Expression and characterization of a novel deoxyribose 5-phosp hate aldolase from Paenibacillus sp. EA001

Abstract

A novel deoC gene was identified from Paenibacillus sp. EA001 isolated from soil. The gene had an open reading frame (ORF) of 663 base pairs encoding a protein of 220 amino acids with a molecular mass of 24.5 kDa. The amino acid sequence was 79% identical to that of deoxyribose 5-phosphate aldolase (DERA) from Geobacillus sp. Y412MC10. The deoC gene encoding DERA was cloned into an expression vector and the protein was expressed in Escherichia coli. The recombinant DERA was purified using Ni-NTA affinity chromatography and then characterized. The optimum temperature and pH of the enzyme were 50°C and 6.0, respectively. The specific activity for the substrate deoxyribose 5-phosphate (DR5P) was 62 μmol/min/mg. The Km value for DR5P was determined to be 145 mM with the kcat value of 3.2×102/s from Lineweaver-Burk plots. The EA001 DERA showed stability toward a high concentration of acetaldehyde (100 mM).