title: Preparation of monomeric B27 Lys destripeptide insulin by intein mediated expression in Escherichia coli.
abstract: 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.
authors.affiliation: Institute of Biology Science and Technology, DongHua University, 2999 North Ren Min Road, Shanghai 201620, China. chenting@dhu.edu.cn
authors.forename: Ting
authors.initials: T
authors.lastname: Chen
authors.forename: Lujuan
authors.initials: L
authors.lastname: Li
authors.forename: Helong
authors.initials: H
authors.lastname: Hao
authors.forename: Yuanhao
authors.initials: Y
authors.lastname: Qiao
mesh_terms: D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D007328:Insulin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: C477145:B27 Lys destripeptide insulin; D007328:Insulin; D011993:Recombinant Fusion Proteins