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Q UORUM S ENSING O DAKLı Y ENILIKLER VE B IYOTEKNOLOJIK U YGULAMALAR Hazırlayan : Murat Kemal AVCI Öğretim Üyesi : Doç. Dr. Zeynep Petek ÇAKAR Aralık-2009.

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... konulu sunumlar: "Q UORUM S ENSING O DAKLı Y ENILIKLER VE B IYOTEKNOLOJIK U YGULAMALAR Hazırlayan : Murat Kemal AVCI Öğretim Üyesi : Doç. Dr. Zeynep Petek ÇAKAR Aralık-2009."— Sunum transkripti:

1 Q UORUM S ENSING O DAKLı Y ENILIKLER VE B IYOTEKNOLOJIK U YGULAMALAR Hazırlayan : Murat Kemal AVCI Öğretim Üyesi : Doç. Dr. Zeynep Petek ÇAKAR Aralık-2009 Maslak MBG 617 B IYOTEKNOLOJIDE S ON G ELIŞMELER İ STANBUL T EKNIK Ü NIVERSITESI M OLEKÜLER B IYOLOJI VE G ENETIK & B IYOTEKNOLOJI

2 İ ÇERIK … Quorum sensing mekanizması... Araştırma alanları... Günümüzdeki uygulamalar ve potansiyeller.... Sonuç ve Öneriler… Murat Kemal AVCI_İTÜ_MOBGAM

3 1. M IKROORGANIZMALAR VE B IZ Murat Kemal AVCI_İTÜ_MOBGAM

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5 Vücudumuzdaki Mikroorganizma habitatları: 1. Baş derisi 2. Göz 3. Kulak 4. Ağız 5. Deri - Göğüs - Kollar - Eller - Bacaklar - Ayaklar 6. İç organlar - Mide - Barsaklar VS.... Murat Kemal AVCI_İTÜ_MOBGAM

6 İnsanları Etkileyen Diğer Faktörlerle Mikroorganizmaların Etkileşimleri Murat Kemal AVCI_İTÜ_MOBGAM

7 S ONUÇ OLARAK.... İ LIŞKININ YÖNLENDIRILMESI Tatlı dil (Atasözü) Mikroorganizmaların Dili... QUORUM SENSİNG (BAKTERİCE) Bir ETKİLEŞİM-İLİŞKİ mevcut.... Murat Kemal AVCI_İTÜ_MOBGAM

8 Q UORUM S ENSING Mikroorganizmaların, ortamdaki hücre konsantrasyonuna bağlı olarak bir takım sinyal molekülleri aracılığıyla aralarında iletişim kurup, davranışlarını/gen ifadelerini populasyon çapında düzenlemelerini sağlayan bir iletişim mekanizmasıdır. Quorum : Yeterli sayı Sense : Hissetmek, algılamak Murat Kemal AVCI_İTÜ_MOBGAM

9 “BAKTERİCE” Q UORUM S ENSING Sözcükler = Kimyasal Moleküller (Autoinducer Peptides, AIPs) Türe özgü davranış (FENOTİP) - Biyofilm oluşumu - Ekstraselüler enzim üretimi - Toksin salgılama - Antibiyotik üretimi - Kommensal-Patojen Dönüşümü v s Murat Kemal AVCI_İTÜ_MOBGAM

10 1980 DEN G ÜNÜMÜZE YILLARYAYIN SAYISI o KARMA TOPLAM :5789 TOPLAM :3194 QUORUM SENSİNG LİTERATURE in SCİENCEDIRECT from 1980 to 2010 Murat Kemal AVCI_İTÜ_MOBGAM

11 M EKANIZMANıN T EMEL E LEMANLARı...  Mikroorganizma Türleri - Bakteriler - Algler - Mayalar - Küfler - Protistler  Genler  Preoteinler  Sinyal Molekülleri - AHL türevleri - Oligopeptitler - Furan türevleri  Algılama sistemleri - Gram (-) - Gram (+)  Davranış/Fenotip/Ürün Murat Kemal AVCI_İTÜ_MOBGAM

