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

doi:10.7503/cjcu20150161

Abstract

Abstract:

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 supramole-cular 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 β-sheet structure come from the week interactions between Fmoc groups. The kinetics behavior, optimum temperature and pH studies showed the typical enzymatic cha-racteristics 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.

Key words: Artificial enzyme, Hydrolase, Peptide assembly, Nanofiber

CLC Number:

O629.8

TrendMD:

LÜ Yuqi, WANG Mengfan, QI Wei, SU Rongxin, HE Zhimin. Artificial Hydrolase Based on Short Peptides Self- and Co-assembly Nanofiber^†[J]. Chem. J. Chinese Universities, 2015, 36(7): 1304.

Figures/Tables 8

Fig.1 Activities of monomers(a), SA-H(b), SA-D(c), SA-S(d), CoA-HS(e), CoA-HD(f), CoA-DS(g) and CoA-HSD(h)

Fig.2 TEM images of SA-H(A), CoA-HS(B), CoA-HD(C) and CoA-HSD(D)

Fig.3 Fluorescence emission spectrum of SA-HConcentration of SA-H/(mol·L-1): a. 1; b. 0.01.

Fig.4 CD spectrum(A) and FTIR spectrum(B) of SA-H

Fig.5 Lineweaver-burk plots for the hydrolysis reaction catalyzed by SA-H and natural enzyme

Table 1 Kinetic parameters of SA-H andnatural enzyme

System	K_m /(mmol·L^-1)	K_cat /(mmol·L^-1·min^-1·mg^-1)	10³ K_cat/K_m(min^-1·mg^-1)
SA-H	20.03	0.014	6.99
Natural enzyme	16.39	0.006	3.66

Fig.6 Effects of temperature(A) and pH(B) on activity of SA-H(a) and PPL(b)Relative activity(%) was calculated by assuming the activity at optimal temperature or pH as 100%.

Fig.7 Effects of metal ions on the activity of SA-HRelative activity(%) was calculated by assuming the activity of SA-H without metal ions as 100%. a. SA-H; b. Fe2+; c. Cn3+; d. Mg2+; e. Ni2+; f. Co2+; g. Cu2+; h. Ba2+; i. Ca2+; j. Zn2+.

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