Cloning, Expression and Purification of Arylsulfate Sulfotransferase from Eubacterium A-44
| Autor: | Yang Jin Hyun, Keun Sook Lee, Dong-Hyun Kim, Bo Mi Kim, Kyoichi Kobashi |
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| Rok vydání: | 2007 |
| Předmět: |
Sequence analysis
Molecular Sequence Data Parabens Tyramine Pharmaceutical Science Molecular cloning medicine.disease_cause Polymerase Chain Reaction Chromatography Affinity Homology (biology) Substrate Specificity Industrial Microbiology Bacterial Proteins Sequence Analysis Protein Escherichia coli medicine Eubacterium Amino Acid Sequence Cloning Molecular Peptide sequence Gene Library Pharmacology Expression vector Base Sequence biology Sequence Analysis RNA General Medicine biology.organism_classification Arylsulfotransferase Molecular biology Recombinant Proteins Open reading frame Biochemistry Tyrosine bacteria Transformation Bacterial Sequence Alignment |
| Zdroj: | Biological and Pharmaceutical Bulletin. 30:11-14 |
| ISSN: | 1347-5215 0918-6158 |
| Popis: | A gene (astA) encoding arylsulfate sulfotransferase (ASST), which transfers a sulfate group from phenolic sulfate esters to phenolic acceptors, was cloned from a Eubacterium A-44 genomic library. The probe (1.5 kb fragment) for the astA gene was prepared from the PCR product of the primers produced using two internal amino acid sequences of ASST, which had been purified from Eubacterium A-44. The astA gene was cloned into the pKF3 vector. Its sequence revealed a 1863 bp open reading frame (ORF) encoding a protein containing 620 amino acids with a secretary signal peptide, and showed 91% homology (identity) to Eubacterium rectale IIIH previously reported. The cloned astA gene was expressed under the T7 promoter of the expression vectors, pET-39b(+) and pET-26b(+), in Escherichia coli BL21 (DE3), and the expressed ASSTs were purified using His Bind column chromatography. The specific activities of the purified ASSTs were 25.6 micromol/min/mg and 37.1 micromol/min/mg, respectively. |
| Databáze: | OpenAIRE |
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