Polimer kaplı paslanmaz çelik yüzeye enzim immobilizasyonunun denizel biyofilm tabakası oluşumu üzerine etkisinin incelenmesi
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Dosyalar
Tarih
2023
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Hatay Mustafa Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Katı- sıvı etkileşiminin olduğu yüzeylerde mikroorganizmalar zamanla biyofilm oluşturmaktadır. Biyofilm oluşumunun neden olduğu ekonomik kaybı önlemek amacıyla tarih boyunca çeşitli boyalar ve kaplamalar kullanılmıştır. Ancak kullanılan tekniklerin çoğu, biyosit etkilerini ağır metallerin çevreye salınımıyla sağlamakta ve çevre kirliliğine sebep olmakla kalmayıp ekosistemi tehdit etmektedir. Bu nedenle çevre dostu, düşük maliyetli, uzun süreli kullanıma uygun yeni tekniklerin üretilmesi önem kazanmaktadır. Bu çalışmada, metal üzerinde deniz kaynaklı biyofilm oluşumunun engellenmesine odaklanılmıştır. Bu amaçla, antibiyofilm etkileri bilinen anilin ve o-anisidin monomerleri kullanılarak ilk aşamada 304 paslanmaz çelik üzerine sodyum okzalat, sodyum salisilat, sodyum molibdat, sodyum p-toluen sülfonat ve amonyum okzalat ortamlarında, dönüşümlü voltametri tekniği kullanılarak polimer film sentezleri gerçekleştirilmiştir. Mersin Balıkçı Barınağı'nda 12 gün boyunca denize bırakılan 304 paslanmaz çelik kuponların üzerindeki kaplamaların antibiyofilm etkileri kristal viyole ile boyama yöntemiyle spektrofotometrik olarak karşılaştırılmıştır. Amonyum okzalat ve sodyum salisilat ortamlarında sentezlenen polianilin (PANİ) ve poli(o-anisidin) (POA) filmlerin antibiyofilm performanslarının diğer elektrolit ortamda sentezlenen kaplamalara göre daha yüksek olduğu belirlenmiştir. Denizel ortamda gelişen biyofilmlerin ağırlıklı olarak protein, karbohidrat, lipit ve nükleik asit içerdikleri gözönünde bulundurularak, PANİ ve POA sentez ortamlarına α-amilaz, DNAz, glukoz oksidaz, α-kimotripsin, lipaz ve pektinaz enzimleri ayrı ayrı ve farklı kombinasyonlarda eklenerek kaplama matriksine enzimlerin tekli ve çoklu immobilizasyonları sağlanmıştır. Enzim ile modifiye edilmiş polimer kaplı yüzeylerin özellikleri antibiyofilm performansları, kristal viyole boyama, floresans mikroskobu, taramalı elektron mikroskobu (SEM), enerji dağıtıcı x-ışını spektroskopisi (EDX) ile karakterize edilmiştir. Beklenilenin aksine, tek bir enzim immobilize edilmiş kaplamalar, ko-immobilize enzim içeren kaplamalara göre daha yüksek antibiyofilm etki göstermiştir. Her aşamada en yüksek antibiyofilm özellik gösteren enzimsiz, tek enzimli ve ko-immobilize enzimli kuponlar denizde bekeltildikten sonra yüzeylerindeki biyofilmin karbohidrat, protein ve bakteri miktarları belirlenmiş; yüzeye tutunan bakteri türleri MALDİ-TOF MS analizi ile araştırılmıştır.
On surfaces where solid-liquid interactions occur, microorganisms form a biofilm over time. Various paints and coatings have been used throughout history to prevent economic loss caused by biofilm formation. However, most of the techniques which have been used provide their biocide effects with the release of heavy metals to the environment and by this they do not only cause environmental pollution, but also threaten the ecosystem. For this reason, it is important to develop new ideas that are environmentally friendly, low-cost and suitable for long-term use. In this study, we focused on marine biofilm formation on metal. For this purpose, aniline and o-anisidine monomers with known antibiofilm effects were used. In the first stage, these polymers were synthesized separately on 304 stainless steel in sodium oxalate, sodium salicylate, sodium molybdate, sodium p-toluene sulfonate and ammonium oxalate electrolytes using cyclic voltammetry technique. Antibiofilm effects of the coatings were quantitatively compared with crystal violet (CV) staining method after immersed in the sea for 12 days in Mersin Fisherman's Shelter. The subsequent studies were carried out with ammonium oxalate and sodium salicylate medium, which showed higher antibiofilm activity compared to the other electrolytes. Considering that the biofilms formed in the marine environment mainly contain protein, carbohydrates, lipids and nucleic acids, α-amylase, DNAse, glucose oxidase, α-chymotrypsin, lipase and pectinase enzymes are added separately and in different combinations to the PANi and POA synthesis media, and the enzymes are added to the coating matrix as single and multiple immobilizations were performed. The properties of enzyme-modified polymer-coated surfaces were characterized by their antibiofilm performance, crystal violet staining, fluorescence microscopy, scanning electron microscope (SEM), energy dispersive x-ray spectroscopy (EDX). Contrary to expectations, coatings with a single enzyme immobilized showed higher antibiofilm activity than coatings with co-immobilized enzyme. After the enzyme-free, single-enzyme and co-immobilized crush coupons with the highest antibiofilm properties at each stage were kept in the sea, the carbohydrate, protein and bacterial amounts of the biofilm on their surfaces were investigated, and MALDI-TOF MS analysis was performed to determine the bacterial species attached to the surface.
On surfaces where solid-liquid interactions occur, microorganisms form a biofilm over time. Various paints and coatings have been used throughout history to prevent economic loss caused by biofilm formation. However, most of the techniques which have been used provide their biocide effects with the release of heavy metals to the environment and by this they do not only cause environmental pollution, but also threaten the ecosystem. For this reason, it is important to develop new ideas that are environmentally friendly, low-cost and suitable for long-term use. In this study, we focused on marine biofilm formation on metal. For this purpose, aniline and o-anisidine monomers with known antibiofilm effects were used. In the first stage, these polymers were synthesized separately on 304 stainless steel in sodium oxalate, sodium salicylate, sodium molybdate, sodium p-toluene sulfonate and ammonium oxalate electrolytes using cyclic voltammetry technique. Antibiofilm effects of the coatings were quantitatively compared with crystal violet (CV) staining method after immersed in the sea for 12 days in Mersin Fisherman's Shelter. The subsequent studies were carried out with ammonium oxalate and sodium salicylate medium, which showed higher antibiofilm activity compared to the other electrolytes. Considering that the biofilms formed in the marine environment mainly contain protein, carbohydrates, lipids and nucleic acids, α-amylase, DNAse, glucose oxidase, α-chymotrypsin, lipase and pectinase enzymes are added separately and in different combinations to the PANi and POA synthesis media, and the enzymes are added to the coating matrix as single and multiple immobilizations were performed. The properties of enzyme-modified polymer-coated surfaces were characterized by their antibiofilm performance, crystal violet staining, fluorescence microscopy, scanning electron microscope (SEM), energy dispersive x-ray spectroscopy (EDX). Contrary to expectations, coatings with a single enzyme immobilized showed higher antibiofilm activity than coatings with co-immobilized enzyme. After the enzyme-free, single-enzyme and co-immobilized crush coupons with the highest antibiofilm properties at each stage were kept in the sea, the carbohydrate, protein and bacterial amounts of the biofilm on their surfaces were investigated, and MALDI-TOF MS analysis was performed to determine the bacterial species attached to the surface.
Açıklama
Anahtar Kelimeler
Biyokimya, Biochemistry
