Microbes,

the living Machine

Speaker:-
Md. Mesbah Uddin
Khulna University
Bangladesh
Potential Microbes
Bacteria e.g. Zymomonas,
Clostridium, etc.
Fungi e.g. Saccharomyces,
Kluyveromyces, etc.
 High
 sugarproducer
Cellulase consumption rate

Desirable Traits
High specific growth rate
High salt tolerance (acetate)
High volumetric productivity
High Ethanol Tolerance
for Bioethanol Production
High specific productivity
Thermotolerance
C5/C6 Cofermentation
High shear tolerance
Minimal nutrient requirements

 Amenable to scale-up
Entner-Doudoroff pathway
Generally Regarded as Safe (GRAS)
Available gene transfer system
Yeast= Saccharomyces
 Ethanol is inhibitory at high concentrations and the alcohol
tolerance of yeast is critical for high yield.
 As the concentration increases, the growth rate is first
reduced, whereas at higher concentrations the biosynthesis of
ethanol itself is inhibited.
 Yeast is more sensitive to endogenously produced ethanol
than to added from external sources to the fermentation
system.
 Growth generally ceases at 5% ethanol ( v of ethanol in v of
water=vv) and the production rate is reduced to zero at 6-10%
(vv).
Zymomonas mobilis
 Zymomonas mobilis is a bacterium belonging to the
genus Zymomonas.
 It is notable for its bioethanol-producing capabilities,
which surpass yeast in some aspects.
 It was originally isolated from alcoholic beverages
like the African palm wine, the Mexican pulque, and
also as a contaminant of cider and beer in European
countries.
 Z. mobilis degrades sugars to pyruvate using the
Entner-Doudoroff pathway. The pyruvate is then
fermentated to produce ethanol and carbon dioxide
as the only products (analogous to yeast).
The advantages of Z. mobilis over
S.cerevisiae with respect to
producing bioethanol:
 higher sugar uptake and ethanol yield,
 lower biomass production,
 higher ethanol tolerance,
 does not require controlled addition of
oxygen during the fermentation,
 amenability to genetic manipulations.
Comparison of Kinetic Parameter:
Zymomonas vs. Saccharomyces
 Ethanol formation rate (g/g.h) 2.9
times higher
 Growth rate (m) 2.4 times higher
 Glucose uptake rate 2.6 times higher
Advantages of Zymomonas
 The bacterium zy.. Has in recent years come under
increasing study because it has a number of
potential advantages:
 Osmotic tolerance to higher sugar concentration
(up to 400 g/l)
 Relatively higher ethanol tolerance ( up to 130 g/l)
 Higher specific growth rate than yeast ( growth rate
m of 0.27 compared to 0.13 for yeast; lab study)
 Carbohydrate metabolism by ED pathway, where
only one mole of ATP is produced per mole of
glucose used, thus reducing the amount of glucose
that is converted to biomass rather than ethanol.
 Zymomonas mobilis
 • Advantages:
 – Natural fermentative microorganism (GRAS)
 – Near theoretical ethanol yield from glucose
 – Reduced yield loss to biomass formation
 – No oxygen requirement
 – Tolerant to inhibitors in hydrolysates
 – High ethanol tolerance
 – Fermentation at low pH
 – Grows at high sugar concentrations
 – High specific productivity
 • Limitations
 – Narrow substrate utilization range
 In the ethanol fermenting bacterium
Zymomonas mobilis hopanoids may
have a role in adaptation of cell
membranes to ethanol accumulation
and to temperature changes which
influence membrane functions.
Diploptene, a hopanoid compound

Cholesterol, a sterol
compound.
Ethanol Fermentation
in Zymomonas
(ED Pathway)
Yeast using molasses as substrate

Batch Process 2.0 g/l.h

Continuous Process 3.35 g/l.h

Immobilized cells 28.6 g/l.h

under continuous
conditions
The Immobilized process has already been
placed in large-scale production in Japan