Digestion of Casein by Trypsin

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Presentation transcript:

Digestion of Casein by Trypsin ■ Proteases Enzymes: Enzymes that catalyze the hydrolysis of pepide bonds of proteins.

Digestion of Casein by Trypsin ■ Exoproteases: liberate amino acids by breaking peptide bonds starting from the end of the polypeptide chain. -H2O

Digestion of Casein by Trypsin ■ Endoproteases: break down peptide bonds from the interior of the polypeptide chain. e.g. Trypsin and Pepsin.

Trypsin A endoprotease enzyme found in the pancreatic juice which catalyzes the hydrolysis of peptide bonds formed from the carboxyl groups of basic amino acids, L-Arginine L-Lysine

Pepsin is an endoprotease found in the stomach which catalyzes the hydrolysis of peptide bonds between aromatic amino acids; Phenylalanine Tyrosine Tryptophan

Casien Free amino groups *Principle Trypsin Casien Free amino groups ~NH3 + H.CHO ~NH.CH2O + H (monomethylol)

Tube # 1 2 3 4 30 min. 37°C 5 min at 3000 rpm Casien (ml) 1 1 1 1 60 30 15 45 5 10 20 25 35 40 50 55 1% 0.8% 0.6% 0.05% 30 min. 37°C Casien (ml) 1 1 1 1 Trypsin (ml) 0.2 0.2 0.2 0.2 TCA 12% (ml) 1 1 1 1 5 min at 3000 rpm

0.1N NaOH 1 Va.a. VNaOH 2 Va.a. VNaOH 3 Va.a. VNaOH 4 Va.a. VNaOH 1% 2 drops ph.ph. Supernatant VNaOH 5 ml 40% HCOOH 2 Va.a. 0.8% VNaOH 3 0.6% Va.a. VNaOH 4 0.5% Va.a. VNaOH

* Calculations ( N × V ) a.a. = ( N `× V `) NaOH Na.a. = ( N `× V `) NaOH Va.a. Samar A. Damiati