Submitted by:
2013-20358
Submitted to:
03 March 2017
PAMANTASAN NG LUNGSOD NG MAYNILA
University of the City of Manila
College of Engineering and Technology
Department of Chemical Engineering
TABLE OF CONTENTS
Stoichiometry 3
Bioreactors 3
Elemental Balances 10
Degree of Reduction 14
Measuring Yields 15
Stoichiometry
Stoichiometry is founded on the law of conservation of mass where the total mass
of the reactants equals the total mass of the products leading to the insight that the
relations among quantities of reactants and products typically form a ratio of positive
integers. This means that if the amounts of the separate reactants are known, then the
amount of the product can be calculated. Conversely, if one reactant has a known quantity
and the quantity of product can be empirically determined, then the amount of the other
reactants can also be calculated.
The term stoichiometry was first used by Jeremias Benjamin Richter in 1792
when the first volume of Richter's Stoichiometry or the Art of Measuring the Chemical
Elements was published. The term is derived from the Greek words stoicheion "element"
and metron "measure".
Introduction to Bioreactors
Biochemical engineering, also biotechnology engineering or bioprocess engineering,
is a branch of chemical engineering that mainly deals with the design and construction of
unit processes that involve biological organisms or molecules, such as bioreactors. Its
applications are in the petrochemical industry, food, pharmaceutical, biotechnology, and
water treatment industries.
BIOREACTOR
any manufactured or engineered device or system that supports a biologically active
environment.
a bioreactor is a vessel in which a chemical process is carried out which involves
organisms or biochemically active substances derived from such organisms. This
process can either be aerobic or anaerobic.
commonly cylindrical
ranging in size from litres to cubic metres
often made of stainless steel
may also refer to a device or system meant to grow cells or tissues in the context of
cell culture. These devices are being developed for use in tissue engineering or
biochemical engineering.
On the basis of mode of operation, a bioreactor may be classified as batch, fed batch
or continuous (e.g. a continuous stirred-tank reactor model).
Organisms growing in bioreactors may be submerged in liquid medium or may be
attached to the surface of a solid medium. Submerged cultures may be suspended or
immobilized. Suspension bioreactors can use a wider variety of organisms, since special
attachment surfaces are not needed, and can operate at much larger scale than immobilized
cultures. However, in a continuously operated process the organisms will be removed from
the reactor with the effluent. Immobilization is a general term describing a wide variety of
cell or particle attachment or entrapment. It can be applied to basically all types of
biocatalysis including enzymes, cellular organelles, animal and plant cells.
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. This observation for E. coli is also
found to be generally true for other cellular organisms.
Table 2 shows that, in different microbes, the carbon content varies from 46-50%,
hydrogen from 6-7%, nitrogen 8-14%, and oxygen from 29-35%. These are small
variations and the variations appear to depend on substrate and growth conditions. For
many engineering calculations, it is reasonable to consider cell as a chemical species having
the formula
CH1.8O0.5N0.2
1. Determine the molecular formula of Aerobacter Aerogenes provided the dry cell
composition (% mass) from Table 2.
Given:
C H O N
48.7% 7.3% 21.1% 13.9%
Required: MFAerobacter Aerogenes
Solution:
Basis: 100g AA
1𝑚𝑜𝑙 1𝑚𝑜𝑙
𝑛𝑐 = 48.7𝑔 ( ) = 4.0550 𝑚𝑜𝑙 𝐶 𝑛𝑂 = 21.1𝑔 ( ) = 1.3188 𝑚𝑜𝑙 𝑂
12.01𝑔 16𝑔
1𝑚𝑜𝑙 1𝑚𝑜𝑙
𝑛𝐻 = 7.3𝑔 ( ) = 7.2277 𝑚𝑜𝑙 𝐻 𝑛𝑁 = 13.9𝑔 ( ) = 0.9921 𝑚𝑜𝑙 𝑁
1.01𝑔 14.01𝑔
C4.0550H7.2277O1.3188N0.9921
Divide all subscripts by the subscript of the dominant element, ie. Carbon, therefore:
CH1.78O0.33N0.24
2. Determine the molecular formula of yeast - Saccharomyces Cerevisiae provided the dry
cell composition (%mass) from Table 2.
Given:
C H O N
47.0% 6.5% 31.0% 7.5%
Required: MFSaccharomyces Cerevisiae
Solution:
Basis: 100g SC
1𝑚𝑜𝑙 1𝑚𝑜𝑙
𝑛𝑐 = 47𝑔 ( ) = 3.9134 𝑚𝑜𝑙 𝐶 𝑛𝑂 = 31𝑔 ( ) = 1.9375 𝑚𝑜𝑙 𝑂
12.01𝑔 16𝑔
1𝑚𝑜𝑙 1𝑚𝑜𝑙
𝑛𝐻 = 6.5𝑔 ( ) = 6.4356 𝑚𝑜𝑙 𝐻 𝑛𝑁 = 7.5𝑔 ( ) = 0.5353 𝑚𝑜𝑙 𝑁
1.01𝑔 14.01𝑔
C3.9134H6.4356O1.9375N0.5353
Divide all subscripts by the subscript of the dominant element, ie. Carbon, therefore:
CH1.64O0.50N0.14
3. Determine the molecular formula of Eschericia Coli provided the dry cell composition
(%mass) from Table 2.
Given:
C H O N
50% 8% 20% 14%
Element Composition below 5% which includes S, Ca, N, etc. are neglected.
