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Contents
Ⅰ. Introduction 9
Ⅱ. Materials and Methods 14
1. Strain and medium 14
2. Composition of defined medium and culture condition 14
3. Seed starter 14
4. Production of zooglan 15
5. Isolation of zooglan 15
6. Purification of zooglan 16
7. Molecular weight analysis of zooglan 18
8. Thin layer chromatography(TLC) analysis 19
9. Gamma irradiation 19
10. Total cholesterol assay 19
11. Cell line culture for in vitro assay 21
12. Cytotoxicity assay 21
13. Measurement of quinone reductase activity 23
14. Measurement of glutathione S-transferase activity 24
Ⅲ. Results and Discussion 26
1. Effects of various fermentation factors on the production of zooglan 26
1) Effect of fermentation time 26
2) Inoculation size of starter culture 26
3) Effect of glucose concentration 30
4) Effect of rifampicin concentration 30
5) Effect of various carbon sources 33
6) Effect of cell density in starter culture 35
2. Recovery and purification of zooglan from Z. ramigera 115SLR 37
3. Molecular weight analysis of zooglan 40
4. Gamma irradiation on zooglan 42
1) Modification of zooglan by gamma irradiation 43
2) Thin layer chromatography of gamma irradiated zooglan 45
5. Functional properties of zooglan 48
1) Comparison of cholesterol adsorption by zooglan and food polysaccharides 48
2) Effect of zooglan concentration on total cholesterol adsorption 55
3) Effect of zooglan on phase2 enzyme activity 57
① Cytotoxicity 58
② Quinone reductase activity 60
③ Glutathione S-transferase activity 60
Ⅳ. Conclusions 64
Ⅴ. References 66
abstract 76
Table 1. Commercial and industrial uses of biopolymers 10
Table 2. Comparison of the productivity and yield of zooglan according to fermentation time 27
Table 3. Comparison of zooglan productivity according to C/N ratio 29
Table 4. Purification of zooglan from Z. ramigera 115SLR by the addition of NaOH and serial dilution 38
Table 5. Structural characteristics of polysaccharides 50
Table 6. Cytotoxicity of zooglan according to concentration 59
Figure 1. Proposed repeating unit of zooglan produced by Zoogloea ramigera 115 12
Figure 2. Schematic diagram for purification and analysis of zooglan produced by Z. ramigera 115SLR in defined medium 17
Figure 3. Schematic diagram for total cholesterol measurement using by total cholesterol kit 20
Figure 4. Schematic diagram for measuring cholesterol adsorption rate by polysaccharide 22
Figure 5. Effect of glucose concentration on the zooglan production in defined medium by Z. ramigera 115SLR 31
Figure 6. Effect of rifampicin on the zooglan production in defined medium by Z. ramigera 115SLR 32
Figure 7. Effects of different carbon source on zooglan production 34
Figure 8. Zooglan production according to the cell density of starter culture 36
Figure 9. Effect of NaOH treatment on the precipitation of zooglan with an equal volume of isopropanol 39
Figure 10. Molar mass distribution of zooglan produced from defined medium with/without rifampicin for 48 h 41
Figure 11. A gamma irradiated 0.1% (w/v) zooglan solution 44
Figure 12. Thin layer chomatograms of gamma irradiated zooglan solution (0.1%, w/v) 46
Figure 13. Comparison of passing time though capillary column in capillary viscometer of gamma radiated zooglan solution 47
Figure 14. In vitro cholesterol adsorption capacity according to the pH and types of exopolysaccharide 49
Figure 15. Changes in cholesterol adsorption in vitro according to the concentration of sodium chloride and zooglan 52
Figure 16. In vitro cholesterol adsorption capacity according to the concentration of zooglan and pH value 56
Figure 17. Relative quinone reductase activity by zooglan (n=6) 61
Figure 18. Relative glutathione S-transferase activity by zooglan (n=3) 62
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