(228ek) Bioactive BDNF Expression Using Brevibacillus Choshinensis

Neurotrophins are essential growth factors in the growth, differentiation and survival of neuronal cells, playing a wide variety of roles in the central and peripheral nervous systems. Recombinant neurotrophins are used for research and therapeutic purposes, however, the cost of neurotrophins, in particular brain-derived neurotrophic factor (BDNF), can be limiting to both researchers and clinicians. The cost of BDNF is related to difficulties in the production of protein with the correct cystine-knot configuration. Here we investigated the recombinant expression of BDNF in the Brevibacillus choshinensis, exploring the effects of nutrients, temperature, and culture time on expression of bioactive BDNF. Of the variables examined, we found that lower temperature (25°C vs. 30°C) had the greatest impact on the expression bioactive BDNF. This proof-of-concept study demonstrates that we can produce ~200 μg/L of bioactive BDNF. Further, BDNF was purified in a single step. This presentation will discuss the use of B. choshinensis to produce BDNF as well as methods to further optimize expression.