12 M ADALYONUN İ KI Y ÜZÜ KommensalPatojen Duyarlı A ürünü X formu Masum Dost Dirençli B ürünü Y ürünü Canavar Düşman QS t1 t2 t3 Zaman OD600 Eksponansiyal Faz [YOĞUNLUĞA BAĞLI] Murat Kemal AVCI_İTÜ_MOBGAM

13 D AVRANıŞıN M OLEKÜLER K ONTROLÜNE B IR Ö RNEK...  ORGANİZMA : Staphylococcus epidermidis Staphylococcus aureus  GEN : agrA, agrC, agrD, agrB (agr operonu)  PROTEİNLER : AgrA, AgrC, AgrD, AgrB, delta toksin  ALGILAMA SİSTEMİ : İki bileşenli algılama sistemi  FENOTİP/ÜRÜN : Kommensal-Patojen dönüşümü/Toksin Üretimi/Biyofilm oluşumu Murat Kemal AVCI_İTÜ_MOBGAM

14 Şekil : Staphylococcal agr sistemi. A) İki ayrı promotor (P2 ve p3) ve bunların transkriptleri. P2 operonu 4 gen içeri; agrA,B,C ve D. B) Agr D’nin açık yapısında autoinducer peptidin (AIP) amino asit dizisi görülmektedir (koyu harfler). Bu peptid AgrC’ye bağlanarak metabolik yolun aktivasyonunu sağlayan ligand olarak iş görür. C) AgrCnin açık yapısında beş transmembran heliksi görülmektedir. S9-rf1/52 Murat Kemal AVCI_İTÜ_MOBGAM

15 Şekil. : Staphylococ’larda accessory gene regulator (agr) sistemi. P2 promotoru sinyal mekanizmasını kodlar (RNAII aracılığıyla) P3 promotoru ise agr lokusunun effektörü olarak hareket eder (RNAIII tarafından kodlanır). [J. M. Yarwood and P. M. Schlievert, The Journal of Clinical Investigation, December 2003, Volume 112, Number 11] Murat Kemal AVCI_İTÜ_MOBGAM 15

16 A GR SISTEMI INAKTIF OLDUĞUNDA Şekil : Agr sisteminin inaktif olması kommensal olmayı teşvik eder. [Yarwood and Schlievert (2003)’den uarlanmıştır, Çizim: M.K.AVCI] Virülans Faktörleri (Ekzo proteinler) Hücre duvarıyla ilişkili proteinler ETKİ (+)(-) - Alfa-toksin - Beta toksin - Delta toksin - Serin proteaz - DNAase - Fibrinolizin - Enterotoksin B - TSST-1 - Protein A - Koagülaz - Fibronektin bağlanma proteini vs… Murat Kemal AVCI_İTÜ_MOBGAM 16

17 A GR SISTEMI A KTIF OLDUĞUNDA Şekil.20 :Agr sisteminin aktif olması patojen olmayı teşvik eder. [Yarwood and Schlievert (2003)’den uarlanmıştır, Çizim: M.K.AVCI] Virülans Faktörleri (Ekzo proteinler) - Alfa-toksin - Beta toksin - Delta toksin - Serin proteaz - DNAase - Fibrinolizin - Enterotoksin B - TSST-1 Hücre duvarıyla ilişkili proteinler - Protein A - Koagülaz - Fibronektin bağlanma proteini vs… ETKİ (-)(+) 17 Murat Kemal AVCI_İTÜ_MOBGAM

18 Q UORUM S ENSING C ROSS T ALK  Türler arası  Cinsler arası  Prokaryot-Ökaryot Ör: S. epidermdis ve S. aureus Murat Kemal AVCI_İTÜ_MOBGAM

19 U YGULAMA A LANLARı Tıp  Yapay Kalp kapakçıkları, implent materyaller Veterinerlik  Mastitis.... Ziraat / Botanik  Bitkisel hastalıklar... Biyoteknoloji Malzeme Mühendisliği  Biyo-uyumlu materyaller / ürünler Medikal Teknolojileri Moleküler Biyoloji Biyoremediasyon  Atık arıtımı... Fermentasyon Endüstriyel Mikrobiyoloji Murat Kemal AVCI_İTÜ_MOBGAM

