4.

1 Material Balance
4.1.1 Introduction
Material quantities as they pass through processing operations can be described by material
balances. Material balances are the basis of process design. A material balance taken over the
complete process will determine the quantities of raw materials required and products
produced. Balances over individual process units set the process stream flows and
compositions. Such balances are statements on the conservation of mass. Similarly, energy
quantities can be described by energy balances, which are statements on the conservation of
energy. If there is no accumulation, what goes into a process must come out. It is true for
continuous operation over any chosen time interval.
Material balances are fundamental to the control of processing, particularly in the control of
yields of the products. The increasing cost of energy has caused the industries to examine
means of reducing energy consumption in processing. Energy balances are used in the
examination of the various stages of a process, over the whole process and even extending over
the total production system from the raw material to the finished product.
The general conservation equation for any process system can be written as:
Material out = Material in + Generation - Consumption – Accumulation
For a steady-state process the accumulation term will be zero. If there is no chemical reaction
the steady-state balance reduces:
Material out = Material in
Time basis might be chosen as a basis for calculation when results are to be presented in tone/y
[1].
4.2. Annual production schedule:
𝑑𝑟𝑦 𝑚𝑎𝑡𝑡𝑒𝑟 𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑖𝑜𝑛 (𝑡𝑜𝑛) 𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑖𝑜𝑛
= (1 − 𝑕𝑢𝑚𝑖𝑑𝑖𝑡𝑦)
𝑦𝑒𝑎𝑟 𝑦𝑒𝑎𝑟
𝑑𝑟𝑦 𝑚𝑎𝑡𝑡𝑒𝑟 𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑖𝑜𝑛 (𝑡𝑜𝑛) 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑦𝑒𝑎𝑠𝑡 𝑑𝑟𝑦 𝑚𝑎𝑡𝑡𝑒𝑟
(%) =
𝑦𝑒𝑎𝑟 𝑡𝑜𝑡𝑎𝑙 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑦𝑒𝑎𝑠𝑡 𝑑𝑟𝑦 𝑚𝑎𝑡𝑡𝑒𝑟
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑎𝑡𝑐𝑕𝑒𝑠 𝑡𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑎𝑡𝑐𝑕𝑒𝑠
= 𝑦𝑒𝑎𝑠𝑡 𝑑𝑟𝑦 𝑚𝑎𝑡𝑡𝑒𝑟 𝑓𝑟𝑎𝑐𝑡𝑖𝑜𝑛 ×
𝑦𝑒𝑎𝑟 𝑦𝑒𝑎𝑟
𝑦𝑒𝑎𝑠𝑡 𝑑𝑟𝑦 𝑚𝑎𝑡𝑡𝑒𝑟 𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑖𝑜𝑛
𝑦𝑒𝑎𝑟
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑎𝑡𝑐𝑕𝑒𝑠
𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑖𝑜𝑛(𝑘𝑔) 𝑦𝑒𝑎𝑟
𝑚𝑎𝑡𝑐𝑕 (1 − 𝑙𝑜𝑠𝑠𝑒𝑠)
After making the calculations, we obtain the following results (Table 5).
Table 5 - Annual production schedule

annual dry
matter
annual production
production
production per matches
yeast type number of
(ton/year) __________
matches (ton)
(ton/year) %
(matches/year)

F.F.B 44100 77.78% 431.5 30.660

F.F.C 3780 22.22% 123.3 30.660

Total 17010 100.00% 554.8 -

4.3. Stoichiometry of the FFC growth reaction

There are different compositions in the literature for FFC yeast. We opted for the composition
indicated in [23], as it constitutes an average of values presentedpreviously by other authors
(see Table 6).
Table 6 - FFC yeast composition

mass molecular mol/100g dry mol/mol of

composition(%) weight yeast carbon

(g/mol)
Element
____________ ____________ ____________ ____________

A PM B=A/PM D=B/B(Carbon)

carbon 46.0 12.011 3.83 1.00

oxygen 23.0 15.999 2.00 0.52

nitrogen 8.5 14.007 0.61 0.16

hydrogen 6.0 1.008 5.95 1.55

ashes 7.5 - - -

TOTAL 100.0 - - -
The ashes are made up of several mineral salts, each representing, per se,
a negligible portion in the establishment of the stoichiometric equation.
Although cell growth encompasses many chemical reactions, the end result can be
be represented by a single equation, which involves only the most metabolism products
representative. This always includes the cell formula (Table 6, column D), depending on
the other terms of the available nutrients and energy sources. The general form (for
any microorganism) from the aerobic growth equation (Equation 3), using the
ammonia as a nitrogen source, is:
OO2 + CHHSOOSNNS + aNH3 → YbCHhbOobNnb + ƩYpiCHhpiOopi + wH2O + cCO2 (Equation 3)
o, a, Yb, Ypi, w and c are the stoichiometric coefficients for O2, NH3, biomass, the ith
metabolic product, H2O and CO2, respectively. The indices s, b and pi refer to the
substrate, biomass i th product, respectively.[27]
In the specific case of FFC yeast, Ypi can be considered null, i.e., there is noformation of
products in quantities that justify being considered in these calculations.
All the others are different from zero, but yet unknown. To determine them it is necessary to
bear in mind the following points:
i) Composition of molasses
The composition of molasses allowed is shown in Table 7.
The quantity of fermentable (reducible) sugars is counted in terms of the mass of
sugars, which is obtained after its inversion by the invertase enzyme (secreted by the
yeast).
Table 7 – Molasses composition [23]

Constituent Beet molasses Cane molasses

Total amount of reducible 47-58 50-58

sugar after
inversion(%mass)

Total amount of 0.2-2.8 0.1-0.6

nitrogen(%mass)
a-amino nitrogen, N(%mass) 0.36 0.03

Phosphorus, P2O5(%mass) 0.02-0.07 0.01-0.08

Calcium. CaO(%mass) 0.15-0.7 0.15-1.0

Magnesium, MgO(%mass) 0.01-0.1 0.25-0.8

Potassium, K2O(%mass) 2.2-5.0 0.8-2.3

Zinc, Zn(%mass) 30-50 5-20

Total amount of carbon, 28-34 28-33

C(%mass)

