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![Page 1: On the mathematical modelling of a batch fermentation process using interval data · PDF file · 2014-10-14On the mathematical modelling of a batch fermentation process using interval](https://reader031.vdocument.in/reader031/viewer/2022030502/5aaf5bc27f8b9a5d0a8d541c/html5/thumbnails/1.jpg)
September, 21–26, 2014, Wurzburg SCAN 2014
On the mathematical modelling of a batch
fermentation process using interval data
and verification methods
Svetoslav Markov
September 21, 2014
Co-authors: R. Alt, V. Beschkov, S. Dimitrov and M. Kamburova
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September, 21–26, 2014, Wurzburg SCAN 2014
Contents
1. Classical cell growth models
2. Two-phase cell growth models based on reaction schemes
3. Numerical experiments on real interval data: model verification
4. Open problems and conclusion remarks
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September, 21–26, 2014, Wurzburg SCAN 2014
Microbial growth (cell growth)
Microorganisms — procariotic, eucariotic
Procariotic — bacterial cells
Cells transform nutrient substrates (via their enzymes) into products
Products = intermediate metabolic subtances + final excrements
In addition, (some) cells divide and the cell population grows
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September, 21–26, 2014, Wurzburg SCAN 2014
Microbial growth—Malthusian growth model
Malthusian growth model, Malthus 1810–1825:
dx/dt = kx, x(0) = x0 x(t) = x0ekt
Xk−→ X +X reproduction
More realistic assumption: S +Xk−→ X +X leading to
dx/dt = ksx
exponential unlimited growth
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September, 21–26, 2014, Wurzburg SCAN 2014
Microbial growth—Verhulst-Pearl model
Verhulst-Pearl model (1838):
dx/dt = kx− (k/a)x2 = (k/a)x(a− x), x(0) = x0
x(t) =a
1 + be−kt, b = (a− x0)/x0
S +Xk−→←−k−1
X +X reproduction !
dx/dt = ksx− k−1x2, x(0) = x0
if s = const then same as above
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September, 21–26, 2014, Wurzburg SCAN 2014
Microbial growth - phases
(A) lag phase: During lag phase, bacteria adapt themselves togrowth conditions. It is the period where the individual bacteria arematuring and not yet able to divide. During the lag phase of thebacterial growth cycle, synthesis of RNA, enzymes and othermolecules occurs.
(B) log phase: Exponential or log phase is a period characterized bycell doubling. The number of new bacteria appearing per unit time isproportional to the present population. If growth is not limited,doubling will continue at a constant growth rate so both the numberof cells and the rate of population increase doubles with eachconsecutive time period.
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September, 21–26, 2014, Wurzburg SCAN 2014
Microbial growth - phases
(C) stationary phase: During stationary phase, the growth rateslows as a result of nutrient depletion and accumulation of toxicproducts. This phase is reached as the bacteria begin to exhaust theresources that are available to them. This phase is a constant valueas the rate of bacterial growth is equal to the rate of bacterial death.
From: http://en.wikipedia.org/wiki/Bacterial growth
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September, 21–26, 2014, Wurzburg SCAN 2014
Bio-reactors: three types
−→ � −→[S +X −→ P +X] batch
S −→ [S +X −→ P +X] fed-batch
S −→ [S +X −→ P +X] −→ P,X continuous
Dynamics of S needed
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September, 21–26, 2014, Wurzburg SCAN 2014
Bio-reactors: Monod model—functiom µ
ds/dt = −αµ(s)x [+F (sin − s)]dx/dt = µ(s)x− kdx [−Fx]
x = x(t) — the biomass
s = s(t) — substrate concentration
kd — decay constant (death rate)
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September, 21–26, 2014, Wurzburg SCAN 2014
Monod model—functiom µ
µ(s) = µmaxs
Ks + s
µ(s) = µmaxs
Ks + s+ s2/Ki
over 30 expressions for µ collected in :
M. Gerber, R. Span: An Analysis of Available Mathematical Modelsfor Anaerobic Digestion of Organic Substances for Production ofBiogas, proc. IGRC, Paris 2008.
