Azocasein Substrate for Determination of Proteolytic Activity: Reexamining a Traditional Method Using Bromelain Samples
Joint Authors
Coêlho, Diego F.
Saturnino, Thais Peron
Fernandes, Fernanda Freitas
Silveira, Edgar
Tambourgi, Elias Basile
Mazzola, Priscila Gava
Source
Issue
Vol. 2016, Issue 2016 (31 Dec. 2016), pp.1-6, 6 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2016-01-27
Country of Publication
Egypt
No. of Pages
6
Main Subjects
Abstract EN
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 optimum at 45°C and 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.
American Psychological Association (APA)
Coêlho, Diego F.& Saturnino, Thais Peron& Fernandes, Fernanda Freitas& Mazzola, Priscila Gava& Silveira, Edgar& Tambourgi, Elias Basile. 2016. Azocasein Substrate for Determination of Proteolytic Activity: Reexamining a Traditional Method Using Bromelain Samples. BioMed Research International،Vol. 2016, no. 2016, pp.1-6.
https://search.emarefa.net/detail/BIM-1099049
Modern Language Association (MLA)
Coêlho, Diego F.…[et al.]. Azocasein Substrate for Determination of Proteolytic Activity: Reexamining a Traditional Method Using Bromelain Samples. BioMed Research International No. 2016 (2016), pp.1-6.
https://search.emarefa.net/detail/BIM-1099049
American Medical Association (AMA)
Coêlho, Diego F.& Saturnino, Thais Peron& Fernandes, Fernanda Freitas& Mazzola, Priscila Gava& Silveira, Edgar& Tambourgi, Elias Basile. Azocasein Substrate for Determination of Proteolytic Activity: Reexamining a Traditional Method Using Bromelain Samples. BioMed Research International. 2016. Vol. 2016, no. 2016, pp.1-6.
https://search.emarefa.net/detail/BIM-1099049
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1099049