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Journals / Türk Biyokimya Dergisi / 2013 / Cilt: 38 - Sayı: 3

Structural stability of commercial ficin under different denaturing conditions

Ticari fisinin yapısal stabilitesine farklı denature edici koşulların etkisi

Nabilah Abdul Aleem SİDEK
Adyani Azizah Abdul HALİM
Saad TAYYAB
Habsah Abdul KADİR
Pages
319–328
Publication Date
2022-11-05
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

CF showed greater stability at acidic pH than MFF.Conclusion: We conclude that CF is more structurally resistant than MFF against chemical and acid denaturations.

📌 3 Key Takeaways

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Objective & Hypothesis:Aim: To study the conformational stability of commercial ficin (CF) in the presence of guanidine hydrochloride (GdnHCl), urea, ethanol or at acidic pH and compare it with that reported for the major ficin fraction (MFF) obtained by purification of CF.Methods: Far-UV and near-UV CD spectral signals, intrinsic fluorescence, acrylamide quenching and enzymatic activity were used to study the effects of chemical denaturants and acidic pH on CF.
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Key Findings & Data:Loss of all enzymatic activity was observed at 4 M GdnHCl.
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Conclusion & Impact:CF showed greater stability at acidic pH than MFF.Conclusion: We conclude that CF is more structurally resistant than MFF against chemical and acid denaturations.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

Aim: To study the conformational stability of commercial ficin (CF) in the presence of guanidine hydrochloride (GdnHCl), urea, ethanol or at acidic pH and compare it with that reported for the major ficin fraction (MFF) obtained by purification of CF.Methods: Far-UV and near-UV CD spectral signals, intrinsic fluorescence, acrylamide quenching and enzymatic activity were used to study the effects of chemical denaturants and acidic pH on CF. The data were analyzed using two-state hypothesis, if required. Results: GdnHCl produced complete loss of secondary and tertiary structures of the protein. Loss of all enzymatic activity was observed at 4 M GdnHCl. CF showed structural resistance against 9 M urea and 50 % ethanol. Significant differences in emission maximum, acrylamide quenching, denaturation transition and enzymatic activity were noted between CF and MFF treated with different denaturants. CF showed greater stability at acidic pH than MFF.Conclusion: We conclude that CF is more structurally resistant than MFF against chemical and acid denaturations.

Özet

Amaç: Bu çalışmanın amacı ticari fisinin (CF) yapısal stabilite çalışmalarını, guanidin hidroklorür (GdnHCl), üre, etanol varlığında veya asidik pH'da yapmak ve CF saflaştırılması ile elde edilen major fisin fraksiyonu (MFF) ile karşılaştırmaktır.Metot: Uzak-UV ve yakın-UV CD spektral sinyalleri, içsel floresan, akrilamid su verme ve enzimatik aktivite metotları denatüre edici kimyasalların ve asidik pH'nın CF üzerindeki etkilerini incelemek için kullanılmışlardır.Bulgular: GdnHCl proteinin ikincil ve üçüncül yapıların tam kaybına sebep oldu. Enzimatik aktivite tam kaybı 4 M GdnHCl gözlemlendi. CF, 9 M üreye ve% 50 etanole karşı yapısal direnç gösterdi. Maksimum emisyonda, akrilamid su vermede, denatürasyon geçişinde ve enzimatik aktivitedeki önemli farklılıklar, farklı denatüranlar ile muamele edilen CF ve MFF arasında kaydedildi. Daha asidik pH da CF, MFF'Den daha fazla stabilitesini korudu.Sonuç: CF'nin kimyasal ve asit denatürantlara karşı MFF'den daha dayanıklı bir yapıya sahip olduğu sonucuna varıldı.

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Cite This Article

SİDEK, N. A. A., HALİM, A. A. A., TAYYAB, S., KADİR, H. A. (2022). Structural stability of commercial ficin under different denaturing conditions. Türk Biyokimya Dergisi, Cilt: 38 - Sayı: 3, 319–328. https://ajindex.com/articles/621289

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