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Introduction

What are Microorganisms?

Microorganisms or microbes are single-celled or multi-cellular organisms which are too small to be

viewed by our naked eye which maybe bacteria, archaea, protozoa, and some algae and fungi.

What is Microbial Growth?

Microbial growth is the asexual reproduction, cell division, of most microorganisms (bacteria) into

two daughter cells, in a process called binary fission, which means that there will be no sex cells

involved. Providing no mutational event occurs, the resulting daughter cells are genetically identical

to the original cell. Growth implies that all chemical components of the cell increase with the same

speed and after a certain time this leads to increase in cell number, which causes increase in size or

number of the individuals (Biomine.skelleftea).


Cell Composition and Elemental Balance

Although there are many different biological species, it turns out that a very large fraction of the mass

these biological species are made of a few elements carbon, oxygen, nitrogen and hydrogen; and these

elements are among the most abundantly found elements on earth.

Cells primarily are made up of water. Typically 70 percent of a cell’s mass is water and the remaining

is dry matter. Therefore it is conventional to express cell composition on a dry basis. . The

microorganism Escherichia coli are widely used in genetic engineering. Typical elements found

in Escherichia coli are given below:

Table 1 Elemental Composition of E. coli (after Stanier et al)

Element % Dry Weight


Carbon, C 50
Oxygen, O 20
Nitrogen, N 14
Hydrogen, H 8
Phosphorus, P 3
Sulfur, S 1
Potassium, K 1
Sodium, Na 1
Calcium, Ca 0.5
Magnesium, Mg 0.5
Cl 0.5
Fe 0.2
others 0.3
Nearly half of the dry matter in cells is carbon and the elements carbon, oxygen, nitrogen and

hydrogen total up to about 92% of the total mass of the cell in a dry weight basis. This observation for

E. coli is also found to be generally true for other cellular organisms.

Also, a microorganism can contain up to 30 – 60% protein, 5 – 30 % carbohydrates, 5 – 10 % lipids,

1% DNA, 5 -15% RNA, and can contain Ash made up of P, K, Mg2+ and etc.
A microorganism can synthesize complex molecules from smaller units, this process is called

anabolism. Small units to amino acid to protein to sugars to carbohydrates to fatty acids to lipids to

nucleotides to DNA and RNA and so on and so forth (Arifin Y., 2014).

The sum of all of these reactions gives the anabolic reaction.

(…) C-source + (…) N-source + (…) P-source + (…) O-source → CHαOβNδ + (…) H2O + (…) CO2

A one mole of a biological material can compose 1 gram of carbon, C, such as CHON assuming no

cellular products are formed ather than water and carbon dioxide.

A simple biological conversion of a carbohydrate (CHmOn) with the presence of ammonia (NH3) and

oxygen (O2) to water (H2O), carbon dioxide (CO2) and a cell CHαOβNδ has the reaction given below:

CHmOn + aO2+ bNH3---> cCHαOβNδ + dH2O + eCO2

Where: one mole of unknown carbohydrate (ChmOn) is given and an unknown number (b) of cellular

material CHαOβNδ is formed.

Perform elemental balance CHONS + RQ.

C : 1=c+e

H : m + 3b = cα + 2d

O : n + 2a = cβ + d + 2e

N : b = cδ

S :

RQ : e/a (respiratory quotient)


Having five equations and five unknown (a, b, c, d and e) and with a measured value of RQ, the

equations above can be solved to determine the stoichiometric coefficients of the elements in the

reaction.

For many engineering calculations, it is reasonable to consider cell as a chemical species having the

formula CH1.8O0.5N0.2. This engineering approximation is a good starting point for many quantitative

analyses while a more carefully formulated empirical formula based on proximate analysis may be

necessary for complete material flow analysis. The cell molecular weight for the above cell formula is

12+1.8 + 0.5(16) +0.2 (14) = 24.6 (gatewaycoalition.org).

Example:

Suppose we want to produce 10 g of cells using glucose as a carbon source. What is the minimum

amount of glucose that would be needed?

Solution:

Assume cell composition as CH1.8O0.5N0.2 and glucose is Glucose is C6H12O6.

MW of glucose is 180 and cell MW is 12+1.8 + 0.5(16) +0.2 (14) = 24.6.

