting step in a fermentation process. At high

temperatures, the rate of bioreaction might become higher than the diffusion rate, and diffusion

would then become the rate-limiting step.

Hydrogen-ion concentration (pH) affects the activity of enzymes and therefore the microbial

growth rate. The optimal pH for growth may be different from that for product formation.

Generally, the acceptable pH range varies about the optimum by ±1 to 2 pH units. Different

organisms have different pH optima: the pH optimum for many bacteria ranges from pH = 3 to 8;

for yeast, pH = 3 to 6; for molds, pH = 3 to 7; for plant cells, pH = 5 to 6; and for animal cells, pH

= 6.5 to 7.5. Many organisms have mechanisms to maintain intracellular pH at a relatively constant

level in the presence of fluctuations in environmental pH. When pH differs from the optimal value,

the maintenance-energy requirements increase. One consequence of different pH optima is that the

pH of the medium can be used to select one organism over another.

Dissolved oxygen (DO)- is an important substrate in aerobic fermentations and may be a

limiting substrate, since oxygen gas is sparingly soluble in water. At high cell concentrations, the

rate of oxygen consumption may exceed the rate of oxygen supply, leading to oxygen limitations.

When oxygen is the rate-limiting factor, specific growth rate varies with dissolved-oxygen

concentration according to saturation kinetics; below a critical concentration, growth or respiration

approaches a first-order rate dependence on the dissolved-oxygen concentration.

Above a critical oxygen concentration, the growth rate becomes independent of the dissolved-

oxygen concentration. Oxygen is a growth-rate-limiting factor when the DO level is below the

critical DO concentration.

Dissolved carbon dioxide (DCO2) concentration may have a profound effect on

performance of organisms. Very high DCO2 concentrations may be toxic to some cells. On the

other hand, cells require a certain DCO2 level for proper metabolic functions. The dissolved carbon

dioxide concentration can be controlled by changing the CO2 content of the air supply and the

agitation speed.

High substrate concentrations that are significantly above stoichiometric requirements are

inhibitory to cellular functions. Inhibitory levels of substrates vary depending on the type of cells

and substrate.

STOICHIOMETRIC CALCULATIONS

A material balance on biological reactions can easily be written when the compositions of

substrates, products, and cellular material are known. The law of conservation of mass has been

used to determined unknown quantities entering or leaving bioprocess. Usually, electron–proton

balances are required in addition to elemental balances to determine the stoichiometric coefficients

in bioreactions. All carbon, hydrogen, oxygen, nitrogen and other elements consumed during

growth are incorporated into new cells or excreted as products. Confining to those compounds

taken up or produced in significant quantity, if the only extracellular products formed are 𝐶𝑂2 and

𝐻2 𝑂, we can write the following equation for aerobic cell growth:

𝑏𝐻𝑔 𝑂ℎ 𝑁𝑖 is the chemical formula of the nitrogen source

𝑐𝐶𝐻𝛼 𝑂𝛽 𝑁𝛿 is the chemical ‘formula’ for the dry biomass

a, b, c, d, and e are stoichiometric coefficient

The figure above shows the macroscopic view of metabolism; it ignores the detailed

structure of the system and consider only those components which has net interchange in the

environment, including ATP and NADH. Vitamins and minerals taken up during metabolism is

neglected as it is consumed in very little amount and does not contribute to the stoichiometry and

energetics of reaction. Though this macroscopic view is simple in its approach, it provides a

powerful tool for thermodynamic analysis.

Table 1 – Elemental Composition for Various Microorganisms

Bacteria tend to have slightly higher nitrogen contents (11-14%) than fungi (6.3-9.0%). For

particular species, cell composition depends on the culture conditions and substrate utilized.

However, the results are remarkably similar for different cells and condition; 𝐶𝐻1.8 𝑂0.5 𝑁0.2 can be

used as general formula when composition analysis is not available. The ‘average molecular

weight’ of the biomass feed is 24.6, although 5-10% residual ash is often added into account for

those elements not included in the formula. Coefficients a, b, c, d, and e are obtained and evaluated

using normal procedures for balancing equations and is govern by the set of equations below.

