Peptide trypsin-catalyzed

The peptide shown m green was isolated by trypsin catalyzed hydrolysis and has an ammo acid sequence that completes the remaining overlaps... 

Among proteases, the digestive enzyme trypsin catalyzes the hydrolysis of only those peptide bonds in which the carbonyl group is contributed by either a Lys or an Arg residue, regardless of the length or amino acid sequence of the chain. The number of smaller peptides produced by trypsin cleavage can thus be predicted... 

Figure 12.7 Left dependence of the yield of the a-chymo-trypsin-catalyzed synthesis of the peptides maleoyl-Tyr-D-Leu-NH2 (o) and maleoyl-Tyr-/3-Ala-Gly-OH ( ) in ice upon the relative amount of nucleophile free base (for conditions, see Table 12.7). Right influence of temperature on the yield of the a-chymotrypsin-catalyzed peptide synthesis in ice using maleoyl-L-Tyr methyl ester as acyl donor and H-D-Leu-NH2 (o), H-Gly-Gly-Gly-OH ( ), and H-Ala-Ala-OH (A) as amino components (for conditions, see Table 12.7) (Schuster, 1990).

Cerovsky, V., Free trypsin-catalyzed peptide synthesis in acetonitrile with low water content, Biotechnol. Lett., 12, 899-904, 1990. 

Trypsin-catalyzed hydrolysis gives three peptides and a single amino acid ... 

Cuanidinophenyl ester (OCp), guanidine (H2N-C(NH2)=NH) substituted phenyl ester of N -protected amino acids and peptides that has been used as substrate mimet-ics in trypsin-catalyzed peptide synthesis [F. Bordusa et al., Angew. Chem. Int. Ed. 1997, 36, 2473]. 

Biocatalysis from discovery to application. Springer, Berlin, Heidelbeig, New York, pp 31-57 Ren X, Yu D, Han S et al. (2007) Thermolysis of recombinant Escherichia coli for recovering a thermostable enzyme. Biochem Eng J 33(l) 94-98 Riechmann L, Kasche V (1985) Peptide synthesis catalyzed by the serine proteinases chymotrypsin and trypsin. Biochim Biophys Acta 830(2) 164-72 Robinson PJ, Wheatley MA, Janson JC et al. (1974) Pilot scale affinity chromatography of p-galactosidase. Biotechnol Bioeng 16 1103-1112... 

The well known specificity of proteinases implies the use of specific amino acids (amides, esters) as acyl donors and—seldom—specific amino acid (derivatives) as acceptors in enzymatic peptide bond formation, since the same structural features of RCONHR that influence the rate of hydrolytic cleavage are also involved in the synthesis. Accordingly trypsin is well suited to the formation of a new -Arg-X or -Lys-X bond. As an example the transformation of the -Lys-AlaOH terminus of the B-chain of porcine insulin into -LysThrOBu of human insuhn may be mentioned. C-terminal Ala was removed by means of car-boxypeptidase A, trypsin-catalyzed condensation of the des-alanine peptide with threonine tert. butylester gave 73% of the ester of human insulin [33] (see also... 

MassTag (Lys) [71] and iTRAQ (Lys) [72], aU of which are suitable for MALDI-MS and MS/MS. A simple method, namely trypsin-catalyzed incorporation of 0 into carboxyl groups, fadhtates the comparative analysis of protein and peptide samples by MALDI-MS [73, 74]. 

A group of proteolytic enzymes, including trypsin and chymotrypsin, can be used to catalyze the hydrolysis of specific peptide bonds. Trypsin catalyzes the hydrolysis of peptide bonds formed by the carboxyl groups of Arg and Lys chymotrypsin catalyzes the hydrolysis of peptide bonds formed by the carboxyl groups of Phe, Tyr, and Trp. 

Table 27.3 Cleavage of Specific Peptide Bonds Catalyzed by Trypsin and Chymotrypsin...

Trypsin-catalyzed hydrolysis forms the following two amino acids and two peptides ... 

Trypsin (Section 27 10) A digestive enzyme that catalyzes the hydrolysis of proteins Trypsin selectively catalyzes the cleavage of the peptide bond between the carboxyl group of lysine or arginine and some other amino acid... 

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