PDBsum entry 5zgo

Transferase

PDB id: 5zgo

Contents

Protein chains

(+ 0 more) 178 a.a.

Waters ×155

PDB id:

5zgo

Name:

Transferase

Title:

Crystal structure of aprt2 from thermus thermophilus hb8

Structure:

Adenine phosphoribosyltransferase. Chain: a, b, c, d, e, f. Engineered: yes

Source:

Thermus thermophilus (strain hb8 / atcc 27634 / dsm 579). Organism_taxid: 300852. Strain: hb8 / atcc 27634 / dsm 579. Gene: ttha1614. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

2.60Å R-factor: 0.204 R-free: 0.264

Authors:

N.Kunishima,H.Naitow,Y.Matsuura

Key ref:

J.D.Arco et al. (2019). Structural and functional characterization of thermostable biocatalysts for the synthesis of 6-aminopurine nucleoside-5'-monophospate analogues. Bioresour Technol, 276, 244-252. PubMed id: 30640018 DOI: 10.1016/j.biortech.2018.12.120

Date:

09-Mar-18 Release date: 06-Mar-19

Protein chains

Q5SHW6  (Q5SHW6_THET8) -  Adenine phosphoribosyltransferase from Thermus thermophilus (strain ATCC 27634 / DSM 579 / HB8)

Seq: 178 a.a.

Struc: 178 a.a.

DOI no: 10.1016/j.biortech.2018.12.120

Bioresour Technol 276:244-252 (2019)

PubMed id: 30640018

Structural and functional characterization of thermostable biocatalysts for the synthesis of 6-aminopurine nucleoside-5'-monophospate analogues.

J.D.Arco, E.Pérez, H.Naitow, Y.Matsuura, N.Kunishima, J.Fernández-Lucas.

ABSTRACT

The present work describes the functional and structural characterization of adenine phosphoribosyltransferase 2 from Thermus thermophilus HB8 (TtAPRT2). The combination of structural and substrate specificity data provided valuable information for immobilization studies. Dimeric TtAPRT2 was immobilized onto glutaraldehyde-activated MagReSyn®Amine magnetic iron oxide porous microparticles by two different strategies: a) an enzyme immobilization at pH 8.5 to encourage the immobilization process by N-termini (MTtAPRT2A, MTtAPRT2B, MTtAPRT2C) or b) an enzyme immobilization at pH 10.0 to encourage the immobilization process through surface exposed lysine residues (MTtAPRT2D, MTtAPRT2E, MTtAPRT2F). According to catalyst load experiments, MTtAPRT2B (activity: 480 IU g^-1_biocatalyst, activity recovery: 52%) and MTtAPRT2F (activity: 507 IU g^-1_biocatalyst, activity recovery: 44%) were chosen as optimal derivatives. The biochemical characterization studies demonstrated that immobilization process improved the thermostability of TtAPRT2. Moreover, the potential reusability of MTtAPRT2B and MTtAPRT2F was also tested. Finally, MTtAPRT2F was employed in the synthesis of nucleoside-5'-monophosphate analogues.