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Cross-border network for knowledge transfer and innovative development in wastewater treatment

WATERFRIENDHUSRB/1203/221/196

1st HUSRB Students Meeting

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1st Students Meeting

Waterfriend 1st Students Meeting

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Bioethanol production from cellulosic and hemicellulosic agricultural waste

Dóra VitayBioengineering Bsc. student

Supervisor: prof. Dr. Cecília HodúrCo-supervisor: Marietta Ábel

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Objectives

• How tobacco can be used as a feedstock for bioethanol production

• Cellulose and hemicellulose enzymatic degradation:• With cellulase and cellobiase mixture on different pH• With xylanase

• Possibility of enzyme recycling• Membrane separation• Monitoring the activity of used and separated

enzymes

Tobacco plant

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Materials and methods• Tobacco: (Nicotiana rustica, Nicotiana tabacum)

• Usually lives in tropic or subtropical zone, angiosperm plant

• 1-2 meters high, good adaptibility• Used samples:

• Experimental leaves + vines• By-product remaining vines after manufacturing

process • Grinding • Fermentation in shaking water bath with cellulase (Asp.

niger), cellobiase (Trichoderma reesei) and xylanase enzymes

• Reducing sugar analysis with 3,5-dinitrosalicylic acid (DNSA)

Sample before grinding

Sample after grinding

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Materials and methods• Ultrafiltration: (Millipore)

• 5 KD, PES (poliethersulfone) membrane• 3.5 bar

• Ultrasonic wave generator• Reused enzyme activity inspection

• „filter paper” / avicell test

Shaking water bath Ultrafilrtation equipment with ultrasonic wave generator

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pH comparisontobacco [g] H2O [cm3] cellulase

[cm3] cellulase [Unit] cellobiase [cm3]

Cellobiase [Unit]

BP0 4 50 0 0 0 0BP1 4 50 0.466 391.44 0.386 115.8EX0 4 50 0 0 0 0EX1 4 50 0.372 312.48 0.458 137.4

• Circumstances: • pH = 5.0 and 4.8 • T = 50°C

• Sampling and pH setting in every 24 hours

• Reducing sugar analysis on every sample Experimental tobacco samples after fermentation

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pH comparison

24 48 72 96

-0.050

0.000

0.050

0.100

0.150

0.200

0.250

0.300Experimental pH=5.0 Experimental pH=4.8 By-product pH=5.0By-product pH=4.8

Fermentation time [h]

Suga

r coc

nent

ratio

n [m

g/cm

3Uni

t]

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Results of degradation with xylanaseSample

Xylanase enzym [mg]

Xylanase enzym [Unit]

Tobacco [g]

Water [cm3]

0 0 0

4 50

1 200 200

2 100 100

3 50 50

4 25 25

5 12,5 12,5

Circumstances:• pH = 4.5• T = 30°CSampling and pH setting in every 24 hoursReducing sugar analisys on every sample (

Experimental tobacco samples after fermentation

Experimental tobacco samples during reducing sugar analysis

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Results of degradation with xylanase

24 48 72 960.000

0.200

0.400

0.600

0.800

1.000

1.200

EX1EX2EX3EX4EX5


Spec

ific

suga

r con

cent

ratio

n [m

g/cm

3Uni

t]

24 48 72 960.000

0.200

0.400

0.600

0.800

1.000

1.200

BP1BP2BP3BP4BP5


Spec

ific

suga

r con

cent

ratio

n [m

g/cm

3Uni

t]

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Results of membrane separation• Circumstances:

• 3.5 bar• 350 rpm

• Membrane: 5KD PES• Ultrasound occasionally

By product samples: feed, concentratum, permeatum

Flux equation

Resistance equation

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Results of membrane separation

0 500 1000 1500 2000 250010.000

15.000

20.000

25.000

30.000

35.000

40.000

45.000

50.000

ExperimentalBy-productExperimental + USBy-product + US

Time [s]

J [L/

m2

h]

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Results of membrane separation

RGRF

RM

0.0E+005.0E+121.0E+131.5E+132.0E+132.5E+13

By-product

Experimental

Experimental + US

By-product + USRe

sist

ance

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Waterfriend

Results of enzyme recycling

0 240.000

0.500

1.000

1.500

2.000

2.500

f(x) = 0.067635784971823 ln(x) + 0.294056703447632

f(x) = 0.150499357223816 ln(x) + 1.84252082693657

f(x) = 0.017688843782107 ln(x) + 0.0264798921287508

f(x) = 0.116282035373111 ln(x) + 0.932924082372535

ExperimentalLogarithmic (Experimental)By-productLogarithmic (By-product)Experimental + USLogarithmic (Experimental + US)time [h]

mg

sug

ar/g

dry

mas

s

• Circumstances: • pH = 5.0• T = 50°C

• Sampling and pH setting in every 24 hours

• Reducing sugar analisys on every sample

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Conclusion• The main goals of our experiments were to study the possibilities of the

by-product tobacco usage• As a feedstock for bioethanol production

• First, we used cellulase and cellobiase mixture and xylanase enzymes to hidrolyze the samples

• Both enzymatic ways are able to degrade the tobacco• There was no significant difference between pH 4.8 and 5.0• Xylanase was more effective than cellulase-cellobiase mixture

• Further studies showed, that membrane separation is a possible method to regain the enzymes

• After separation the enzymes keeped their activity, even after the ultrasound treatment, but with lesser activity

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Thank you for your kind attention!

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Presentation/lecture has been produced with the financial assistance of the European Union. The content of the presentation/lecture is the sole responsibility of University of Novi Sad, Faculty of Technology and can under

no circumstances be regarded as reflecting the position of the European Union and/or the Managing Authority.

Acknowledgements

Jelen eredmények megjelenését „Zöld Energia Felsőoktatási Együttműködés (ZENFE)” című, TÁMOP-4.1.1.C-12/1/KONV-2012-0012 azonosítószámú projekt támogatja. A projekt az Európai Unió támogatásával,

az Európai Szociális Alap társfinanszírozásával valósul meg.

A szerzők köszönetet mondanak az Országos Tudományos Kutatási Alapprogram pénzügyi támogatásáért. A projekt azonosító száma: K 105021.

I would like to thank to my supervisor prof. dr. Cecília Hodúr and my co-supervisor Marietta Ábel.

cross-border network for knowledge transfer and innovative development in wastewater treatment...

Documents

xylanase slide

sample slide

fermentation slide

waterfriend materials

waterfriend conclusion

waterfriend objectives

husrb students

sugar analysis slide