Total amount of ash(%mass) 4-11 3.5-7.5

Sulfer, SO3(%mass) 0.3-0.4 -

Biotin(μg/g) 0.01-0.13 0.9-1.8

Calcium pantothenate(μg/g) 40-100 16

Inositol(μg/g) 5000-8000 2.5

Thiamine(μg/g) 1-4 -

pyrodixin(μg/g) 2.3-5.6 -

riboflavin(μg/g) 0-0.75 -

Folic acid(μg/g) 0.21 -

This type of reaction is necessary due to the fact that molasses are made up ofdifferenttypes of
sugars, which have different degrees of assimilation by cells: [22]
Simple sugars: they are monosaccharides, such as fructose, glucose and maltose. They are
isomers of chemical formula C6H12O6 and are directly assimilable.
-More complex sugars (eg sucrose and raffinose), are not directly assimilable by the cells. They
must first be hydrolyzed to monosaccharides outside the cell by the action of the invertase
enzyme.
-Very complex sugars: they are not hydrolyzable by invertase, they are very difficult assimilated.
It should be noted that the production of invertase by Sacharomyces cerevisiae is so fast that
simple and complex sugars are assimilated with the same speed.Another aspect to be taken
into account is that this type of reaction consumes water molecules. ThusTherefore, it cannot
be accounted for due to the lack of knowledge of the composition of theMolasses in each of the
various types of fermentable sugars that make them up.
Bearing in mind the considerations mentioned above, it is immediate that the formulamost
correct chemistry to use for the substrate in Equation 3 of the cells is C6H12O6.
ii) Yield of biological reaction
Different definitions for the yield of the reaction are found in the literature, depending on the
emphasis that one wishes to place on a given product involved in the production of biomassAnd
/ or biomass status (wet or dry basis). In this work one of the most cited isused:[25]

biomass produced
η =
sugar mass(C6H12O6 Conumed)

Since it is assumed that [25]: μ = 0.5

Taking into account what was mentioned in i), Equation 3 can be rewritten as follows form:
C6H12O6 + OO2 + aNH3 → YbCO0.52H0.16H1.55 + cCO2 + wH2O (Equation 4)
The following balances can then be established for the elements:
C - Yb + c = 6 (Equation 5)
H - 1.55 Yb + 2w -3a = 12 (Equation 6)
O - 0.52 Yb + 2c + w -2o = 6 (Equation 7)
N - 0.16 Yb - a = 0 (Equation 8)
From ii),

180𝑔 𝑠𝑢𝑔𝑎𝑟 η biomass

× 𝑔 𝑠𝑢𝑔𝑎𝑟
𝑚𝑜𝑙 𝑠𝑢𝑔𝑎𝑟
𝑌𝑏 =
26.09 ∗ 𝑔 𝑏𝑖𝑜𝑚𝑎𝑠𝑠
𝑚𝑜𝑙 𝑏𝑖𝑜𝑚𝑎𝑠𝑠

* this factor was determined from Table 6, adding the elements of column D after
themultiplication by the respective molecular weight (PM) and adding the masscorresponding
to the ashes (Table 8)[27]
Table 8 - Calculation of the molecular weight of biomass

molecular weight mol/mol g element/mol

(g/mol) in C biomass
Element
____________ ____________ ____________
PM D=B/B(C) E=D×PM

Carbon 12 1 12

oxygen 16 0.52 8.36

nitrogen 14 0.16 2.22

hydrogen 1 1.55 1.55

Total - - 24.13

Bearing in mind that the fixed income is based on experimental resultspublished and not in
theoretical considerations (since only in these can we speak of a formulachemistry for biomass
with a molecular weight easily calculated from it), in thethe determination of biomass has to
take into account the 7.5% of ash that constitute it.
Therefore, we have:
24.13𝑔 𝑏𝑖𝑜𝑚𝑎𝑠𝑠 100% 26.09𝑔 𝑏𝑖𝑜𝑚𝑎𝑠𝑠
× =
𝑚𝑜𝑙 𝑏𝑖𝑜𝑚𝑎𝑠𝑠 92.5% 𝑚𝑜𝑙 𝑏𝑖𝑜𝑚𝑎𝑠𝑠

The equations listed above, 5 to 8 constitute a system of linear equationswhich can be

represented in the matrix form:
𝑌𝑏 6
𝑎 12
× 𝑜 = 6
𝑐 0
𝑤 3.45

Its resolution leads to the following values for stoichiometric coefficients:

Yb = 3.45 c = 2.55
a = 0.551 w = 4.16
o = 2.53
Taking into account the coefficients determined above, the stoichiometric equation can
bewritten as follows:
C6H12O6 + 2.53 O2 + 0.551 NH3 →3.45 CO 0.52 N 0.16 H 1.55 + 2.55 CO2 + 4.16 H2O
(Equation 9)
In mass terms, for later calculations, based on 1 mole of CO 0.52 N 0.16 H1.55, the
stoichiometric values are in Table 9.
Table 9 - Mass stoichiometry of the biological reaction of fresh yeast

component C6H12O6 O2 NH3 CO0.52N0.16H1.55 CO2 H2O

_________ _________ _________ _________ _________ _________ _________
unity (g) (g) (g) (g) (g) (g)

Mass 52.24 23.44 2.72 26.12 32.57 21.71

4.4. Determination of the fraction of beet molasses in the feed streammolasses

The molasses fraction is a parameter that depends essentially on two factors: price
andcomposition.The composition varies depending on where it comes from, such as the time of
year and other factors,however, we fixed it for the calculations, taking into account the range
shown in Table 10.
Table 10 - Concentration of some components in molasses

Constituent beet molasses cane molasses

Total amount of reucible 52.5 54

sugar after inversion
(%mass)

