Screening and study on microorganisms degrading biopolymers in vietnam

Screening and study on microorganisms
degrading biopolymers in Vietnam

Trịnh Thị Vân Anh


Viện Vi sinh vật và Công nghệ Sinh học
Luận văn Thạc sĩ ngành: Công nghệ sinh học; Mã số: 60 42 80
Người hướng dẫn: TS. Nguyễn Quang Huy
Năm bảo vệ: 2010



Keywords. Vi sinh vật; Tạo vòng phân huỷ; Biopolymers; Công nghệ sinh học

Content
Table of contents
INTRODUCTION 1
CHAPTER 1: LITERATURE REVIEW 4
1.1. PLASTIC WASTE POLLUTION 4
1.1.1. Plastic waste pollution in the world 4
1.1.2. Plastic waste pollution in Vietnam 5
1.1.3. Treatment of plastic waste 6
1.1.3.1. Landfill 6
1.1.3.2. Recycling 6
1.1.3.3. Incineration 7
1.2. BIODEGRADABLE PLASTICS 8
1.2.1. Biodegradable plastics 8
1.2.1.1. Poly(Lactic Acid) (PLA) 11
1.2.1.2. Poly(3-Hydroxybutyrate) (PHB) 12
1.2.1.3. Poly(ε-Caprolactone) (PCL) 13

1.2.2. Applications 13
1.2.2.1. Medicine and pharmacy 14
1.2.2.2. Packaging 15
1.2.2.3. Agriculture 15
1.2.2.4. Others fields 16
1.3. THE DEGRADATION OF BIOPOLYMERS 17
1.4. MICROORGANISMS DEGRADING BIODEGRADABLE POLYMERS 19
1.4.1. Microorganisms degrading PLA 19
1.4.2. Microorganisms degrading PHB 22
1.4.3. Microorganisms degrading PCL 23
CHAPTER 2: MATERIALS AND METHODS 25
2.1. MATERIALS 25
2.2. CHEMICALS 25
2.3. EQUIPMENTS 26
2.4. METHODS 26
2.4.1. Isolation of biopolymer-degrading microorganisms 26
2.4.2. Screening biopolymer-degrading microorganisms 27
2.4.3. Identification of biopolymers-degrading strains 27
2.4.3.1. Gram staining method 27
2.4.3.2. Observation under scanning electron microscopy (SEM) 28
2.4.3.3. Extraction of genomic DNA from bacteria 29
2.4.3.4. Amplification of the 16S rDNA PCR reaction 29
2.4.3.5. Agarose gel electrophoresis 30
2.4.3.6. Sequencing 30
2.4.3.7. Effect of culture conditions 31
2.4.3.8. Utilization of of sugars 31
2.4.3.9. Activity of some extracellular enzymes 31
2.4.4. Study degradation of biodegradable polymers by isolated strains 32
2.4.4.1. Growth experiment in the PLA, PHB or PCL containing media 32
2.4.4.2. Measurement of the PLA, PHB or PCL residual weight 32

2.4.4.3. Determination of TOC in culture broth 32
2.4.4.4. Degradation experiment with biopolymer film 34
2.4.4.5. Statistical analysis 34
CHAPTER 3: RESULTS AND DISCUSSION 35
3.1. ISOLATION AND SCREENING PLA, PHB, PCL-DEGRADING ORGANISMS 35
3.2. PLA-DEGRADING MICROORGANISMS 37
3.2.1. Identification of strains G5 and Cz1 37
3.2.1.1 Morphology of strain G5 and Cz1 38
3.2.1.2. 16S rDNA sequencing of strain G5 39
3.2.1.3. Biochemical and physiological characteristics of strains G5 and Cz1 41
3.2.2. PLA degradation by S. thermoflavus G5 and P. citrinium Cz1 44
3.3. PHB-DEGRADING MICROORGANISM 47
3.3.1. Identification of strain B2 47
3.3.1.1. Morphology of strain B2 47
3.3.1.2. Sequencing 16S rDNA gene of strain B2 48
3.3.1.3. Biochemical and physiological characteristics of strain B2 49
3.3.2. PHB degradation by B. gelatini B2 52
3.4. PCL-DEGRADING MICROORGANISM 54
3.4.1. Identification of strain B1 55
3.4.1.1. Morphology of strain B1 55
3.4.1.2. Sequencing 16S rDNA gene of strain B1 56
3.4.1.3. Biochemical and physiological characteristics of strain B1 57
3.4.2. PCL degradation by Br. agri B1 60
3.5. DEGRADATION OF POLYMERS BY ISOLATED STRAINS 61
CONCLUSIONS 64
FURTHER STUDY 65
REFERENCES 66
WEB REFERENCES 73

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