Abstract
Bacillus subtilis is a Gram-positive bacterium that is frequently used in the bioindustry for the production of various proteins, because of its superior protein secretion capacities. To determine optimal conditions for protein secretion by B. subtilis, a quick and sensitive method for measuring protein secretion is crucial. A fast and universal assay is most useful for detecting diverse proteins in a high-throughput manner. In this study, we introduce a split-luciferase-based method for measuring protein secretion by B. subtilis. The NanoBiT system was used to monitor secretion of four different proteins: xylanase A, amylase M, protein glutaminase A, and GFP nanobody. Our findings underscore the split-luciferase system as a quick, sensitive, and user-friendly method.
| Original language | English |
|---|---|
| Article number | 001460 |
| Pages (from-to) | 1-9 |
| Number of pages | 9 |
| Journal | Microbiology (Reading, England) |
| Volume | 170 |
| Issue number | 6 |
| Early online date | 7 Jun 2024 |
| DOIs | |
| Publication status | Published - Jun 2024 |
Funding
MBMJK and BW were supported by an NWO-TTW (17833) grant awarded to JL and LWH. The authors would like to thank Demi van Es for her practical work, Anna van den Berg van Saparoea for the strain construction, Tjeerd van Rij for providing plasmids and materials, Wouter Jong for providing sequences of GFP nanobody and HiBiT, and Thomas Wiegert for providing the pJHS12 plasmid. Figure 1 was created with BioRender.com. MBMJK and BW were supported by an NWO-TTW (17833) grant awarded to JL and LWH.
| Funders | Funder number |
|---|---|
| Anna van den Berg van Saparoea | |
| NWO-TTW | 17833 |
Keywords
- Bacillus subtilis
- heterologous protein production
- NanoBiT
- protein detection toolbox
- protein secretion
- split-luciferase assay