Artificial Hydrolase Based on Short Peptides Self- and Co-assembly Nanofiber

The catalytic triad(Ser/His/Asp) of natural hydrolase were introduced into the peptide segment of 9-fluorenylmethoxycarbonyl-diphenylalanine (Fmoc-FF), 9-fluorenylmethoxycarbonyl-phenylalanine-phenylalanine-histidine(Fmoc-FFH), 9-fluorenylmethoxycarbonyl-phenylalanine-phenylalanine-serine(Fmoc-FFS) and 9-fluorenylmethoxycarbonyl-phenylalanine-phenylalanine-aspartate(Fmoc-FFD) were designed to construct the artificial hydrolase. Based on the self-assembly and co-assembly of Fmoc-peptides, a series of supramolecular nanofibers built from the self-assembly of Fmoc-FFH(SA-H), self-assembly of Fmoc-FFS(SA-S), self-assembly of Fmoc-FFD(SA-D), co-assembly of Fmoc-FFH and Fmoc-FFS(CoA-HS), co-assembly of Fmoc-FFH and Fmoc-FFD(CoA-HD), co-assembly of Fmoc-FFD and Fmoc-FFS(CoA-DS), co-assembly of Fmoc-FFH, Fmoc-FFD and Fmoc-FFS (CoA-HDS) were obtained as artificial hydrolases. SA-H exhibited the highest hydrolysis activity for p-nitrophenyl acetate (PNPA). It was found that the well-ordered structure of nanofiber is important for catalytic activity, and the beta-sheet structure come from the week interactions between. Fmoc groups. The kinetics behavior, optimum temperature and pH studies showed the typical enzymatic characteristics of artificial hydrolase as that of:natural enzyme. In addition, metal ions Ca2+, Ba2+ can activate the artificial hydrolase, whereas Mg2+ Ni2+, Co2+, Cu2+ Zn2+ inhibit the activity.