PRODUCTION, IMMOBILIZATION AND APPLICATIONS OF ANTIMICROBIAL PEPTIDES FROM MICROOGANISMS
Project no. 642 · Status: Kapanmış · 2015
Summary
Antimicrobial peptides (AMPs), also known as host defense peptides, are short and generally positively charged peptides found in a wide variety of life forms from microorganisms to humans. Most AMPs have the ability to kill microbial pathogens directly, whereas others act indirectly by modulating the host defense systems. Against a background of rapidly increasing resistance development to conventional antibiotics all over the world, efforts to bring AMPs into clinical use are accelerating. Several AMPs are currently being evaluated in clinical trials as novel anti-infectives, but also as new pharmacological agents to modulate the immune response, promote wound healing, and prevent post-surgical adhesions. In this study, it is aimed that produce and purify AMP from different microorganisms and develop biomaterial with antimicrobial properties via immobilization of purified AMPs to calcium alginate. For this purpose, AMP production from Lactobacillus pentosus (DSM 16366), Bacillus subtilis (DSM 347), B. cereus, Pseudomonas aeruginosa (ATCC 27853) and B. subtilis (ATCC 6653) was performed. Optimization studies of AMP production were carried out. In the optimization studies, carbon source type and amount, nitrogen source type and amount, pH and temperature parameters were examined. AMP production was monitored every two hours by applying antibacterial test to the samples taken from the media. Disc diffusion method was used in the antibacterial test stage. AMP purification was performed from the microorganism (L. pentosus (DSM 16366)) which indicated the highest antibacterial activity as a result of optimization of AMP production. The purified AMP was covalently immobilized to calcium alginate beads using 1-ethyl-3- (3-dimethylaminopropyl) cabodiimide (EDC). As a result of the thesis, the product which was aimed to be developed was subjected to antibacterial test as it was designed as antibacterial. As a result of the antibacterial test, microbial growth was not observed in the region where AMP immobilized calcium alginate bead was applied. As a result, AMP production from microorganisms, purification of the produced AMP and immobilization of purified AMP to calcium alginate beads were achieved successfully and an antibacterial biomaterial was obtained. The product obtained as a result of the thesis has the potential to be used in order to prevent microbial contamination especially in open wounds by taking advantage of the use of alginate as a wound dressing.