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

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논문

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

학술지

Protein expression and purification

저자명

Chen, Ting; Li, Lujuan; Hao, Helong; Qiao, Yuanhao

초록

<P><B>Highlights</B></P><P>&#x025BA; The monomeric insulin B27 DTrI precursor (MIP) was prepared by intein mediated expression in <I>Escherichia coli</I>. &#x025BA; The B27K-DTrI insulin was obtained by tryptic digestion, further purified by HPLC and analyzed by mass spectrometry. &#x025BA; The molecular mass of purified B27K-DTrI was consistent with the theoretical value.</P> <P><B>Abstract</B></P><P>The B27K-DTrI insulin (human insulin with B28&ndash;30 removed and B27 Thr replaced by Lys) was reported to have superior monomeric property with 80% insulin activity <I>in vivo</I>. 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 <I>Escherichia coli</I>. 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.</P>

발행연도

2011

ISSN

1046-5928

ISSN

1096-0279

80

1

페이지

pp.152-156

주제어

B27 Lys DTrI insulin; B27K-DTrI precursor; Intein-mediated expression

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1 2023-12-11

논문; 2011-11-01

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