Tăng cường hoạt tính sinh học của hoạt động oxy hóa Amoni bởi Graphene Oxide và Titanium Nanotubes

國立交通大學

環境工程研究所

碩士論文

利用氧化石墨烯及鈦酸鹽奈米管增強厭氧氨

氧化菌活性之研究

Bio-augmentation of anaerobic ammonium oxidation

activity supported on graphene oxide and titanium

nanotubes

研究生：屈氏青玄(Khuat Thi Thanh Huyen)

指導教授：董瑞安教授 (Dr. Ruey-an Doong)

林志高教授 (Dr. Jih-Gaw Lin)

中華民國106年 7 月

利用氧化石墨烯及鈦酸鹽奈米管增強厭氧氨氧化菌活性之研究

Bio-augmentation of anaerobic ammonium oxidation activity

supported on graphene oxide and titanium nanotubes

研究生： Student：Thi Thanh Huyen Khuat

指導教授： Advisor：Ruey-an Doong

Jih-Gaw Lin

國 立 交 通 大 學

環 境 工 程 研 究 所

碩 士 論 文初稿

A Thesis

Submitted to Institute of Environmental Engineering

College of Engineering

National Chiao Tung University

in Partial Fulfillment of the Requirements

for the Degree of

Master of Science

in

Environmental Engineering

July, 2017

Hsinchu, Taiwan

中 華 民 國 106年 7 月

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Bio-augmentation of anaerobic ammonium oxidation activity supported on graphene oxide

and titanium nanotubes

Student：Thi Thanh Huyen Khuat Advisor：Ruey-an Doong

Jih-Gaw Lin

Institute of Environmental Engineering

National Chiao Tung University

ABSTRACT

Nitrogen removal from wastewater is gaining worldwide attention because of the potential

threat of nitrogen species to the environment. Anaerobic ammonium oxidation (Anammox) is a

promising method which can convert ammonium ion directly to nitrogen gas. However,

Anammox still faces the difficulty in the enhancement of slow growth rate of Anammox

microorganisms. Herein, we have systematically investigated the effect of graphene oxide (GO)

and titanium nanotube (TNTs) on the Anammox biomass for nitrogen removal. TNTs, the 1-

dimensional negatively charged nanomaterials show little effect on the enhancement of

Anammox growth, presumably attributed to the strong repulsive force between TNTs and

Annamox bacteria. GO makes a remarkable impact on the bacterial growth. After the batch

incubation of 5h, the Anammox activity in the presence of 0.1 g/L GO is enhanced 14.2 % when

compares with that in the absence of nanomaterial. The continuous experiment also proves the

applicability of using GO as an effective support to improve nitrogen removal. GO utilization can

enhance Anammox activity by 30% compares with that of the normal reactor after 113d of

incubation. In the GO induced reactor, the effluent concentrations of ammonium and nitrite

decrease steadily and reach the steady state after 80d of incubation, while it takes 127 d in the

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reactor without GO to achieve the steady state. These results clearly demonstrate the feasibility of

utilizing GO as the support for Anammox bacteria to shorten the start-up incubation time by rapid

acclimation as well as to enhance the activity of Anammox bacteria.

Keywords: Graphene oxide, Anammox, nitrogen removal, start-up

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ACKNOWLEDGEMENT

Foremost, I would like to express my sincere gratitude to my both advisors Prof. Ruey-an

Doong and Prof. Jih-Gaw Lin for their infinite guidance, patience, generous encouragement and

supports to me during my two years of studying at National Chiao Tung University. Their great

insight and profound knowledge was of great help for me to finish my work step by step. I

appreciated all the opportunities my professors offered me by giving me intellectual freedom in

my work, supporting my attendance at various conferences, engaging me in new ideas, and

demanding a high quality of work in all my endeavors.