Phosphorus P2O5(%mass) 0.045 0.045

Biotin(μg/g) 0.07 1.35

There are different criteria that could be adopted, based on the compositions of themolasses,
to determine the fraction of sugarcane molasses in the stream. In this work we chosefor taking
vitamin biotin as a determining component, which, as already mentioned,assumes a
fundamental role in the cell phone. Thus, bearing in mind the following points:
  Composition of molasses in biotin and sugar (Table 7)
 Yeast composition in biotin equal to 1.6 μg/g [22]
 Yield of biological reaction, η = 0.5
Therefore, we proceed to calculate:
Biotin present in molasses = b
b = biotin that the cell needs× η
1.6 × 10−6 𝑘𝑔 𝑏𝑖𝑜𝑡𝑖𝑛 0.5 𝑘𝑔 𝑦𝑒𝑎𝑠𝑡
𝑏= ×
𝑘𝑔 𝑏𝑒𝑒𝑡 𝑚𝑜𝑙𝑎𝑠𝑠𝑒𝑠 𝑘𝑔 𝑠𝑢𝑔𝑎𝑟

0.8 × 10−6 𝑘𝑔 𝑏𝑖𝑜𝑡𝑖𝑛

𝑏=
𝑘𝑔 𝑜𝑓 𝑠𝑢𝑔𝑎𝑟

where k is the fraction of beet molasses present in the molasses stream:

𝑘𝑔 𝑏𝑖𝑜𝑡𝑖𝑛 𝑘𝑔 𝑏𝑖𝑜𝑡𝑖𝑛
𝑘 × 0,07 × 1006 1.35 × 10−6
𝑘𝑔 𝑏𝑒𝑒𝑡 𝑚𝑜𝑙𝑎𝑠𝑠𝑒𝑠 𝑘𝑔 𝑜𝑓 𝑐𝑎𝑛𝑒 𝑚𝑜𝑙𝑎𝑠𝑠𝑒𝑠
+ (1 − 𝑘) ×
𝑘𝑔 𝑠𝑢𝑔𝑎𝑟 𝑘𝑔 𝑠𝑢𝑔𝑎𝑟
0.525 0.54
𝑘𝑔 𝑏𝑒𝑒𝑡 𝑚𝑜𝑙𝑎𝑠𝑠𝑒𝑠 𝑘𝑔 𝑐𝑎𝑛𝑒 𝑚𝑜𝑙𝑎𝑠𝑠𝑒𝑠
𝑘𝑔 𝑏𝑖𝑜𝑡𝑖𝑛
= 0.8 × 10−6
𝑘𝑔 𝑜𝑓 𝑠𝑢𝑔𝑎𝑟
𝑘 × 0.07 × 10−6 1.35 × 10−6
+ (1 − 𝑘) × = 0.8 × 10−6
0.525 0.54

Solving the equation, we get: k = 0.72

The obtained ratio (k) has a value close to that mentioned in [22] (80%)
4.5. Mass balance to the preparation section of the culture medium:
In this sub-chapter, calculations will be carried out in order to satisfy the need molasses for the
production of fresh yeast.
4.5.1. Balance of the need for molasses:
The molasses requirement is calculated taking into account the production of yeast and Yield.
The consumer units of molasses are the 1st, 2nd and 3rd stages of propagation, with a
yield of 0.5 (as previously mentioned in stoichiometric calculations), and the laboratory phase,
with a yield of 0.36 (see Annex I).
Using as basis for calculation = a match.
In the 3rd stage, 30.66 ton of biomass was obtained (see Table 5).
Knowing that at the end of the game the concentration of dry yeast should be 8 to 10%,
we define the value of 8% as an operating parameter, therefore, the mass that we will have in
the end of this stage is:
30.66
𝑇𝑜𝑡𝑎𝑙 𝑚𝑎𝑠𝑠 = = 383.25𝑡𝑜𝑛
0.08

Bearing in mind that in this process there is an octuplication of yeast mass (multiplication of
biomass equal to 8), as previously mentioned, we have toinitial mass of the stage (inoculum) is
1/8 of the final mass:
30660
𝑖𝑛𝑜𝑐𝑢𝑙𝑢𝑚 𝑚𝑎𝑠𝑠 = = 3833𝑘𝑔
8

Therefore, the amount of dry yeast that is produced will be: dry yeast
30.66𝑡𝑜𝑛 − 3.833𝑡𝑜𝑛 = 26.827 𝑡𝑜𝑛 𝑜𝑓 𝑑𝑟𝑦 𝑦𝑒𝑎𝑠𝑡
Bearing in mind the definition of income, and assuming the value of the same 0.5, as already
was mentioned earlier:
26.827𝑡𝑜𝑛 26.827
𝑟𝑒𝑞𝑢𝑖𝑟𝑒𝑑 𝑠𝑢𝑔𝑎𝑟 𝑚𝑎𝑠𝑠 = = = 53.654𝑡𝑜𝑛
η 0.5
Since, what is produced in the previous stage is the inoculums of the next one, and that the
multiplication and yeast concentration at the end are the same in all stages, can identical
calculations must be carried out for the stages upstream of the 3rd (results compiled in Table
11).
In the laboratory phase, only the third stage consumes significant molasses. At this stage, the
give factor and the yeast concentration at the end are the same in all stages, the calculations to
be carried out and the biomass multiplication is 950 [25] and the yield, as mentioned
previously, it is 0.36. The method of calculating the quantity of sugar needed for this step is the
same as that performed for the 3rd stage of the laboratory phase (see Table 11).
Table 11 - Sugar requirements for the different stages of the process

mass(dry base) final(kg) inoculum(kg) produced(kg) sugar(kg)

