Trypsin-catalyzed peptide synthesis withm-guanidinophenyl andm-(guanidinomethyl)phenyl esters as acyl donor component
- 87 Downloads
Two series of inverse substrates,m-guanidinophenyl andm-(guanidinomethyl)phenyl esters derived fromN-(tert-butyloxycarbonyl)amino acid, were prepared as an acyl donor component for trypsin-catalyzed peptide synthesis. The kinetic behavior of these esters toward tryptic hydrolysis was analyzed. They were found to couple with an acyl acceptor such asl-alaninep-nitroanilide to produce dipeptide in the presence of trypsin.Streptomyces griseus trypsin was a more efficient catalyst than the bovine trypsin. Within the enzymatic peptide coupling methods, this approach was shown to be advantageous, since the resulting peptides are resistant to the enzymatic hydrolysis.
KeywordsAmino acid esters Inverse substrate Kinetics of tryptic hydrolysis Protease catalysis Peptide synthesis
Unable to display preview. Download preview PDF.
- Bieth J, Weremath CG (1970) The action of elastase onp-nitroanilide substrates. Biochem Biophys Res Commun 53: 383–390Google Scholar
- Brot FE, Bender ML (1969) Use of the specificity constant ofα-chymotrypsin. J Am Chem Soc 91: 7187–7191Google Scholar
- Chase T, Jr., Shaw E (1967)p-Nitrophenylp′-guanidinobenzoate-HCI: a new active site titrant for trypsin. Biochem Biophys Res Commun 29: 508–514Google Scholar
- Itoh K, Sekizaki H, Toyota E, Fujiwara N, Tanizawa K (1996) Application of inverse substrates to trypsin-catalyzed peptide synthesis. Bioorg Chem 24: 59–68Google Scholar
- Kawai M, Nagai U (1982) Upon DNP-aromatic rule. Effect of chromophore exchange on the CD spectra ofN-DNP-arylalkylamines. Bull Chem Soc Jpn 55: 1327–1328Google Scholar
- Nakano M, Tanizawa K, Nozawa M, Kanaoka Y (1980) Efficient tryptic hydrolysis of aryl esters with a cationic center in the leaving group. Further characterization of “inverse substrates”. Chem Pharm Bull 28: 2212–2216Google Scholar
- Nakatsuka T, Sasaki T, Kaiser ET (1987) Peptide segment coupling catalyzed by the semisynthetic enzyme thiolsubtilisin. J Am Chem Soc 109: 3808–3810Google Scholar
- Ryan TJ, Fenton JW, II, Chang T-1, Feinman RD (1977) Specificity of thrombin: evidence for selectivity in acylation rather than binding forp-nitrophenylα-amino-p-toluate. Biochemistry 15: 1337–1341Google Scholar
- Schellenberger V, Jakubke H-D (1991) Protease-catalyzed kinetically controlled peptide synthesis. Angew Chem Int Ed Engl 30: 1437–1449Google Scholar
- Sekizaki H, Itoh K, Toyota E, Tanizawa K (1997) Enzymatic coupling of α, α-dialkyl amino acids using inverse substrates as acyl donors. Tetrahedron Lett 38: 1777–1780Google Scholar