20 T ARTıŞMA VE Ö NERILER ...???? √ √ √ Murat Kemal AVCI_İTÜ_MOBGAM

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22 REFERANSLAR 1. Bakke, R., Trulear, M.G., Robinson, J.A., Characklis,W.G., Activity of Pseudomonas aeruginosa in biofilms: steady state. Biotechnol. Bioeng. 26, 1418–1424. [BİOF-15/rf] 2. Costerton, J.W., Cheng, K.J., Geesey, G.G., Ladd, T.I., Nickel, J.C., Dasgupta, M., Marrie, T.J., Bacterial biofilms in nature and disease. Annu. Rev. Microbiol. 41, 435– Blaschek, H. P., Wag, H. H., Agle, M. E., Biofilms in the Food Environment. IFT Press, Blackwell Publishing (Book) 2007 Chapter 1, s.3 4. Prof. Dr. Menemşe GÜMÜŞDERELİOĞLU, Müge DESTAN. Biyofilmler. BİTOTEK (2003) YIL: 3 SAYI: Cerca, N. et al., Quantitative analysis of adhesion and biofilm formation on hydrophilic and hydrophobic surfaces of clinical isolates of Staphylococcus epidermidis. Res. Microbiol May ; 156 (4): 506–514. [QS-222] 6. Galdbart, J.O. et al., Phenotypic and Genomic Variation among Staphylococcus epidermidis Strains Infecting Joint Prostheses. Journal Of Clinical Microbiology, May 1999, p. 1306–1312 Vol. 37, No. 5. [QS-252] 7. Gross, M. et al., Key Role of Teichoic Acid Net Charge in Staphylococcus aureus Colonization of Artificial Surfaces. Infectıon and Immunity, May. 2001, p. 3423–3426 Vol. 69, No. 5. [QS-274] 8. Rupp, M. E. et al. Characterization of the Importance of Staphylococcus epidermidis Autolysin and Polysaccharide Intercellular Adhesin in the Pathogenesis of Intravascular Catheter–Associated Infection in a Rat Model. The Journal of Infectious Diseases 2001;183:1038–42. [QS-275] 9. Briandet, R., Determination of the van der Waals, electron donor and electron acceptor surface tension components of static Gram-positive microbial biofilms. Colloids and Surfaces B: Biointerfaces 21 (2001) 299–310. [BİOF-23] 9. Saldarriaga Fernández, I. C., et al., In vitro and in vivo comparisons of staphylococcal biofilm formation on a cross- linked poly(ethylene glycol)-based polymer coating. Acta Biomaterialia xxx (2009) xxx–xxx. [BİOF-F35] 10. van der Borden, A. J., Electric Current-Induced Detachment of Staphylococcus epidermidis Biofilms from Surgical Stainless Steel. Applıed And Envıronmental Mıcrobıology, Nov. 2004, p. 6871–6874. [QS-227] 11. Anderson, J.M. and Marchant R. E., Biomaterials: factors favoring colonization and infection. In: Waldvogel FA, Bisno AL, editors. Infections associated with indwelling medical devices. Washington DC: ASM Press; p. 89–110. [BİOF-F35/rf.3] Murat Kemal AVCI_İTÜ_MOBGAM

