Mohamed Mosaad Abo El-Gheit

MSc. Student, Applied Microbiology, SCU,

Ismailia, Egypt
mhmd.aboelgheit@gmail.com

Contents:

Biofuel
Types of Biofuel
1st and 2nd generation of Bioethanol
Lignocellulosic Biomass in Egypt
Composition of Lignocellulose
Pretreatment of Lignocellulose
Microbial Enzymatic Hydrolysis
Bioprocessing of Biomass
Cellulosic Activities in Actinomycetes
Microbial Consortia

Biofuel:
Energy from newly-growing plant

sources
CO2-neutral
alternative source of energy to the
current traditional sources e.g. gasoline

CO2 Neutral?

1st Generation of Bioethanol

BRAZIL

sugarcane
(sucrose)

extract

Sugars
ferment

ethanol

USA

(starch)

Sugars

Hydrolyze
(enzymes)
Cosgrove; 2005

ethanol

ferment

Types of Biofuel
Solid animal wastes and agricultural residues can be

used as sources of energy by direct burning (primitive way)

Organic wastes

Liquid

Direct burning

Heat energy

Bioethanol C2H5OH ( fermentation of

sugar)
Biodiesel (by saturation of vegetable

oils)
Gas e.g. methane and biogas derived from organic

wastes by anaerobic digestion

nd
2

Generation of Bioethanol

Cosgrove; 2006

Lignocellulosic Biomass
Agricultural Residues:

Source: Quantitative appraisal of biomass resource and their

energy potential in Egypt; 2013

Lignocellulosic Biomass:
Energy crops: plants which grow at low cost, to make

biofuel.

Composition of Lignocellulose
Cellulose

Cellulose (6 carbon sugars)

Hemicelluloses
Lignin

Lignin (phenols)
Hemicellulose
(both 5 and 6 carbon sugars)
(need modified microbe to
convert to ethanol)

Extractives
Ash

Chapple, 2006; Ladisch, 1979, 2006

Pretreatment
break down the

shield formed by
lignin and
hemicellulose
Open the fiber
structure
reduce the degree
of polymerization
of cellulose.
Source: Overview of biomass pretreatment
for cellulosic ethanol production; 2009

 Pretreatment has been viewed as one of the most

expensive processing steps within the conversion of

biomass to fermentable sugar
Pretreatment methods maybe: physical, chemical or
biological
Biological:
Adv. : no chemicals, no energy requirements, mild
environmental conditions
Disadv.: slow, the activity of the microorganisms maybe
not specific to lignin only!

Pretreated Lignocellulose
What is Pretreated Biomass?
increased surface area,
solubilization of cellulose,
redistribution of cellulose and lignin
Cellulose 35-50%
Hemicellulose 20- 35%
Lignin 5-30%

Microbial cellulose utilization fundamental and biotechnology; 2002

Enzymatic Treatment
Pretreated
Lignocellulose

cellulose
hemicellulose

Enzymatic
Hydrolysis

Pentoses and
hexoses + lignin and
lignin degradation

glucose
glucose + xylose+ other
C5 and C6 sugars

Microbial cellulose utilization fundamental and biotechnology; 2002

Microbial Enzyme system:

Substrate cellulose + hemicellulose
Enzymes:
endoglucanases: cut at random internal sites

along the cellulose/hemicellulose chain

exoglucanases: act at reducing and nonreducing
ends
beta-glucosidase: break betaglucoside bond to
form glucose

Enzyme system
Cellulose

Endogluconase

Oligosaccharides (<10)

Exoglucanase
Cellobiose
+ glucose

Beta-glucosidase

glucose

Microbial cellulose utilization fundamental and biotechnology; 2002

Lignocellulosic Activities of
Actinomycetes
According to Lynd et al (2002) there is a considerable

concentration of cellulytic capabilities among

Actinomyceltales.
Actinomycetes are well known for their ability to
decompose complex molecules, particularly
lignocellulose components
Micromonospora spp and Strptomyces spp are well
known for their decomposition ability on Biomass

Actinomycetes and cellulytic

activities
Speices

Growth Temp

M. chalcea

mesophilic

S. roseflavus

mesophilic

S. reticuli

Mesophilic

Thermobifidia fusca

Thermophilic

Kibdelosporanguim Philippinenses

mesophilic

Most of actinomycete species can be isolated from both soil and

water.

Bioprocessing of cellulosic Biomass

Steps (mediated events):

Cellulase production
2) Hydrolysis of
cellulose/hemicellulose
3) Fermentation of cellulose
hydrolysis products e.g.
glucose
4) Fermentation of
hemicellulose hydrolysis
products other than
glucose e.g. xylose
1)

biomass

fuel

Microbial cellulose utilization fundamental and biotechnology; 2002

Bioprocessing of cellulosic Biomass

SHF: Separated
Hydrolysis and
Fermentation
SSF: Simultaneous
Saccharification and
Fermentation
SSCF: Simultaneous
Saccharification and
Cofementation
CBP: Consolidate
Bioprocessing
This diagram shows the capability of consolidation or separation of
mediate events (steps) of bioprocessing of Biomass
Source: Microbial cellulose utilization fundamental and biotechnology; 2002

Consolidated Bioprocessing CBP

In which all bioprocessing steps are combined together

as one process
Biomass processing technology has exhibited a trend
toward increasing consolidation over time
Advantages Efficiency + Economically effective
CBP organisms:
Single organism
Community of organisms( symbiotic consortium)

(which is more efficient???)

Symbiotic Consortium
A community of organisms
i.e 2 or more organisms living in association
Symbiosis may be : mutualism, commensalism, o

antagonism
Types:
Natural consortuim

Genetically

Engineered consortuim
Recombined natural capabilities
i.e. ecological approaches

Natural Consortium
The main problem doesnt accumulate high levels

of biofuel why?
Biofuel molecules are molecules of energy
Biofuels represents an a pportunity for a new consortia

member (organism) to exploit

Natural consortia tend to thermodynamically free
energy of molecules till the lowest level

Be overcome by engineering consortia

Models of microbial interactions in

a consortuim (dual culture)

Sequential utilization
2 oranisms M1 and M2
The fuel molecule (F1) is

considered a waste product of

M1. However, it is degraded by
M2 as source of energy e.g.
commensalism
No accumlation of fuel
molecules

Co-utilization
M1 & M2 are competing to

utilize the substrate , producing

fuel molecules
Competitive symbiosis i.e.
controlled by inhibitors
/activatiors
Fuel considered waste product
of both organsims
There is accumulation of fuel

Substrate transformation
M1 acts on substrate converting

it to a form that can be utilized

by M2
e.g. pretreatment of
lignocellulosic material
mutualsim

Product transformation
M1 produces fuel products as

waste product
M2 act on fuel to convert it into
an alternative fuel
Look like sequential utilization.
However, the fuel molecules are
converted to alternative fuel ,
not completely utilized