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Determination and optimization of a strong promoter element from Bacillus amyloliquefaciens by using a promoter probe vector

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Abstract

Objective

To construct a promoter probe vector, pBE-bgaB, to screen strong promoters from Bacillus amyloliquefaciens.

Results

266 colonies containing active promoter elements from the genomic DNA of B. amyloliquefaciens were identified. Among these, promoter P41 exhibited the strongest β-Gal activity in Escherichia coli and B. amyloliquefaciens. Sequence analysis showed that promoter P41 contained P ykuN , a ykuN gene encoding flavodoxin. Optimization of the ribosome-binding site from P41 to P382 improved β-Gal activity by ~ 200%.

Conclusion

A new strong promoter for protein expression and genetic engineering of Bacillus species.

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Acknowledgements

This work was supported by the Science and Technology Planning Project of Guangdong Province (Grant Nos. 2016A050503016, 2016A010105004 and 2013B010404007), the Science and Technology Planning Project of Guangzhou City (Grant No. 201510010191) and the National High-tech R&D Program (863 Program) (Grant No. 2014AA021304). The authors thank Professor Ying Lin and Professor Shuangyan Han in South China University of Technology for kindly providing plasmid contained xyn gene.

Supplementary information

Supplementary Table 1—Bacterial strains and plasmids used in this study.

Supplementary Table 2—Primers used in this study.

Supplementary Fig. 1—Construction of the promoter probe plasmid pBE-bgaB.

Supplementary Fig. 2—Construction a series of expression plasmids.

Author information

Authors and Affiliations

  1. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, Guangdong, People’s Republic of China

    Yuling Liao, Bin Wang, Yanrui Ye & Li Pan

  2. Star Lake Bioscience Co., Inc, Zhaoqing, 526060, Guangdong, People’s Republic of China

    Yuling Liao

  3. Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, Guangzhou, 510006, People’s Republic of China

    Yanrui Ye

Authors
  1. Yuling Liao
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  2. Bin Wang
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  3. Yanrui Ye
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  4. Li Pan
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Corresponding author

Correspondence to Yanrui Ye.

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Liao, Y., Wang, B., Ye, Y. et al. Determination and optimization of a strong promoter element from Bacillus amyloliquefaciens by using a promoter probe vector. Biotechnol Lett 40, 119–126 (2018). https://doi.org/10.1007/s10529-017-2449-4

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  • DOI: https://doi.org/10.1007/s10529-017-2449-4

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