C Balance: w=c+d

H Balance: x + bg = cα + 2e

O Balance: y + 2a + bh = cβ + 2d + e

N Balance: z + bi = cδ

Another additional information is needed to complete the five unknowns in the equation,

and is represented by the parameter respiratory quotient (RQ):

𝑚𝑜𝑙𝑒𝑠 𝐶𝑂2 𝑝𝑟𝑜𝑑𝑢𝑐𝑒𝑑 𝑑

𝑅𝑄 = =

𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑂2 𝑐𝑜𝑛𝑠𝑢𝑚𝑒𝑑 𝑎

EXAMPLE:

Production of single-cell protein from hexadecane is described by the following reaction equation:

C16H34 + aO2 + bNH3 cCHl.66O0.27N0.20 + dCO2 + eH2O

where CHl.66O0.27N0.20 represents the biomass. If RQ= 0.43, determine the stoichiometric

coefficients.

Solution:

(1) C balance: 16 = c + d

(2) H balance: 34 + 3b = 1.66c + 2e

(3) O balance: 2a = 0.27c + 2d + e

(4) N balance: b = 0.20c

(5) RQ: 0.43 = d/a.

We must solve this set of simultaneous equations. Solution can be achieved in many different

ways; usually it is a good idea to express each variable as a function of only one other variable, b

is already written simply as a function of c in (4); let us try expressing the other variables solely in

terms of c. From C balance,

d = 16 – c

From the RQ,

d

a= =2.326d

0.43

Combining the new formed equations gives an expression for a in terms of c only:

a = 2.326(16 - c)

a =37.22 - 2.326c

Substituting (4) into (2) gives:

34 + 3(0.20c) = 1.66c + 2e

34 = 1.06c + 2e

e = 17-0.53c

Substituting (8), (6) and (9) into (3) gives:

2(37.22 - 2.326c) = 0.27c + 2(16 - c) + (17 - 0.53c)

25.44 = 2.39c

c = 10.64

Using this result for c in (8), (4), (6) and (9) gives:

a = 12.48

b = 2.13

d = 5.37

e = 11.36

Check that these coefficient values satisfy Eqs (1)-(5). The complete reaction equation is:

C16H34 + 12.48O2 + 2.13NH3 10.64CHl.66O0.27N0.20 + 5.37CO2 + 11.36H2O

DEGREE OF REDUCTION

Elemental balances provide no insight into the energetics of the reaction. Consequently,

the concept of degree of reduction has been developed and used for proton–electron balances in

bioreactions. Available electrons refer to the number or electrons available for transfer to oxygen

in combustion of a substance to 𝐶𝑂2, 𝐻2 𝑂 and nitrogen-containing compounds.

Degree of reduction γ – number of equivalents of available electrons per gram

atom C

The degree of reduction of any element in a compound is equal to the valence of this

element. Therefore, for substrate 𝐶𝑤 𝐻𝑥 𝑂𝑦 𝑁𝑧 , the number of available atoms is 4w + x – 2y – 3z

and the degree of reduction for the substrate, 𝛄𝑆 , is (4w + x – 2y – 3z)/w. Degree of reduction for

𝐶𝑂2, 𝐻2 𝑂, and 𝑁𝐻3 is zero. The following are examples of how to calculate the degree of reduction

for substrates.

Methane (𝐶𝐻4 ): 4w + x – 2y – 3, 1(4) + 4(1) = 8 γ = 8/1 = 8

Glucose (𝐶6 𝐻12 𝑂6): 4w + x – 2y – 3 6(4) + 12(1) + 6(-2) = 24 γ = 24/6 = 4

Ethanol (𝐶2 𝐻5 𝑂𝐻): 4w + x – 2y – 3 2(4) + 6(1) + 1(-2) = 12 γ = 12/2 = 6

The degrees of reduction of biomass is 𝛾𝑏 = 4𝑤 + 𝛼 − 2𝛽 − 3𝛿.

In a balanced growth equation, number of available electrons is conserved by the virtue

of the fact that amounts of each chemical element are conserved. The available-electron balance

equation, as ammonia as nitrogen source, is:

𝑤γ𝑠 − 4𝑎 = 𝑐γ𝐵

where: γ𝑠 and γ𝐵 are the degrees of reduction for substrate and product respectively.

