Abstract
Three DHAP-dependent aldolases, rhamnulose-1-phosphate aldolase (Rham-IPA), fuculose-1-phosphate aldolase (Fuc-1PA) and tagatose-1,6-diphosphate aldolase (TDPA) have been cloned and overexpressed in Escherichia coli using two different expression vectors: pTrcHis for the expression of Rham-1PA and Fuc-1PA and pRSET for the expression of TDPA. In each case the recombinant enzyme is synthesized as a fusion protein with a hexahistidine tag on the N-terminus. The three enzymes have been purified in only one step by chelation affinity chromatography. The effects of cultivation temperature and concentration of inducer have been studied in order to optimize the expression of the recombinant proteins and to avoid the formation of inclusion bodies.
| Original language | English |
|---|---|
| Pages (from-to) | 945-953 |
| Number of pages | 9 |
| Journal | Bioorganic and Medicinal Chemistry |
| Volume | 3 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - 1995 Jul |
| Externally published | Yes |
ASJC Scopus subject areas
- Biochemistry
- Molecular Medicine
- Molecular Biology
- Pharmaceutical Science
- Drug Discovery
- Clinical Biochemistry
- Organic Chemistry
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Garcia-Junceda, E., Shen, G. J., Sugai, T., & Wong, C. H. (1995). A new strategy for the cloning, overexpression and one step purification of three DHAP-dependent aldolases: Rhamnulose-1-phosphate aldolase, fuculose-1-phosphate aldolase and tagatose-1,6-diphosphate aldolase1. Bioorganic and Medicinal Chemistry, 3(7), 945-953. https://doi.org/10.1016/0968-0896(95)00077-T