Abstract

Bu çalışmada Aspergillus'a ait izolat LC3'ün ekstrasellüler proteinleri, yeni antimikrobiyal polipeptit (AMP) keşfi için saflaştırıldı ve ardından Staphylococcus aureus (ATCC 25923) ve Metisiline dirençli S. aureus'a (MRSA) karşı antimikrobiyal aktivite açısından test edildi. Antimikrobiyal aktivitenin polipeptit kaynaklı olduğu tripsin/proteinaz K testi ile belirlendi ve bu proteinin jel overlay testi ile yaklaşık 11 kDa'lık bir protein olduğu gözlendi. Saflaştırılmış AMP molekülünün S. aureus ATCC 25923 ve MRSA'ya karşı minimum inhibisyon konsantrasyonu sırasıyla 8 µg/ml ve 32 µg/ml’dir ve AMP molekülü doğrulandı. ITS sekans analizi, LC3 izolatının Bioedit sekans birleştirme programı kullanılarak Aspergillus niger olarak tanımlandığını gösterdi, sekans, MK332597 erişim numarasıyla GenBank veri tabanına kaydedildi. Bu sonuçlar, saflaştırılmış AMP molekülünün S. aureus'un neden olduğu enfeksiyonlarda kullanılma potansiyeline sahip olduğunu göstermektedir.

ISOLATION OF A NOVEL ANTIMICROBIAL POLYPEPTIDE FROM AN Aspergillus niger ISOLATE

Abstract

In this study the extracellular proteins from the isolate LC3 belonging to Aspergillus were purified for new antimicrobial polypeptide (AMP) discovery and then tested for antimicrobial activity against Staphylococcus aureus (ATCC 25923) and Methicillin-resistant S. aureus (MRSA). Antimicrobial activity was determined by the trypsin/proteinase K assay, which was polypeptide-based, and it was observed that this protein was a protein of about 11 kDa by gel overlay assay. The minimum inhibitory concentration of purified AMP molecule against S. aureus ATCC 25923 and MRSA was 8 µg/ml and 32 µg/ml, respectively and the AMP molecule was confirmed. ITS sequence analysis showed that isolate LC3 was identified as Aspergillus niger, using the Bioedit sequence assembly program. The sequence was deposited with the GenBank database with accession number MK332597. The results indicate that the purified AMP molecule has the potential to be used in infections caused by S. aureus.

Keywords

• Antimicrobial polypeptide
• Aspergillus niger
• Gel overlay assay
• Staphylococcus aureus.

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