Required: MFEschericia Coli
Solution:
Basis: 100g EC
1𝑚𝑜𝑙 1𝑚𝑜𝑙
𝑛𝑐 = 50𝑔 ( ) = 4.1632 𝑚𝑜𝑙 𝐶 𝑛𝐻 = 8𝑔 ( ) = 7.9208 𝑚𝑜𝑙 𝐻
12.01𝑔 1.01𝑔
1𝑚𝑜𝑙 1𝑚𝑜𝑙
𝑛𝑂 = 20𝑔 ( ) = 1.25 𝑚𝑜𝑙 𝑂 𝑛𝑁 = 14𝑔 ( ) = 0.9993 𝑚𝑜𝑙 𝑁
16𝑔 14.01𝑔
C4.1632H7.9208O1.25N0.9993
Divide all subscripts by the subscript of the dominant element, ie. Carbon, therefore:
CH1.90O0.30N0.24
4. Determine the molecular formula of Candida Utilis, which feeds on glucose, provided the
dry cell composition (%mass) from Table 2.
Answer: CH1.83O0.56N0.20
5. Determine the molecular formula of Candida Utilis, which feeds on ethanol, provided the
dry cell composition (%mass) from Table 2.
Answer: CH1.84O0.55N0.20
6. 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? Assume cell composition as
CH1.8O0.5N0.2 (MW = 24.6)
1 10
𝑀𝑜𝑙𝑒𝑠 𝑜𝑓 𝑔𝑙𝑢𝑐𝑜𝑠𝑒 𝑛𝑒𝑒𝑑𝑒𝑑 = 𝑥
6 24.6
Therefore,
1 10
𝑀𝑖𝑛𝑖𝑚𝑢𝑛 𝑔𝑙𝑢𝑐𝑜𝑠𝑒 𝑛𝑒𝑒𝑑𝑒𝑑 (𝑚𝑎𝑠𝑠) = 𝑥 𝑥 180
6 24.6
𝑀𝑖𝑛𝑖𝑚𝑢𝑛 𝑔𝑙𝑢𝑐𝑜𝑠𝑒 𝑛𝑒𝑒𝑑𝑒𝑑 (𝑚𝑎𝑠𝑠) = 12.2𝑔
In the above example, we have assumed that all of the carbon found in substrate
(glucose) is incorporated into cell mass. This does not happen as the cell needs to
"oxidize" or respire some of the carbon to produce energy for biosynthesis and
maintenance of cellular metabolic machinery. In addition cells may produce
extracellular products that accumulate in the broth. Hence we can represent growth as:
Cells + (C source, N source, others) + O2 → More Cells + Extracellular Biomass + CO2 + H2O
The medium is the "food" for the cell. It serves as a source for all elements needed
by the cell to grow (or biosynthesis) and for product formation. The compounds carbon
dioxide and water on the product side of the reaction above result from oxidation of
glucose in the medium.
Since the cellular material contains C, N, P, S, K, Na, Ca, etc, the medium must be
formulated to supply these elements in the appropriate form.
If we neglect the "others" and assign stoichiometric coefficient for each of the
species in the above equation on the basis of one mole of glucose (C-source) consumed, we
re-write the above as:
Where-in ammonia represents the nitrogen source. We will refer to this reaction as
growth reaction.
Elemental Balances:
During the growth of cells, substrates that provide energy and raw materials are
needed for the synthesis of additional cell mass. Generally, in a biochemical process, the cell
environment should contain elements required in order to form additional cell mass and
the free energy from the substrate consumed should exceed the free energy of cells and
metabolic products formed.
In order to obtain the amount of every element that took part in a growth process,
the cellular content of all elements should be known. These include carbon, nitrogen,
oxygen and hydrogen (C, N, O and H respectively).
One mole of biological materials is defined as the amount containing 1 gram atom of
carbon. Assumption: No extracellular products other than water and carbon dioxide are
produced.
We have five equations and five unknowns (a, b, c, d, e). With a measured value of
RQ, these equations can be solved to determine the stoichiometric coefficients. Elemental
balances provide no insight into the energetics of a reaction. The concept of degree of
reduction has been developed and used for proton-electron balances in bioreactions.
Degree of reduction (γ) for organic compounds is defined as the number of equivalents of
available electrons per gram atom C.
The method of obtaining a balanced growth equation is very much similar to the
equation describing chemical reactions. One needs to establish a chemical formula for dry
cell material if the elemental composition of a particular strain growing under conditions is
known, the ratios of subscripts in the empirical cell formula can be determined.
Consider an aerobic growth which the products are cells, carbon dioxide and water,
and the initial components such as carbon and nitrogen sources; CHxOy and HlOmNn
respectively the equation can be written as follows:
with CH1:66O0:27N0:20 represent the biomass. Given that RQ = 0.43, determine the
stoichiometric coefficients a, b, c, d and e.
SOLUTION:
RQ = d/a = 0.43
C : 16 = c + d
H : 34 + 3(0.20c) = 1.66c +2e
34 = 1.06c + 2e
O : 2(d/0.43) = 0.27c + 2d + e
0.27c – 2.6157d + e
Thus,
Given:
C6H12O6 + aO2+ bNH3 → cCH1.64O0.50N0.14 + dCO2 + eH2O
Respiratory Quotient: d/b = 0.70
Equation 2 &4:
Equation 1, 6 & 7:
Note that CO2, H2O and NH3 the degree of reduction is zero
Measuring Yields
It is stressed that stoichiometric yield coefficient are ratios of
conversion rates (rx is given as C•mol X m3 reactor per h; r1 in mol i m3 per h).
There are five unknown stoichiometric coefficients for which five conservation
constraints can be formulated. C-conservation H-conservation O-conservation N-
conservation and Charge-conservation
Thus we see that Yax = 1/1.857 C-mol X / mol O2. Also Ycx =1/10.53 C-mol X/ mol HCO3