________ ________ ________
A B A-B

3th stage 30659.697 3832.462 26827.235 53654.470

2th stage 3832.462 479.058 3353.404 6706.809

1th stage 479.058 59.882 419.176 838.351

3th step(lab) 59.882 0.063 59.819 119.638

Total - - - 61319.268

The total amount of molasses that is needed is calculated on the basis of its sugar content:
𝑛𝑒𝑐𝑒𝑠𝑠𝑎𝑟𝑦 𝑚𝑎𝑠𝑠 𝑜𝑓 𝑠𝑢𝑔𝑎𝑟
𝑚𝑜𝑙𝑎𝑠𝑠𝑒𝑠 𝑚𝑎𝑠𝑠 =
𝑠𝑢𝑔𝑎𝑟 𝑓𝑟𝑎𝑐𝑡𝑖𝑜𝑛 𝑜𝑓 𝑚𝑜𝑙𝑎𝑠𝑠𝑒𝑠

Swing to molasses filter: in this operation, remove the solids that the molasses contains, and
dilute it sufficiently so that can be handled and transported.
Since it is a continuous operation, the calculations are made considering 1 hour of operation as
the basis for calculation.
The flow rate of the molasses inlet current is calculated as follows:
𝑘𝑔
𝑛𝑒𝑒𝑑 𝑓𝑜𝑟 𝑚𝑜𝑙𝑎𝑠𝑠𝑒𝑠(𝑚𝑎𝑡𝑐 𝑕
)
𝑡𝑕𝑒 𝑓𝑙𝑜𝑤 𝑚𝑜𝑙𝑎𝑠𝑠𝑒𝑠 𝑖𝑛𝑙𝑒𝑡 𝑐𝑢𝑟𝑟𝑒𝑛𝑡 = 𝑕𝑜𝑢𝑟
𝑡𝑖𝑚𝑒 𝑜𝑓 𝑚𝑎𝑡𝑐𝑕(𝑚𝑎𝑡𝑐 𝑕
)

According to the values indicated in Table 7 and Table 10, and taking into account thecontent of
solid (insoluble) impurities [22] in molasses, it is possible to fix its composition ininert (soluble
impurities).
Table 12 - Composition of beet molasses

sugar impurities solid other nutrients inert

52.5% 32% 8% 7.5%

The flow rate of each component of the M1 stream is obtained by making the product of the
flow rate by the composition of each component.
The values of these flows are shown in Table 1.
The efficiency of the filter is very high [22], and for this reason it can be assumed that 99.9%of
solid impurities leave the solid effluent (ESC). It is also considered, that this leaves with 5%
humidity and that there is no loss of sugar.
Thus, in the ESC chain, there are:

𝑠𝑜𝑙𝑖𝑑 𝑖𝑚𝑝𝑢𝑟𝑖𝑡𝑖𝑒𝑠 𝑡𝑕𝑎𝑡 𝑐𝑜𝑚𝑒 𝑜𝑢𝑡 𝑖𝑛 𝐸𝑆𝐶

= 𝐹𝑖𝑙𝑡𝑒𝑟 𝑒𝑓𝑓𝑖𝑐𝑖𝑒𝑛𝑐𝑦 × 𝑡𝑜𝑡𝑎𝑙 𝑎𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝑠𝑜𝑙𝑖𝑑 𝑖𝑚𝑝𝑢𝑟𝑖𝑡𝑖𝑒𝑠

The current that flows from the filter to the molasses tank M2 contains:
Total amount of impurities-impurities that come out of the filter = Amount of impurities that
follow in M2
Bearing in mind that, as previously mentioned, the current ES1 is constituted by solid impurities
with 5% humidity, the flow of this current can be calculated:
𝑖𝑚𝑝𝑢𝑟𝑖𝑡𝑖𝑒𝑠 𝑡𝑕𝑎𝑡 𝑐𝑜𝑚𝑒 𝑜𝑢𝑡 𝑖𝑛 𝐸𝑆𝐶
𝑓𝑙𝑜𝑤 𝑟𝑎𝑡𝑒 𝑜𝑓 𝐸𝑆𝐶 =
(1 − 0.05)

Therefore, it is possible to calculate the water flow in the ES1 stream:

𝐸𝑆𝐶 𝑤𝑎𝑡𝑒𝑟 𝑓𝑙𝑜𝑤 = 𝑡𝑜𝑡𝑎𝑙 𝑓𝑙𝑜𝑤 − 𝑓𝑙𝑜𝑤 𝑜𝑓 𝐸𝑆𝐶 𝑖𝑚𝑝𝑢𝑟𝑖𝑡𝑖𝑒𝑠

With the exception of the flow of solid impurities and water, all partial flow rates of thecurrent
M2 are the same as current M1 (Table 1).It is assumed that a 5% dilution of molasses is
sufficient, the total flow rate of the molasses is calculatedcurrent M2, FM2:
𝐹𝑀2𝑖
𝐹𝑀3 =
(1 − 𝑋𝑀2𝑖
Being:
𝐹𝑀2𝑖 - Flow rate of component i in current M2
𝑋𝑀2𝑎 - Water fraction in stream M2
The water flow, FM2i, is from: 𝐹𝑀𝑠𝑎 = 𝐹𝑀2 × 𝑋𝑀3𝑎 ,being the composition of the current
𝐹𝑀 2𝑖
M2: 𝑋𝑀2𝑖 = , 𝑋𝑀2𝑖 being the fraction of component i in the current M2.
𝐹𝑀2
The water flow of the AP1 stream, FAP1, is determined by making a balance to the water:
𝐹𝐴𝑃 1 = 𝐹𝑀 3𝑎 + 𝐹𝐸𝑆𝐶
Balance to molasses sterilizer:
The function of the sterilizer at this stage is to eliminate the microorganisms that are in
themolasses chain.In this operation, the objective of diluting sugar to 40% [22] (mass) is also
achieved. The objective of this operation is to prepare the culture medium, both in terms of
composition andtemperature (303.15 K), to be used in the propagation section.
This operation is carried out quickly, assuming that there are no sugar losses due
tocaramelization. [2]
The calculation base is 1 hour.
The AP2 stream is made up of water only, so the partial flows of the M3 and
M4 are the same, except for water.
𝐹𝑀4𝑎𝑐
Current flow M4, FM4, is calculated by: 𝐹𝑀4 = , FM4ac being the flow of fermentable
𝑋 𝐹𝑀 4
sugars in chain M4, FM4ac the fraction of fermentable sugars in chainM4.
The water flow in the M4 stream is necessary to complete FM4.
The composition of the M4 current is obtained by the quotient between partial and total flows.
The flow rate of current AP2, FAP2, is then calculated by making an overall water balance:
𝐹 𝐴𝑃2 = 𝐹𝑀4𝑎𝑔 − 𝐹𝑀3𝑎𝑔 , with 𝐹𝑀4𝑎𝑔 being the water flow in stream 4, and FM3ag being the
water flowin chain 3.
Table 3 compares the values of flow rates and current compositions in the
sterilizer.

