Fermentation

Kinetics of Yeast Growth

and Production

Introduction

Fermentation can be defined as an energy yielding process where yeast

converts organic molecules (such as sugar) into energy, carbon dioxide
or/and ethanol depending on the respiration pathway.
Yeast can respire in anaerobically and aerobically.
However, yeast gets more energy from aerobic respiration, but in the
absence of oxygen it can continue to respire anaerobically, though it does
not get as much energy from the substrate. Yeast produces ethanol when it
respires anaerobically and ultimately the ethanol will kill the yeast (find
out why is yeast continue to produce ethanol even the last is an inhibitor).

C6H1206 2 CH3CH2OH + 2 CO2 + 2 ATP

C6H1206 + 6O2 6CO2 + 6H2O + 16-18 APT

When the feed substrate to the reactor is not

monosaccharide e.g. sucrose (C12H22O11), yeast
enzyme cause glycosidic bond to break in a process
called hydrolysis

Industrial and Commercial Applications

Food Industry
~ Beer
~ Bread
~ Cheese
~ Wine
~ Yogurt

Pharmaceutical Industry
~ Insulin
~ Vaccine Adjuvants

Energy
~ Fuel Ethanol

Objective
To find the kinetics of the system by using
Nonlinear Regression (guess for k s and m)
The Sum of the Least Squares and the
Lineweaver-Burk Plot methods in order to
determine the parameters m and ks
To determine the yield coefficient and to
project min. and max. amount yeast cell mass,
carbon dioxide and ethanol produced

Experimental Set Up
Apparatus
pH probe
D-oxygen
probe

Bioreactor

Mixer
Temperature
sensor
YSI 2700
Biochemistry
Analyzer

pH meter
Sampling
device

Experimental: Procedure
Using Biochemistry Analyzer and Spectrophotometer
to measure and make calibration curves for sugar and
yeast cell concentrations
Reactant initial concentration

dextrose/or sucrose 25 g/L

yeast 3 g/L
volume reactant solution 2 L

Initial conditions & assumptions

Initial Conditions

2 L of solution
50 g sugar
pH around 5.0
Temperature around 28-30C

Assumptions

the bioreactor content is

well mixed and has a constant medium volume at a certain
initial conditions
Temperature is constant
pH maintained at optimal pH of 3.00
All reactants or nutrients present in excess except for sugar
substrate.

Theory

In ideal fermentation process in which the growing cells are

consuming the substrate (sugars), and producing more cells
according to the following scheme.
rsx
rx

Cells (x)

rsx = rate of substrate consumption

rx = rate of cell growth
s = substrate concentration
x = cell concentration
P = ethanol concentration (in anaerobic case)

Cells (x)
P

Theory
dx
rx
dt

x Biomass

dCx
rx
dt

The plot showing the trends for yeast cell growth over time

Theory continue

Yeast Growth occurs in 4 stages

Lag phase, yeast mature and acclimate to environment (no growth occurs)
The exponential growth section, the rate of reaction follows first order kinetics

dx
rx x
dt

During the deceleration phase, a large number of parameters, each with saturation effects,
have an effect on the kinetics of yeast growth (such as substrate and waste concentrations)
S O P
dx
rx
mx
...
rx ( s) x


dt
K s S K o O K p P
The growth rate is ruled by the limiting substrate concentration (sugar)

dx
rx
mx
dt

The final equation, often referred to as the Monod equation, looks very similar to the
Michaelis-Menten equation.

( s)

S
s

ss

s
s s

ks = the Monod constant (g/L)

m = a maximum specific growth reaction rate (min-1)

Stationary phase, no growth occurs due to high waste concentration or compleate substrate
consuming

Lineweaver-Burk Rearrangement
ss
( s) m

s
s s

k s ks 1 1
1
s

( s)
ms m s m

Nonlinear Regression
ss
dx
rx
mx

dt
ks ss

1.

Define Model

2.

Solve for Rpredicted (dx/dt)

(calculate dx/dt from the polynomial equation fitted
to the curve x(t)

3.

Make initial guess for ks and m

(m is the max. specific growth rate can be achieved
when S >> ks
ks is saturation constant or the value of limiting
substrate conc. S at which s equal to the half of m
Minimize (R-Rpredicted)2 using solver function in
Excel by varying ks and m

Yield Coefficient Determination

dx x xo
Yx

ds so s
s

dP P P0
Yp

ds s0 s
s

Ratio of cell or Ethanol concentration to substrate concentration.

Knowing Yx/s will give you an idea for how much additional
yeast cell mass, on average, is produced for a given amount of
sugar substrate consumed.
As well allowed you to calculate a lower bound on the
experimental stoichiometric coefficient, , and therefore to
calculate ranges for ethanol and CO2 production.
(Yeast Cell) + C6H12O6 (CO2 + CH3CH2OH) + (Yeast Cells)

Error in Lineweaver-Burk
Parameters

Error in ks and m relative to error in slope and y-intercept of

linear fit
Random Error in y values:

y
i i
n 2

sy
x

STDEV of slope:

sb

sy

x x

STDEV of y-intercept:

sa sy

xi

n xi x

Lower Bound on
(stoichiometric coefficient)
(Yeast Cell) + C6H12O6 (CO2 + CH3CH2OH) + (Yeast Cells)
Where, theoretically, = 2.

Assume all yeast generated is attributable only to sugar

complete consumption

Conservation of mass requires that the remaining product be

equimolar amounts CO2 and ethanol