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Type: Artigo de Periódico
Title: Azocasein Substrate For Determination Of Proteolytic Activity: Reexamining A Traditional Method Using Bromelain Samples
Author: Coelho
DF; Saturnino
TP; Fernandes
FF; Mazzola
PG; Silveira
E; Tambourgi
Abstract: Given the importance of protease's worldwide market, the determination of optimum conditions and the development of a standard protocol are critical during selection of a reliable method to determine its bioactivity. This paper uses quality control theory to validate a modified version of a method proposed by Charney and Tomarelli in 1947. The results obtained showed that using azocasein substrate bromelain had its optimumat 45 degrees Cand pH 9 (Glycine-NaOH 100 mM). We also quantified the limit of detection (LoD) and limit of quantification (LoQ) in the above-mentioned optimum (0.072 and 0.494 mg.mL(-1) of azocasein, resp.) and a calibration curve that correlates optical density with the amount of substrate digested. In all analysed samples, we observed a significant decrease in response after storage (around 17%), which suggests its use must be immediately after preparation. Thus, the protocol presented in this paper offers a significant improvement, given that subjective definitions are commonly used in the literature and this simple mathematical approach makes it clear and concise.
Subject: Ananas-comosus
Citation: Biomed Research International. HINDAWI PUBLISHING CORP, n. 8409183, p. .
Rights: aberto
Identifier DOI: 10.1155/2016/8409183
Date Issue: 2016
Appears in Collections:Unicamp - Artigos e Outros Documentos

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