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September, 21–26, 2014, Wurzburg SCAN 2014
Classical cell growth models
A batch mode chemostat/bioreactor
ds/dt = −αµ(s)x, (1)
dx/dt = µ(s)x− kdx, (2)
with s(0) = s0 > 0, x(0) = x0 > 0
Monod function:
µ(s) = µmaxs
Ks + s, Ks ≥ 0 (3)
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September, 21–26, 2014, Wurzburg SCAN 2014
Classical unstructured models with product
A classical model for batch cultivation of bacteria cells involving thedynamics of s, x and p
ds/dt = − δµxdx/dt = µx (−γx)dp/dt = (α µ+ β)x
(4)
s(0) = s0, x(0) = x0, p(0) = p0 = 0
α, β, δ are specific rate constants
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September, 21–26, 2014, Wurzburg SCAN 2014
Classical unstructured models
The parameter µ is the “specific growth rate” which is (usually) afunction of s. Typical choices for µ are the Monod function
µ(s) = µmax s/(Ks + s) (5)
and the Haldane/Andrews function
µ(s) = µmax s/(Ks + s+Kis2) (6)
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September, 21–26, 2014, Wurzburg SCAN 2014
Experimental data provided
Experimental data provided:
a) for the CGTase production by bacteria Bacteria circulansATCC21783 (data provided by prof. V. Beschkov from the Instituteof Chemical Engineering, BAS);
b) for EPS production from thermophylic bacteria (data provided byprof. M. Kamburova from the Institute of Microbiology, BAS)
Both models provide data for the biomass x and the product p
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September, 21–26, 2014, Wurzburg SCAN 2014
Classical Monod model
The Monod model has been applied to fit real experimental data forEPS production
using first the Monod function and then the Haldane function
s0 = 0.1, x0 = 0.05, µmax = 5., α = 0.1, β = 0, γ = 0.005,Ks = 0.25, Ki = 30, δ = 13
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September, 21–26, 2014, Wurzburg SCAN 2014
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
0 5 10 15 20 25 30
Biom
ass
and
EPS
Time in hrs
Monod function, gamma=0, Fermentor1; 1 l; 600 rpm,Air 0.8:1
Interval experim. biomassinterval experim.EPS
Theor. biomassTheor. substrate
Theor. product EPS
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September, 21–26, 2014, Wurzburg SCAN 2014
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
0 5 10 15 20 25 30
Biom
ass
and
EPS
Time in hrs
Haldane function,ki=30, gamma=0, Fermentor1; 1 l; 600 rpm,Air 0.8:1
Interval experim. biomassinterval experim.EPS
Theor. biomassTheor. substrate
Theor. product EPS
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September, 21–26, 2014, Wurzburg SCAN 2014
Modelling process: graphics via Mathematica
Parameter identification is performed conveniently usingMathematica tools
Slide keys (“sliders”)
http://reference.wolfram.com/language/ref/Slider.html
If no solution sets passing through all intervals are found,
then we abandon the model and look for another model structure
— usually a modified one
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September, 21–26, 2014, Wurzburg SCAN 2014
Out[20]=
T 70
e0 0.06
s0 2.6
k0 0.1
k1 2.62
k2 1.25
XMIN 0
YMIN 0
XMAX 70
YMAX 3.9
0 10 20 30 40 50 60 70
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
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September, 21–26, 2014, Wurzburg SCAN 2014
A two-phase model
Model (4) is a two-phase model proposed in CAMWA [1]
ds/dt = −k1xs− (α+ β)ys,dx/dt = −k1xs+ k2y + αys− kxx,dy/dt = k1xs− k2y + βys− kyydp/dt = γ ∗ s ∗ (x+ y)
(7)
with the initial conditions
s(0) = s0, x(0) = x0, y(0) = y0, p(0) = p0.
s is the substrate, p is the product
x, y are two phases of the biomass
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September, 21–26, 2014, Wurzburg SCAN 2014
A two-phase model
A good approximation of the experimental data using this modelhave been found with the following coefficients:
k1 = 0.5; k2 = 0.5; kx = 0.01; ky = 0.1; α = 3; β = 0.2
and with initial conditions:
s0 = 2; x0 = 5.10−5, y0 = 0, p0 = 0
The above approximation is visualized on the next Figure
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September, 21–26, 2014, Wurzburg SCAN 2014
0
0.5
1
1.5
2
0 5 10 15 20 25
Biom
ass
and
EPS
Time in hrs
Smarkov model 2, Fermentor1; 1litre; 600 rpm,Air 0.8:1, data error=10%
Interval experim. Biomassinterval experim.EPS
Theor. substrateTheor. lag biomass
Theor. active biomassTheor. product EPSTheor. total biomass
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September, 21–26, 2014, Wurzburg SCAN 2014
The basic two-phase model
Reproduction—simple reaction scheme
Y -cells are engaged in formation of products P , which are importantcomponents of newborn cells. Reproduction as part of the productionprocess: a) cell growth is due to reproduction—hence to dividingY -cells; b) Y -cells utilize product P to reproduce
P + Yk3−→ 2Y
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September, 21–26, 2014, Wurzburg SCAN 2014
The basic two-phase model
The dividing mother Y -cells transform into daughter cells plus anewborn cell built by means of product P
S +Xk1−→←−k−1
Yk2−→ P +X,
P + Yk3−→ 2Y,
Many other variants (like replacing Y by X) were checked and waived
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September, 21–26, 2014, Wurzburg SCAN 2014
The basic two-phase model
Applying the mass action law, we obtain the model:
ds/dt = −k1xs+ k−1ydx/dt = −k1xs+ k−1 y + k2ydy/dt = k1xs− k−1 y − k2y + k3pydp/dt = k2y − k3py
(8)
with initial conditions (corresponding to a batch cultivationprocess): s(0) = s0, x(0) = x0, y(0) = y0, p(0) = 0.