The available-electron balance is independent of the complete set of the elemental

balances; if the stoichiometric equation is balanced in terms of each element including H and O,

the electron balance is implicitly satisfied.

Table 2 - Degree of Reduction and Weight of One Carbon Equivalent of One Mole

of Some Substrates and Biomass

BIOMASS YIELD

As cells grow there is, as a general approximation, a linear relationship between the

amount of biomass produced and the amount of substrate consumed. This relationship is

expressed quantitively using biomass yield, 𝒀𝑿𝑺 .

𝑔 𝑐𝑒𝑙𝑙𝑠 𝑝𝑟𝑜𝑑𝑢𝑐𝑒𝑑

𝒀𝑿𝑺 =

𝑔 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒 𝑐𝑜𝑛𝑠𝑢𝑚𝑒𝑑

Large number of factors influences biomass yield, including medium composition, nature

of carbon and nitrogen sources, pH and temperature. Biomass is greater in aerobic than in

aerobic cultures; choice of electron acceptor e.g 𝑂2, nitrate or sulfate, can also have a significant

effect.

When 𝒀𝑿𝑺 is constant throughout growth, its experimentally determined value can be

used to determine the stoichiometric coefficient c expressed in terms of:

𝑐(𝑀𝑊𝑐𝑒𝑙𝑙𝑠)

𝒀𝑿𝑺 =

𝑀𝑊 𝑜𝑓 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒

where MW is molecular weight

‘MW cells’ means biomass formula weight plus any residual ash

However, before applying measured values of 𝒀𝑿𝑺 to evaluate c, we must be sure that the

experimental culture system is well represented by the stoichiometric equation. For example, we

must be sure that substrate is not used to synthesize extracellular products other than 𝐶𝑂2 and 𝐻2 𝑂.

One complication with real cultures is that some fraction of substrate consumed is always used for

maintenance activities such as maintenance of membrane potential and internal pH, turnover of

cellular components and cell motility. These metabolic functions require substrate but do not

necessarily produce cell biomass, 𝐶𝑂2 and 𝐻2 𝑂. For the time being, we will assume that available

values for biomass yield reflect substrate consumption for growth only.

EXAMPLE:

Assume that experimental measurements for a certain organism have shown that cells can convert

two-thirds (wt/wt) of the substrate carbon (alkane or glucose) to biomass.

a. Calculate the stoichiometric coefficients for the following biological reactions:

Hexadecane: C16H34 + aO2 + bNH3 c(C4.4H7.3N0.86O1.2) + dH2O + e CO2

Glucose: C6H12O6 + aO2 + bNH3 c(C4.4H7.3N0.86O1.2) + dH2O + e CO2

b. Calculate the yield coefficients YX/S (g dw cell/g substrate), YX/O2 (g dw cell/g O2) for both

reactions. Comment on the differences.

Solution

a. For hexadecane,

amount of carbon in 1 mole of substrate = 16(12) = 192 g

amount of carbon converted to biomass = 192(2/3) = 128 g

Then,

128 = c(4.4)(12)

c = 2.42.

amount of carbon converted to CO2 = 192 - 128 = 64 g

64 = e (12)

e = 5.33

The nitrogen balance yields

14b = c(0.86)(14)

b = (2.42)(0.86)

b = 2.085

The hydrogen balance is

34(1) + 3b = 7.3c + 2d

d = 12.43

The oxygen balance yields

2a(16) = 1.2c(16) + 2e(16) + d(16)

a = 12.427

For glucose,

amount of carbon in 1 mole of substrate = 72 g

amount of carbon converted to biomass = 72(2/3) = 48 g

Then,

48 = 4.4c(12)

c = 0.909.

amount of carbon converted to CO2 = 72 - 48 = 24 g

24 = 12e

e=2

The nitrogen balance yields

14b = 0.86c(14)

b = 0.782

The hydrogen balance is

12 + 3b = 7.3c + 2d

d = 3.854

The oxygen balance yields

6(16) + 2(16)a = 1.2(16)c + 2(16)e + 16d

a = 1.473

b. For hexadecane,

2.42 (𝑀𝑊)𝑏𝑖𝑜𝑚𝑎𝑠𝑠

𝑌𝑥 =

𝑠 (𝑀𝑊)𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒

2.42 (91.34)

