Assembly of peptide linker to amino acid dehydrogenase and immobilized with metal–organic framework

Abstract

BACKGROUND:The metal-organic framework (MOF) ZIF-8 has the advantages of a large specific surface area and high stability. A new strategy for the assembly of amino acid dehydrogenase with peptide linker and for improving enzyme immobilization is proposed.RESULTS: Peptide linker enhanced enzyme activity after immobilization by 138.7%, and maintained 81.2% of its initial activity after seven recycles. The stability of PheDH_1D02/ZIF-8 was greatly enhanced at 70-80 °C and pH 10-11. The catalytic efficiency of PheDH_1D02/ZIF-8 was also enhanced compared with PheDH_1C1D/ZIF-8 and two free enzymes.CONCLUSIONS: Metal ions coordinated with the peptide linker and reoriented immobilization of enzyme onto the MOF with high enzyme loading capacity were achieved. The mesoporous structure of PheDH_1D02/ZIF-8 facilitates substrate diffusion. Combining the assembly of peptide linker and MOF immobilization is a simple, economical and efficient method for improving dehydrogenase stability.

Design of motifs interfacial interactions by co‐evolved analysis of D‐amino acid dehydrogenase for stability enhancement

“…HPLC analysis with standard solution of L‐phenylalanine and D‐phenylalanine showed that the peak time of L‐phenylalanine was 25 min, and the peak time of D‐phenylalanine was in 33 min. HPLC detection condition for D/L phenylalanine was as follow: Chromatographic column: Philomon Chirex 3126 (D)‐Penicillamine 250 nm × 4.6 nm, 5 μm; mobile phase, 5% (v/v) acetonitrile and 95% (v/v) 2.0 mM, CuSO 4 aqueous solution; flow rate, 0.8 mL/min; detection wavelength, 254 nm 22 …”

Design of Motifs Interfacial Interactions by Co-evolved Analysis of D-amino Acid Dehydrogenase for Stability Enhancement

“…HPLC analysis with standard solution of L-phenylalanine and D-phenylalanine showed that the peak time of L-phenylalanine was 25 min, and the peak time of Dphenylalanine was in 33 min. HPLC detection condition for D/L phenylalanine was as follow: Chromatographic column: Philomon Chirex 3126 (D)-Penicillamine 250 nm*4.6 nm, 5 μm; mobile phase, 5% (v/v) acetonitrile and 95% (v/v) 2.0 mM, CuSO 4 aqueous solution; flow rate, 0.8 mL/min; detection wavelength, 254 nm [22].…”

Laccase–mediator co-immobilized doped-HKUST-1 cellulose composite beads and their application for the biodegradation of carbazole

“…The lamellar structure can provide more attachment points for laccase and facilitate the immobilization of laccase molecules. 34 The sponge-like pore structure is beneficial for the transfer of the substrate components and can improve the reaction rate between laccase and substrate. In addition, the element mapping of the Lac@Ce-GMA-Cu-DA@AS biocatalyst showed that C, N, O, and Cu elements were evenly distributed in its beads.…”