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By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological Research Institute, Mysore. 1

By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Page 1: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

By Abitha.R,

Research Scholar,Indian Institute of Science

Bangalore.

Conducted under the guidance of Dr.R.Sharada,

Scientist,Central Food Technological Research Institute,

Mysore. 1

Page 2: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Contents

Introduction1

Objective of the Study2

Methodologies3

Results and Conclusion4

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Page 3: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Page 4: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Page 5: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

I) Need13 TW/year today26 TW/year by 205039 TW/year by 2100

II) Resources (C neutral)1) Fossil Fuel/Carbon Capture

-25 billion metric tons of CO2/year-Volume of Lake Superior

2) Nuclear-10 TW/year requires 1 new GW fission plant every day for 50 years-Terrestrial uranium would be exhausted in 10 years-Fusion – no sooner than 2040

3) Renewable-Hydroelectric 0.5 TW maximum (UN

estimates)-Tides and oceans <2 TW/year maximum-Geothermal 12 TW/year (but only

fraction extractable)-Wind 2-4 TW/year maximum-Sun 120,000 TW/year (biomass + electricity

<2% today)

More energy from the sun strikes the earth in 1

hour than all of the energy currently

consumed on the planet in 1 year!

Source : Basic Research Needs for Solar Energy Utilization Report of the Basic Energy Sciences Workshop on Solar Energy Utilization, April 18-21, 2005, DOE

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Page 6: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Third Generation Renewable Fuels

Source : Basic Research Needs for Solar Energy Utilization Report of the Basic Energy Sciences Workshop on Solar Energy Utilization, April 18-21, 2005, DOE

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Page 7: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Carbon dioxide Mitigation

Energy from

Waste

Using B.braunii

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Page 8: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Easily available

Easy to culture and grow

Limited area required

Fallow land can be used

Hydrocarbon production

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Page 9: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Botryococcus braunii was isolated from the fresh water ponds in Mamallapuram, Tamil Nadu, India, and was maintained in modified Chu13 media.

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Page 10: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Experiments – An Overview

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Page 11: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Page 12: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

•The spent wash was collected from Mandya Sugar Factory, Mandya (in Jan 2009)

•The Dairy Waste was collected from Mysore- Chamarajnagar Dist. Corporation, Milk Producer’s Society Union Ltd ( Nandini milk processing industry, Mysore) (in Jan 2009)

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Page 13: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Before Treatment After Treatment 13

Page 14: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Culture Conditions

•Control

• 0.5%

• 1%

• 2%

• 3%

• Control• Media sugar and dairy waste • Concentrated sugar and dairy waste• Dilute Sugar and Dairy waste• Centrifuged Sugar waste

•4

•4.5

•5

•5.5

•6

Wastewater CO2 pH

14

•6.5

•7

•7.5

•8

•8.5

Page 15: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Page 16: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Page 17: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Retained and Observed for 30 days

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Page 18: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Collection of samples after every three days

Tests conducted:1.Biomass test (Lichtenthaler (1987))

2.Carbohydrates Test (By Phenol-Sulphuric acid method(Dubois et al., 1956))

3.Protein Test (By Lowry’s Method (Lowry et al., 1951) )

4.Chlorophyll Test (Davies (1976)) 18

Page 19: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Page 20: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Initial and Final CODs were estimated20

Page 21: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Harvesting and Lipid Extraction

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Page 22: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Page 23: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Growth of Algae in Spent Wash and Dairy Wastewater

• Centrifuged concentrated sugar waste has the highest cell density of 0.97g/L

• The Media sugar waste has the second highest cell density of 0.7915 g/L 23

0

0.2

0.4

0.6

0.8

1

1.2

0 3 9 12 15 18 21 24

No. of Days

Bio

ma

ss

in g

/L

control

mdw

ddw

cdw

ccdw

msw

dsw

csw

ccsw

Page 24: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

0

10

20

30

40

50

60

70

Samples

Per

cent

age

of L

ipid

s

Influence of the culture with different concentrations of wastewater on lipid production

• The percentage of lipid is highest (65.56%) in control

•Dairy waste has shown a considerable amount of lipid production (about 58.2%) 24

Page 25: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

0200400600800

1000120014001600

Dairy

wast

ewat

er(In

itial

)Sp

ent w

ash

(Initi

al)

Cont

rol

med

ium

(Fin

al)

Dilu

te s

ugar

wast

e (1

:1)

(Fin

al)

Dilu

te s

ugar

wast

e (2

:1)

(Fin

al)

Med

ia s

ugar

wast

e (F

inal

)Co

ncen

trate

ddi

ary

wast

e(F

inal

)Di

lute

dia

rywa

ste

(Fin

al)

Med

ia d

iary

wast

e (F

inal

)

COD

valu

es in

mg/

L

Initial and Final COD values of the Wastewater before and after Treatment

 

• COD has reduced from 1185 mg/L to an average of 20 mg/L in Dairy waste

• There is a considerable amount of reduction of COD from 1560 mg/L to an average of 650 mg/L in spent wash 25

Page 26: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Growth of Algae in Different Partial Pressures of CO2

• The biomass density is observed to be the highest in control

• 3% Carbon-dioxide has the second highest biomass density (about 2.56 mg/L)

26

00.5

11.5

2

2.53

3.54

4.5

0 3 6 12 18 21 24 27 30

Number of days

To

tal C

hlo

rop

hyl

l in

mg

/LControl

0.50%

1%

2%

3%

Page 27: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Influence of the culture with different partial pressures of CO2 on lipid production

• 2% CO2 has lead to the maximum lipid production (58%) when compared to control (62%).