4.6. Mass balances to the yeast propagation section of the Yeast:

Bearing in mind the aforementioned, the balances are made at the stages ofpropagation and
not to the fermenters that constitute them.
Yeast flows in the inoculums and yeast streams are already known, as well assugar needs (see
Table 11).
The volume of each stage is shown in Annex II.
Balance at the 3rd Propagation Stage (3rd stage fermenter - F3)
The calculation basis is a departure.
Molasses current (M8)
The composition of this current (M8) is known, and it is the same as that of the current M4.
Being the flow rate, FM8, calculated by:
𝐹𝑀8
𝐹𝑀8 =
𝑋𝑀8𝑎𝑐
Therefore, the partial currents are calculated by multiplying the total flow of thecurrent M8 for
its composition. The results are compiled in Table 4.
Ammonia current (NH3)
The requirements of the yeast in ammonia, for each stage, are calculated from the
stoichiometric mass ratio that is in Table 9.
𝑀𝐹3 ×𝑀𝐴𝐸𝐴𝐸
𝐹𝑁𝐻3𝑛 = , being 𝐹𝑁𝐻3𝑛 the need for ammonia (ton), MF3 the yeast mass produced in
𝑀𝐹𝐸
this stage (ton), MAE the stoichiometric ammonia mass (2.72 g / molCO0.52N0.16H1.55) and
MFE the stoichiometric yeast mass (26.12 g / molCO0.52N0.16H1.55).
By setting the ammonia concentration in the NH3 stream to 25%, the flow rate, 𝐹𝑁𝐻3 :
𝐹
𝐹𝑁𝐻3 = 𝑋 𝑁𝐻 3 , with NH3n being the ammonia fraction in the NH3 stream.
𝑁𝐻 3𝑛

Assuming that this current consists only of ammonia and water, 75% of the current will be
water. So, the water flow is calculated will be the difference between the total flow and the
water flow ammonia.
Air inlet current (AR3)
The oxygen requirements for yeast are calculated, for each stage, from thestoichiometric mass
ratio that is in Table 9.
𝑀𝐹3 × 𝑀𝑂𝐸
𝑁𝑂3 =
𝑀𝐹𝐸
NO3 being the need for oxygen (ton) and MOE the stoichiometric oxygen mass (23.44 g / mol
CO0.52N0.16H1.55).
Since, in the dimensioning of fermenters, the percentage ofoxygen absorbed by the yeast in the
fermentation medium is different at each stage and isequal to:
- 12.5% in the first stage
- 31% in the second stage
- 69% in the third stage
Although the oxygen concentration in the fermentation medium is not constant (transient), in a
start all the oxygen that is absorbed is consumed. Therefore, the flow of this, FAR3, is given by:
03
𝐹𝐴𝑅3𝑜 =
0.69
The air flow is calculated, taking into account a composition of 21% in oxygen, and the
remaining 79% nitrogen.
𝐹𝐴𝑅3𝑂
𝐹𝐴𝑅 =
0.21
Since the nitrogen flow in the AR3 stream, FAR3N is: 𝐹𝐴𝑅3𝑁 = 0.79𝐹𝐴𝑅3
Phosphorus current (HP3)
The phosphorus that yeast needs is satisfied by and by the phosphoric acid that it also helps to
maintain the desired pH.
The calculations are made based on P2O5, as it is in the form of this oxide that phosphorus
concentrations in molasses and yeast are tabulated.
𝑁𝑃𝐿 = 𝐹𝑃𝐿 × 𝑀𝐹3 (Equation 10)
𝐹𝑀
8𝑎𝑐
𝑀𝑃 = 𝐹𝑃𝑀 × ( 𝑓𝑎𝑚 ) + 𝐵𝑅𝐸𝐴𝐷 (Equation 11)

Being:
NPL = Need for phosphorus in yeast
FPL = Fraction of phosphorus in yeast
MP = Total mass of phosphorus to be supplied
FPM = fraction of phosphorus in molasses
fam = sugar fraction in molasses
PO = amount of phosphorus (in P2O5) that needs to be added
Equating equations 10 and 11, the value of PO is removed, which is converted to acid
phosphoric:
𝑞𝑢𝑎𝑛𝑡𝑖𝑡𝑦 𝑜𝑓 𝑏𝑟𝑒𝑎𝑑 × 2 × 𝑃𝑀 𝐻3𝑃𝑂4
𝐻𝑃3𝑃 =
𝑃𝑀 𝑃2𝑂5
To avoid large pH gradients inside the fermenters, phosphoric acid isadded diluted to 5%.
Therefore, and in a similar way to the calculation made for theammonia, the flow rates, total
and water in the PH3 stream are calculated.
The results are shown in Table 4.
Sulphuric acid stream (HS3)
PH control is important to avoid contamination, however, it is important thatdo not go down to
values that inhibit optimal growth. The optimum value is in the 4.5 rangeto 6.5 [23], assuming a
pH value of 5. This control is done by addingsulphuric acid.
The compounds that most influence pH are ammonia and phosphoric and sulphuric acids.
In this balance, it is considered that acids and bases completely dissociate: theammonia and
phosphoric acid, because the cells consume nitrogen and phosphorus,respectively, shifting the
balance towards dissociation; sulphuric acid,because their dissociation constants arehigh. [34]

