TECNOLGICO NACIONAL

DE MXICO EN CELAYA
Ingeniera Bioqumica
Ingeniera de Biorreactores

PROYECTO FINAL:

OPTIMIZATION OF BAKERS
YEAST FERMENTATION SIMULATOR

Autores:

Daniel Ortiz Leal

Cecilia Cervantes Briseo

Docente:

MC. Ildefonso Prez Yez

INDEX

1 - ABSTRACT 3

2 - INTRODUCTION 3

3 - BACKGROUND 4

4 - GOALS 8

5 - METHODOLOGY 8

6 - RESULTS AND DISCUSSIONS 12

7 - CONCLUSIONS 16

8 REFERENCES 17

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 1. ABSTRACT

Due to the difficult of making a bioreactor scale-down, a simulation was carried out to
observe the behavior of bakers yeast. The simulator program BY-SIM, compiled in Borland
C++ was used. Based in the default parameters that the program includes, were modified
to improve the yield of production and economics status were optimized. There were two
operational options, using burble column and stirred vessel. In this work, was selected the
second option.
The general aim of this research was find the variables needed to improve the yield of
production, and change them to get the best economics based in the program
configuration.
The kinetics parameter of de microorganism and the geometrical and properties of the tank
were not modified, but only the variables that we can change in a continuous fermentation
process with same characteristics.
The results show that the principles variables involved were oxygen rate, substrate initial
concentration and both in broth as in cooling area and all of them was increase.

2. INTRODUCTION

Bioreactors are vessels or tanks in which whole cells or cell-free enzymes transform raw
materials into biochemical products and/or less undesirable by-products. The microbial
cell itself is a miniature bioreactor; other examples include shake flasks, Petri dishes, and
industrial fermentors. Diagnostic products based on enzymatic reactions, farm silos for
silage fermentations, bread pans with fermenting yeast, and the soil in a Kansas wheat
field may also be viewed as bioreactors. While the bioreactor may be simple or highly
instrumented, the important consideration is the ability to produce the desired product
or result. (Neymann, Wegerhoff, & Engell, 2010)

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From the biochemical engineering point of view, the straightforward way of improving
the economics is to invest in process optimization and control. In industrial practice,
improvements are often achieved by educated trial and error methods, by empirical
methods guided by intuition and experience. (Yuan, Guo, & Bellgardt, 1995)

Process optimization, supervision and control are becoming an increasingly important

issue due to hard competition between companies. Still, the acceptance of model-based
methodologies remains rather limited in the industry, mainly because the benefit/cost
ratio is not clearly attractive for such developments. In the classical model-based
approach, mathematical models are used to describe the relevant bioprocess
mechanisms. ( Freitas Oliveira, 1998)

Most often, mathematical process models are considered to represent the a priori
knowledge. When we speak about models in biochemical engineering and with respect
to process optimization and control, we think of relationships, which describe the basic
aspects of the real process that are most important to process performance. (Cooney,
Wang, & Wang, 1977)

3. BACKGROUND

Automatic process control in industrial fermentation is currently limited to the control of

temperature, pH, and, in a few cases, dissolved oxygen. Many investigators. However,
are now examining more sophisticated control systems, computerized systems that
monitor and analyze data while directly coupled to the fermentation. The primary
objective of many fermentations is productivity, or the conversion yield of product from
the carbon-energy substrate. This goal is especially true for fermentation cost intensive
processes, where fermentation costs exceed recovery costs.

