Cloning, expression, and characterization of a new deoxyribose 5-phosphate aldolase from Yersinia sp. EA015.
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.
Kim
Yong-Mo
YM
Enzyme Fusion Technology Research Team, Molecular Bioprocess Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology, Jeonbuk 580-185, Republic of Korea.
Chang
Young-Hyo
YH
Choi
Nack-Shick
NS
Kim
Yongook
Y
Song
Jae Jun
JJ
Kim
Joong Su
JS
D000079:Acetaldehyde / Q000378:metabolism; D000446:Aldehyde-Lyases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005986:Glyceraldehyde 3-Phosphate / Q000378:metabolism; D006639:Histidine / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D012988:Soil Microbiology; D013696:Temperature; D015007:Yersinia / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification
D011993:Recombinant Fusion Proteins; D005986:Glyceraldehyde 3-Phosphate; D006639:Histidine; D000446:Aldehyde-Lyases; C020019:deoxyribose-phosphate aldolase; D000079:Acetaldehyde