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목차

[표제지 등]=0,1,2

제출문=1,3,2

요약문=3,5,20

목차=23,25,12

제1장 서론=35,37,4

제1절 해양 미세조류=38,40,2

제2절 동정 및 특성=39,41,4

제3절 미세조류의 이용=43,45,5

제4절 세포외다당류=47,49,3

제5절 황 함유 다당류=50,52,2

제6절 세포외다당류의 생산=52,54,2

제7절 전망=54,56,1

제2장 국내외 기술개발 현황=55,57,2

제1절 실용화된 미생물유래 다당류=57,59,4

제2절 국외 EPS 개발 동향=61,63,3

제3절 국내의 EPS 개발 동향=64,66,1

제3장 연구개발수행 내용 및 결과=65,67,2

제1절 연구수행방법=67,69,1

1. Hahella Chejuensis 96CJ10356 세포외다당류의 미백효과=67,69,1

2. 해양미세조류의 세포외다당류=67,69,1

가. 시료채취=67,69,3

나. G. Impudicum의 순수분리=70,72,6

다. 균주 동정=75,77,6

라. G. Impudicum KG03의 생장특성=81,83,2

마. 세포외다당류(p-KG03)의 생리활성=82,84,10

바. 세포외다당류의 생산=91,93,12

사. 세포외다당류의 특성=102,104,9

제2절 연구수행 결과=111,113,1

1. Hahella Chejuensis 96CJ10356 세포외다당류의 미백효과=111,113,1

2. Gyrodinium Impudicum KG03 세포외다당류=111,113,1

가. 균주분리 및 동정=111,113,38

나. p-KG03의 생리활성=149,151,7

다. p-KG03의 생산=156,158,94

라. p-KG03의 특성=250,252,43

제4장 연구개발목표 달성도 및 대외기여도=293,295,2

제1절 연구목표 및 연구내용=295,297,3

제2절 연구목표 및 평가착안점에 입각한 연구개발목표의 달성도=298,300,2

제3절 연구개발성과 현황=300,302,3

제4절 관련분야의 기술발전에의 기여도=303,305,2

제5장 연구개발결과의 활용계획=305,307,2

제1절 연구개발의 결과=307,309,2

제2절 활용계획=309,311,2

제6장 참고문헌=311,313,28

영문목차

[title page etc.]=0,1,14

Summary=13,15,12

Contents=25,27,2

List of Figures=27,29,4

List of Tables=31,33,4

Chapter 1. Introduction=35,37,4

Section 1. Marine Microalgae=38,40,2

Section 2. Identification And Characteristics=39,41,4

Section 3. Application Of Microalgae=43,45,5

Section 4. Exopolysaccharide Of Microalgae=47,49,3

Section 5. Sulfur-Containing Exopolysaccharides=50,52,2

Section 6. Production Of Exopolysaccharides=52,54,2

Section 7. Future Aspects=54,56,1

Chapter 2. State Of The Art=55,57,2

Section 1. Commercialized Microbial Exopolysaccharides=57,59,4

Section 2. Microbial Exopolysaccharides Development In Abroad=61,63,3

Section 3. Domestic Situation For Exopolysaccharides Development=64,66,3

Chapter 3. Contents And Results Of Study=67,69,1

Section 1. Methods=67,69,1

1. Whitening Effect Of Hahella Chejuensis 96CJ10356 Producing Exopolysaccharide=67,69,1

2. Exopolysaccharide Of Gyrodinum Impudicum KG03=67,69,1

A. Sampling Site=67,69,3

B. Axenic Separation Of Gyrodinium Impudicum=70,72,6

C. Identification=75,77,6

D. Cultural Conditions Of Gyrodinium Impudicum KG03=81,83,2

E. Physiological Activity Of P-KG03=82,84,10

F. Production Of Exopolysaccharide=91,93,12

G. Characteristics Of Exopolysaccharide=102,104,9

Section 2. Results=111,113,1

1. Whitening Effect Of Hahella Chejuensis 96CJ10356 Producing Exopolysaccharide=111,113,1

2. Exopolysaccharide Of Gyrodinum Impudicum KG03=111,113,1

A. Separation And Identification Of Strain=111,113,38

B. Physiological Activity Of P-KG03=149,151,7

C. Production Of P-KG03=156,158,94

D. Characteristics Of P-KG03=250,252,43

Chapter 4. Achievement And Contributions=293,295,2

Section 1. Object And Scope Of Research=295,297,3

Section 2. Propulsive Results=298,300,2

Section 3. Presentation Of Results=300,302,3

Section 4. Contribution External=303,305,2

Chapter 5. Suggestion For Application Of The Results=305,307,2

Section 1. Suggestion For Development=307,309,2

Section 2. Application=309,311,4

Chapter 6. References=313,315,26

List of Figures

Fig.3-1. Location Of Sampling Stations For Isolation Of Gyrodinium Impudicum=68,70,1

