Low temperature cultivation ofEscherichia colicarrying a rice lipoxygenase L-2 cDNA produces a soluble and active enzyme at a high level
| Autor: | Daisuke Shibata, Yumiko Shirano |
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| Rok vydání: | 1990 |
| Předmět: |
Molecular Sequence Data
Lipoxygenase DNA Recombinant Biophysics Regulatory Sequences Nucleic Acid medicine.disease_cause Biochemistry Viral Proteins Structural Biology Complementary DNA Gene expression Escherichia coli Genetics medicine T7 RNA polymerase Amino Acid Sequence Cloning Molecular Promoter Regions Genetic Molecular Biology Peptide sequence Ammonium sulfate precipitation Inclusion body chemistry.chemical_classification Base Sequence biology Low temperature cultivation of E. coli Temperature Oryza DNA-Directed RNA Polymerases Cell Biology Molecular biology Enzyme Solubility chemistry biology.protein T7 RNA polymerase promoter medicine.drug |
| Zdroj: | FEBS Letters. 271:128-130 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/0014-5793(90)80388-y |
| Popis: | Using a T7 RNA polymerase promoter system, rice lipoxygenase L-2 cDNA was expressed in E. coli as a fusion protein with 18 amino acid residues at the amino terminal end of the original enzyme. Incubation at 37 degrees C for 3 h in the presence of the inducer resulted in the production of inactive lipoxygenase. However, when induction was carried out at 15 degrees C for 16 h, active lipoxygenase, amounting to 3% of the total soluble protein, was produced. The enzyme was purified by ammonium sulfate precipitation and Mono-Q column chromatography to homogeneity at a yield of 80%. Expression of this protein should permit future site-directed mutagenesis of the gene and crystallization of the enzyme. |
| Databáze: | OpenAIRE |
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