BIODEGRADATION OF POLYCYCLIC AROMATIC HYDROCARBONS IN PETROLEUM OIL ...

339 Pages·2014·5.36 MB·English

by  El Zahar

Aim of the Work

BIODEGRADATION OF
POLYCYCLIC AROMATIC
HYDROCARBONS IN PETROLEUM
OIL CONTAMINATING THE
ENVIRONMENT

Presented by

Abir Moawad Partila

A Thesis Submitted
to
Faculty of Science

In Partial Fulfillment of the
Requirements for
the Degree of
Ph.D. of Science
(Microbiology)

Botany Department
Faculty of Science
Cairo University

(2013)
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Aim of the Work

ABSTRACT

Student Name: Abir Moawad Partila Girgis

Title of the thesis: Biodegradation of Polycyclic Aromatic Hydrocarbon
in petroleum oil contaminating the environment

Degree: Ph. D. (Microbioliogy)
Soil and sludge samples polluted with petroleum waste from Cairo
Oil Refining Company Mostorod, El-Qalyubiah, Egypt for more than 41
years were used for isolation of indigenous microbial communities. These
communities were grown on seven polycyclic aromatic hydrocarbon
compounds .Six isolates (MAM-26, 29, 43, 62, 68, 78) were able to grow on
different concentrations of five chosen PAHs. The best degraders bacterial
isolates MAM-29 and MAM-62 were identified by 16S-rRNA. As
Achromobacterxylosoxidans and Bacillus amyloliqueficiensrespectively.The
most promising bacterial strain Bacillus amyloliqueficiens have been exposed
.
to different doses of gamma radiation to improve its qualities

Keywords: Polycyclic- Aromatic- Hydrocarbon – Biodegradation-Pollution.

Supervisors:
Signature
1- Prof. Dr. Youssry Saleh
2-Prof. Dr. Mervat Aly Abou-State

Prof. Dr. Gamal Fahmy
Chairman of Botany Department

Faculty of Science-Cairo University

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Aim of the Work
APROVAL SHEET FOR SUMISSION

Thesis Title: Biodegradation of Polycyclic Aromatic
Hydrocarbons In Petroleum Oil Contaminating The Environment

Name of candidate: Abir Moawad Partila

This thesis has been approved for submission by the
supervisors:

1- Prof. Dr. Youssry Saleh

Signature:

2- Prof. Dr. Mervat Ali Abou State

Signature:

Prof. Dr. Gamal Fahmy

Chairman of Botany Department
Faculty of Science- Cairo University

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Aim of the Work

TO WHOM IT MAY CONCERN

This Thesis has not been previously submitted for any
degree at this or at any other university.

Signature

Abir Moawad Partila
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Aim of the Work

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Aim of the Work
Acknowledgement
I would like to acknowledge with deep gratitude Prof. Dr.
Youssry Saleh, Professor of Microbiology, Botany Department,
Faculty of Science, Cairo University for valuable advice and his
direct supervision and support during the stages of this work.
From my deep heart my thanks are for Prof. Dr. Mervat
Aly Abou-State Professor of Microbiology Department of
Microbiology, National Center for Radiation Research and
Technology, Nasr City, Cairo for her great active continuous help
in the theoretical and practical parts of this work and for her direct
supervision.
I wish to thanks also Prof. Dr. Nagy Halim Aziz
Professor of Microbiology Department of Microbiology, National
Center for Radiation Research and Technology, Nasr City, Cairo
for his encouragement.
I wish also to express my deepest thanks to every body else
who contributed in any way in this work.
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Aim of the Work

Dedication

To

The Spirit of my Father

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LIST OF CONTENTS

Title Page No.
Introduction ........................................................................................ i
Aim of hte Work ................................................................................. iii
1. Literature review ............................................................................ 1
1.1. Spreading of polycyclic aromatic hydrocarbons
(PAHs) in nature ........................................................................ 1
1.2. Health impact of PAHs .............................................................. 7
1.3. Petroleum oil contamination ...................................................... 23
1.4. Polycyclic aromatic hydrocarbons as constituent of
petroleum contaminants .............................................................. 24
1.5. Microorgnaisms degrading PAHs. ............................................. 34
1.6. Mechanism of PAHs degradation strains .................................... 42
1.7. Pathway for PAHs degradation .................................................. 53
2. Materials and Methods ............................................................ 67
2.1. Materials ................................................................................... 67
2.2. Methods ..................................................................................... 73
3. Results and discussion ............................................................. 80
3.1. Growth of different indigenous bacterial communities
on different PAHs ....................................................................... 80
3.2. Determination of the total bacterial count and the
hydrocarbon degrading bacteria (HDB) found in each
community ................................................................................. 111
3.3. Isolation and determination of the most potent strains
having the ability to degrade different PAHs............................... 113
3.4. Growth and degradation of naphthalene by the most
potent isolated strains ................................................................ 118
3.5. Growth and degradation of phenanthrene by the most
potent isolated strains ................................................................ 135
3.6. Growth and degradation of anthracene by the most
potent isolated strains ................................................................. 155
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LIST OF CONTENTS
(Cont…)

Title Page No.

3.7. Growth and degradation of pyrene by the most potent
isolated strains ............................................................................ 189
3.8. Growth and degradation of benzo-a-anthracene by the
most potent isolated strains ......................................................... 207
3.9. Identification of the most potent PAHs degrading
bacterial strains .......................................................................... 224
3.10. Effect of gamma radiation on the viability of Bacillus
amyloliquefaciens ...................................................................... 230
3.11. Selection of the hyper PAHs degrading bacterial
mutant ....................................................................................... 232
3.12. Pathway of B. amyloliquefaciens for degradation of
PAHs compounds. ...................................................................... 236
Summary ............................................................................................ 256
References .......................................................................................... 260
Arabic Summary ................................................................................ –––

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LIST OF FIGURES

Fig. No. Title Page No.
Figure (1): Proposed pathway for the degradation of
naphthalene by Pseudomonas putida.......................... 55
Figure (2): Proposed pathway for the degradation of
naphthalene by Streptomyces griseus ......................... 55
Figure (3): Proposed pathway for the degradation of
phenanthrene by Sphingomonas sp............................. 58
Figure (4): Postulated metabolic pathway of PAH-
degradation in aerobic bacteria .................................. 58
Figure (5): Proposed phenanthrene degradation
pathways by the managrove enriched
bacterial consortium .................................................. 59
Figure (6): Proposed pathway for the degradation of
phenanthrene by pleurotus ostreatus .......................... 60
Figure (7): Proposed pathway for the degradation of
anthracene by Aspergillus fumigatus .......................... 62
Figure (8): Proposed pyrene degradation pathways by
the mangrove enriched bacterial
consortium ................................................................. 64
Figure (9): Proposed pathway for the degradation of
pyrene by Mycobacterium flavescens .......................... 65
Figure (10): Proposed pathway for the degradation of
pyrene by Mycobacterium sp. strain PYR-
1 ................................................................................ 65
Figure (11): Proposed pathway for the degradation of
pyrene by Aspergillus niger SK 93/7 ......................... 66
Figure (12): Proposed pathway of benzo[a]anthracene
degradation by the ligninolytic fungus
Irpex lacteus .............................................................. 67
Figure (13): Proposed pathways for the degradation of
[B-a-Anth.] by Mycobacterium sp. strain
RJGII-135 based on isolated metabolites ................... 67
Figure (14): Sampling site map....................................................... 69
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