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Use of Brevibacillus choshinensis for the production of biologically active brain-derived neurotrophic factor (BDNF)

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Abstract

Brain-derived neurotrophic factor (BDNF) is a member of the neurotrophin family critical for neuronal cell survival and differentiation, with therapeutic potential for the treatment of neurological disorders and spinal cord injuries. The production of recombinant, bioactive BDNF is not practical in most traditional microbial expression systems because of the inability of the host to correctly form the characteristic cystine-knot fold of BDNF. Here, we investigated Brevibacillus choshinensis as a suitable expression host for bioactive BDNF expression, evaluating the effects of medium type (2SY and TM), temperature (25 and 30 °C), and culture time (48–120 h). Maximal BDNF bioactivity (per unit mass) was observed in cultures grown in 2SY medium at extended times (96 h at 30 °C or >72 h at 25 °C), with resulting bioactivity comparable to that of a commercially available BDNF. For cultures grown in 2SY medium at 25 °C for 72 h, the condition that led to the greatest quantity of biologically active protein in the shortest culture time, we recovered 264 μg/L of BDNF. As with other microbial expression systems, BDNF aggregates did form in all culture conditions, indicating that while we were able to recover biologically active BDNF, further optimization of the expression system could yield still greater quantities of bioactive protein. This study provides confirmation that B. choshinensis is capable of producing biologically active BDNF and that further optimization of culture conditions could prove valuable in increasing BDNF yields.

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Acknowledgements

The authors thank Professor Michael Garavito for his initial suggestion to work with B. choshinensis and his insights in culturing and preparing protein from B. choshinensis, the members of the Cellular and Biomolecular Laboratory, specifically Kwasi Adu-Berchie and Ian Drobish, for their intellectual support, and Professor R. Mark Worden for use of his plate reader for AlamarBlue readouts. Financial support for this work was provided, in part, by Michigan State University, the National Science Foundation (CBET 1510895 and 1547518), and the National Institutes of Health (GM079688, GM089866, and CA176854).

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  1. Sarah Thorwall

    Present address: Department of Chemical & Environmental Engineering, Bourns College Of Engineering, University of California Riverside, Riverside, CA, 92521, USA

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  1. Department of Chemical Engineering and Materials Science, Michigan State University, 428 S. Shaw Lane, Room 2100, Engineering Building, East Lansing, MI, 48824-1226, USA

    Phillip A. Angart, Rebecca J. Carlson, Sarah Thorwall & S. Patrick Walton

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  1. Phillip A. Angart
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Correspondence to S. Patrick Walton.

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Angart, P.A., Carlson, R.J., Thorwall, S. et al. Use of Brevibacillus choshinensis for the production of biologically active brain-derived neurotrophic factor (BDNF). Appl Microbiol Biotechnol 101, 5645–5652 (2017). https://doi.org/10.1007/s00253-017-8273-x

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