5ZGO

Crystal structure of APRT2 from Thermus thermophilus HB8

Summary for 5ZGO

• Entry DOI: 10.2210/pdb5zgo/pdb
• Descriptor: Adenine phosphoribosyltransferase (2 entities in total)
• Functional Keywords: phosphoribosyltransferase, thermostable, enzyme, nucleic acid, transferase
• Biological source: Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
• Total number of polymer chains: 6
• Total formula weight: 113472.81

Authors

Kunishima, N.,Naitow, H.,Matsuura, Y. (deposition date: 2018-03-09, release date: 2019-03-06, Last modification date: 2023-11-22)

Primary citation

Arco, J.D.,Perez, E.,Naitow, H.,Matsuura, Y.,Kunishima, N.,Fernandez-Lucas, J.
Structural and functional characterization of thermostable biocatalysts for the synthesis of 6-aminopurine nucleoside-5'-monophospate analogues.
Bioresour. Technol., 276:244-252, 2019

PubMed 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, activity recovery: 52%) and MTtAPRT2F (activity: 507 IU g, 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.

PubMed: 30640018
DOI: 10.1016/j.biortech.2018.12.120

Experimental method

X-RAY DIFFRACTION (2.6 Å)