Protein preparation, crystallization and preliminary X-ray analysis of Polygonum cuspidatum bifunctional chalcone synthase/benzalacetone synthase

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2013-08, Vol.69 (8), p.871-875
• Main Authors: Lu, Heshu, Yang, Mingfeng, Liu, Chunmei, Lu, Ping, Cang, Huaixing, Ma, Lanqing
• Format: Article
• Language: English
• Subjects: Acyltransferases - chemistry; Acyltransferases - genetics; Amino Acid Sequence; Aquatic plants; Barium; Barium chloride; Butanones - chemistry; chalcone synthase/benzalacetone synthase; Crystallization; Crystallization Communications; Crystals; Diffraction; E coli; Escherichia coli; Fallopia japonica - enzymology; Fallopia japonica - genetics; Homogeneity; Metabolites; Molecular Sequence Data; Plant Proteins - chemistry; Plant Proteins - genetics; Polygonum cuspidatum; Proteins; Secondary metabolites; X-Ray Diffraction; X-rays
• ISSN: 1744-3091; 1744-3091; 2053-230X
• DOI: 10.1107/S1744309113017004

The chalcone synthase (CHS) superfamily of type III polyketide synthases (PKSs) generate the backbones of a variety of plant secondary metabolites. An active bifunctional chalcone synthase/benzalacetone synthase (CHS/BAS) from Polygonum cuspidatum was overexpressed in Escherichia coli as a C‐terminally polyhistidine‐tagged fusion protein, purified to homogeneity and crystallized using polyethylene glycol 4000 as a precipitant. The production of well shaped crystals of the complex between PcPKS1 and benzalacetone was dependent on the presence of sorbitol and barium chloride as additives. The crystals belonged to the orthorhombic space group P212121, with unit‐cell parameters a = 80.23, b = 81.01, c = 122.89 Å, and diffracted X‐rays to at least 2.0 Å resolution.