Fig.3-2. Procedure Of Axenic Culture In Gyrodinium Impudicum Strain KG03=71,73,1

Fig.3-3. Procedure For Algal Wash By Phototaxsis Using Algal Separation Tube=74,76,1

Fig.3-4. Schematic Diagram Of The ALBTPR(Air-Lift Ballon Type Photobioreactor) System Used For P-KG03 Production=101,103,1

Fig.3-5. Procedure For Purification Of p-KG03=104,106,1

Fig.3-6. The Effects Of EPS-R On Tyrosinase Inhibition Activity=112,114,1

Fig.3-7. Relative Survival Ratio Of Gyrodinium Impudicum Strain KG03 After Antibiotic Treatments=120,122,1

Fig.3-8. Photography Of Strain KG03 With Light Microscopy=125,127,1

Fig.3-9. The Photography Of KG03 With Scanning Electron Microscopy=126,128,1

Fig.3-10. The Photography Of Strain KG03 With Transmission Electron Microscopy=129,131,1

Fig.3-11. The Chromatogram Of Strain KG03 Fatty Acids With Gas Chromatography=133,135,1

Fig.3-12. The Chromatogram Of Strain KG03 Amino Acids Compositions=135,137,1

Fig.3-13. PCR Products Of 5.8S And ITS Region Amplified From Strain KG03=139,141,1

Fig.3-14. Sequence Alignment Of The ITS-I, ITS-2 And 5.8S rDNA Region=140,142,1

Fig.3-15. PCR Products Of 18S Amplified From Strain KG03=141,143,1

Fig.3-16. Neighbor-Joining Tree Based On Nearly Complete 18S rDNA Sequences Showing Relationship Between Strain KG03=142,145,1

Fig.3-17. Effect Of Ph, Temperature, Naci And Light Intensity On The Growth Of Gyrodinium Impudicum Strain KG03=147,149,1

Fig.3-18. Effects Of Sea-Salt Components On The Growth Of Gyrodinium Impudicum KG03=148,150,1

Fig.3-19. Dose Response Curve For The Fraction Of P-KG03=152,154,1

Fig.3-20. Cytotoxicity Of P-KG03 Stimulated Macrophages Against B16 Melanoma Cells=153,155,1

Fig.3-21. The Effect Of p-KG03 On Production Of TNF-a (A) And Nitrite (B) By Macrophages=154,156,1

Fig.3-22. The Effect Of Extracts On IL-1(a) And IL-6 (b) Production By Macrophages=155,157,1

Fig.3-23. Effect Of The Different Inculum Size On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=157,159,1

Fig.3-24. Effect Of The Different Light Intensity On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03=160,162,1

Fig.3-25. Effect Of Different Period Of Light On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=164,166,1

Fig.3-26. Effect Of Different Culture Temperature On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03=168,170,1

Fig.3-27. Effect Of Different Initiated Ph On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03=171,173,1

Fig.3-28. Effect Of Different Salinity On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03=175,177,1

Fig.3-29. Effect Of NaCI 100 mM On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=178,180,1

Fig.3-30. Effect Of CaCl₂ㆍ2H₂0 200mM On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03(이미지 참조)=181,183,1

Fig.3-31. Effect Of Different MgCI₂ㆍ6H₂O On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=184,186,1

Fig.3-32. Effect Of Na₂SO₄ On The Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=187,189,1

Fig.3-33. Effect Of Na₂NO₃ On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=190,192,1

Fig.3-34. Effect Of NH₄Cl On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03=194,196,1

Fig.3-35. Effect Of Nah₂PO₄ On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=197,199,1

Fig.3-36. Effect Of NaHCO₃ On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=200,202,1

Fig.3-37. Effect Of FeCl₂ㆍ6H₂O On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=205,207,1

Fig.3-38. Effect Of MnCI₂ㆍ4H₂O On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=208,210,1

Fig.3-39. Effect Of Trace Metal Solution On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=211,213,1

Fig.3-40. Effect Of Fe:Mn Mixture On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=214,216,1

Fig.3-41. Effect Of Vitamin B12 Onthe Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03(이미지 참조)=217,219,1

Fig.3-42. Effect Of Biotin On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03=220,222,1

Fig.3-43. Effect Of f/2 Vitamin Solution On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03=223,225,1

Fig.3-44. Effect Of Inositol On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03=226,228,1