NH3 + H2O ↔ 𝑁𝐻4++ 𝑂𝐻 − Kd = 1.8 × 10−2

𝐻3𝑃𝑂4 ↔ 𝐻2𝑃𝑂4− + 𝐻 + Kd = 1.7.6 × 10−3

𝐻2𝑃𝑂4− ↔ 𝐻𝑃𝑂42− + 𝐻 + 𝐾𝑑 = 6.3 × 10−8

𝐻𝑃𝑂42− ↔ 𝑃𝑂43− + 𝐻 + Kd = 4.4 × 10−8
𝐻2𝑆𝑂4 ↔ 𝐻𝑆𝑂4− + 𝐻 + Kd = high
𝐻𝑆𝑂42− ↔ 𝑆𝑂42− + 𝐻 + Kd = 1.2 × 10−2
Taking stock, the following equation is obtained:

2 × [𝐻2 𝑆𝑂4 ] + 3 × [𝐻3 𝑃𝑂4 ] − 1 × [𝑁𝐻3 ] = 10−5

Transforming concentrations (mol / L) into mass (ton):
2 × 𝑚(𝐻2 𝑆𝑂4 ) 3 × 𝑀(𝐻3 𝑃𝑂4 ) 𝑀(𝑁𝐻3 )
+ − = 10−5
𝑉 × 𝑃𝑀𝑆 𝑉 × 𝑃𝑀𝑃 𝑉 × 𝑃𝑀𝑁
Being:
V = volume of liquid at the end of propagation in the 3rd stage (m3)
Pmi = molecular weight of compound i (kg / mol)
m (i) = mass of compound i (ton)
Taking into account the reason given for the phosphoric acid stream, sulphuric acid is
added to 5%.
Adopting the same strategy previously used in determining the flow rate of the
current of phosphoric acid, partial and total water flow rates (values presented in Table 4)
Yeast output current (L3)
From this current, the yeast flow and composition [23], 8% (mass), are already known. IT'S the
total flow per departure is also known.
To determine the other partial flows of this current, it was admitted that the composition of
thecurrent L3 is the same as the inoculum current (I3). Making a global mass balance,identical
for solid and inert impurities.
For example:
𝐹𝐿1𝑖 𝐹𝐿1𝑖
= (Equation 12)
𝐹𝐼3 𝐹𝐿1

𝐹𝐿1𝑖 = 𝐹𝐼3𝑖 + 𝐹𝑀8𝑖 (Equation 13)

Being:
FI3i = inert flow in current I3
FI3 = total current flow I3
FL1i = inert flow in L1 current
FL1 = total flow of current L1
FM8i = inert flow in the M8 current
Solving equations 12 and 13 simultaneously, the values of FL3i and FI3i are obtained.
For solid impurities it is done in exactly the same way as for inert ones.
The flow of water in this stream cannot be calculated stoichiometrically, although figure in the
biomass growth equation, due to the hydrolysis of sugars. Thus therefore, the flow of water in
this stream is determined by difference.
The composition of the L1 current is determined by the quotient between the partial flows and
the total flow (Table 4).
Yeast inlet current (I3)
The flow rate of aggregates and solid impurities was determined in the description of the
calculations ofprevious current. The yeast flow and the total are already known. The
composition of thisyeast current is equal to L1 current (current I3 is the output current of the
2ndphase). The only calculation that is not determined is that of water, as in the current
above, it is also calculated by difference.
The composition is obtained in the same way, dividing the partial flows of the current I3 by
theits total flow. The results are compiled in Table 4.
Water flow (AP5)
This chain has the purpose of maintaining the optimum operating conditions, in terms
ofviscosity and concentration. To calculate the flow of this current, it is necessary to make
aoverall mass balance to all fermenter streams, then, by differencethe value of this current is
determined.
Balance at the 2nd stage of propagation (2nd stage fermenter - F2)
Bearing in mind that the decisions taken for this stage are the same as those described for the
3rdpropagation stage, both flow rates and current stream compositionsfermenters are
calculated using the same strategy and, therefore, theirdescription. However, it should beborne
in mind that the fraction of oxygen absorbed is different (31%), as already mentioned in the
calculations made for the third stage.
The results obtained for this fermenter are summarized in Table 5.
Balance to the 1st stage of propagation (1st stage fermenter - F1)
At this stage, the same decisions made in the previous stages are assumed. The parameters
that assume different values are the fraction of oxygen absorbed (12.5%) and thepH, which is
maintained at 4.5, as a precautionary measure against contamination. [23]
The results obtained in this unit are shown in Table 6.

4.7. Mass Balances to the Separation and Purification Section of Yeast

4.7.1. Balance to yeast centrifuge (CL)
With this unit, the objective is to concentrate and wash the current that leaves the 3rd stage.
This balance is the same for both yeasts, given that the composition of the current here treated
is the same in both cases.
The basis of calculation in this unit is a departure.
To perform the calculations, we admit the following:
- There are 2.5% losses;
- The solid impurities all come out in the L2 stream;
- The ratio of inert mass to water mass is the same in both currents that leave of this unit (both
currents are in equilibrium);
-The wash water flow (current AP6) is the same as the wash flow (current L2).
The yeast concentration is 20% in the L2 stream [23].
Making a mass balance to yeast, and taking into account the considered efficiency, we have to

𝐹𝐿2𝑙 = 𝑒𝑓 × 𝐹𝐿1𝑙
𝐹𝐸𝐿2𝑓 = (1 − 𝑒𝑓) × 𝐹𝐿1𝑙

Being:
FL2l - yeast flow in L2 stream
FEL2l - yeast flow in the EL2 stream
ef - centrifuge efficiency (0.975)
FL1l - yeast flow in stream L1
The calculation of the current L2, FL2, is done through the flow rate of the yeast current and
composition in the aforementioned stream:
𝐹𝐿
𝐹𝐿2 = 𝑋 2𝑙 , 𝑋𝐿2𝑙 being the yeast fraction in stream L2
𝐿2𝑙

