Production of Biologically Active Mature Brain-derived Neurotrophic Factor in Escherichia coli

概要

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A gene coding for a brain-derived neurotrophic factor (BDNF) obtained from rat BDNF cDNA was used to construct an expression plasmid, pTRSBDNF. It contained a BDNF gene that was fused, in frame, to a region encoding the β-lactamase signal peptide. The E. coli HB101 harboring pTRSBDNF produced 18mg/liter of mature protein. The E. coli cells produced a larger amount of BDNF than that of human nerve growth factor (hNGF), which was produced using the same expression system. The E. coli cells harboring pTRSBDNF were sonicated and centrifuged to obtain a supernatant and precipitate. The BDNF purified from the supernatant containing 20% of the total BDNF had high biological activity (EC _<50> of 30pg/ml) against neurons of chick dorsal root ganglia. On the other hand, the BDNF puritied from the precipitate had low biological activity (EC_<50> of 2ng/ml) and incorrect disultide bonds.
社団法人日本農芸化学会の論文
1995-09-23