Moles of cells to be grown = 10/24.6

Since glucose has 6 moles of carbon per mole of glucose,

Moles of glucose needed = (1/6) (12/24.6)

Therefore, min glucose needed = (1/6) (12/24.6) (180) = 12.2g


Degree of Reduction (γ)

It is defined as the number of equivalents of available electrons per grams of atom (Yaacob M. N. D.,

2017).

Thermodynamically, energy is also needed for cell maintenance. And elemental balances provide no

insight into the energetics of a reaction. Here, catabolism enters. Catabolism generates energy for

anabolism and cell maintenance. It is consist of electron donor couple and electron accepter couple.

Example,

Glucose + (…) O2 → (...)HCO3- + H2O


Donor couple: glucose and HCO3-

Acceptor couple: O2 and H2O

Glucose → (...)HCO3- + (...)Ethanol

Donor couple: glucose and HCO3-

Acceptor couple: CO2 and Ethanol

The catabolism of these reactions produces Gibbs energy. Concepts of catabolism have been used for

proton – electron balance in bio-reactions.

Degree of reduction is defined as the number of equivalents of available electrons per gram of atom.

Degree of reduction for some key elements

Element γi
Carbon, C 4
Hydrogen, H 1
Oxygen, O -2
Nitrogen, N -3
Sulfur, S 6
Fe 3
+ charge -1
- charge 1
NH4+ as N-source -3
N2 as N-source 0
NO3- as N-source 5
For example:

Glucose (C6H12O6): 6x4 + 12x1 + 6x(-2)=24

γ = 24/6 = 4

Methane (CH4): 1x4 + 4x1 = 8

γ = 8/1 = 8

Ethanol (C2H5OH): 2x4 + 6x1 + 1x(-2) = 12

γ = 12/2 =6

γ - balance is used to calculate stoichiometry. It follows from the conservation relations (C, H, O, N,

charge, etc.) by eliminating the unknown stoichiometric coefficient for reference compounds. It

relates biomass, substrate/donor, acceptor, product (H2O, H+, HCO3-, N – Source are always absent).

Ex: Catabolism of glucose to ethanol in anaerobic culture

-C6H12O6 + a C2H6O + bCO2 + cH2O + dH+

γ glucose = 24

γ ethanol = 12

γ balance = -24 + 12a = 0

Therefore, a = 2

From C, O and charge conservation b, c, and d will follow.

Thus: -C6H12O6 + 2 C2H6O + 2 CO2


Yield Coefficients

Cell mass and product formation by microorganisms can be described quantitatively by yield

coefficients expressed as the mass of cells or product formed per unit mass of substrate consumed,

Yx/s, and Yp/s, for cells and product, respectively (J. Hong, 1988). And the above value is often called

cell yield, growth yield or yield coefficient. With the yield coefficients, the material balance equations

for cells, substrate, and product can be straightforwardly formulated.

Let's consider the overall stoichiometric equation for growth and production:

sS + nN + oO2 → X + pP + wH2O + eCO2

Where: S, carbon source

N, nitrogen source

X, cell mass

P, product

And s, n, o, p, w and e are stoichiometric coefficients.


Theoretical yield coefficients can be determined from the above stoichiometry when the chemical

formula for S, N, X and P are already known.

The cell mass yield coefficient and the product yield coefficient are

Yx/s= Mx / sMs

Yp/s = pMp / sMs

Where: Mx, Mp, and Ms are the molecular weights of cell mass, product and carbon source

References:

http://wiki.biomine.skelleftea.se/wiki/index.php/Microbial_growth

Stoichiometry of Cellular Growth

http://www.gatewaycoalition.org/files/hidden/react/ch2/2_0f.htm

KINETICS, MICROBIAL GROWTH

http://people.math.gatech.edu/~weiss/uploads/5/8/6/1/58618765/panikov_kinetics_microbial_gro

wth.pdf

Stoichiometry of Microbial Growth and Product Formation

http://slideplayer.com/slide/3404226/

Fermentation ppt

https://www.slideshare.net/pkdsshukla/fermentation-ppt-42822875

BIOPROCESS ENGINEERING – BIOPROCESS ANALYSIS THROUGH CALORIMETRY

AND BIOTHERMODYNAMICS

https://www.eolss.net/sample-chapters/C17/E6-58-04.pdf