The flow rate of solid impurities in this stream is equal to the flow rate of impurities in the
streamL1.
The numerical value of the flow rate AP, as already mentioned, is equal to the flow rate of the
current L2.
There is still a need to calculate the flow rates for water and aggregates in both streams.
Therefore, the ratio between them is constant:
𝑀𝑇𝐴𝐸 (𝐹𝐿1𝑎 + 𝐹𝐴1 )
=
𝑀𝑇𝐼𝐸 𝐹𝐿1𝑖
Being:
MTAE - total body of water entering this unit
MTIE - total mass of aggregates entering this unit
FL1a - water flow in stream L1
FL1i - inert flow in L1 current
AND,
𝑀𝑇𝐴𝐸 𝐹𝐿2𝑎
=
𝑀𝑇𝐼𝐸 𝐹𝐿2𝑖
Being:
FL2a - water flow of L2 stream
FL2i - L2 current inert flow
And, making a balance to the current L2, we have:

𝐹𝐿2𝑎 + 𝐹𝐿2𝑖 = 𝐹𝐿2 − 𝐹𝐿2𝐿 − 𝐹𝐿2𝑖𝑚

By balancing water and aggregates throughout the unit, the flow rate can be calculatedof these
in the EL1 current.
The compositions of both output currents are calculated by dividing each of the partial flows by
total flow. The results are summarized in Table 7.
4.7.2. Swing to rotary bar yeast filter:
This unit is the first that is distinguished in the production of both yeasts and aims toobjective
to concentrate the yeast.
It is located after the fresh yeast tank, operating in a continuous regime.
We admit, like the centrifuge that:
- The solid impurities all come out in the L4 chain;
- The mass ratio of inert / mass of water is the same in both currents that leave this
unity (are in chemical equilibrium);
- The wash water flow (current AP7) is the same as the wash flow L2.
The basis of calculation: 1 hour
The flow rate of current L3, FL3, is:
𝐹𝐿2 × 𝑓𝑏
𝐹𝐿3 =
𝑡
Being:
fb - fraction of bar yeast to be produced
t - time and a start
The composition of this current is the same as that of the L2 current and the partial flows
aredetermined by multiplying the total flow rate of the L3 current by its composition.
Bearing in mind that this unit is the last one where losses are considered, the flowof yeast in
the output stream L4, FL4l, is necessary to satisfy the annual production intended:
 𝑃𝐿𝐹𝐵
𝐹𝐿4𝑙 =
𝑡𝑎𝑝
Being:
PLFB - Yeast Bar Production
tap - annual production time
𝐹𝐿
Yield is calculated by: 𝐹𝐿4𝑙
3𝑙

At the end of this operation, the yeast has practically the composition of the finished
product,only the additive mixture is missing, which constitute 0.15% of the final mass [23].
The composition of the current L4, χL4l, in yeast, is thus determined, taking into account the
final composition and the fraction of additives to be added:
𝑋𝐹𝐵
𝑋𝐿4𝑙 =
(1 − 𝑓𝑎)
Being:
𝜒𝐹𝐵 - yeast composition in yeast bar (finished product)
fraction of additives in baking powder (finished product)
Since the flow rate and the yeast fraction in the L4 current are known, it is possible to calculate
the total flow of current L4, FL4:
𝐹𝐿4𝑙
𝐹𝐿4 =
𝑋𝐿4𝑙
This flow rate is the same as the current AP7, as we admitted at the beginning.
The calculation of the flow of aggregates and water, in both outlet streams is carried out from
the same way as that performed in the centrifuge. The results are compiled in the Table 8.
Swing to mixer (unit M)
It is in this unit that the additives are mixed in the yeast, being subsequently
homogenized.
The composition of additives in yeast bars, as already mentioned, is 0.15% [23].
Calculation basis is 1 hour.
The flow rates that make up the L5 current are the same as those of the L4 current, with the
exception of the additions.
The total flow of current L5, FL5, is then determined by:
(𝐹𝐿5𝑙 + 𝐹𝐿5𝑎 + 𝐹𝐿5𝑖 + 𝐹𝐿5𝑖𝑠)
𝐹𝐿5 =
(1 − 𝑓𝑎𝑑)
Being:
FL5l - yeast flow in L5 stream
FL5a - water flow in L5 stream
FL5i - inert flow in L5 current
FL5is - flow of solid impurities in L5 stream
fad - fraction of additives in the final yeast bar
The flow rates of additives in the AD and L5 currents are calculated as follows: FL5 x% of
additions.
The composition of the chain is determined by the same methodology already used for other
currents in previous units. The results are grouped in Table 9.
5. Energy Balances
5.1. Introduction to energy balances [26]
In the process design, energy balances are made to determine the requirements for
process energy: the necessary heating, cooling and motive power. At the
operation of a factory, an energy balance will show the pattern of utilization of the
and will suggest areas for conservation and savings.
The general energy conservation equation is given by:
∆E = -∆ (U + pV + K + P) + Q + w
Where of the first member means the difference between the final state and the initial state
(inters) and the  of the second term means the difference (or balance) between the
contributions from all outgoing streams and contributions from all streams entrance. K
represents kinetic energy, P represents potential energy, U represents internal energy, Q
represents heat and W represents work.
What is despised:

 The potential energy (∆P), since the difference in elevation between the input currents
and output is negligible;
 The kinetic energy (∆K), because the speed of entry and exit of the currents is
approximately equal;
 The work (W), because the work done by the system is very small;
Therefore,∆E = -∆U + Q, and in steady state: ∆H = Q