23 12. Costerton J.W., Introduction to biofilm. International Journal of Antimicrobial Agents 11 (1999) 217–221. [BİOF-F22] 13. Rohde, H. et al., Structure, function and contribution of polysaccharide intercellular adhesin (PIA) to Staphylococcus epidermidis biofilm formation and pathogenesis of biomaterial-associate dinfections. European Journal of Cell Biology (2009) 1-9. [BİOF-S7] 14. Sutherland, I. W., The biofilm matrix – an immobilized but dynamic microbial environment (Review). TRENDS in Microbiology, May 2001, Vol.9 No.5 [BİOF-S13] 15. Van Houdt, R. et al., Biofilm formation and cell-to-cell signalling in gram-negative bacteria isolated from a food processing environment. Journal of Applied Microbiology, 2004, 96, [QS-463] 16. Blaschek, H. P., Wag, H. H., Agle, M. E., Biofilms in the Food Environment, (Book) Chapter 5, s.106) 17. Monds, R. and O’Toole, G. A., The developmental model of microbial biofilms: ten years of a paradigm up for review. Trends in Microbiology (2009) Vol.17 No.2 [BİOF-24] 18. Lee, J.. et al, Structure and Function of the Escherichia coli Protein YmgB: A Protein Critical for Biofilm Formation and Acid-resistance. J. Mol. Biol. (2007) 373, 11–26. [BİOF-S6] 19. Mack, D. et.al, Mechanisms of biofilm formation in Staphylococcus epidermidis and Staphylococcus aureus: functional molecules, regulatory circuits, and adaptive responses (REVIEW). International Journal of Medical Microbiology 294 (2004) 203–212. [QS-201] 20. Kozitskaya, S. et al., Clonal Analysis of Staphylococcus epidermidis Isolates Carrying or Lacking Biofilm-Mediating Genes by Multilocus Sequence Typing. Journal Of Clinical Microbiology, Sept. 2005, Vol. 43, No. 9 p. 4751–4757. [QS-220] 21. Zhang, Y.-Q. et al., Genome-based analysis of virulence genes in a non-biofilm-forming Staphylococcus epidermidis train (ATCC 12228). Molecular Microbiology, (2003) (6), 1577–1593. [QS-245] 21. Arevalo-Ferroa, C. et al., Biofilm formation of Pseudomonas putida IsoF: the role of quorum sensing as assessed by proteomics. Systematic and Applied Microbiology 28 (2005) 87–114. [BİOF-F17] 22. Cucarella, C. et al. Bap, a Staphylococcus aureus Surface Protein Involved in Biofilm Formation. Journal Of Bacterıology, May-2001, Vol. 183, No. 9, p. 2888–2896. [QS-256] Murat Kemal AVCI_İTÜ_MOBGAM

24 24. Hansen, S. K. Et al., Evolution of species interactions in a biofilm community. NATURE| Vol 445| 1 February [BİOF-6] 25. Kumar, C. G. and Anand, S.K., Significance of microbial biofilms in food industry: a review. International Journal of Food Microbiology 42 (1998) 9–27. [BİOF-15] 26. Melchior, M.B. et. al, Biofilms: A role in recurrent mastitis infections? (Review) The Veterinary Journal, 171 (2006) 398–407. [BİOF-16] 27. Morikawa, M, Beneficial Biofilm Formation by Industrial Bacteria Bacillus subtilis and Related Species. Journal Of Bıoscıence And Bıoengıneerıng. Vol. 101, No. 1, 1– [BİOF-19] 28. Senior, K., Navigating the hidden depths of biofilms. Infectious Diseases Vol 4 April [BİOF-20] 29. Jefferson, K. K., What drives bacteria to produce a biofilm?, MiniReview, FEMS Microbiology Letters 236 (2004) 163–173. [BİOF-F18] 30. Hansen, S.K., Evolution of species interactions in a biofilm community, NATURE, Vol 445|, 1 February [BİOF-6] 31. Kennedy, P., et al., Burns, biofilm and a new appraisal of burn wound sepsis, B U R N S( ) 1-8. [BİOF-11] 32. Huq, A., Biofilms in water, its role and impact in human disease transmission. Current Opinion in Biotechnology 2008, 19:244–247. [BİOF-12] 33. Stephens, C., Microbiology: Breaking Down Biofilms. Current Biology, Vol. 12, R132–R134, February 19, [BİOF-26] 34. Matsukawa, M., Biofilms: From Bench to the Bedside. Speakers' Abstracts/International Journal of Antimicrobial Agents 26S (2005) S1-S63 [BİOF-27] 35. Ramey, B. E., Biofilm formation in plant–microbe associations, Current Opinion in Microbiology 2004, 7:602– 609. [BİOF-28] 36. Shapiro, J.A., Thinking about bacterial populations as multicellular organisms. Annu. Rev. Microbiol. (1998) 52, 81–104. [BİOF-F18/rf.26] 37. Federle, M.J. and Bassler, B.L., Interspecies communication in bacteria. J. Clin. Invest. (2003) 112, 1291– [BİOF-F18/rf.32] Murat Kemal AVCI_İTÜ_MOBGAM

