Application of Prodigiosin Produced by Serratia sp. PDGS120915 Isolated from Stream Water
- Alternative Title
- 하천으로부터 분리한 Serratia sp. PDGS120915 균주로부터 생산된 Prodigiosin의 응용
- Abstract
- Serratia spp are gram-negative bacteria, classified in the large family of Enterobacteriaceae. Serratia are opportunistic human, plant and insect pathogens and have been isolated from soil, water, air, foodstuff, plant surface and animals. Another characteristic feature of Serratia is the production of cell associated red color pigment called prodigiosin.
Prodigiosin, a non-diffusible red pigment, is a secondary metabolite formed by the enzymatic condensation of 2-methyl-3-amylpyrrole and 4-methoxy-2,2`-bipyrrole-5-carboxyaldehyde, leading to a tripyrrole derivative, 2-methyl-3-amyl-6-methoxyprodigiosene. The pigment has no defined role in the physiology of producing strains, but have been reported to have antifungal, antibacterial, algicidal, antiprotozoal, antimalarial activities, immunosuppressive and anticancer activities.
In this study, a bacterial strain PDGS120915 producing a red pigment was isolated from slightly contaminated stream water in Busan, Korea. It was identified as a strain of Serratia sp. based on 16S rDNA gene sequence analysis and physiological characteristics. The prodigiosin was extracted directly using acidified ethanol and characterized. The optimal conditions for pigment production was 25℃, pH 7 and 0% NaCl concentration for 14 h. Also the treatment of carbon source and nitrogen sources such as fructose and beef extract showed increased prodigiosin production. In case of mixed all them, prodigiosin production yield was increased over 5-folds compared with wild type. The purified prodigiosin showed antimicrobial activities against intestinal pathogens such as Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Salmonella typhimurium, Pseudomonas aeruginosa and Vibrio parahaemolyticus, and also inhibited for growth of Methicillin-resistant Staphylococcus aureus (MRSA). Furthermore, purified prodigiosin have synergistic activity with aminoglycoside (amikacin, tobramycin) against intestinal pathogens and with β-lactam antibiotics (ampicillin, penicillin) against MRSA. The purified prodigiosin showed algicidal activity against dinophyceae, Alexandrium catenella, Gymnodinium impudicum and Cochlodinium polykrikoides. In addition, it showed a broad range of algicidal activities against raphidophyceae (Chattonella marina, Heterosigma akashiwo), coscinodiscophyceae (Skeletonema costatum) and bacillariophyceae (Nitzchia pungens). The optimum concentration of this activity was observed at 5 ppb. Additionally, algicidal activity increased rapidly after 9 h and showed more than 80% algicidal activity at 12 h. This research suggested that the prodigiosin purified from bacterium Serratia sp. PDGS120915 could be a potential bio-agent for the prevention and control of public health.
- Issued Date
- 2016
- Awarded Date
- 2016. 2
- Type
- Dissertation
- Publisher
- 부경대학교 대학원
- Affiliation
- 부경대학교 대학원
- Department
- 대학원 미생물학과
- Advisor
- 김영태
- Table Of Contents
- List of Tables V
List of Figures VI
List of Abbreviation VII
Abstract VIII
Part I. General Introduction of Serratia and Prodigiosin 1
Chapter 1. Overview of the Genus Serratia and Prodigiosin 2
1. Introduction of Serratia 3
2. Ecology 3
3. Compounds produced by Serratia 4
4. Prodigiosin production conditions 6
5. Prodigiosin structure 8
6. Prodigiosin biosynthetic clusters 8
7. Effects of prodigiosin 9
8. References 15
Part II. Application of Prodigiosin Produved by Serratia sp. PDGS120915 Isolated from Stream Water 19
Chapter 2. Isolation and Characterization of Serratia sp. PDGS120915 Isolated from Stream Water which Produces Prodigiosin 20
1. Abstract 21
2. Introduction 22
3. Materials and Methods 22
3.1. Isolation and identification of bacterial strains 22
3.2. Extraction of prodigiosin 23
3.4. Estimation of prodigiosin 23
3.4. Optimization of prodigiosin production 24
4. Results and Discussion 25
4.1. Isolation and identification of a prodigiosin producing microorganism 25
4.2. Optimum culture conditions of Serratia sp. PDGS120915 25
4.3. Characteristics of pigment 29
4.4. Effects of culture media for the production of prodigiosin 29
4.5. Effects of medium composition for the prodigiosin production 33
5. References 36
Chapter 3. Antimicrobial Activity of Prodigiosin from Serratia sp. PDGS120915 Against Intestinal Pathogens 39
1. Abstract 40
2. Introduction 41
3. Materials and Methods 41
3.1. Bacterial strains and medium 41
3.2. Antimicrobial activity against intestinal pathogens 42
3.3. Synergistic effect between extract and antibiotics 42
4. Results and Discussion 44
4.1. Antimicrobial activity 44
4.2. Determination of MIC and MBC 44
4.3. Determination of FIC indices 44
5. References 49
Chapter 4. Anti-MRSA properties of prodigiosin from Serratia sp. PDGS120915 52
1. Abstract 53
2. Introduction 54
3. Materials and Methods 54
3.1. Bacterial strains and medium 54
3.2. Disc diffusion assay 55
3.3. Measurement of MIC 55
3.4. Isolation and identification of anti-MRSA substance producing microorganisms 55
3.5. Synergistic effect between extract and β-lactams against MRSA 55
4. Results and Discussion 57
4.1. Anti-MRSA activity of Serratia sp. PDGS120915 57
4.5. Synergistic effects between prodigiosin and β-lactams against MRSA 57
5. References 61
Chapter 5. Algicidal effect of Serratia sp. PDGS120915 against harmful dinophyceae algae 63
1. Abstract 64
2. Introduction 65
3. Materials and Methods 65
3.1. Algal cultures 66
3.2. Bacterial strain 66
3.3. Characterization of algicidal effect 66
3.6. Mode of activity 76
3.7. Analysis of algicidal activity 67
4. Results 68
4.1. Algicidal activity of Serratia sp. PDGS120915 against harmful algae 68
4.2. Interaction between algicidal activity and bacterial growth 68
4.3. Algicidal activity according to prodigiosin concentration 68
4.4. The algicidal range of Serratia sp. PDGS120915 68
5. Discussion 74
6. References 76
Part III. Appendex 81
Chapter 6. Isolation of Lactic Acid Bacteria Showing Anti-Oxidative and Probiotic Activities from Kimchi and Infant Feces 82
1. Abstract 83
2. Introduction 84
3. Materials and Methods 85
3.1. Isolation of microorganisms 85
3.2. Identification of microorganisms 85
3.3. Acid and bile salt tolerance 85
3.4. Cell surface hydrophobicity 86
3.5. Hemolytic activity and enzymatic activity 86
3.6. Antimicrobial activity 86
3.7. Antibiotic susceptibility assay 87
3.8. Determination of DPPH scavenging activity 87
3.9. Determination of ABTS radical 88
3.10. Determination of superoxide radical scavenging activity 88
4. Results 89
4.1. Isolation and identification of lactic acid bacteria 89
4.2. Resistance to bile salts and acid 89
4.3. Cell surface hydrophobicity 89
4.4. Hemolysis test and enzymatic activity test 92
4.5. Inhibit the growth of pathogenic bacteria 92
4.6. Antibiotic susceptibility 92
4.7. Each strain culture filtrate scavenged DPPH and ABTS radicals 92
4.8. Superoxide radical scavenging activity 96
5. Discussion 102
6. References 105
Korean Summary 110
- Degree
- Doctor
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