5.2. Energy balances - Introduction to calculations

The calculations necessary to carry out energy balances include the following
equations:
𝑄 = 𝐹 × 𝑐𝑝 × (𝑇 − 𝑇𝑟𝑒𝑓) (equation 14)
𝑖𝑛 = 𝑜𝑢𝑡 (equation 15)
𝑖𝑛 = 𝑄𝑖 + 𝐶𝐹 + 𝐶𝐺 (equation 16)
𝑜𝑢𝑡 = 𝑄𝑜 + 𝐶𝑅 (equation 17)
Being:
F - current flow mass
Cp - current (or component) heat capacity
Q - current enthalpy
In- total energy entering the unit
out- total energy coming out of the unit
Qi - energy of the currents that enter the system, obtained through Equation 14
Qo - energy of the currents leaving the system, obtained through Equation 14
T - chain temperature
Tref - reference temperature
CF - heat supplied from outside
CG - heat generated by the system
CR - heat removed from the system
In these balances, in addition to calculating the energy content of the currents, the values for
the heat generated and exchanged are determined. The following are not yet chosen:
exchange systems or utility needs (cooling water). At the However, whatever the choice, they
will have to provide the power values determined in these swings.
The mixing heat is supposed to be zero, that is, the mixtures are considered ideal.
It is defined for reference temperature 30ºC, as it is a value that simplifies the calculations.
For the pressure, it is assumed as a reference value of 1 atm, to which the entire process
occurs, except compressed air streams that have higher values, in order to be able to
overcome the pressure of the height of the liquid inside the fermenters. Given that
fermenters are not yet dimensioned, the pressure is not yet known, in a first approach, the air
pressure is equal to 1 atm. The states of considered are as follows:
- Air (oxygen, carbon dioxide and nitrogen) - gaseous state;
- Molasses and yeast - solid state;
- Water, ammonia (25%), sulphuric acid (5%) and phosphoric acid (5%) - liquid state.
The ambient temperature is assumed to be 20ºC. Therefore, we admit that the currents
of water, molasses, acids and additives are all at this temperature.
Bearing in mind that in the whole process the temperature is not significantly different
the ambient temperature, heat losses in the units are disregarded.

5.3. Energy balances to the culture medium preparation section

5.3.1. Swing to molasses filter (FB unit)
In this unit, the enthalpy values of the input currents are calculated by applying the equation
14.
Calculation basis: 1 hour
Bearing in mind that this unit does not generate heat (enthalpy of the null mixture) and
molasses is stored at room temperature, there is no energy exchange with therefore, the
temperature of the output currents M2 and ESC are the same as those of the input.
We admit that the heat capacity of current M2 is equal to that of M1, since the composition
this does not change significantly.
Applying equation 14, to the M2 current we determine its energy content:
𝐸𝑀2 = 𝐶𝑢𝑟𝑟𝑒𝑛𝑡 𝑚𝑎𝑠𝑠 𝑀2 × 𝐶𝑝 × (𝑇 − 𝑇𝑟𝑒𝑓)
Substituting equations 16 and 17 in equation 15, the energy flow of ESC is calculated,
EESC:

𝐸𝑀1 + 𝐸𝐴𝑃1 = 𝐸𝑀2 + 𝐸𝐸𝑆𝐶

The results of the calculations performed for this unit are compiled in the
Table 10.

5.3.3. Swing to molasses sterilizer (unit in)

In this unit, molasses are preheated with the stream of molasses already sterilized.
Then, they are heated up to 416.15K with steam to sterilize. After pre-heat the incoming
molasses, water is added to cool them down to the temperature at which fermentation takes
place, 303.15K.
Calculation basis in this unit is from a start.
The energy contents of currents AP2 and M4 are determined using equation 14 the supplied
heat, CF, is calculated by substituting equation 16 in equation 15. The results obtained are
compiled in Table 12.

5.4. Balance to the yeast propagation phase

5.4.1. Balance to the 3rd propagation stage (unit F3)
 In these units, it is intended that the operation is as isothermal as possible, at 303.15K, with all
the interest that the inoculum and final temperatures are equal to 303.15K, either for reasons
of adapting the yeast to the medium or for procedural issues. During this operation, a large
amount of heat is generated. It is therefore necessary cool the fermentation medium. The
inoculum is always at the medium temperature, 303.15K, as it comes from another fermenter.
The air stream, which is at room temperature, when compressed, heats up, allowing it to if it
reaches the temperature of the medium. The gaseous stream leaving these units does not
suffer neither heating nor cooling in relation to the one that enters, as it is equal to the medium
temperature. Calculation basis in this unit is a departure. The energy content of all currents is
determined using equation 5. The heat formed (CF) is:
𝐶𝐹 = 𝐻𝑅 × 𝐿𝐹
Where:
HR - heat of the reaction at 30ºC, 3870 kcal / kg yeast produced [25]
LF - yeast dough produced (kg)
The heat to be removed, CR, is determined, substituting equations 8 and 5 in equation 6,
obtaining
if:
𝐶𝑅 = 𝐸𝑁𝐻3 + 𝐸𝐻𝑃3 + 𝐸𝐻𝑆3 + 𝐸𝐴𝑃3 + 𝐶𝐹
The results of these balance sheets are compiled in Table 13.

5.4.2. Balances at the 1st and 2nd stages of propagation (units F1 andF2)
In these stages the calculations are made similar to those made for the 3rd stage.
Tables 14 and Table 15 summarize the values obtained for these two fermenters.

5.5. Energy balance to the separation and purification section of the

yeast
5.5.1. Balance to yeast centrifuge (CL unit)
The only sources of heat in this unit are the input currents.
It is assumed that the output currents (L2 and EL1) are in thermal equilibrium, that is
that is, they are at the same temperature.
Basis of calculation: one item
The energy flow of the current of the current AP6 is determined using equation 5.
Substituting equation 14 in equations 16 and 17 and these, in turn, in equation 15,
the temperature of the output currents is determined. Bearing in mind that the capacity
heating is a function of temperature, the resolution must be made through a processiterative:
- a temperature is assumed, which is the average of the inlet flows;
- consult the value of Cp [36]
- solve the following equation in order of T: 𝑄𝑖 = 𝐹 × 𝐶𝑝(𝑇) × (𝑇 − 𝑇𝑟𝑒𝑓 )

a new value is read for Cp

The energy content of the output currents is calculated by applying equation 14 to each
one of them.
The results are compiled in Table 16.

5.5.2. Swing to heat exchanger

The purpose of this unit is to cool the yeast stream down to the temperature of
276.15K [22]

5.5.3:Balance to the rotary filter:

The results are shown in table ().