Characterization and application of endogenous phase-dependent promoters in Bacillus subtilis
- 942 Downloads
Bacillus subtilis as an important host has been widely used in synthetic biology, metabolic engineering, and production of industrial enzymes. To fully take advantage of this organism, 114 endogenous putative promoters were measured with a green fluorescent protein reporter and four classes of phase-dependent promoters (class I: exponential phase; class II: middle-log and early stationary phases; class III: lag-log and stationary phases; class IV: stationary phase) with different strengths were identified. The transcriptional strengths ranged from 0.03 to 2.03-fold of that of the commonly used strong promoter P43. On this basis, the temperature (for instance P bltD , P ydaD , and P gerBC ) and pH (such as P abrB , P ydjO , and P opuE ) inducible phase-dependent promoters were further identified and characterized. In comparison, P abrB (class I), P spoVG (class II), and P lytR (class III) achieved the highest expression levels of esterase, keratinase, and alkaline polygalacturonate lyase, respectively. The constructed phase-dependent promoter library should have great application potentials for enzyme production, metabolic engineering, and synthetic biology.
KeywordsBacillus subtilis Microbial cell factory Phase-dependent promoter Synthetic biology
This work was financially supported by the National Natural Science Foundation of China (31670092), the Natural Science Foundation of Jiangsu Province (BK20141107), a grant from the Key Technologies R&D Program of Jiangsu Province, China, (BE2014607), and Program for Changjiang Scholars and Innovative Research Team in University (No. IRT_15R26).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Bongers RS, Veening JW, Van Wieringen M, Kuipers OP, Kleerebezem M (2005) Development and characterization of a subtilin-regulated expression system in Bacillus subtilis: strict control of gene expression by addition of subtilin. Appl Environ Microbiol 71(12):8818–8824CrossRefPubMedPubMedCentralGoogle Scholar
- Daszczuk A, Dessalegne Y, Drenth I, Hendriks E, Jo E, van Lente T, Oldebesten A, Parrish J, Poljakova W, Purwanto AA, van Raaphorst R, Boonstra M, van Heel A, Herber M, van der Meulen S, Siebring J, Sorg RA, Heinemann M, Kuipers OP, Veening J-W (2014) Bacillus subtilis biosensor engineered to assess meat spoilage. ACS Synth Biol 3(12):999–1002CrossRefPubMedGoogle Scholar
- Gerosa L, Kochanowski K, Heinemann M, Sauer U (2013) Dissecting specific and global transcriptional regulation of bacterial gene expression. Mol Syst Biol 9(1):395–408Google Scholar
- Keren L, Zackay O, Lotan-Pompan M, Barenholz U, Dekel E, Sasson V, Aidelberg G, Bren A, Zeevi D, Weinberger A (2013) Promoters maintain their relative activity levels under different growth conditions. Mol Syst Biol 9(1):2367–2367Google Scholar
- Nicolas P, Mader U, Dervyn E, Rochat T, Leduc A, Pigeonneau N, Bidnenko E, Marchadier E, Hoebeke M, Aymerich S, Becher D, Bisicchia P, Botella E, Delumeau O, Doherty G, Denham EL, Fogg MJ, Fromion V, Goelzer A, Hansen A, Hartig E, Harwood CR, Homuth G, Jarmer H, Jules M, Klipp E, Le Chat L, Lecointe F, Lewis P, Liebermeister W, March A, Mars RA, Nannapaneni P, Noone D, Pohl S, Rinn B, Rugheimer F, Sappa PK, Samson F, Schaffer M, Schwikowski B, Steil L, Stulke J, Wiegert T, Devine KM, Wilkinson AJ, van Dijl JM, Hecker M, Volker U, Bessieres P, Noirot P (2012) Condition-dependent transcriptome reveals high-level regulatory architecture in Bacillus subtilis. Science 335(6072):1103–1106CrossRefPubMedGoogle Scholar
- Ploss TN, Reilman E, Monteferrante CG, Denham EL, Piersma S, Lingner A, Vehmaanpera J, Lorenz P, van Dijl JM (2016) Homogeneity and heterogeneity in amylase production by Bacillus subtilis under different growth conditions. Microb Cell Fact 15Google Scholar
- Rajkumar AS, Liu G, Bergenholm D, Arsovska D, Kristensen M, Nielsen J, Jensen MK, Keasling JD (2016) Engineering of synthetic, stress-responsive yeast promoters. Nucleic Acids ResGoogle Scholar
- Toymentseva AA, Schrecke K, Sharipova MR, Mascher T (2012) The LIKE system, a novel protein expression toolbox for Bacillus subtilis based on the liaI promoter. Microb Cell Factories 11(1):–13Google Scholar