Protocol detail

Enzymatic Protein Substrate Specificity

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The substrate specificity of UcB5 against natural protein substrates was studied at 0.5% (w/v) concentration following the procedure of Peng et al.^{14 (link)}. These include casein, elastin, hemoglobin, fibrin, gelatin, fibrinogen, collagen, mucin, IgG, and serum albumin. This was determined against several natural proteins. One unit (1 U) of proteolytic activity was calibrated as the quantity of enzyme that releases the comparable to 1 µmol of the amino acid tyrosine per milliliter per minute under the standard conditions of the assay.

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Kotb E., El-Nogoumy B.A., Alqahtani H.A., Ahmed A.A., Al-shwyeh H.A., Algarudi S.M, & Almahasheer H. (2023). A putative cytotoxic serine protease from Salmonella typhimurium UcB5 recovered from undercooked burger. Scientific Reports, 13, 3926.

Publication 2023

Amino acid Assay Casein Collagen Elastin Enzyme Fibrin Fibrinogen Gelatin Hemoglobin Mucin Proteins Proteolytic Releases Serum albumin Tyrosine

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Variable analysis

independent variables

Substrate type (casein, elastin, hemoglobin, fibrin, gelatin, fibrinogen, collagen, mucin, IgG, serum albumin)

dependent variables

Proteolytic activity (amount of enzyme that releases 1 μmol of the amino acid tyrosine per milliliter per minute under the standard assay conditions)

control variables

Substrate concentration (0.5% (w/v))
Assay procedure (following Peng et al. (2002))
Assay conditions (standard)

controls

The authors do not explicitly mention any positive or negative controls.

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