CH465 Biochemical

Engineering
Spring 2017
Monday April 24

Mixed Cultures

Chapter 16 Shuler and Kargi

On reserve in the library
 The Rest of the Semester
Lab Report on Fermentation Experiment
Due 11 PM this Wednesday
Submit pdf file via Moodle; only one report per group should be
submitted
Group Evaluation
Due 11 PM this Friday
Submit via Moodle; all must respond
HW Set 8
Due in class this Friday
Final Exam
Monday May 8, 11:45AM 2:45PM
SC362
Will cover adsorption, chromatography, genetic engineering:
science and reactors (HW sets 7 and 8)
 Mixed Cultures
Interacting Species
Food fermentations
Cheese, yogurt manufacturing
Biological waste water treatment
Recombinant cultures
 Interactions in Mixed
Cultures
Competition
Each population competes for the same
substrate
Negative effects on both populations
Neutralism
Neither population is affected by the
presence of the other
One example: yogurt starter strains of
Streptococcus and Lactobacillus in a
chemostat
 Interactions in Mixed
Cultures
Mutualism
Interaction is essential to the survival of both
species
Example: phenylalanine-requiring strain of
Lactobacillus and a folic-acid requiring strain of
Streptococcus. Exchange of growth factors
produced by partner organisms helps each to
grow; pure cultures exhibit no growth
Example: aerobic bacteria and photosynthetic
algae. Bacteria use O2 and carbohydrate; algae
convert CO2 to carbohydrates and liberate O2 in
the presence of sunlight.
 Interactions in Mixed
Cultures
Commensalism
One population is positively affected by
the presence of the other; the 2nd
population is unaffected by the presence
of the 1st.
Mechanisms:
2nd population produces a required nutrient
or growth factor for the 1st population
Example: production of H2S by Desulfovibrio.
H2S is used by sulfur bacteria
 Interactions in Mixed
Cultures
Commensalism
Mechanisms:
2nd population removes a substance from the
medium that is toxic to the 1st population
Example: removal of lactic acid by the
fungus Geotrichium candidum which allows
for the growth of Streptococcus lactis. Lactic
acid is produced by Streptococcus lactis .
Used in cheesemaking.
 Interactions in Mixed
Cultures
Amensalism
One population is negatively affected by
the presence of the other; the 2nd
population is unaffected by the presence
of the 1st.
Mechanisms:
2nd population produces a toxic substance
that inhibits the growth of the 1st population
Example: production of antibiotics by certain
molds that inhibit the growth of others.
 Interactions in Mixed
Cultures
Amensalism
Mechanisms:
2nd population removes essential nutrients
from the media, negatively affecting the
growth of the 1st population
 Interactions in Mixed
Cultures
Predation and Parasitism
One population benefits at the expense
of the other
Predation
ingestion of prey by the predator
Common in aerobic waste water treatment
reactors
Parasitism
Host, usually the larger organism, is
damaged by the parasite
 Mixed Culture Example
Shuler and Kargi, problem 16.1
1L batch fermenter with 5 g/L of glucose in
medium. Glucose is the growth rate limiting
nutrient for a mixed population of two bacteria
(E. coli and Axotobacter vinelandii).

A. vinelandii is 5 times larger than E. coli. The

inoculum for the fermenter is 0.03 g/L E. coli (1
x 108 cells/mL) and 0.15 g/L A. vinelandii (1 x
108 cells/mL).
 Mixed Culture Example
Shuler and Kargi, problem 16.1

The specific growth rates for the two

organisms are: 1.0 hr s 1
EC 0.05 hr 1
0.01 g / L s
0.5 hr 1 s
AV 0.1 hr 1
0.02 g / L s

What do these terms represent?

Yield coefficients are:

YEC 0.5 g dw/ g glucose
YAV 0.35 g dw/ g glucose

What is g dw ?
 Mixed Culture Example
Shuler and Kargi, problem 16.1

What will be the ratio of A. vinelandii to

E. coli when all of the glucose is
consumed?
s (time) = ?
xEC (time) = ?
x AV (time) ?

What is expected?
 Mixed Culture Example
Shuler and Kargi, problem 16.1

Balance on E. coli and A. vinelandii:

d xEC
EC xEC
dt
d x AV
AV x AV
dt

EC and AC are not constant with time. Why?

