Expression and characterization of a novel 2-deoxyribose-5-phosphate aldolase from Haemophilus influenzae Rd KW20

Mi Hee Woo, Min Soo Kim, Namhyun Chung, Joong Su Kim

Research output: Contribution to journalArticle

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Abstract

A codon-optimized 2-deoxyṟibose-5-phosphate aldolase (DERA) gene from Haemophilus influenzae Rd KW20 was synthesized and expressed in Escherichia coli, and the biochemical properties of its product were investigated. DERA was purified using affinity chromatography and characterized using 2-deoxyribose-5-phosphate as the substrate. Specific activity of the recombinant DERA was 34.1 Umg−1. The pH and temperature optima were 7.5 and 40°C, respectively. Additionally, the recombinant enzyme retained stability up to temperatures below 50°C. Maximal enzyme activity was observed in presence of 300 mM of acetaldehyde. The apparent Km and Vmax of purified enzyme towards 2-deoxyribose-5-phosphate were 0.14 mM and 70.42 μmol min−1 mg−1 and towards 2-deoxy-D-ribose were 24.77 mM and 1.94 μmol min−1 mg−1, respectively. For synthesis of statin intermediates, the bioconversion process for production of (3R, 5S)-6-chloro-2,4,6-trideoxy-erythro-hexose from chloroacetaldehyde and acetaldehyde using the recombinant DERA was studied and this process took 3 h for maximal conversion. This recombinant DERA could be potentially applied in the production of (3R, 5S)-6-chloro-2,4,6-trideoxy-erythro-hexose.

Original languageEnglish
Pages (from-to)655-660
Number of pages6
JournalJournal of the Korean Society for Applied Biological Chemistry
Volume57
Issue number5
DOIs
Publication statusPublished - 2014 Jan 1

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deoxyribose-phosphate aldolase
Fructose-Bisphosphate Aldolase
Hexoses
Acetaldehyde
Haemophilus influenzae
Affinity chromatography
Hydroxymethylglutaryl-CoA Reductase Inhibitors
Enzyme Stability
Bioconversion
Temperature
Ribose
Enzyme activity
Enzymes
Affinity Chromatography
Codon
Escherichia coli
Genes
Phosphates
Substrates
2-deoxyribose 5-phosphate

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Expression and characterization of a novel 2-deoxyribose-5-phosphate aldolase from Haemophilus influenzae Rd KW20. / Woo, Mi Hee; Kim, Min Soo; Chung, Namhyun; Kim, Joong Su.

In: Journal of the Korean Society for Applied Biological Chemistry, Vol. 57, No. 5, 01.01.2014, p. 655-660.

Research output: Contribution to journalArticle

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