α-Chymotrypsin catalyzed oligomerization of the "dipeptide lego" KL-ethyl ester (OEt) in aqueous media triggers a rapid sol-gel transition due to formation of alternating (KL)x. Resulting mixed chain oligomers, at alkaline pH, self-assemble into β-sheets. Thereafter, intermolecular backbone hydrogen bonding between peptides causes formation of physically entangled nanofibrillar networks.

The direct oligomerization of l-lys-OMe by bromelain catalysis gave oligo­(l-lys) with DPavg ∼ 3.6 and dispersity ∼ 1.1. Since higher chain length oligo­(l-lys) with lower dispersity values and one reactive amine for selective conjugation would be beneficial, we explored protease-catalyzed oligomerization of Nε-protected l-lys monomers where Nε-groups included tert-butoxycarbonyl (Boc) or benzyloxycarbonyl (Z) groups. By using Nε-protected l-lys monomers, oligopeptide side-chains are hydrophobic-neutral which should dramatically alter enzyme kinetic and binding constants relative to nonprotected l-lys. Schechter and Berger’s conceptual model guided our choice of papain as the protease catalyst. Papain-catalyzed oligomerization of Nε-Boc-l-Lys-OMe gave products with DPavg values that were pH dependent and varied from 4.7 ± 0.2 to 7.5 ± 0.1. Similarly, oligo­(Nε-Z-l-Lys) synthesis was pH dependent, and DPavg values varied from 4.3 ± 0.2 to 5 ± 0.2. Oligo­(Nε-Boc/Z-l-Lys) that precipitates from reaction media had a low dispersity (∼1.01). The relatively smaller Nε-Boc group should increase propagating chain solubility enabling oligopeptides to reach higher DPavg values prior to their precipitation. Since papain-catalyzed oligomerizations of Nε-Boc/Z-l-Lys proceeded slowly at 0.54 mg/mL, higher enzyme concentrations were studied. By increasing the enzyme concentration in oligomerizations from 0.54 to 1.62 mg/mL for 3 h reactions, the %-yield and DPavg of oligo­(Nε-Z-l-lys) increased from 24 ± 0 to 88 ± 2 and 4.1 ± 0.7 to 5.7 ± 0.1, respectively. Furthermore, at 1.89 mg/mL papain, the %-yield of oligo­(Nε-Z-l-lys) increased with time reaching 91% in 2 h. Acetonitrile at 20%-by-volume was a useful cosolvent that increased the oligopeptide yield and DPavg relative to reactions run in pure buffer.