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Cloning, expression, and characterization of an aldehyde dehydrogenase from Escherichia coli K-12 that utilizes 3-Hydroxypropionaldehyde as a substrate

  • Ji-Eun Jo
  • Subramanian Mohan Raj
  • Chelladurai Rathnasingh
  • Edwardraja Selvakumar
  • Woo-Chel Jung
  • Sunghoon ParkEmail author
Biotechnologically Relevant Enzymes and Proteins

Abstract

3-Hydroxypropionaldehyde (3-HPA), an intermediary compound of glycerol metabolism in bacteria, serves as a precursor to 3-Hydroxypropionic acid (3-HP), a commercially valuable platform chemical. To achieve the effective conversion of 3-HPA to 3-HP, an aldH gene encoding an aldehyde dehydrogenase in Escherichia coli K-12 (AldH) was cloned, expressed, and characterized for its properties. The recombinant AldH exhibited broad substrate specificity for various aliphatic and aromatic aldehydes. AldH preferred NAD+ over NADP+ as a cofactor for the oxidation of most aliphatic aldehydes tested. Among the aldehydes used, the specific activity was highest (38.1 U mg−1 protein) for 3-HPA at pH 8.0 and 37 °C. The catalytic efficiency (k cat) and the specificity constant (k cat/K m) for 3-HPA in the presence of NAD+ were 28.5 s−1 and 58.6 × 103 M−1 s−1, respectively. The AldH activity was enhanced in the presence of disulfide reductants such as dithiothreitol (DTT) or 2-mercaptoethanol, while several metal ions, particularly Hg2+, Ag+, Cu2+, and Zn2+, inhibited AldH activity. This study illustrates that AldH is a potentially useful enzyme in converting 3-HPA to 3-HP.

Keywords

Aldehyde dehydrogenase aldH 3-Hydroxypropionaldehyde dehydrogenase 3-Hydroxypropionic acid Enzyme characterization Escherichia coli 

Notes

Acknowledgments

This work was financially supported by the Korean Ministry of Commerce, Industry, and Energy (Grant No. 10028380-2006-11). Dr. SM Raj is grateful to the Brain Korea 21 program (Pusan National University) for financial assistance. The authors wish to express their gratitude to Prof. Watanabe (Kyoto University, Japan) for providing the pQE-80L vector.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Ji-Eun Jo
    • 1
  • Subramanian Mohan Raj
    • 1
  • Chelladurai Rathnasingh
    • 1
  • Edwardraja Selvakumar
    • 1
  • Woo-Chel Jung
    • 1
  • Sunghoon Park
    • 1
    Email author
  1. 1.Department of Chemical and Biochemical EngineeringPusan National UniversityBusanRepublic of Korea

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