25 38. Voung, C., Otto, M.,2000. Staphylococcus epidermidis infections. Microbs and Infections 4 (2002) [QS-241] 39. Vacheethasanee K., and Marchant, R. E., Surfactant polymers designed to suppress bacterial (Staphylococcus epidermidis) adhesion on biomaterials. Student Research Award in the Ph.D. Degree Candidate Category, World Biomaterials Congress 2000, Kamuela, HI, May 15-20, [QS-271] 40. Milohanic, E., et al., The autolysin Ami contributes to the adhesion of Listeria monocytogenes to eukaryotic cells via its cell wall anchor. Molecular Mibrobiology (2001) 39 (5), [QS-308] 41. Patti, J.M., (1994) MSCRAMM-mediated adherence of microorganisms to host tissues. Annu. Rev. Microbiol. 48, 585– 617. [BİOF-F18/rf.20] 42. Mazmanian, S.K. et al., Sortase-catalysed anchoring of surface proteins to the cell wall of Staphylococcus aureus, Mol. Microbiol. 40 (2001) 1049–1057. ) [QS-276] 43. Hussain, M., A 140-kilodalton extracellular protein is essential for the accumulation of Staphylococcus epidermidis strains on surfaces, Infect. Immun. 65 (1997) 519–524. [QS-241/rf23] 44. Vuong, C., Polysaccharide intercellular adhesin (PIA) protects Staphylococcus epidermidis against major components of the human innate immune system. Cellular Microbiology (2004) 6 (3), 269–275. [QS-244] 45. Arciola, C.R., Presence of icaA and icaD Genes and slime production in a collection of staphylococcal strains from catheter-associated infections, J. Clin. Microbiol. 39 (2001) 2151–2156. [S9-rf1/31] 46. Chamberlain, N.R., and Brueggemann, S.A., Characterisation and expression of fatty acid modifying enzyme produced by Staph. epidermidis, J. Med. Microbiol. 46 (1997) 693–6 [S9-rf1/35] 47. Ronner, U., Husmark, U., Henriksson, A., Adhesion of Bacillus species in relation to hydrophobicity. J. Appl. Bacteriol. 69, 550– Dewanti, R., Wong, A.C.L., Influence of culture conditions on biofilm formation by Escherichia coli O157:H7. Int. J. Food Microbiol. 26, 147– Collins, E.B., Hardt, P., Inhibition of Candida albicans by Lactobacillus acidophilus. J. Dairy Sci. 63, 830– B. Gottenbos, H.C. van der Mei, H.J. Busscher, Initial adhesion and surface growth of Staphylococcus epidermidis and Pseudomonas aeruginosa on biomedical polymers, J. Biomed. Mater. Res. 50 (2000) 208– Kumamoto, C. A., Candida Biofilms, Curr. Op. İn Micbio. 2002, 5: Rinker, K.D., Kelly, R.M., Growth physiology of the hyperthermophilic archaeon Thermococcus litoralis: Development of a sulfur-free defined medium, characterization of an exopolysaccharide and evidence of biofilm formation. Appl. Environ. Microbiol. 62, 4478–4485. [BİOF-15/rf.198] Murat Kemal AVCI_İTÜ_MOBGAM

26 53. Rogers, J., Dowsett, A.B., Keevil, C.W., A paint incorporat ing silver to control mixed biofilms containing Legionella pneumophila. J. Ind. Microbiol. 15, 377–382. BİOF-15/rf.201] 54. Stern, N.J., Kazmi, S.U., Campylobacter jejuni. In: Doyle, M.P. (Ed.), Foodborne Bacterial Pathogens. Marcel Dekker, New York, USA, pp. 71–110. [BİOF-15/rf.219] 55. Wirtanen, G., Mattila-Sandholm, T., Measurement of biofilm of Pediococcus pentosaceus and Pseudomonas fragi on stainless steel surfaces. Coll. Surf. B: Biointerfaces 2, 33–39. [BİOF-15/rf.241] 56. Masako, K., A novel method to control the balance of skin microflora, Part 1. Attack on biofilm of Staphylococcus aureus without antibiotics. Journal of Dermatological Science (2005) 38, 197—205. [BİOF-C2] Murat Kemal AVCI_İTÜ_MOBGAM

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"Q UORUM S ENSING O DAKLı Y ENILIKLER VE B IYOTEKNOLOJIK U YGULAMALAR Hazırlayan : Murat Kemal AVCI Öğretim Üyesi : Doç. Dr. Zeynep Petek ÇAKAR Aralık-2009." indir ppt

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