site > International journal > (2011) Production and solid-phase refolding of human glucagon-like peptide-1 using recombinant Escherichia coli. Protein Expression and Purification.

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작성일 : 12-07-17 18:36

(2011) Production and solid-phase refolding of human glucagon-like peptide-1 using recombinant Escherichia coli. Protein Expression and Purification.

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http://www.sciencedirect.com/science/article/pii/S1046592811000738 [1042]

Production and solid-phase refolding of human glucagon-like peptide-1 using recombinant Escherichia coli.

Sung-Gun Kim, So-Yeon Shin, Yong-Cheol Park, Chul-Soo Shin and Jin-Ho Seo
Protein Expression and Purification. 78(2): 197-203, 2011.08.01
(SCI I/F: 1.587)

Humanglucagon-likepeptide-1 (GLP-1), an incretin hormone with pharmaceutical potential in treating type 2 diabetes mellitus is known to be rapidly degraded when expressed in Escherichiacoli. For the efficient production of the intact GLP-1 using a recombinantE. coli system, a fusion protein of GLP-1 was designed to be composed of the 6-lysine tag, ubiquitin and GLP-1 (K6UbGLP-1) in a row. A fed-batch fermentation of recombinantE. coli BL21(DE3)/pAPK6UbGLP-1 was carried out in a pH-stat feeding strategy and resulted in 11.3 g/L of K6UbGLP-1 as a form of inclusion body. Solid-phaserefolding of K6UbGLP-1 inclusion body using a cation exchanger of the SP Sepharose FF led to a refolding yield over 90% even at 5.2 mg protein/mL resin. On-column cleavage of the refolded K6UbGLP-1 with ubiquitin-specific protease 1 gave an authentic form of GLP-1. Instrumental analyses using mass spectrometry and reverse-phase HPLC showed that the recombinant GLP-1 released from K6UbGLP-1 was identical to the standard GLP-1.

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