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 language | English |
|---|---|
| Pages (from-to) | 655-660 |
| Number of pages | 6 |
| Journal | Journal of the Korean Society for Applied Biological Chemistry |
| Volume | 57 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2014 Jan 1 |
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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 journal › Article
}
TY - JOUR
T1 - Expression and characterization of a novel 2-deoxyribose-5-phosphate aldolase from Haemophilus influenzae Rd KW20
AU - Woo, Mi Hee
AU - Kim, Min Soo
AU - Chung, Namhyun
AU - Kim, Joong Su
PY - 2014/1/1
Y1 - 2014/1/1
N2 - 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.
AB - 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.
KW - 2-deoxyribose-5-phosphate aldolase
KW - Haemophilus influenzae
KW - statin intermediates synthesis
UR - http://www.scopus.com/inward/record.url?scp=84909646287&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84909646287&partnerID=8YFLogxK
U2 - 10.1007/s13765-014-4231-9
DO - 10.1007/s13765-014-4231-9
M3 - Article
AN - SCOPUS:84909646287
VL - 57
SP - 655
EP - 660
JO - Applied Biological Chemistry
JF - Applied Biological Chemistry
SN - 2468-0834
IS - 5
ER -