Home >Documents >STUDY OF REMOVAL BTX BY PSEUDOMONAS PUTIDA IN CD8325) mohd   STUDY OF REMOVAL BTX BY...

STUDY OF REMOVAL BTX BY PSEUDOMONAS PUTIDA IN CD8325) mohd   STUDY OF REMOVAL BTX BY...

Date post:08-Mar-2019
Category:
View:212 times
Download:0 times
Share this document with a friend
Transcript:

III

STUDY OF REMOVAL BTX BY PSEUDOMONAS

PUTIDA IN BIOREACTOR

MOHD ASHRAF BIN ABDUL RAHMAN

Thesis submitted in partial fulfilment of the requirements

for the award of the degree of

Bachelor of Chemical Engineering

Faculty of Chemical & Natural Resources Engineering

UNIVERSITI MALAYSIA PAHANG

JULY 2013

MOHD ASHRAF BIN ABDUL RAHMAN (2013)

VIII

ABSTRACT

Benzene, toluene and xylene isomers (BTX) are the major components of gasoline which it

is used widely as solvents and feedstock by the chemical industry. It became a common

environmental contaminant problem that represents a serious threat to groundwater resources

and to public health, being both toxic and relatively water soluble compared to other

petroleum constituents. The biological treatment that use microorganism such as

Pseudomonas putida has been selected because the ability to reduce the hazard of BTX. The

objective of this research is to remove the BTX from wastewater using P. putida in a

bioreactor. Before starting the experiment, P. putida is prepared by the growth it inside

nutrient broth at a suitable temperature and concentration. The bacteria are put in to ratio of

benzene, toluene, xylene, benzene-toluene, benzene-xylene, toluene-xylene and BTX

solution inside simple conical flask. But the only BTX sample is only running in the

fermenter for comparing the growth of bacteria in a conical flask with the growth of bacteria

in fermenter. The bacterial growth will be analyzed using UV-Vis Spectrophotometer while

concentration BTX after degradation is analyzed by HPLC. From the data obtain, the growth

rate of bacteria in toluene sample is higher than other solution. While the growth rate of

bacteria in BTX of conical flask is higher than the growth rate of bacteria in BTX of the

fermenter. From overall experiment, Toluene is achieved the highest percent of removal

compare to percent removal of benzene and xylene. Thus; removal of BTX by bacteria will

be a potential biological treatment which it can solve our environmental pollutants in the

future.

IX

ABSTRAK

Benzena, toluena dan isomer xilena (BTX) adalah komponen utama petrol mana ia digunakan

secara meluas sebagai pelarut dan bahan mentah oleh industri kimia. Ia menjadi masalah

pencemaran alam sekitar yang biasa merupakan ancaman serius kepada sumber air bawah

tanah dan kesihatan awam, yang kedua-dua toksik dan agak larut dalam air berbanding

dengan juzuk petroleum lain. Rawatan biologi yang menggunakan mikroorganisma seperti

Pseudomonas putida telah dipilih kerana keupayaannya untuk mengurangkan bahaya BTX.

Objektif kajian ini adalah untuk menghapuskan BTX dari air sisa menggunakan P. putida di

dalam bioreaktor. Sebelum memulakan eksperimen, P. putida disediakan oleh pertumbuhan

ia di dalam sup nutrien pada suhu yang sesuai dan penumpuan. Bakteria dimasukkan ke

dalam kepada nisbah benzena, toluena, xilena penyelesaian, benzena-toluena, benzena-

xilena, toluena-xilena dan BTX di dalam kelalang kon yang mudah. Tetapi satu-satunya BTX

sampel hanya berjalan dalam fermenter untuk membandingkan pertumbuhan bakteria dalam

satu kelalang dengan pertumbuhan bakteria dalam fermenter. Pertumbuhan bakteria akan

dianalisis dengan menggunakan spektrofotometer UV-Vis manakala BTX tumpuan selepas

kemusnahan dianalisis oleh HPLC. Dari data yang diperolehi, kadar pertumbuhan bakteria

dalam sampel toluene adalah lebih tinggi daripada penyelesaian lain. Walaupun kadar

pertumbuhan bakteria dalam BTX kelalang kon adalah lebih tinggi daripada kadar

pertumbuhan bakteria dalam BTX daripada fermenter itu. Dari eksperimen keseluruhan,

Toluena dicapai peratus penyingkiran tertinggi berbanding dengan peratus penyingkiran

daripada benzena dan xilena. Oleh itu, penyingkiran BTX oleh bakteria akan menjadi

rawatan biologi yang berpotensi yang ia boleh menyelesaikan pencemaran alam sekitar kita

pada masa hadapan.

.

X

TABLE OF CONTENTS

.

