pISSN 2288-6982
eISSN 2288-7105




BioDesign 2024; 12(2): 19-22

Published online June 30, 2024

© Korean Society for Structural Biology

Purification, crystallization, and X-ray crystallographic analysis of D-allulose epimerase from Leucobacter ruminantium

Min-Woo Heo1, Ji-Won Kim2, Won-Heong Lee1,* and Jeong-Sun Kim2,*

1Department of Bioenergy Science and Technology, Chonnam National University, Gwangju 61186, Republic of Korea
2Department of Chemistry, Chonnam National University, Gwangju 61186, Republic of Korea

Correspondence to:,

Received: March 8, 2024; Accepted: March 27, 2024


D-allulose epimerase (DAE) catalyzes the C-3 epimerization reaction that converts d-fructose to d-allulose. The preliminary structural study on the annotated DAE from Leucobacter ruminantium (LrDAE) was performed to understand the interaction between enzyme and substrate or product, which may provide structural background to design mutants for higher production of d-allulose from d-fructose than the wild-type and other enzymes. For this, the LrDAE encoding gene was cloned and expressed in Escherichia coli. The purified LrDAE protein was crystallized from the precipitant composed of 0.3 M Magnesium acetate, 0.1 M Sodium citrate (pH 5.6), and 13% (w/v) polyethylene glycol 8000. Diffraction data was collected to 3.0 Å resolution. The crystal belongs to the primitive orthorhombic P212121 space group with unit-cell parameters a = 69.58 Å, b = 117.65 Å, c = 150.47 Å, and α = β = γ = 90°. There are four LrDAE molecules in the asymmetric unit.