Summary
Hydrolytic enzymes (proteases, penicillin amidase) can be used also for synthetic purposes. The interest to use these enzymes for synthetic purposes as the semisynthesis of β-lactam antibiotics and peptides has increased recently.
This enzyme catalyzed synthesis of C—N (peptide) bonds can be performed under equilibrium of kinetically controlled conditions.
The latter is studied here for transpeptidations or transfer of β-lactam side chains to 6-amino penicillanic acid using activated side chains (esters, amides). In this enzyme catalyzed reaction a transient maximum concentration (kinetically controlled maximum) much larger than the final equilibrium concentration of the desired product (peptide, β-lactam) can be obtained.
The yield at this maximum depends on the ratio of the deacylation rate constants of the acyl-enzyme with the nucleophiles and H2O. The particle size-, ionic-strength-, pH- and nucleophile structure-dependence of this ratio has been experimentally studied for free and immobilised biocatalysts. The data were found to correlate well with theoretical predictions.
The kinetically controlled maximum is compared with the equilibrium controlled synthesis as an endpoint for enzyme catalyzed semisynthetic procedures.
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Abbreviations
- Ac-Tyr, Ac-Tyr-O-Et, Ac-Tyr-O-Me:
-
N-acetyl-L-tyrosine (ethyl-or methylester)
- Ac-Ty-Tris:
-
N-acetyl-L-Tyrosine-Tris
- 7-ACA:
-
7-aminocephalosporanic acid
- 7-ADCA:
-
7-aminodesacetoxycephalosporanic acid
- AMP:
-
ampicillin
- 6-APA:
-
6-aminopenicillanic acid
- BP:
-
benzyl-penicillin
- CT:
-
α-chymotrypsin (E.C. 3.4.21.1)
- Lys-O-Me:
-
L-lysine-methyl-ester
- PA:
-
penicillin amidase (E.C. 3.5.1.11)
- PAA:
-
phenyl-acetic-acid
- PAG:
-
phenyl-acetyl-glycine
- PG:
-
D-phenyl-glycine
- PG-O-Me:
-
D-phenyl-glycine-methyl-ester
- Tris:
-
tris(hydroxymethyl)methylamine
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Kasche, V., Galunsky, B., Haufler, U., Zöllner, R. (1983). Kinetically VS. Equilibrium-Controlled Synthesis of C—N Bonds in β-Lactams and Peptides with Free and Immobilized Biocatalysts. In: Lafferty, R.M. (eds) Enzyme Technology. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-69148-5_24
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