𝑌𝑥/𝑠 = = 0.98 𝑔 𝑑𝑤 𝑐𝑒𝑙𝑙𝑠/𝑔 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒

226

2.42 (𝑀𝑊)𝑏𝑖𝑜𝑚𝑎𝑠𝑠

𝑌𝑥 =

𝑂2 12.43(𝑀𝑊)𝑂2

2.42 (91.34)

𝑌𝑥 = = 0.557 𝑔 𝑑𝑤 𝑐𝑒𝑙𝑙𝑠/𝑔 𝑂2

𝑂2 12.43 (32)

For glucose,

0.909 (91.34)

𝑌𝑥/𝑠 = = 0.461 𝑔 𝑑𝑤 𝑐𝑒𝑙𝑙𝑠/𝑔 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒

180

0.909 (91.34)

𝑌𝑥 = = 1.76 𝑔 𝑑𝑤 𝑐𝑒𝑙𝑙𝑠/𝑔 𝑂2

𝑂2 1.473 (32)

The growth yield on more reduced substrate (hexadecane) is higher than that on partially

oxidized substrate (glucose), assuming that two-thirds of all the entering carbon is incorporated in

cellular structures. However, the oxygen yield on glucose is higher than that on the hexadecane,

since glucose is partially oxidized.

PRODUCT STOICHIOMETRY

Consider formation of an extracellular product 𝐶𝑗 𝐻𝑘 𝑂𝑙 𝑁𝑚 during growth. Then the

chemical equation for the whole process will be:

𝐶𝑤 𝐻𝑥 𝑂𝑦 𝑁𝑧 + 𝑎𝑂2 + 𝑏𝐻𝑔 𝑂ℎ 𝑁𝑖 → 𝑐𝐶𝐻𝛼 𝑂𝛽 𝑁𝛿 + 𝑑𝐶𝑂2 + 𝑒𝐻2 𝑂 + 𝑓𝐶𝑗 𝐻𝑘 𝑂𝑙 𝑁𝑚

Where f is the stoichiometric coefficient of the product. This is usually provided as another

experimentally determined yield coefficient, the product yield from substrate, 𝒀𝑷𝑺 .

𝑔 𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑠 𝑓𝑜𝑟𝑚𝑒𝑑 𝑓(𝑀𝑊 𝑝𝑟𝑜𝑑𝑢𝑐𝑡)

𝑌𝑃𝑆 = =

𝑔 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒 𝑐𝑜𝑛𝑠𝑢𝑚𝑒𝑑 (𝑀𝑊 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒)

The same with 𝒀𝑿𝑺 , we must be sure that the experimental system used to measure YPS

does not hold if product formation is not directly linked with growth. In these cases, independent

reaction equations must be used to describe growth and product synthesis.

THEORETICAL OXYGEN DEMAND

Oxygen demand is an important parameter in bioprocessing as oxygen is often the limiting

substrate in aerobic fermentations. Oxygen demand is represented by the stoichiometric coefficient

a. Oxygen requirement is related directly to the electrons available for transfer to oxygen; the

oxygen demand can therefore be derived from an appropriate electron balance. When product

synthesis occurs, the electron balance is:

𝑤γ𝑆 − 4𝑎 = 𝑐γ𝐵 + 𝑓𝑗γ𝑃

where γ𝑃 is the degree of reduction of the product. Rearranging gives:

1

𝑎 = (𝑤γ𝑆 − 𝑐γ𝐵 − 𝑓𝑗γ𝑃 )

4

It means that if we know which organism(γ𝐵 ) substrate (𝑤 𝑎𝑛𝑑 𝛾𝑆 ) and product

(𝑗 𝑎𝑛𝑑 γ𝑃 ) are involved in cell culture, and the yields of biomass (c) and product (f), we can

quickly calculate the oxygen demand. Of course we could also determine a by solving for all the

stoichiometric coefficients.

