Solid state fermentation (SSF) derived cellulase for saccharification of the green seaweed Ulva for bioethanol production Solid state

Solid state fermentation (SSF)derived cellulase for
saccharification of the green
seaweed Ulva for bioethanol
production
Nitin Trivedi, C.R.K. Reddy, Ricardo
Radulovich, Bhavanath Jha.


1.
Introduction
Cellulose

Cellulose, a structural component
of plant biomass, is the most
abundant feedstock used for the
production of alternative liquid
fuels, mainly bioethanol.
In which Cellulose from algae
has high Carbohydrate content,
not mixed with impurities such
as Lignin, Hemi-Cellulose and
Pectin, so it is easy to purify
Cellulose


1.
Introduction
Hydrolysis

CHEMICAL HYDROLYSIS

ENZYMATIC HYDROLYSIS

Acid hydrolysis results in the
production of some non-sugar byproducts

Enzymatic
hydrolysis
indeed presents a green
approach


1.
Introduction
Objectives

In this study, we isolated
microbial strains that
Cellulaseproduce
through
Solid
State Fermentation (SSF)
and using to
product Bioethanol
product Bioethanol


2. Methods

2.1.
Microorganism

Cellulase
positive
CMC (1,5%)
then Lugol’s
Iodine
isolate
d

Degraded
Ulva

C.
sphaerospermu
m

The molecular
identification of the
fungal strain was carried
out by 18S rDNA
sequancing


2. Methods

2.2. Collection of algal
sample

Washed and
dried

Then
grind
U. fasciata

Powdered seaweed


2. Methods

2.3. SSF and optimization of parameters for Cellulase
production

Cellulase production
was optimized with:
- Moisture
- Incubation period
- pH
- temperature
250 mL Erlenmeyer
flasks containing 10g of
powdered seaweed

Mineral salt
pH 5

Fugal spore
suspension

Flasks were incubated at room
temperature for 6 days

2. Methods

2.4. Enzyme extraction and
assay

Suspend
ed
By Sodium
Acetate
buffer
Fermented
substrate

Filtered

Enzyme activity
assay:
- FPase
- CMCase

Cold
Centrifuged

The clear
supernatant

Enzyme stability
assay:

- pH
- temperature


2. Methods

2.5. Hydrolysis of algal biomass through SSF-derived
cellulase
SSF-derived
Cellulase

Cellulose from U.
fasciata

Reducing sugar

Spectrophotometric
lly using DNS
method

ORANGE


2. Methods

2.5. Hydrolysis of algal biomass through SSF-derived
cellulase

SSF-derived
Cellulase

Reducing sugar

Cellulose from U.
fasciata

Enzyme
dosage

Hydrolysis
temperature

OPTIMIZATION OF
pH

Incubation
period

Spectrophotometric
lly using DNS
method


2. Methods

2.5. Hydrolysis of algal biomass through SSF-derived
cellulase
SSF-derived
Cellulase

Reducing sugar

Cellulose from U.
fasciata

Qualitatively
analyzed using TLC

Spectrophotometric
lly using DNS
method


2. Methods
2.6. Fermentation of algal
hydrolysate

Saccharomyces
cerevositae
MTCC No. 180

Reducing sugar
(from algal hydrolysate)

Bioethanol

Using GC-MS and DNS method to
analyzed Ethanol yield and residual
reducing sugars

3. Results

3.1. Optimization of SSF for enzyme
production

Inoculated fungus with 60% moisture
content, incubated for 4 days at 25 °C
and pH 4, showed optimum enzyme
production.


3. Results

3.2. Effect of pH and temperature on cellulase activity and
stability

Enzyme activity, found to be optimal at pH 4 and 40 °C


3. Results

3.3. Hydrolysis of algal biomass through SSF-derived
cellulase

Optimization of algal
biomass with:
-

10

U/g

Enzyme

dosage
-

24h

period
- pH 4
- 40ºC

incubation

Enzyme dosage (U/g)


3. Results
3.3. Hydrolysis of algal biomass through SSF-derived
cellulase

The presence of Glucose in the
hydrolysate, determined with TLC

Standard
Glucose

Algal

Cellulose

Algal
Biomass


3. Results
3.4. Fermentation of algal
hydrolysate


4. Conclusions
The optimal conditions SSF is a dry algal
substrate containing inoculated fungus with
60% moisture content, incubated for 4 days
at 25 °C and pH 4
The optimal conditions for fermenting
aldal biomass are 10 U/g enzymes,
incubation period of 24 h, hydrolysis
temperature of 40 °C and pH 4

The optimum ethanol yield was
found to be 0.44g with 93.81%
conversion efficiency after 12 h of
fermentation

15
%

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