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 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, a structural component of plant biomass, is the most abundant feedstock used for the production of alternative liquid fuels, mainly bioethanol Cellulose In which Cellul.

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, a structural component

of plant biomass, is the most abundant feedstock used for the production of alternative liquid fuels, mainly bioethanol

Cellulose

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

Acid hydrolysis results in the

production of some non-sugar

by-products

Hydrolysis

indeed presents a green approach

In this study, we isolated

1

Introduction

Objectives

microbial strains that

produce

State Fermentation (SSF) and using to

product Bioethanol product Bioethanol

2 Methods

Degraded Ulva

2.1

Microorganism

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

sequancing

C

sphaerospermu m

CMC (1,5%) then Lugol’s Iodine

isolate d

Cellulase positive

2 Methods

2.2 Collection of algal

sample

Washed and dried

U fasciata

Then grind

Powdered seaweed

2 Methods

2.3 SSF and optimization of parameters for Cellulase

production

250 mL Erlenmeyer

flasks containing 10g of

powdered seaweed

Fugal spore suspension Mineral salt

pH 5

Cellulase production was optimized with:

- Moisture

- Incubation period

- pH

- temperature

Flasks were incubated at room temperature for 6 days

2 Methods

2.4 Enzyme extraction and

assay

Fermented

substrate

Filtered

The clear supernatant

Cold Centrifuged

Suspend ed

By Sodium Acetate buffer

Enzyme activity assay:

- FPase

- CMCase

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

Cellulose from U

fasciata

Reducing sugar

Spectrophotometric lly using DNS method

OPTIMIZATION OF

Enzyme dosage

Incubation period

Hydrolysis temperature

pH

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

Qualitatively analyzed using TLC

2 Methods

2.6 Fermentation of algal

hydrolysate

Saccharomyces cerevositae

MTCC No 180

Bioethanol Reducing sugar

(from algal hydrolysate)

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

Enzyme dosage (U/g)

Optimization of algal

biomass with:

dosage

period

- pH 4

- 40ºC

3 Results

3.3 Hydrolysis of algal biomass through SSF-derived

cellulase

The presence of Glucose in the

hydrolysate, determined with TLC

Algal Cellulose

Algal Biomass

Standard Glucose

3 Results

3.4 Fermentation of algal

hydrolysate

4 Conclusions

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

%

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

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