Preparation of monomeric B27 Lys destripeptide insulin by intein mediated expression in Escherichia coli

Ting Chen; Lujuan Li; Helong Hao; Yuanhao Qiao

doi:10.1016/j.pep.2011.06.002

Preparation of monomeric B27 Lys destripeptide insulin by intein mediated expression in Escherichia coli

Protein Expr Purif. 2011 Nov;80(1):152-6. doi: 10.1016/j.pep.2011.06.002. Epub 2011 Jun 25.

Authors

Ting Chen¹, Lujuan Li, Helong Hao, Yuanhao Qiao

Affiliation

¹ Institute of Biology Science and Technology, DongHua University, 2999 North Ren Min Road, Shanghai 201620, China. chenting@dhu.edu.cn

PMID: 21708268
DOI: 10.1016/j.pep.2011.06.002

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Escherichia coli / genetics*
Gene Expression
Humans
Insulin / analogs & derivatives*
Insulin / chemistry
Insulin / genetics
Insulin / isolation & purification
Molecular Sequence Data
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / isolation & purification

Substances

B27 Lys destripeptide insulin
Insulin
Recombinant Fusion Proteins