İpek atıksularından geri kazanılan serisin proteininin karakterizasyonu

Year 2015, Volume: 72 Issue: 3, 219 - 234, 01.09.2015

Gökşen Çapar Seylan Saniye Aygün

Abstract

Amaç: Bu çalışmanın amacı, ipek atıksularından geri kazanılan serisin proteininin özelliklerini belirlemektir. Yöntem: Orta Doğu Teknik Üniversitesi Mühendislik Bilimleri Bölümü’nde 2007-2008 yılları arasında ipek atıksuyundan membran teknolojisi ile serisin proteini geri kazanılmıştır. Protein karakterizasyon çalışması Ankara Üniversitesi Su Yönetimi Enstitüsü’nde 2012 yılında tamamlanmıştır. Geri kazanılan protein moleküler ağırlık, nem ve kül içeriği, elementel analiz ve amino asit kompozisyonu yönünden incelenmiştir. Proteinin saflaştırılması için diyaliz işlemi uygulanmıştır. Serisin kozadan hidrotermal işlemle ekstrakte edilmiştir. Geri kazanılan proteinin farklı pH değerlerinde suda çözünürlüğü belirlenmiştir. Protein tanımlanması için 2-D jel elektroforez ve MALDI-TOF analizleri kullanılmıştır. Bulgular: Geri kazanılan serisinin molekül ağırlık aralığı 40-176 kDa olarak tespit edilmiş, en yüksek fraksiyonun ise 86-96 kDa aralığında ve %79-97 oranında olduğu bulunmuştur. Geri kazanılan serisin, biyomalzeme ve membran yapmaya uygun olan yüksek-molekül ağırlıklı serisin olarak sınıflandırılmıştır. Nem ve kül içerikleri %2,8-3,9% ve %11,3-14,4 olarak bulunmuştur. Geri kazanılan serisinin elementel kompozisyonunda, C, H, N içerikleri sırasıyla %36,7-45,3; %5,4-8,8 ve %10,2-16,8 olarak bulunmuştur. Geri kazanılan serisinin özellikleri, bu çalışmada kullanılan referans serisine ve literatürde yer alan diğer serisin örneklerine oldukça benzer çıkmıştır. Çözelti pH’sı, geri kazanılan serisinin suda çözünürlüğünü belirgin bir derecede etkilemiş; bu nedenle etanolün, asitlere kıyasla daha uygun bir çökeltme ajanı olduğu gözlenmiştir. Bazı amino asitler uygulanan işlemler sırasında kaybedilmiş olsa da amino asit kompozisyonu kabul edilebilir bulunmuştur. İpek atıksuyundan geri kazanılan serisin, %28,9’luk serin içeriği ile yüksek nem tutma potansiyeline sahiptir. Sonuç: İpek atıksuyundan geri kazanılan serisin biyomedikal, kozmetik ve ilaç endüstrilerinde potansiyel uygulamalar için umut verici bir hammaddedir

Keywords

Amino asit, moleküler ağırlık, serisin, ipek, geri kazanım

Characterization of sericin protein recovered from silk wastewaters

Abstract

Objective: This study aims to determine the characteristics of sericin protein recovered from silk wastewaters. Method: Sericin protein was recovered from silk wastewaters by membrane technology in Engineering Sciences Department of the Middle East Technical University between 2007 and 2008. The protein characterization study was completed in Ankara University Water Management Institute in 2012. The recovered protein was characterized in terms of molecular weight, moisture and ash contents, elemental and amino acid compositions. Dialysis was adopted to purify the protein. Sericin was extracted from native cocoons via hydrothermal processing. The solubility of recovered sericin samples at various pH values was determined. 2-D gel electrophoresis and MALDI-TOF analyses were used for protein identification. Results: The molecular weight range of recovered sericin was found as 40-176 kDa, with 86-96 kDa at the highest fraction of 79-97%. The recovered sericin was classifed as high-molecular weight sericin, which is suitable for making biomaterials and membranes. Moisture and ash contents were found as; 2.8-3.9% and 11.3-14.4%. In terms of elemental composition, C, H, N contents of recovered sericin were determined as; 36.7-45.3%, 5.4-8.8% and 10.2-16.8%, respectively. Properties of recovered sericin were quite similar to the reference sericin used in this study and those reported in literature. Solution pH significantly influenced the solubility of recovered sericin, so ethanol was observed to be a better precipitation agent than the acids used. Although some amino acids were lost during processing, the amino acid composition was acceptable. Sericin recovered from silk wastewater, with a serine content of 28.9%, had the potential of high moisture absorption. Conclusion: Sericin recovered from silk wastewaters is a promising raw material for potential applications in biomedical, cosmetics and pharmaceutical industries

Keywords

Amino acid, molecular weight, sericin, silk, recovery

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