Preparation of monomeric B27 Lys destripeptide insulin by intein mediated expression in Escherichia coli
| Autor: | Ting Chen, Lujuan Li, Helong Hao, Yuanhao Qiao |
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| Rok vydání: | 2011 |
| Předmět: |
Molecular mass
Insulin medicine.medical_treatment Recombinant Fusion Proteins Molecular Sequence Data Gene Expression Biology medicine.disease_cause High-performance liquid chromatography Molecular biology chemistry.chemical_compound Monomer chemistry Biochemistry In vivo medicine Escherichia coli Humans Amino Acid Sequence Intein Digestion Biotechnology |
| Zdroj: | Protein expression and purification. 80(1) |
| ISSN: | 1096-0279 |
| Popis: | The B27K-DTrI insulin (human insulin with B28–30 removed and B27 Thr replaced by Lys) was reported to have superior monomeric property with 80% insulin activity in vivo. It has potential use as a new fast-acting analog of insulin. We cloned the monomeric insulin B27 DTrI precursor (MIP) into the pTWIN1 vector, and prepared by intein mediated expression in Escherichia coli. After tryptic digestion, the MIP was converted to B27K-DTrI insulin. The product was purified by HPLC. The mass spectrometry showed that the molecular mass of purified B27K-DTrI was consistent with the theoretical value. |
| Databáze: | OpenAIRE |
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