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To achieve effective control, one must continuously assess the product and residual
substrate concentrations. (Cooney, Wang, & Wang, 1977)
In the production of biomass by either batch or continuous culture, the major
manufacturing expense is the cost of the carbon-energy source. With the rapidly
fluctuating, and generally increasing, cost of raw materials, a high efficiency of conversion
of the carbon source to biomass is critical to a successful process. When batch culture is
used for production, as in the case of bakers yeast, one must also maintain a high
volumetric productivity to get the most product per unit of capital investment. This
constant maintenance of a high yield and high productivity for each batch is usually
difficult. (Y. WANG, L. COONEY, & I. C. WANG, 1977)

In industrial practice, process control of baker's yeast production is performed usually

based on the on-line measurement of respiration ratio or ethanol concentration in
fermenter with the aim of maximization of yield. while other empirical dispatching
decisions are carried out to maintain a high productivity. Clearly, the profiles of molasses
feeding rate for maximal yield may be contradictory with that for maximal productivity. In
this case, a general objective function, named as the profit function, may be used for
global process optimization. The profit function is defined as the gross profit of a process
divided by the production period For a given yeast plant. the gross profit may be
evaluated by material and energy balances. Maximizing of the profit function will
guarantee the whole plant at its global optimum. Meanwhile, the yield and productivity
indices will be optima11y coordinated. In fact, for many other industrial processes,
maximization of profit function is also a generic target. (Guo & Bellgardt, 1995)

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With exception of cooling limitation problems usually found in large bioreactors, the most
frequently encountered limitation to growth in aerobic fermentations is dissolved oxygen
in the broth. Oxygen has a very low solubility in fermentation media in comparison to
other typical substrates. Therefore, it must be continuously supplied, usually by aeration.
The mass transfer capacity between gas and liquid phases is a central problem in aerobic
bioprocesses design and operation.
The mathematical description of the oxygen transfer rate from the gas phase into the
liquid phase is based on general concepts of mass transfer theory. (R. Omstead, 1989)

The mathematical description of microorganisms growth kinetics is a critical issue in

bioprocess modelling. Quite often kinetic models are based on unstructured and
nonsegregated cell models. Unfortunately, in many cases, such models are not accurate
enough to solve the problems in study. The other critical issue is related to the
identification of kinetic parameters. Parameter identification requires a careful and
expensive experimental planning. As such, there is a clear incentive to develop algorithms
for state estimation and parameters estimation while avoiding the knowledge of the
underlying kinetic model. ( Freitas Oliveira, 1998)

The need of a kinetics model constitutes a key point in this discussion. The reaction rates
are most often a very complex relation of the operating conditions and of the state of the
process. The construction of a suitable kinetics model may constitute a very difficult task,
if not an impossible one. As such, there is a clear incentive to design monitoring and
control algorithms for bioprocesses with a minimal modelling of the kinetics. (Guo &
Bellgardt, 1995)

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The building and use of mathematical models based on observed data is for long
accepted as a basic scientific methodology. Models may be of a more or less formal
character, but they have the basic feature that they attempt to link observations together
into some pattern.

With the progress in digital technology, and thinking of bringing the theory into practice,
computational modelling and model-based applications have emerged and are
recognized as areas of great priority. (R. Omstead, 1989)

The conventional approach for process modelling is based on the balance equations for
mass, energy, and, if necessary, momentum and population. This form of modelling
requires further knowledge about reaction kinetics, thermodynamic, transport and
physical properties. (Leo, C. P., & Soares, F. O.,2002)

Real processes in the chemical, biochemical and food industry are in their clear majority
non-linear MIMO systems (Multiple Input Multiple Output). Their dynamics and control
are difficult to study both for theoretical and practical reasons. In many instances
experiments with real industrial processes are not carried out for reasons of economy and
safety. Often on-line measurements are not available or simply they are too expensive.
The simulation procedure represents an important tool to understand clearly the bakers
yeast fermentation process. (Leo, C. P., & Soares, F. O., 2002)

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 4. GOALS

GENERAL: To find the correct variable that, in a real process are adjustable, to improve
the yield of production in a reactor among the simulation in the program BYSIM compiled
in Borland C++
SPECIFIC: To determinate the values on the variables involves and adjustable that give
the optimal economics status in the program BYSIM.

5. METHODOLOGY

Based in the analysis descripted in the article Two different strategies for bakers yeast
fermentation process simulation and State estimation of a fed-batch baker's yeast
fermentation about the differential equations that explains the behavior of the ethanol,
biomass, oxygen rate, carbonic dioxide generation and glucose into the reactor is used
to understand how, mathematically, the variables are involved. As follow, this ED are in
present in the code of the program.