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0

10

20

30

40

50

60

70

Control 0.5% (v/v) 1% (v/v) 2% (v/v) 3% (v/v)

Lipi

ds in

per

cent

age

Page 28: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Influence of the culture with pH on the lipid production

• pH of 5.5 and 8 are seen to be optimum pH for the maximum lipid production

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05

101520253035404550

pH 4 pH 4.5 pH 5 pH 5.5 pH 6.0 pH 6.5 pH 7 pH 7.5 pH 8 pH 8.5

Lipi

d in

Per

cent

age

•The final pH of all the samples after 30 days was found to be 6.3 – 7.2 after the inoculation period

Page 29: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Page 30: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

• The sugar industries and dairy industries discharge their wastewater directly in the river and municipal sewers respectively. The usage of the wastewater for the production of hydrocarbon can open up new opportunities of generating energy from waste.

• The growth of algae has proved to improve with the increase in the partial pressure of CO2 on the culture media which shows that the algae has high CO2 mitigation capacity. Converting a 100-MW thermal power plant from using coal to using liquid fuel derived from B.braunii can reduce CO2

emissions by 1.5 X 105 tons/yr (Sawayama et al., 1999).

• The algae has , further, been able to grow in various pH and has successfully neutralized a wide range of pH.

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Page 31: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

Source: http://www.freewebs.com/renata17/Biodiesel%20oil.JPG

CO2

Was

tewat

er

Biofuel

Page 32: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Banerjee, A., Sharma, R., Yusuf, C., Banerjee, U.C., 2002. Botryococcusbraunii: a renewable source of hydrocarbons and other chemicals.Crit. Rev. Biotechnol. 22, 245–279.

Dayananda, C., Sarada, R., Bhattacharya, S., Ravishankar, G.A., 2005.EVect of media and culture conditions on growth and hydrocarbonproduction by Botryococcus braunii. Process Biochem. 40,3125–3131.

Florencio F.J. and Vega J.M. 1983. Utilization of nitrate, nitritem and ammonium by Chlamydomonas reinhardtii. Planta 158: 288–293. Fogg G.E. 1966. Algal Cultures and Phytoplankton Ecology. University of London Press, University of Wisconsin Press, London, Madison, Milwaukee, pp. 44–46. Garbisu C., Gil J.M., Bazin M.J., Hall D.O. and Serra J.L. 1991. Removal of nitrate from water by foam-immobilize Phormidium laminosum in batch and continuous-flow bioreactors. J. appl. Phycol. 3: 221–234.

Page 33: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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Largeau C., Casadevall E., Berkaloff C. and Dhamelincourt P. 1980. Sites of accumulation and composition of hydrocarbons in Botryococcus braunii. Phytochem. 19: 1043–1051. Lee S.J., Yoon B.D. and Oh H.M. 1998. Rapid method for the determination of lipid from the green alga Botryococcus braunii. Biotechnol. Techniq. 12: 553–556.

Lowry O. H., Rosebrough N. I., Farr A. L. and Randall R. J. (1951) Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265-275. Martínez M.E., Sánchez S., Jiménez J.M., Yousfi F.E. and Muñoz L. 2000. Nitrogen and phosphorus removal from urban wastewater by the microalga Scenedesmus obliquus. Biores. Tech. 73: 263–272.

M. Dubois, K.A. Gilles, J.K. Hamilton, P.A. Reebers, F. Smith, Anal.Chem. 28 (1956) 350–356. Okada S., Murakama M. and Yamaguchi K. 1997. Hydrocarbon production by theYayoi, a new strain of the green microalga Botryococcus braunii. Appl. Biochem. Biotechnol. 67: 79–86.

P. Rao, T.N. Pattabiraman, Reevaluation of the phenol–sulfuric acid reaction for the estimation of hexoses and pentoses, Anal. Biochem. 181 (1989) 18–22. Sawayama S., Minowa T., Dote Y. and Yokoyama S. 1992. Growth of the hydrocarbon-rich microalga Botryococcus braunii in secondarily treated sewage. Appl. Microbiol. Biotechnol. 38: 135– 138.Wolf F.R., Nonomura A.M. and Bassham J.A. 1985. Growth and branched hydrocarbon production in a strain of Botryococcus braunii (Chlorophyta). J. Phycol. 21: 388–396. Yamaguchi K., Nakano H. and Murakami M. 1987. Lipid composition of a green alga, Botryococcus braunii. Agric. biol. Chem. 51: 493–498.

Page 34: By Abitha.R, Research Scholar, Indian Institute of Science Bangalore. Conducted under the guidance of Dr.R.Sharada, Scientist, Central Food Technological

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