pISSN 2288-6982
eISSN 2288-7105




BioDesign 2021; 9(3): 51-54

Published online September 30, 2021

© Korean Society for Structural Biology

Purification, crystallization, and X-ray crystallographic analysis of Acetolactate synthase from Hydrogenobacter thermophilus

Ji-Won Kim and Jeong-Sun Kim*

Department of Chemistry, Chonnam National University, Gwangju 61186, Republic of Korea

Correspondence to: *

Received: August 26, 2021; Revised: September 13, 2021; Accepted: September 13, 2021


Acetolactate synthase (ALS) catalyzes the chemical reaction in plants and microorganisms that converts two pyruvate molecules to an acetolactate molecule and a carbon dioxide. To confer change on its substrate specificity for one carbon chemicals, structural study was initiated. For this, the recombinant ALS protein from Hydrogenobacter thermophilus(HtALS) was expressed in Escherichia coli, and homogeneously purified. The purified HtALS protein was crystallized from the precipitant composed of 0.3 M Ammonium sulfate, 0.1 M Sodium cacodylate (pH 6.5), and 18% (w/v) polyethylene glycol 4000. Diffraction data was collected to 2.4 Å resolution. The crystal belongs to the primitive orthorhombic P212121 space group with unit-cell parameters a = 75.24 Å, b = 76.62 Å, c = 194.69 Å, and α = β = γ = 90°. There are two HtALS molecules in the asymmetric unit.