Fig.3-45. Effect Of 2,4-D On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=229,231,1

Fig.3-46. Effect Of IBA On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=232,234,1

Fig.3-47. Effect Of NAA On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=235,237,1

Fig.3-48. Effect Of BAP On The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03=238,240,1

Fig.3-49. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With M-KG03=241,243,1

Fig.3-50. Photography Of 2 L-ALBTPR(Air-Lift Ballon Type Photobioreactor) For P-KG03 Production=244,246,1

Fig.3-51. Effect Of 1ℓ ALBTPR With Air 50 cc/m In. On The Cell Growth And P-KG03 Production Of Gyrodinium Impudicum KG03=245,247,1

Fig.3-52. Effect Of CO₂ With ALBTPR On The Cell Growth And P-KG03 Production Of Gyrodinium Impudicum KG03=247,249,1

Fig.3-53. Gel Filtration Of The p-KG03=251,253,1

Fig.3-54. TLC Chromatogram Of The Hydrolystate Of p-KG03=254,256,1

Fig.3-55. Chromatogram Of The Hydrolysate In p-KG03=255,257,1

Fig.5-56. Estimation Of Molecular Weight Of p-KG03 Using GPC=256,258,1

Fig.3-57. SEM Photograph Of p-KG03 (X 15,000)=258,260,1

Fig.3-58. The Chromatogram Of Amino Acids Compositions In p-KG03=259,261,1

Fig.3-59. Characteristics Of Thermal Decomposition Of p-KG03 Using TGA=264,266,1

Fig.3-60. Differential Scanning Calorimeter Thermogram Of p-KG03=265,267,1

Fig.3-61. UV Spectrum Of p-KG03=272,274,1

Fig.3-62. Infra-Red Absorption Spectrum Of p-K03=273,275,1

Fig.3-63. Intrinsic Viscosity Of p-KG03 Fluid Calculated By Reduced Viscosity Equation(Huggins Equatio)=274,276,1

Fig.3-64. Relationship Between Shear Stress And Shear Rate Of p-KG03 1.0 %=278,280,1

Fig.3-65. Relationship Between Shear Stress And Shear Rate Of p-KG03 At The Different Concentration=281,283,1

Fig.3-66. Relationship Between Shear Stress And Shear Rate Of p-KG03 0.5% At Various Concentration Of Nacl=283,285,1

Fig.3-67. Relationship Between Shear Stress And Shear Rate Of p-KG03 0.5% At Various Concentration Of CaCI₂=284,286,1

Fig.3-68. Relationship Between Shear Stress And Shear Rate Of p-KG03 0.5% At Various pH=285,287,1

Fig.3-69. Relationship Between Shear Stress And Shear Rate Of p-KG03 0.5% Under Heat Treatment Of High Temperatures=287,289,1

Fig.3-70. Relationship Between Shear Stress And Shear Rate Of p-KG03 0.5% At Various Temperature=288,290,1

Fig.3-71. Relationship Between Shear Stress And Shear Rate Of p-KG03 0.5% Mixture With Gellan Gum=290,292,1

Fig.3-72. Relationship Between Shear Stress And Shear Rate Of p-KG03 0.5% Mixture With Xanthan Gum=291,293,1

List of Tables

Table3-1. A List Of Chemical Components Of f/2 Media For Culture Of Gyrodinium Impudicum=69,71,1

Table3-2. A List Of Chemical Components Of Szobell Medium For Symbiotic Bacteria With Gyrodinium Impudicum KG03=72,74,1

Table3-3. The Primer Sequences To Identify ITS, 5.8S rDNA And 18S rDNA Of Strain KG03=79,81,1

Table3-4. The Chemical Compositions Of Artificial Seawater For Omission Method Of Gyrodinium Impudicum Strain KG03=83,85,1

Table3-5. Test Conditions Of Antiviral Evaluation With p-KG03=87,89,1

Table3-6. List Of Standard Compounds For Test Antiviral Activity Against p-KG03=88,90,1

Table3-7. The Chemical Compositions Of M-KG03 Media For The Production Of p-KG03=100,102,1

Table3-8. Summary Of The Environmental Parameters In Sampling Site For Isolation Of Gyrodinium Impudicum=115,117,1

Table3-9. Morphological Characterizations Of Assorted Symbiotic Bacteria In Gyrodinium Impudicum KG03=117,119,1

Table3-10. The Survival Types Of Symbiotic Bacteria With Gyrodinium Impudicum Strain KG03 After Antibiotic Treatments=118,120,1

Table3-11. Comparison Of Cell Growth And Carbohydrate Production With Strain KG03, KG09 And GJ01=122,124,1

