Investigation of the effects of selected drugs on the lactonase activity of the paraoxonase enzyme

Öz

Paraoxonase 1 (PON1) is a member of the PON family, which possesses essential functions in numerous metabolic processes due to its detoxification and antioxidant properties. In addition, through its lactonase activity, PON1 hydrolyzes lactone-based signaling molecules produced by bacteria, thereby inhibiting their quorum sensing (QS) mechanisms. One of the most critical outcomes of these mechanisms is biofilm formation, a clinically significant problem. Therefore, the lactonase activity of PON1 makes it a potential antibiofilm agent. In this study, hPON1 was purified from human serum for the first time using Sepharose 4B–L-tyrosine-1-naphthylamine gel synthesized by our team, employing hydrophobic interaction chromatography. The inhibitory effects of Rifamycin and Teicoplanin on the enzyme’s lactonase activity were investigated in vitro, and both antibiotics were found to significantly inhibit the enzyme. The calculated IC₅₀ values were 42.81 µg/mL for Rifamycin and 42.89 µg/mL for Teicoplanin. These findings suggest that, in addition to their bactericidal effects, these antibiotics may also weaken hPON1-mediated antibiofilm mechanisms.In this study, hPON1 was purified from human serum for the first time using Sepharose 4B–L-tyrosine-1-naphthylamine gel synthesized by our team, employing hydrophobic interaction chromatography. The inhibitory effects of Rifamycin and Teicoplanin on the enzyme’s lactonase activity were investigated in vitro, and both antibiotics were found to significantly inhibit the enzyme. The calculated IC₅₀ values were 42.81 µg/mL for Rifamycin and 42.89 µg/mL for Teicoplanin. These findings suggest that, in addition to their bactericidal effects, these antibiotics may also weaken hPON1-mediated antibiofilm mechanisms.

Anahtar Kelimeler

• hPON1
• lactonase
• quorum sensing
• biofilm
• hydrophobic interaction chromatography
• rifamycin
• teicoplanin

Bazı ilaçların paraoksonaz enziminin laktonaz aktivitesi üzerindeki etkilerinin araştırılması

Öz

Paraoksonaz 1 (PON1), detoksifikasyon ve antioksidan özellikleri ile birçok metabolik süreçlerde önemli fonksiyonlara sahip PON ailesinin bir üyesidir. Ayrıca PON1, laktonaz aktivitesi aracılığıyla bakteriler tarafından üretilen lakton yapısındaki sinyal moleküllerini hidrolize ederek onların quorum sensing (QS) mekanizmalarının inhibisyonuna sebep olur. Bu mekanizmaların en kritik çıktılarından biri, klinik açıdan ciddi bir problem olan biyofilm oluşumudur. Bu nedenle PON1’in laktonaz aktivitesi, enzimi potansiyel bir anti-biyofilm ajanı haline getirmektedir. Bu çalışmada, ilk kez tarafımızca sentezlenen Sepharose 4B–L-tirozin-1-naftilamin jeli kullanılarak insan serumundan hPON1 enzimi hidrofobik etkileşim kromatografisi ile saflaştırılmıştır. Enzimin laktonaz aktivitesi üzerine Rifamisin ve Teikoplanin etken maddelerinin inhibisyon etkileri in vitro olarak araştırılmış ve her iki antibiyotiğin de enzimi belirli düzeyde inhibe ettiği görülmüştür. Elde edilen IC₅₀ değerleri; Rifamisin için 42.81 µg/mL, Teikoplanin için ise 42.89 µg/mL olarak hesaplanmıştır. Bulgular, bu antibiyotiklerin bakterisidal etkilerinin yanı sıra hPON1 aracılı antibiyofilm mekanizmalarını da zayıflatabileceğini düşündürmektedir.

Anahtar Kelimeler

• hPON1
• laktonaz
• quorum sensing
• biyofilm
• hidrofobik etkileşim kromatografisi
• rifamisin
• teikoplanin

Kaynakça

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