1 / 16

Production of Rhamnolipids by Pseudomonas chlororaphis , a Nonpathogenic Bacterium

Production of Rhamnolipids by Pseudomonas chlororaphis , a Nonpathogenic Bacterium. Nereus W. Gunther IV , Alberto Nuñez, William Fett and Daniel K.Y. Solaiman Applied And Environmental Microbiology 71 (2005) 2288-2293 報告學生：陳佳. Rhamnolipids. 生物介面活性劑 清潔除污性能

keegan
Download Presentation

Production of Rhamnolipids by Pseudomonas chlororaphis , a Nonpathogenic Bacterium

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Production of Rhamnolipids by Pseudomonas chlororaphis, a Nonpathogenic Bacterium Nereus W. Gunther IV , Alberto Nuñez, William Fett and Daniel K.Y. Solaiman Applied And Environmental Microbiology 71 (2005) 2288-2293 報告學生：陳佳

  2. Rhamnolipids • 生物介面活性劑 • 清潔除污性能 • Pseudomonas aeruginosa、 Pseudomonas putida 皆具有致病力 (Anaissie, 1987、Macfarlane, 1991)

  3. 實驗流程 33株菌株接種至液態培養基 Minaeral salt-CTAB-methylene blue agar method觀察有無產生rhamnolipids 增加產量，以DCAT 11 tensiometer測量其表面張力 純化rhamnolipids TLC及HPLC/MS分析rhamnolipid種類 比較三菌株之rhamnolipid

  4. 菌株 • P. aeruginosa strains PG201, PG201-RhlA::Tn5, and NRRL B-800 • P. fluorescens strains ATCC 17397, ATCC 17824, ATCC 17577, and ATCC 17816 • P. chlororaphis strains ATCC 17812, NRRL B-14869, NRRL B-30761, and ATCC 9446 • P. putida strains ATCC 17527, W4P396, ATCC 17391, and ATCC 12633 • P. marginalis strains ATCC 10844 and HT041B • P. tolaasii strains ATCC 33618, ATCC 14340, C8, and C9 • P. syringae strain Cit7 • P. flavescens strain B62 • P. oleovorans strains NRRL B-778, NRRL B-14682, and NRRL B-14683 • P. resinovorans strain NRRL B-2649 • P. stutzeri strain ATCC 17588 • P. corrugata strains 388, ATCC 29756, and VC1-VC7 • Ralstonia eutropha strain • R. solanacearum strain

  5. Kay’s minimal medium • Kay’s minimal medium(液態)：0.3% NH4H2PO4, 0.2% K2HPO4, 0.2% glucose, 0.5mg/liter FeSO4, 0.1%MgSO4 • 培養條件：37℃，250 rpm

  6. 實驗流程 33株菌株接種至液態培養基 Minaeral salt-CTAB-methylene blue agar method觀察有無產生rhamnolipids 增加產量，以DCAT 11 tensiometer測量其表面張力 純化rhamnolipids TLC及HPLC/MS分析rhamnolipid種類 比較三菌株之rhamnolipid

  7. Mineral salt-CTAB-methylene blue agar plate method • Mineral salts medium 添加 200μg/ml cetyltrimethylammonium bromide, 5 μg/ml methylene blue, and 1.5% agar • Mineral salts medium：per liter, 0.7g KH2PO4, 0.9g Na2HPO4,2g NaNO3, 0.4g MgSO4˙7H2O, 0.1g CaCl2‧2H2O, 2 ml of trace elements (per liter, 2g FeSO4 ,1.5g MnSO4‧H2O, 0.6g (NH4)6Mo7O24‧4H2O • 30 ℃下培養24~48小時 • 移至4℃下培養數天

  8. glass pipette(10-ml )加熱 • 接種10-μl菌液

  9. 結果 • 陽離子介面活性劑及甲基藍與rhamnolipids結合呈藍紫色 • P. chlororaphis strain NRRL B-30761，30 ℃下呈現弱的正反應 • P. chlororaphis strain NRRL B-30761於4℃下仍呈正反應

  10. 實驗流程 33株菌株接種至液態培養基 Minaeral salt-CTAB-methylene blue agar method觀察有無產生rhamnolipids 增加產量，以DCAT 11 tensiometer測量其表面張力 純化rhamnolipids TLC及HPLC/MS分析rhamnolipid種類 比較三菌株之rhamnolipid

  11. ‧培養基表面積與體積比越大，產量 越高 ‧培養120小時，有最大產量 ‧23℃時，有最大產量

  12. 實驗流程 33株菌株接種至液態培養基 Minaeral salt-CTAB-methylene blue agar method觀察有無產生rhamnolipids 增加產量，以DCAT 11 tensiometer測量其表面張力 純化rhamnolipids TLC及HPLC/MS分析rhamnolipid種類 TLC及HPLC/MS分析rhamnolipid種類 比較三菌株之rhamnolipid

  13. 結果 • 將P. chlororaphis strain NRRL B-30761與市面販賣P.aeruginosa純化過的rhamnolipids做比較 • 不同分子的極性不相同 • TLC：P. chlororaphis的確具有rhamnolipids • HPLC： P. chlororaphis分泌的rhamnolipids無觀測到di-rhamnolipids

  14. 實驗流程 33株菌株接種至液態培養基 Minaeral salt-CTAB-methylene blue agar method觀察有無產生rhamnolipids 增加產量，以DCAT 11 tensiometer測量其表面張力 純化rhamnolipids TLC及HPLC/MS分析rhamnolipid種類 比較三菌株之rhamnolipid

  15. 結論 • 工業上以P. chlororaphis生產rhamnolipids可消除致病菌株產生毒性的問題 • 以glucose為碳源並改變萃取方式後P. chlororaphis最大rhamnolipids產量可達1 g/liter和P. aeruginosa利用glucose為碳源產量差不多(1.0至1.6 g/liter) • 靜置培養可節省能源

More Related