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Construction and application of a dual promoter system for efficient protein production and metabolic pathway enhancement in Bacillus licheniformis.
J Biotechnol. 2020 Mar 20; 312:1-10.JB

Abstract

Promoter plays the critical role in regulating gene transcription, and dual-promoter has received the widespread attentions due to its high efficiency and continuity, here, we want to construct an efficient dual-promoter for protein production and metabolic pathway enhancement. Firstly, our results indicated that P43 promoter efficiently transcribed at logarithmic period, while the σB-type promoters (PylB, PgsiB, PykzA) were active at stationary phase. Then, several dual promoters were constructed by coupling these σB-type promoters with P43, and the attained dual-promoter PykzA-P43 showed the best performance, which led to 1.72-, 3.46- and 1.85-fold increases of green fluorescence intensity, red fluorescence intensity and α-amylase activity, compared with those of the recognized strong promoter P43, respectively. Furthermore, α-amylase activity was further increased to 389.65 U/mL by 32.20 % via optimizing sigma factor binding sites (-10 and -35 boxes) of PykzA-P43, attaining the optimized dual promoter Pdual3. Finally, Pdual3 was applied in metabolic pathway enhancement, and the yields of Poly γ-glutamic acid, acetoin and 2, 3-butanediol were respectively improved by 82.01 %, 17.09 % and 99.39 %. Our results indicated that dual-promoter significantly enhanced gene expression, and this study provided an energetic dual-promoter Pdual3 for efficient protein production and metabolic pathway enhancement in Bacillus licheniformis.

Authors+Show Affiliations

State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, China.State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, China.State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, China.State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, China.State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, China.State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, China; Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao, China.State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, China.Hubei Provincial Key Laboratory of Industrial Microbiology, Key Laboratory of Fermentation Engineering (Ministry of Education), School of Food and Biological Engineering, Hubei University of Technology, Wuhan, 430068, Hubei, China.State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, China. Electronic address: mel212@126.com.State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, Wuhan, 430062, China. Electronic address: maxin@hubu.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32119878

Citation

Rao, Yi, et al. "Construction and Application of a Dual Promoter System for Efficient Protein Production and Metabolic Pathway Enhancement in Bacillus Licheniformis." Journal of Biotechnology, vol. 312, 2020, pp. 1-10.
Rao Y, Cai D, Wang H, et al. Construction and application of a dual promoter system for efficient protein production and metabolic pathway enhancement in Bacillus licheniformis. J Biotechnol. 2020;312:1-10.
Rao, Y., Cai, D., Wang, H., Xu, Y., Xiong, S., Gao, L., Xiong, M., Wang, Z., Chen, S., & Ma, X. (2020). Construction and application of a dual promoter system for efficient protein production and metabolic pathway enhancement in Bacillus licheniformis. Journal of Biotechnology, 312, 1-10. https://doi.org/10.1016/j.jbiotec.2020.02.015
Rao Y, et al. Construction and Application of a Dual Promoter System for Efficient Protein Production and Metabolic Pathway Enhancement in Bacillus Licheniformis. J Biotechnol. 2020 Mar 20;312:1-10. PubMed PMID: 32119878.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Construction and application of a dual promoter system for efficient protein production and metabolic pathway enhancement in Bacillus licheniformis. AU - Rao,Yi, AU - Cai,Dongbo, AU - Wang,Hao, AU - Xu,Yuxiang, AU - Xiong,Shijie, AU - Gao,Lin, AU - Xiong,Min, AU - Wang,Zhi, AU - Chen,Shouwen, AU - Ma,Xin, Y1 - 2020/02/28/ PY - 2019/12/14/received PY - 2020/02/20/revised PY - 2020/02/28/accepted PY - 2020/3/3/pubmed PY - 2020/3/3/medline PY - 2020/3/3/entrez KW - Bacillus licheniformis KW - Dual-Promoter KW - Metabolic pathway enhancement KW - Protein expression SP - 1 EP - 10 JF - Journal of biotechnology JO - J. Biotechnol. VL - 312 N2 - Promoter plays the critical role in regulating gene transcription, and dual-promoter has received the widespread attentions due to its high efficiency and continuity, here, we want to construct an efficient dual-promoter for protein production and metabolic pathway enhancement. Firstly, our results indicated that P43 promoter efficiently transcribed at logarithmic period, while the σB-type promoters (PylB, PgsiB, PykzA) were active at stationary phase. Then, several dual promoters were constructed by coupling these σB-type promoters with P43, and the attained dual-promoter PykzA-P43 showed the best performance, which led to 1.72-, 3.46- and 1.85-fold increases of green fluorescence intensity, red fluorescence intensity and α-amylase activity, compared with those of the recognized strong promoter P43, respectively. Furthermore, α-amylase activity was further increased to 389.65 U/mL by 32.20 % via optimizing sigma factor binding sites (-10 and -35 boxes) of PykzA-P43, attaining the optimized dual promoter Pdual3. Finally, Pdual3 was applied in metabolic pathway enhancement, and the yields of Poly γ-glutamic acid, acetoin and 2, 3-butanediol were respectively improved by 82.01 %, 17.09 % and 99.39 %. Our results indicated that dual-promoter significantly enhanced gene expression, and this study provided an energetic dual-promoter Pdual3 for efficient protein production and metabolic pathway enhancement in Bacillus licheniformis. SN - 1873-4863 UR - https://www.unboundmedicine.com/medline/citation/32119878/Construction_and_application_of_a_dual_promoter_system_for_efficient_protein_production_and_metabolic_pathway_enhancement_in_Bacillus_licheniformis L2 - https://linkinghub.elsevier.com/retrieve/pii/S0168-1656(20)30048-1 DB - PRIME DP - Unbound Medicine ER -
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