Lysozyme Cross-Linked Antibodious Destroyers Against Staphylococcus Aureus

Abstract

In the present study, we have provided a strategy to block staphylococcal protein A SpA which acts as a viru- lence factor and increase lysozyme activity against the staphylococcal infection. For this purpose, polymeric antibodious nano IgG has synthesized according to photosensitive micro emulsion technique and developed some binding parts on nano IgG for lysozyme. The bioconjugation method based on aminoacid monomer-prote- in cross-linking applying photosensitation and conjugation approach ANADOLUCA on micro and nano-struc- tures by ruthenium-chelate based monomers has used. The antibacterial activities of polymeric nano-IgG and lysozyme cross-linked polymeric nano-IgG have investigated against to Staphylococcus aureus. The effects on S. aureus have determined using dilution plate method and turbidity method. In addition, the effects on S. au- reus of lysozyme cross-linked polymeric nano-IgG have determined using confocal laser scanning microscopy. The biocompatibility of nanobioconjugates has determined by flow cytometry. The results have shown that nanobioconjugates could be used as an effective mimic and antimicrobial agents.

Keywords

• Staphylococcus aureus,IgG,bioconjugation

Staphylococcus Aureus’a Karşı Lizozim Çapraz Bağlı Antibadi Destroyerler

Abstract

B u çalışmada, stafilokokal enfeksiyonlara karşı lizozim aktivitesini arttıran ve virulans faktör gibi davranan stafilokokal protein A SpA bloklamak için bir strateji geliştirmiş bulunmaktayız. Bu amaç için, polimerik antibadimsi nano IgG destroyerler, fotosensitif mikro emülsiyon tekniğine göre sentezlenmiş ve nano IgG üzerinde lizozimler için bazı bağlanma bölgeleri geliştirilmiştir. Aminoasit-monomer çapraz bağlı fotosensitizasyon ve konjugasyon yaklaşımını ANADOLUCA mikro ve nano yapılara rutenyum-şelat tabanlı monomerlerle uygulama esasına dayanan biyokonjugasyon yöntemi kullanılmıştır. Staphylococcus aureus’a karşı polimerik nano IgG ve lizozim çapraz bağlı polimerik nano IgG’nin antibakteriyel etkileri araştırılmıştır. S. aureus’a etkiler dilüsyon pleyt yöntemi ve türbiditi yöntemine göre belirlenmiştir. Buna ek olarak, lizozim çapraz-bağlı nano-IgG’nin S. aureus ‘a etkisi konfokal lazer taramalı mikroskop ile belirlenmiştir. Nanobiyokonjugatların biyo uyumluluğu akış sitometrisi ile belirlenmiştir. Sonuçlar, nanobiyokonjugatların etkili mimik ve antimikrobiyal ajanlar olarak kullanılabilirliğini göstermiştir

Keywords

• Staphylococcus aureus,IgG,biyokonjugasyon

References

1. 1. G.D. Wright, Bacterial resistance to antibiotics: enzymatic degradation and modification, Advanced Drug Delivery Reviews, 57 (2005) 1451-1470.
2. 2. R. Satishkumar, A. Vertegel, Charge‐directed targeting of antimicrobial protein‐nanoparticle conjugates, Biotechnology and Bioengineering, 100 (2008) 403-412.
3. 3. S.C. Yadav, A. Kumari, R. Yadav, Development of peptide and protein nanotherapeutics by nanoencapsulation and nanobioconjugation, Peptides, 32 (2011) 173-187.
4. 4. E. Gustafssona, S. Karlssona, J. Oscarsson, P. Sogard, P. Nilsson, S. Arvidson, Mathematical modelling of the regulation of spa (protein A) transcription in Staphylococcus aureus, International Journal of Medical Microbiology 299, (2009) 65-74.
5. 5. M. Askarian, A. Zeinalzadeh, A. Japoni, A. Alborzi, Z.A. Memish, Prevalence of nasal carriage of methicillin-resistant Staphylococcus aureus and its antibiotic susceptibility pattern in healthcare workers at Namazi Hospital, Shiraz, Iran International Journal of Infectious Diseases, 13 (2009) 241-247.
6. 6. R. Saginur, K.N, Suh, Staphylococcus aureus bacteraemia of unknown primary source: Where do we stand? International Journal of Antimicrobial Agents, 32 (2008) 2-25.
7. 7. N. Palmqvist, G.J. Silverman, E. Josefsson, A. Tarkowski, Bacterial cell wall-expressed protein A triggers supraclonal B-cell responses upon in vivo infection with Staphylococcus aureus, Microbes and Infection, 7 (2005) 1501-1511.
8. 8. L. Pan, F. Yue, J. Miao, L. Zhang, J. Li, Molecular cloning and characterization of a novel c-type lysozyme gene in swimming crab Portunus trituberculatus, Fish & Shellfish Immunology, 29 (2010) 286-292.

9. 9. M. Khalili, A. Liwo, H. A. Scheraga, Kinetic Studies of Folding of the B-domain of Staphylococcal Protein A with Molecular Dynamics and a United-residue (UNRES) Model of Polypeptide Chains, J Mol Biol., 355 (2006) 536-547.
10. 10. T. J. Foster, Immune Evasion by Staphylococci, Nature Publishing Group 3, (2005) 948-958.
11. 11. K. Greenhalgh, E. Turos, In vivo studies of polyacrylate nanoparticle emulsions for topical and systemic applications, Nanomedicine: Nanotechnology, Biology, and Medicine, 5 (2009) 46-54.
12. 12. J.S. Kim, E. Kuk, K.N. Yu, J.H. Kim, S.J. Park, H.J. Lee, S.H. Kim, Y.K. Park, Y.H. Park, C.Y. Hwang, Y.K. Kim, Y. S.Lee, D.H. Jeong, M.H. Cho, Antimicrobial effects of silver nanoparticles, Nanomedicine: Nanotechnology, Biology, and Medicine, 3 (2007) 95-101.
13. 13. A. Nanda, M. Saravanan, M. Phil, Biosynthesis of silver nanoparticles from Staphylococcus aureus and its antimicrobial activity against MRSA and MRSE, Nanomedicine: Nanotechnology, Biology, and Medicine, 5 (2009) 452-456.
14. 14. R. Say, Photosensitive amino acid-monomer linkage and bioconjugation applications in life sciences and biotechnology. World Intellectual Property Organization Patent Scope. 2011,www. wipo.int/patentscope/search/en/WO2011070402 (Pub. No.:WO/2011/070402; Int. Appl. No.: PCT/ IB2009.055707).
15. 15. M. Liu, S. Zhang, Z. Liu, H. Li, A. Xu, Gene, 367 (2006) 110-117.
16. 16. I. Fedtke, F. Gotz, A. Peschel, Bacterial evasion of innate host defenses–the Staphylococcus aureus lesson, International Journal of Medical Microbiology, 294 (2004) 89-194.
17. 17. A. Bera, R. Biswas, S. Herbert, E. Kulauzovic, C. Weidenmaier, A. Peschel, F. Gotz, Influence of wall teichoic acid on lysozyme resistance in Staphylococcus aureus, Journal of Bacteriology, 189 (2007) 280-283.
18. 18. R. Say, R. Keçili, Ö. Biçen, F. Yılmaz, D. Hür, A. Denizli, A. Ersöz, A novel nanoprotein particle synthesis: Nanolipase, Process Biochemistry, 46 (2011) 1688- 1692.
19. 19. M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Analytical Biochemistry, 72 (1976) 248-254.