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Refolding and Structural Characteristic of TRAIL/Apo2L Inclusion Bodies from Different Specific Growth Rates of Recombinant Escherichia coli
Author(s) -
Kang Hui,
Sun AiYou,
Shen YaLing,
Wei DongZhi
Publication year - 2007
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp060238c
Subject(s) - inclusion bodies , recombinant dna , escherichia coli , chemistry , protease , apoptosis , trypsin , ligand (biochemistry) , microbiology and biotechnology , wild type , biophysics , biochemistry , biology , enzyme , receptor , gene , mutant
TNF‐related apoptosis‐inducing ligand (TRAIL/Apo2L) was produced mainly as inclusion bodies (IBs) by recombinant Escherichia coli with a temperature‐inducible expression system. The yield of TRAIL type 2 IBs at higher preinduction specific growth rate (μ = 0.15 h ‐1 ) was higher than that of TRAIL type 1 IBs at lower preinduction specific growth rate (μ = 0.05 h ‐1 ). With the same optimized refolding protocols, two types of IBs exhibited different refolding features. Refolded type 1 IBs had higher recovery of more than 80% compared with type 2 IBs (57–63%). By the measurements of fluorescence and CD spectroscopy, type 1 TRAIL IBs dissolved by urea appeared to be a closer secondary structure to the native TRAIL than type 2. Furthermore, with trypsin treatment, the striking decrease in stability of type 1 IBs against protease digestion cannot be attributed to their small size particles observed by scanning electron microscope and probably depend on different protein structure properties between the two IBs. Different properties of inclusion bodies were mainly influenced by different physiological states of the cells just prior to the induction.