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September, 21–26, 2014, Wurzburg SCAN 2014
The basic two-phase model
Respectively, assuming k−1 = 0 we obtain the simpler model:
ds/dt = −k1xsdx/dt = −k1xs+ k2ydy/dt = k1xs− k2y + k3pydp/dt = k2y − k3py
(9)
induced by the reaction scheme:
S +Xk1−→ Y
k2−→ P +X,
P + Yk3−→ 2Y.
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September, 21–26, 2014, Wurzburg SCAN 2014
0
0.5
1
1.5
2
0 5 10 15 20
Bio
mass
and E
PS
mg/m
l
Time in hrs
Fermentor 1litre, 500 rpm, Air 0.8:1
Interval experim. Biomassinterval experim.EPS
X cells Y cells
Product EPS Total biomass
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September, 21–26, 2014, Wurzburg SCAN 2014
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
0 5 10 15 20
Bio
mass
and E
PS
mg/m
l
Time in hrs
Fermentor, 1litre; 100 rpm, Air 0.8:1
Interval experim. Biomassinterval experim.EPS
X cellsY cells
Product EPS Total biomass
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September, 21–26, 2014, Wurzburg SCAN 2014
0
0.5
1
1.5
2
0 5 10 15 20
Bio
mass
and p
roduct
mg/m
l
Time in hrs
Aeribacillus pallidus 418, Fermentor 1 litre
X -cells Y - cells
Product EPSTotal biomass x+y
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September, 21–26, 2014, Wurzburg SCAN 2014
Comparison between Monod model and our basic model
Proposition. Monod model is a special case of our basic Model underthe assumption that favourable conditions are present (the biomass xand the product p are nearly constant)
Proof — in BIOMATH 2/2 paper
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September, 21–26, 2014, Wurzburg SCAN 2014
Our basic model and Henri reaction scheme
Our model makes use of Victor Henri reaction scheme in enzymekinetic, when identifying free enzymes with X-cells and boundedenzymes with Y -cells, following the familiar stoichiometric-likescheme for the kinetics of enzymes with just one active site:
S +Xk1−→←−k−1
Yk2−→ P +X,
wherein k1, k−1, k2 are rate constants.
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September, 21–26, 2014, Wurzburg SCAN 2014
Open math problems induced by cell growth kinetics
P1. To approximate in Hausdorff metric the interval step function bymeans of the X-solution (or the X + Y -solution) of the basiccell-growth system
P2. To approximate in Hausdorff metric the interval Dirac functionby means of the y-solution of the basic cell-growth system
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September, 21–26, 2014, Wurzburg SCAN 2014
Conclusion remarks
Basic biological processes—such as cell growth—still need adequatemodels
For the model construction and parameter identification the followingtools prove to be useful:
Interval approach—very useful both in model construction andparameter identification
Contemporary computational tools—CAS Mathematica
Reaction schemes—very helpful when establishing the modelstructure of the biological process
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September, 21–26, 2014, Wurzburg SCAN 2014
Conclusion: achievement
It has been demonstrated that classical Monod type models fail tomodel adequately cell growth in the transision periods (lag-log,log-stationary)
Proposed two-phase models provide more flexibility and can be verywell fitted to real data
Our proposed two-phase models are built on simple principles relatedto enzyme kinetic, implied by reaction schemes. This contributes tomore clarity in the biological meaning
All parameters in the proposed two-phase models are numeric andhave clear biological meanings as specific rate constants (notdepending on the concentrations involved)
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September, 21–26, 2014, Wurzburg SCAN 2014
Bastin-Dochain theory
Bastin-Dochain “reaction scheme”
S +Xν(s)−→P +X
“Applying MAL” we get:
ds/dt = −ν(s)sx
putting ν(s) = 1/(Ks + s) one obtains Monod model
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September, 21–26, 2014, Wurzburg SCAN 2014
References
[1] Alt, R., S. Markov, Theoretical and computational studies ofsome bioreactor models, Computers and Mathematics withApplications 64 (2012), 350–360.
[2] Markov, S., Biomathematics and interval analysis: a prosperousmarriage. In: M. D. Todorov, Ch. I. Christov, Eds., AIP Conf.Proc. 1301, Amitans2010, AIP, 2010, 26–36.
[3] Markov, S., Cell Growth Models Using Reaction Schemes: BatchCultivation, Biomath 2/2 (2013), 1312301.
[4] Radchenkova, N., M. Kambourova, S. Vassilev, R. Alt, S.Markov, On the mathematical modelling of EPS production by athermophilic bacterium, BIOMATH 3/1 (2014), 1407121
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Figure 1: Rue Monod in Lion SCAN 2010