PMID- 28466111 OWN - NLM STAT- MEDLINE DCOM- 20180118 LR - 20180118 IS - 1432-0614 (Electronic) IS - 0175-7598 (Linking) VI - 101 IP - 14 DP - 2017 Jul TI - Use of Brevibacillus choshinensis for the production of biologically active brain-derived neurotrophic factor (BDNF). PG - 5645-5652 LID - 10.1007/s00253-017-8273-x [doi] AB - 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 degrees 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 degrees C or >72 h at 25 degrees C), with resulting bioactivity comparable to that of a commercially available BDNF. For cultures grown in 2SY medium at 25 degrees C for 72 h, the condition that led to the greatest quantity of biologically active protein in the shortest culture time, we recovered 264 mug/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. FAU - Angart, Phillip A AU - Angart PA AD - Department of Chemical Engineering and Materials Science, Michigan State University, 428 S. Shaw Lane, Room 2100, Engineering Building, East Lansing, MI, 48824-1226, USA. FAU - Carlson, Rebecca J AU - Carlson RJ AD - Department of Chemical Engineering and Materials Science, Michigan State University, 428 S. Shaw Lane, Room 2100, Engineering Building, East Lansing, MI, 48824-1226, USA. FAU - Thorwall, Sarah AU - Thorwall S AD - Department of Chemical Engineering and Materials Science, Michigan State University, 428 S. Shaw Lane, Room 2100, Engineering Building, East Lansing, MI, 48824-1226, USA. AD - Department of Chemical & Environmental Engineering, Bourns College Of Engineering, University of California Riverside, Riverside, CA, 92521, USA. FAU - Patrick Walton, S AU - Patrick Walton S AD - Department of Chemical Engineering and Materials Science, Michigan State University, 428 S. Shaw Lane, Room 2100, Engineering Building, East Lansing, MI, 48824-1226, USA. spwalton@egr.msu.edu. LA - eng PT - Journal Article DEP - 20170502 PL - Germany TA - Appl Microbiol Biotechnol JT - Applied microbiology and biotechnology JID - 8406612 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Culture Media) RN - 0 (Recombinant Proteins) RN - 48TCX9A1VT (Cystine) SB - IM MH - Animals MH - Brain-Derived Neurotrophic Factor/*biosynthesis/genetics/isolation & purification/*pharmacology MH - Brevibacillus/genetics/*metabolism MH - Cell Proliferation/drug effects MH - Culture Media/chemistry MH - Cystine/chemistry MH - Mice MH - NIH 3T3 Cells MH - Neurons/chemistry/metabolism MH - Recombinant Proteins/chemistry/genetics/isolation & purification/metabolism MH - Temperature OTO - NOTNLM OT - BDNF OT - Bioactive OT - Brain-derived neurotrophic factor OT - Brevibacillus choshinensis OT - Cystine-knot protein OT - Neurotrophin OT - Recombinant EDAT- 2017/05/04 06:00 MHDA- 2018/01/19 06:00 CRDT- 2017/05/04 06:00 PHST- 2017/01/16 00:00 [received] PHST- 2017/03/23 00:00 [accepted] PHST- 2017/03/09 00:00 [revised] PHST- 2017/05/04 06:00 [pubmed] PHST- 2018/01/19 06:00 [medline] PHST- 2017/05/04 06:00 [entrez] AID - 10.1007/s00253-017-8273-x [pii] AID - 10.1007/s00253-017-8273-x [doi] PST - ppublish SO - Appl Microbiol Biotechnol. 2017 Jul;101(14):5645-5652. doi: 10.1007/s00253-017-8273-x. Epub 2017 May 2.