MAXIMUM POSSIBLE YIELD

The fractional allocation of available electrons in the substrate can be written as:

4𝑎 𝑐γ𝐵 𝑓𝑗γ𝑃

1= + +

𝑤γ𝑆 𝑤γ𝑆 𝑤γ𝑆

The first term on the right-hand side is the fraction of available electrons transferred from

substrate to oxygen, the second term is the fraction of available electrons transferred to biomass,

and the third term is the fraction of available electrons transferred to product. This relationship can

be used to obtain upper bounds for the yields of biomass and product from substrate.

Let us define CB as the fraction of available electrons in the substrate transferred to

biomass:

𝑐γ𝐵

𝜁𝐵 =

𝑤γ𝑆

In the absence of product formation, if all available electrons were used for biomass

synthesis, 𝜻𝑩 would equal unity. Under these conditions, the maximum value of the

stoichiometric coefficient c is:

𝑤γ𝑆

𝑐𝑚𝑎𝑥 =

γ𝐵

𝑐𝑚𝑎𝑥 can be converted to a biomass yield with mass units using 𝑌𝑋𝑆 , as it is equal to γ𝐵 .

Therefore, even if we do not know the stoichiometry of growth, we can quickly calculate an upper

limit for biomass yield from the molecular formula for substrate and product. If the composition

of the cells is unknown, γ𝐵 can be taken as 4.2 corresponding to the average biomass formula

𝐶𝐻1.8 𝑂0.5 𝑁0.2. Maximum biomass yield can be expressed in terms of mass (𝑌𝑋𝑆,𝑚𝑎𝑥 ) or as number

of C atoms in the biomass per substrate C atom consumed (𝑐𝑚𝑎𝑥 /𝑤 ). These quantities are

sometimes known as thermodynamic maximum biomass yields.

Likewise, the maximum possible product yield in the absence of biomass synthesis can be

determined,

𝑤γ𝑆

𝑓𝑚𝑎𝑥 =

𝑗γ𝑃

EXAMPLE:

The chemical reaction equation for respiration of glucose is:

C6H12O6 + 6O2 6CO2 + 6H2O.

Candida utilis cells convert glucose to CO2 and H2O during growth. The cell composition is

CH1.84O0.55N0.2 plus 5% ash. Yield of biomass from substrate is 0.5 g g- 1. Ammonia is used as

nitrogen source.

(a) What is the oxygen demand with growth compared to that without?

(b) C. utilisis also able to grow with ethanol as substrate, producing cells of the same composition

as above. On a mass basis, how does the maximum possible biomass yield from ethanol compare

with the maximum possible yield from glucose?

Solution:

Molecular weights: glucose = 180; ethanol = 46

MW of biomass is (25.44 + ash); since ash accounts for 5% of the total weight, 95% of the total =

25.44. Therefore, MW of biomass is equal to

25.44

𝑀𝑊 = = 26.78

0.95

If given γS for glucose is 4.00; γS for ethanol is 6.00,

γB =[4(1)+1(1.84)-2(0.55)-3(0.2)]=4.14

a. YXS == 0.5 g g-1 Converting this mass yield to a molar yield:

0.5 g biomass 180 g glucose 1 gmol biomass gmol biomass

YXS = ( )( )( ) = 3.36 =c

g glucose 1 gmol glucose 26.78 g biomass gmol glucose

1

a = [6(4)-(3.36)(4.14)] = 2.52

4

Therefore, the oxygen demand for glucose respiration with growth is 2.5 gmol O2 per gmol glucose

consumed. By comparison with the chemical reaction equation for respiration, this is only about

42% that required in the absence of growth.

b. Using the data above, the maximum possible yield for glucose is

6(4.00)

cmax = =5.80

4.14

Converting this to a mass basis:

5.80 g biomass 1 gmol glucose 26.78 g biomass g biomass

YXS,max = ( )( )( ) = 0.86

gmol glucose 180 g glucose 1 gmol biomass g glucose

For ethanol,

2(6.00)

cmax = =2.90

4.14

Converting this to a mass basis:

2.90 gmol biomass 1 gmol ethanol 26.78 g biomass g biomass

YXS,max = ( )( )( ) = 1.69

gmol glucose 46 g ethanol 1 gmol biomass g ethanol

Therefore, on a mass basis, the maximum possible amount of biomass produced per gram ethanol

consumed is roughly twice that per gram glucose consumed.

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