Besides my advisors, I would like to thank the rest of my thesis committee: Prof. Shen-Yi

Chen and Prof. Ji-Doong Gu for spending their valuabe time to help me to evaluate my thesis

defense and give me some suggestion to enrich my work.

My deep gratitude also goes to fellow labmates in ECCL, including Akhilesh, Rama,

Amber, Bin and Anh for their kindness and help in training me and for many valuable advices

that given to me whenever I get stuck with my research. I would be remiss if I did not thank to

Dr. Ankan, from his keen scientic insight and his ability to put complex ideas into simple terms to

broaden and stimulated my research interest. I would also like to thank to Prof. Lin labmates for

their help and supporting, including Mak, Jolin, Dai, and Henry. Iam also fortunate to meet a

great Taiwanese friend who always give me a hand without any hesitation whenever I need help.

Special thanks to Shila for supporting me during two years in Taiwan. I woud also like to

acknowledge to all other fellow labmates in ECCL for their assistances in many aspects that I can

not list them all here. I learnt a lot from all of you and all of you are helpful to color my stay in

Taiwan.

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I would also like to express my gratitude to Institute of Environmental Engineering,

National Chiao Tung University for all the opportunities they have given me over the past two

years.

Last but not the least, I would like to thank my family for their unconditional love and

support they gave me during these years.

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Content Index

Chapter 1. Introduction .................................................................................................................1

1.1. Motivation............................................................................................................................. 1

1.2. Objective............................................................................................................................... 2

Chapter 2. Background and theory ..............................................................................................4

2.1 Nitrogen cycle........................................................................................................................ 4

2.2 Anaerobic ammonium oxidation ........................................................................................... 5

2.3 Cell structure and metabolism of Anammox ......................................................................... 6

Figure 2.2 Anammox cell with different compartments and three surrounding membranes.7

2.4 Reactor system used for enrichment of Anammox bacteria .................................................. 9

2.5 Nanomaterials...................................................................................................................... 12

2.5.1 Definition ...................................................................................................................... 12

2.5.2 Properties...................................................................................................................... 12

2.6. Graphene material............................................................................................................... 12

2.6.1 Concepts and properties................................................................................................ 12

2.6.2 Bacterial interaction with Graphene Oxide and surfaces.............................................. 15

2.6.3 Anammox bacteria interact with different Graphene family materials......................... 19

2.7 Titanium nanotubes (TNTs)................................................................................................. 20

2.7.1 Concepts and properties................................................................................................ 20

2.7.2 Photochemical Properties and Photocatalytic Functions .............................................. 23

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2.7.3 Parameters influencing the morphology of titanate nanotubes..................................... 25

2.7.4 Bacteria and cell interaction with titanium nanotubes.................................................. 28

2.8 Interaction between bacteria and surface............................................................................. 29

Chapter 3. Materials and methods..............................................................................................31

3.1. Synthesis............................................................................................................................. 31

3.1.1 Synthesis of GO ............................................................................................................ 31

3.1.2 Synthesis of TNT .......................................................................................................... 33

3.2 Instrumentations................................................................................................................... 34

3.2.1 X-ray diffraction (XRD)................................................................................................ 34

3.2.2 Raman spectroscopy...................................................................................................... 34

3.2.3 Fourier transform infrared spectroscopy (FTIR)........................................................... 35

3.2.4 Transmission electron microscope (TEM).................................................................... 35

3.2.5 Scanning electron microscope (SEM)........................................................................... 35

3.2.6 Brunauer–Emmett–Teller (BET)................................................................................... 36

3.2.7 UV-visible spectrometer (UV-Vis)............................................................................... 37

3.3 Specific Anammox activity test........................................................................................... 37

3.3.1 Materials and Equipemnt .............................................................................................. 37

3.3.2 Theory of Specific Anammox Activity (SAA) test....................................................... 38

3.4 Long-term effects of GO on Anammox activity.................................................................. 39

3.4.1 Microorganisms and feeding medium........................................................................... 39