5.1 Kinetic model

Yeast growth is characterized by three metabolic pathways

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where S represents glucose; O oxygen; X biomass; E ethanol; C carbon dioxide and
, , : specific growth rates for the three pathways. In the sequel X, S, E, O, C mean
concentrations.

The metabolic pathways of fermentative growth on glucose and oxidative growth on

ethanol are competitive. This competition is governed by the respiratory capacity of the
cells. If the instantaneous oxygen uptake capacity exceeds the oxygen need for total
respiratory glucose uptake, then, all sugar uptakes follows the respiratory pathway (1) with
the remaining oxygen being spent on ethanol respiratory uptake (3).
Otherwise, if the instantaneous oxygen uptake capacity is not enough, then, part of
glucose uptake follows the respiratory pathway (1) while the remaining follows the
fermentative pathway (2).

The kinetics equations for bakers yeast growth, considered as Monod equations, are
determined as follows. The total specific growth rate, is the sum of the growth rates for
the three pathways.
= + +
The specific growth rates, , can be related to the corresponding substrate fluxes, q,
and yield coefficients (Table 1), Y, by

where /

and /

represent the yield coefficients of biomass in glucose in the oxidative

and fermentative phases, respectively; /

is the yield coefficient of biomass in ethanol

in the oxidative phase in ethanol. As ethanol uptake is influenced by the priority of glucose
uptake, which functions as an inhibitor, the specific growth rate on ethanol can be
described as

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where is the maximal specific growth rate, Ki is the inhibition parameter and KE is
the saturation parameter. However, this equation holds true only if there is an available
respiratory capacity of the cells.

The glucose uptake, qs, is slightly different because it follows two metabolic pathways:
oxidative and fermentative

The glucose, , and oxygen, 0 , uptake follow a Monod kinetics, respectively

where max , is the maximal specific glucose uptake rate, Ks and Ko are saturation
parameters and 0 , is the maximal specific oxygen uptake rate.

Two situations may occur: excess of oxygen that implies no fermentative growth of
biomass; lack of oxygen and consequently excess of glucose that implies no respiratory
growth on ethanol.

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Two auxiliary equations must be added to the equations for the estimation of the specific
growth rate on ethanol, defined as:

5.2 Mechanistic model

The mechanistic model for the fed-batch fermentation is obtained from mass balances
for all components, considering that the reactor is well mixed. Furthermore, it is assumed
that the yield coefficients (Ys) are constant and the dynamics of the gas phase can be
neglected.
Then the set of differential model equations is:
* Mass balance for the biomass

* Mass balance for sugar

where Sf is the substrate concentration in the initial feed.

* Mass balance for the ethanol

* Mass balance for the oxygen

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* Mass balance for the carbon dioxide

The accumulation of the working volume during the fed-batch process is represented by:

The dilution rate (ratio feed/volume), D, is defined by:

The gas transfer rates are given by:

where are overall mass transfer coefficients for oxygen and carbon dioxide and O*
and C* are the corresponding equilibrium concentrations.

6. RESULTS AND DISCUSSIONS

Figure 1 show the initial values in the reactor configuration.

Figure 1 Default Reactor configuration

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Figure 2 show the initial parameters about every yield and coefficients.

Figure 2 Default kinetic Coefficients

Figure 3 show the initial values for each concentration in different feeds.

Figure 3 Default initial concentration and feed flow concentration

Figure 4 show the economics status given the simulation.

Figure 4 Economics of simulation.

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Figure 5 show the plot that describes the behavior of Ethanol, Glucose, Biomass and
Oxygen and the total volume into the tank. Also, we se the final results of production with
default configuration.

Figure 5 Plot 2D of the simulation.