SUPERVISORS DECLARATION.IV

STUDENTS DECLARATION.....V

DEDICATION..VI

ACKNOWLEDGEMENT...VII

ABSTRACT...VIII

ABSTRAKIX

TABLE OF CONTENTS...X

LIST OF FIGURES....XIII

LIST OF TABLE.XV

LIST OF ABBREVIATIONSXVI

LIST OF ABBREVIATIONS...XVII

1 INTRODUCTION..1

1.1 Motivation and statement of this problem.1

1.2 Objectives..........2

1.3 Scope of this research2

1.4 Main contribution of this work..............2

1.5 Organization of this thesis.3

2 LITERATURE REVIEW..................4

2.1 Overview...4

2.2 BTX...................4

2.2.1 Benzene5

XI

2.2.2 Toluene.7

2.2.3 Xylene...........9

2.3 Pseudomonas putida..11

2.4. Shake Flask Culture....15

2.4.1 Advantages.15

2.4.1 Disadvantages.15

2.5 Bioreactor....16

2.5.1 Fermenter...16

2.5.1.1 Advantages.18

2.5.1.2 Disadvantages.18

2.6 Summary...19

3 METHODOLOGY.20

3.1 Overview..20

3.2 Preparation of culture medium.20

3.3 Preparation of stock solution (BTX) ...22

3.4 Preparation of Acid and Base for Fermenter.22

3.5 Preparation Sample...23

3.6 Monitoring Growth of Bacteria.25

3.7 Analysis Method...25

3.8 Summary...27

4 RESULT AND DISCUSSION..28

4.1 Overview.28

4.2 Pseudomonas putida Growth Curve in Shake Flask...28

XII

4.3 Pseudomonas Putida Growth Curve in Fermenter.31

4.4 Comparison Between Growth Rate of Pseudomonas Putida in Shake Flask Culture

with Growth Rate of Pseudomonas Putida in Fermenter.33

4.5 Standard Curve of Benzene, Toluene and Xylene......34

4.6 Percent Removal BTX....37

4.7 Summary.....39

5 CONCLUSION..40

5.1 Conclusion..40

5.2 Recommendation40

6 REFERENCE.42

7 APPEENDICES.49

XIII

LIST OF FIGURES

Figure 2.1: Major products derived from BTX (US DOE, 2012 and Anne, 2013)..4

Figure 2.3: Benzene Structure (EPA, 1988).6

Figure 2.5: Toluene Structure (HSDB, 2008)..8

Figure 2.7: o-xylene, m-xylene and p-xylene structure (HSDB, 2009).10

Figure 2.8: Pseudomonas aeruginosa (Dr. Howard Porter, 2012).13

Figure 2.9: Pseudomonas putida (Fekete et al., 2009....14

Figure 2.10: Shake Flask....15

Figure 2.11 (a): Head Plate of Fermenter (Fauzan et al, 2012).17

Figure 2.11 (b): Fermenter Body Parts (Fauzan et al, 2012).18

Figure 3.1: Overall Process BTX Removal by P. putida in Bioreactor....20

Figure 3.2: Preparation of Bacteria....21

Figure 3.3: Preparation of Stock Solution of BTX.22

Figure 3.4: Process of Preparation of 0.1 mole of Acid and Base..22

Figure 3.6: The sample of Conical Flask is running in Incubator Shaker..24

Figure 3.7: The Sample inside Fermenter..24

Figure 3.8: UV-Vis Spectrometer..25

Figure 3.9: The sample in Microbiological Centrifuge..25

Figure 3.10: The Flow Process in make Analyze the Sample....26

Figure 3.12: HPLC Equipment..27

Figure 4.1: Graph of Log (OD600) Against Time for Sample in Shake Flask...28

Figure 4.3: Graph of Log (OD600) Against Time for Sample in Fermenter..32

XIV

Figure 4.5: The Graph of Growth Bacteria Against Time of Shake Flask Culture and

Fermenter..33

Figure 4.6: Standard Curve of Benzene35

Figure 4.7: Standard Curve of Toluene..35

Figure 4.8: Standard Curve of Xylene...36

XV

LIST OF TABLES

Table 2.2: Chemical and Physical Properties of benzene (Fischer Scientific, 2008).5

Table 2.4: Chemical and Physical Properties of Toluene (HSDB, 2008)..7

Table 2.6: Chemical and Physical Properties of m-xylene, o-xylene, and p-xylene (Reynas,

2009).9

Table 2.7: Type of P. putida strain and its function.....11

Table 3.5: The Ratio of Sample Used..23

Table 3.11: Specific Parameter of HPLC..26

Table 4.2 : Summary of Specific Growth Rate of Sample29

Table 4.4: Summary of Specific Growth Rate of Sample.32

Table 4.9: Summary Comparison BTX Treatment Result36

XVI

LIST OF ABBREVIATIONS

m micrometer

% percent

oC degree Celsius

atm Atmospheric pressure

g/ml gram per milliliters

g/l gram per liters

hr hour

I liter

mg/l milligram per liters

pH potential Hydrogen

XVII

LIST OF ABBREVIATIONS

Specific Growth rate

ASTDR Agency for Toxic Substances and Disease Registry

BTX Benzene, Toluene and Xylene

DOE Department of Environment

EPA Environment Protection Agency

HCI Hydrochloric Acid

HSDB Hazardous Substances Data Bank

HPLC High Performance Liquid Chromatography

MIDA Malaysian Investment and Development Agency

NaOH Sodium Hydroxide

Click here to load reader

Embed Size (px)
Recommended