Protein Expression and Purification 2009-12-01

Cloning, expression, and characterization of a new deoxyribose 5-phosphate aldolase from Yersinia sp. EA015.

Yong-Mo Kim, Young-Hyo Chang, Nack-Shick Choi, Yongook Kim, Jae Jun Song, Joong Su Kim, Yong-Mo Kim, Young-Hyo Chang, Nack-Shick Choi, YongOok Kim, Jae Jun Song, Joong Su Kim

Index: Protein Expr. Purif. 68(2) , 196-200, (2009)

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Abstract

A new deoC gene encoding deoxyribose 5-phosphate aldolase (DERA) was identified in Yersinia sp. EA015 isolated from soil. The DERA gene had an open reading frame (ORF) of 672 base pairs encoding 223 amino acids to yield a protein of molecular mass 24.8 kDa. The amino acid sequence was 94% identical to that of DERA from Yersinia intermedia ATCC 29909. DERA was over-expressed in Escherichia coli and purified using Ni-NTA affinity chromatography. The specific activity was 137 micromol/min/mg. The Michaelis constant (k(m) value) of DERA was 9.1 mM. DERA was optimally active at pH 6.0 and 50 degrees C. DERA was tolerant to a high concentration (300 mM) of acetaldehyde.