Table3-12. Comparison Of Cell Size Of Strain KG03 With Gymnodium Sp=127,129,1

Table3-13. The Proximate Chemical Compositions Of Strain KG03=132,134,1

Table3-14. The Compositions Of Fatty Acids In Strain KG03=134,136,1

Table3-15. The Compositions Of Amino Acids In Strain KG03=136,138,1

Table3-16. Neighbor-Joining Tree Based On Nearly Complete 18S rDNA Sequences Showing Relationship Between Strain KG03=143,145,1

Table3-17. Characterizations Of Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With Different Culture Types=146,148,1

Table3-18. Antiviral Activity Of p-KG03=151,153,1

Table3-19.The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With Inoculum Size Of 1 x 10³Cells/㎖=158,160,1

Table3-20. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03 With 150 ΜE/㎡/S Of Light Intensity=161,163,1

Table3-21. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With 16:8(L:D) Of Light Period=165,167,1

Table3-22. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With Culture Temperature Of 20℃=169,171,1

Table3-23. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03 With pH 8=172,174,1

Table3-24. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03 With Salinity Of 30 ‰=176,178,1

Table3-25. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With NaCI 100 mM=179,181,1

Table3-26. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With CaCI₂ㆍ2H₂0, 200 ΜM=182,184,1

Table3-27. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With MgCI₂.6H₂O 20 mM=185,187,1

Table3-28. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With Na₂S0₄ 100 ΜM=188,190,1

Table3-29. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With NaNO₃ 100 ΜM=191,193,1

Table3-30. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With NH₄Cl 75ΜM=195,197,1

Table3-31. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With NaH₂PO₄ 200 ΜM=198,200,1

Table3-32. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With NaHCO₃ 50 ΜM=201,203,1

Table3-33. Changes Of Gyrodinium Impudicum Strain KG03 Cell Growth And P-KG03 Production With Macro-Elements=203,205,1

Table3-34. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With FeCI₂ㆍ6H₂O 25ΜM=206,208,1

Table3-35. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With Mncl₂ㆍ4H₂O 0.25ΜM=209,211,1

Table3-36. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With f/2 Trace Metal Solution 0.25 ㎖ Addition=212,214,1

Table3-37. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With Fe : Me =10:0.1 ΜM=215,217,1

Table3-38. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03 With Vitamin B12 0.75 ㎍/ℓ(이미지 참조)=218,220,1

Table3-39. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03 With Biotin 5.0 ㎍/ℓ=221,223,1

Table3-40. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03 With f/2 Vitamin Solution 2 ㎖/ℓ=224,226,1

Table3-41. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum Strain KG03 With Inositol 200 ㎎/ℓ=227,229,1

Table3-42. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With 2.4-D 0.075 ㎍/㎖=230,232,1

Table3-43. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With IBA 0.075 ㎍/㎖=233,235,1

Table3-44. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With NAA 0.075 ㎍/㎖=236,238,1

Table3-45. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With BAP 0.2 ㎍/㎖=239,241,1

Table3-46. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With M-KG03 Media=242,244,1

Table3-47. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 With 1ℓ ALBTPR (Air 50 cc/min)=246,248,1

Table3-48. The Cell Growth And Carbohydrate Production Of Gyrodinium Impudicum KG03 In 1ℓ ALBTPR With CO₂ 1%=248,250,1

Table3-49. Summary Of Purification Steps Of The p-KG03 From Gyrodinium Impudicum KG03=252,254,1

Table3-50. The Composition Of Amino Acids In p-KG03=260,262,1

Table3-51. Elemental Analysis Of p-KG03=261,263,1

Table3-52. Water Activity Values (Aw) And Water Content Of Various Polysaccharide=262,264,1

Table3-53. Color Reactions Of The p-KG03=266,268,1

Table3-54. Solubility Test Of p-KG03 In Various Solvents=268,270,1

Table3-55. Flocculating Activity Of p-KG03=269,271,1

Table3-56. Emulsion Stability Test Of The p-KG03 And The Other Exopolysaccharides=270,272,1

Table3-57. Viscosity For The Concentration Of P-KG03 Fluid Calculated By Several Equation At 25℃=275,277,1

Table3-58. Comparison Of Chain Stiffness Parameter(B) p-KG03 With Other Polysaccharide=276,278,1

Table3-59. Consistency Index(K) And Flow Behavior Index(N) With Different Concentrations Of p-KG03=279,281,1

Table3-60. Consistency Index(K) And Flow Behavior Index(N) Of p-KG03 And Other Polysaccharides=280,282,1

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