Balance on glucose:
ds 1 d xEC 1 d x AV

dt YEC d t YAV d t
 Mixed Culture Example
Shuler and Kargi, problem 16.1

Approximate derivatives as discrete

differences:
d xEC xEC xEC ,i 1 xEC ,i 1.0 si
EC xEC ,i 0.05 xEC ,i
dt t ti 1 ti 0.01 si

d x AV x AV x x AV ,i 0.5 si
AV ,i 1 AV x AV ,i 0.10 x AV ,i
dt t ti 1 ti 0.02 si

d s s si 1 si 1 1.0 si 1 0.5 si
0.05 x EC ,i 0.10 x AV ,i
d t t ti 1 ti YEC 0.01 si YAV 0.02 si

t0 0, s0 5g/L, xEC ,0 0.03 g/L, x AV ,0 0.15 g/L

 Mixed Culture Example
Shuler and Kargi, problem 16.1

Approximate derivatives as discrete

differences:
d xEC xEC xEC ,i 1 xEC ,i 1.0 si
EC xEC ,i 0.05 xEC ,i
dt t ti 1 ti 0.01 si

d x AV x AV x x AV ,i 0.5 si
AV ,i 1 AV x AV ,i 0.10 x AV ,i
dt t ti 1 ti 0.02 si

d s s si 1 si 1 1.0 si 1 0.5 si
0.05 x EC ,i 0.10 x AV ,i
d t t ti 1 ti YEC 0.01 si YAV 0.02 si

ti 1 ti 0.1 hr
Mixed Culture Example
Shuler and Kargi, problem 16.1
6

3
concentration (g/L) glucose
E coli
2 A. vinelandii

0
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5

time (hour)

@ t 4.4 hours, s 0, xEC 1.6 g/L, x AV 0.83 g/L

xEC x AV = 1.9
 Mixed Culture Example
Shuler and Kargi, problem 16.1
2

0
0 2 4 6
d concentration/dt (g/L hr) -1 glucose
E coli
-2 A. vinelandii

-3

-4

time (hour) 1
0.9
0.8
0.7
0.6
0.5
specific growth rates (g/L)
0.4 E coli
0.3 A. vinelandii
0.2
0.1
0
0 2 4 6

time (hour)
 Mixed Culture Example
Shuler and Kargi, problem 16.1
What if we assume = constant?

d xEC
EC xEC
dt xEC xEC ,0 exp EC t
d x AV
AV x AV x AV x AV ,0 exp AV t
dt

ds 1 d xEC 1 d x AV 1 1
EC xEC ,0 exp EC t AV xAV ,0 exp AV t
dt YEC d t YAV d t YEC YAV

Integrate:
1 1
s s0 xEC ,0 exp EC t 1 x AV ,0 exp AV t 1
YEC YAV
Mixed Culture Example
Shuler and Kargi, problem 16.1

4
glucose
3 E coli
concentration (g/L)
A. vinelandii
2
glucose - w constant mu
1 E coli w constant mu
A vinelandii w constant mu
0
0 1 2 3 4 5
-1

time (hour)

@ t 4.2 hours, s 0, xEC 1.6 g/L, x AV 0.80 g/L

xEC x AV = 2.0
 Mixed Culture Example
Shuler and Kargi, problem 16.1
1L batch fermenter with 5 g/L of glucose in
medium. Glucose is the growth rate limiting
nutrient for a mixed population of two bacteria
(E. coli and Axotobacter vinelandii).

A. vinelandii is 5 times larger than E. coli. The

inoculum for the fermenter is 0.03 g/L E. coli (1
x 108 cells/mL) and 0.15 g/L A. vinelandii (1 x
108 cells/mL).

What mechanism is at play here??