Abstract
A novel Bacillus thuringiensis (Bt) bacteriocin BtCspB, active against a food-borne pathogen Bacillus cereus, was identified and purified by a traditional four-step chromatographic process with low yield (44.5 µg/L) in our lab previously. The aim of this study was to dramatically increase its yield by heterologous expression of BtCspB. The BtCspB gene from Bt BRC-ZYR2 was successfully heterologously expressed in Escherichia coli BL21 (DE3). Affinity chromatography was used to obtain the pure BtCspB up to 20 mg/L. The purified BtCspB showed a MIC value of 12.5 µg/mL and a MBC value of 50.0 µg/mL against Bacillus cereus ATCC 10987. The bacteriocin activity of BtCspB against B. cereus ATCC 10987 was further directly detected in a gel-overlay assay. The anti-B. cereus activity, however, was lower than the bacteriocin purified by the traditional four-step chromatographic process probably because of structural modifications. Compared with the traditional method, the yield of the bacteriocin by heterologous expression increased by 449 times, and the purification step was dramatically simplified, which laying a foundation for the industrial production of this novel cold-shock protein-like bacteriocin BtCspB active against B. cereus.
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Acknowledgements
The authors thank Gale A. Kirking at English Editorial Services, s.r.o. for language correction and additional assistance in improving the text. The authors thank Xiaoyu Su, the teachers and the students of the Biopesticide Research Center for their help.
Funding
This project was supported by the National Key R&D Program of China (No. 2017YFD0200400; 2017YFE0121700; 2017YFE0122000), National Natural Science Foundation of China (No. 31672084), Fuzhou Health and Family Planning Science and Technology Project (No. 2018-S-wt7) and by institutional support RVO: 60077344 of the Biology Centre CAS, Institute of Entomology.
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XJ, JY, HF, YC carried out the experiments; XJ wrote the draft manuscript; IG, TH, JP, LZ, XP edited the manuscript; TH, XG designed the project. All authors read and approved the final manuscript.
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Jin, X., Yao, J., Fan, H. et al. Heterologous expression and purification of BtCspB, a novel cold-shock protein-like bacteriocin from Bacillus thuringiensis BRC-ZYR2. World J Microbiol Biotechnol 35, 23 (2019). https://doi.org/10.1007/s11274-019-2595-z
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DOI: https://doi.org/10.1007/s11274-019-2595-z