Now, changing the values and making different probes, were found the next changes for
optimize the economics status
Broth Temperature| From: 30C to 33C
Cooling Temperature| From: 1.5C to 4.0C
Substrate initial| From 2.5E+03 to 3.5E+03 Cmol/m3
Substrate feed| From 1.E+03 to 1.2E+03 Cmol/m3
Oxygen initial| From 1.50E+03 to 3.0E+03 Cmol/m3
Oxygen feed| From 1.50E-01 to 2.50E-01 Cmol/m3

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Figure 6 show the plot after the modifications of initial values

Figure 6 Plot 2D of second run of program

Figure 7 show the economics with the modifications

Figure 7 Economics status after modifications.

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In the first run of program with default configuration in the parameters and values, we
see in the plot that glucose decreasing exponentially, and volume has a similar, but inverse
behavior. However, the ethanol suffers a spontaneous decreasing after 9.6 hours of the
fermentation process. We see to that oxygen suffers an oscillation period around ten 10
hours, just after the decreasing of ethanol.

In the second plot of the process, we see that the consumption of glucose is lower, but
the increasing of volume still with good inclination. The oxygen suffers a rapid decreasing
in the first fraction of hour, then still a good level with a low decreasing. Its observed that
ethanol is in an exponential period of growth as the volume too. The biomass has a step
by step growth, keeping a regular level in the total volume.

About the economics, is observed an improve on the status but obviously there are thing
we need to considerate to make this changes in real life. For example, the initial and the
feed of glucose was increase, but the same happened to the yield and the production.
According to the modifications to the feed of oxygen, we going to spend more electrical
energy. Just after a global balance of entries and outs of money will show the rentability
of make those changes in a real process. However, in the simulator we see the
optimization of economics and the improve of global yield.
7. CONCLUSIONS
The optimization of Bakers Yeast simulator was done. We obtained best result changing
the initial values, but to recommend the changes in a real process depends of and deeper
study in economic engineering. We see that the variables that concerns to the production
yield in a fermentation process is glucose dilution in the feed, how this is continuously
reposed to keep the conditions and how ethanol is present in the process and this affect
the production. Is need a huge quantity of oxygen to make favorable a respiratory growth
on glucose .

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 8. REFERENCES

Freitas Oliveira, R. M. (1998). Supervision, Control and Optimization of Biotechnological

Processes. (R. Neubert, Ed.) Germany.

Cooney, C. L., Wang, H. Y., & Wang, D. I. (January de 1977). Computeraided material
balancing for prediction of fermentation parameters. Biotechnology and
Bioengineering, 19(1), 55-67. doi:10.1002

Duro Vieira, ., Stupiello Andrietta, M., & Andrietta, S. R. (2013). Yeast biomass production:
a new approach in glucose-limited feeding strategy. Brazilian Journal of
Microbiology, 44(2), 551-558. doi:1678-4405

Guo, S. R., & Bellgardt, J. Q. (1995). SIMULATION OF PROFIT OPTIMIZATION FOR

INDUSTRIAL BAKER'S. IFAC Computer Applications in Biotechnology, 251-254.

Neymann, T. C., Wegerhoff, S., & Engell, S. (July de 2010). Modelling and Simulation of
Budding Yeast Cultures. (1. I. Biotechnology, Ed.) IFAC Proceedings Volumes, 461-
4666. doi:10.3182/20100707

R. M., D. (Agosto de 1983). State Estimation of a Fed-Batch Baker's Yeast Fermentation. IFAC
Proceedings Volumes, 16(14), 201-211. doi:10.1016

R. Omstead, D. (1989). Computer Control of Fermentation Processes. (C. Press, Ed.)

doi:084935496X, 9780849354960

Y. WANG, H., L. COONEY, C., & I. C. WANG, D. (1977). Computer-Aided Baker's Yeast
Fermentations. BIOTECHNOLOGY AND BIOENGINEERING, XIX(1), 69-86.
doi:10.1002

Yuan, J. Q., Guo, S. R., & Bellgardt, K. H. (1995). SIMULATION AND PROFIT ESTIMATION
FOR BAKER'S YEAST. IFAC Computer Applications in Biotechnology., 171-176.

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