To enhance rhamnolipids production in Pseudomonas aeruginosa, an optimization strategy based on selectively blocking the metabolic bypass that competed precursors with rhamnolipids biosynthesis pathway, containing exopolysaccharide (Psl and Pel) and polyhydroxyalkanoates (PHA) synthesis pathways.
Blocking the synthesis of Psl and PHA by genes knockout, both mutants P. aeruginosa SG ∆pslAB and P. aeruginosa SG ∆phaC1DC2 can grow normally in fermentation medium and increase the production of rhamnolipids by 21% and 25.3%, respectively. While blocking the synthesis of Pel, the cell growth of the mutant strain P. aeruginosa SG ∆pelA was inhibited, thus its production yield of rhamnolipids was also decreased by 39.8%. In addition, simultaneously blocking the synthesis of Psl and PHA, a double mutant strain P. aeruginosa SG ∆pslAB ∆phaC1DC2 was constructed. Rhamnolipids production was significantly increased in strain SG ∆pslAB ∆phaC1DC2 by 69.7%.
Through selectively blocking metabolic bypasses, increasing the amount of glycosyl and fatty acid precursors can significantly enhance rhamnolipids production in P. aeruginosa.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Abdel-Mawgoud AM, Hausmann R, Lépine F, Müller MM, Déziel E (2011) Rhamnolipids: detection, analysis, biosynthesis, genetic regulation, and bioengineering of production. In: Soberón-Chávez G (ed) Biosurfactants: from genes to applications. Springer, Berlin, Heidelberg, pp 13–55
Byrd MS et al (2009) Genetic and biochemical analyses of the Psl exopolysaccharide reveal overlapping roles for polysaccharide synthesis enzymes in Psl and LPS production. Mol Microbiol 73:622–638
Choi MH, Xu J, Gutierrez M, Yoo T, Cho Y-H, Yoon SC (2011) Metabolic relationship between polyhydroxyalkanoic acid and rhamnolipid synthesis in Pseudomonas aeruginosa: comparative 13C NMR analysis of the products in wild-type and mutants. J Biotechnol 151:30–42
Colvin KM, Gordon VD, Murakami K, Borlee BR, Wozniak DJ, Wong GCL, Parsek MR (2011) The Pel polysaccharide can serve a structural and protective role in the biofilm matrix of Pseudomonas aeruginosa. PLoS Pathog 7(1):e1001264
Friedman L, Kolter R (2004) Genes involved in matrix formation in Pseudomonas aeruginosa PA14 biofilms. Mol Microbiol 51:675–690
Gutiérrez-Gómez U, Soto-Aceves MP, Servín-González L, Soberón-Chávez G (2018) Overproduction of rhamnolipids in Pseudomonas aeruginosa PA14 by redirection of the carbon flux from polyhydroxyalkanoate synthesis and overexpression of the rhlAB-R operon. Biotechnol Lett 40:1561–1566
He C, Dong W, Li J, Li Y, Huang C, Ma Y (2017) Characterization of rhamnolipid biosurfactants produced by recombinant Pseudomonas aeruginosa strain DAB with removal of crude oil. Biotechnol Lett 39:1381–1388
Ma KY, Sun MY, Dong W, He CQ, Chen FL, Ma YL (2016) Effects of nutrition optimization strategy on rhamnolipid production in a Pseudomonas aeruginosa strain DN1 for bioremediation of crude oil. Biocatal Agric Biotechnol 6:144–151
Madison LL, Huisman GW (1999) Metabolic engineering of poly(3-hydroxyalkanoates): from DNA to plastic. Microbiol Mol Biol Rev 63:21–53
Müller MM, Kügler JH, Henkel M, Gerlitzki M, Hörmann B, Pöhnlein M, Syldatk C, Hausmann R (2012) Rhamnolipids-next generation surfactants? J Biotechnol 162:366–380
Neto DC, Meira JA, de Araújo JM, Mitchell DA, Krieger N (2008) Optimization of the production of rhamnolipids by Pseudomonas aeruginosa UFPEDA 614 in solid-state culture. Appl Microbiol Biotechnol 81(3):441
Nordin N, Zakaria MR, Effendi Halmi M, Ariff A, Mohd Zawawi R, Wasoh H (2013) Isolation and screening of high efficiency bio-surfactant-producing bacteria Pseudomonas sp. J Biochem Microbiol Biotechnol 1:25–31
Qi Q, Rehm BHA, Steinbüchel A (1997) Synthesis of poly(3-hydroxyalkanoates) in Escherichia coli expressing the PHA synthase gene phaC2 from Pseudomonas aeruginosa: comparison of PhaC1 and PhaC2. FEMS Microbiol Lett 157:155–162
Schäfer A, Tauch A, Jäger W, Kalinowski J, Thierbach G, Pühler A (1994) Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 145:69–73
Shah MUH, Sivapragasam M, Moniruzzaman M, Yusup SB (2016) A comparison of recovery methods of rhamnolipids produced by Pseudomonas aeruginosa. Procedia Eng 148:494–500
Soberón-Chávez G, Lépine F, Déziel E (2005a) Production of rhamnolipids by Pseudomonas aeruginosa. Appl Microbiol Biotechnol 68:718–725
Soberón-Chávez G, Aguirre-Ramírez M, Sánchez R (2005b) The Pseudomonas aeruginosa RhlA enzyme is involved in rhamnolipid and polyhydroxyalkanoate production. J Ind Microbiol Biotechnol 32:675
Zhao F, Cui Q, Han S, Dong H, Zhang J, Ma F, Zhang Y (2015a) Enhanced rhamnolipid production of Pseudomonas aeruginosa SG by increasing copy number of rhlAB genes with modified promoter. RSC Adv 5:70546–70552
Zhao F, Zhang J, Shi R, Han S, Ma F, Zhang Y (2015b) Production of biosurfactant by a Pseudomonas aeruginosa isolate and its applicability to in situ microbial enhanced oil recovery under anoxic conditions. RSC Adv 5:36044–36050
Zhao F, Liang X, Ban Y, Han S, Zhang J, Zhang Y, Ma F (2016) Comparison of methods to quantify rhamnolipids and optimization of oil spreading method. Tenside Surfact Det 53(3):243–248
This work was financially supported by the National Natural Science Foundation of China (31700117), the China Postdoctoral Science Foundation (2017M621292) and the Science and Technology Service Network Initiative (KFJ-STS-ZDTP-064).
Supplementary Table S1—Shows all bacterial strains and plasmids in this study
Supplementary Table S2—Shows the primer pairs used to construct mutants SG ∆pslAB, SG ∆pelA and SG ∆phaC1DC2
Conflict of interest
The authors declare that they have no conflict of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Lei, L., Zhao, F., Han, S. et al. Enhanced rhamnolipids production in Pseudomonas aeruginosa SG by selectively blocking metabolic bypasses of glycosyl and fatty acid precursors. Biotechnol Lett (2020). https://doi.org/10.1007/s10529-020-02838-9
- Metabolic bypass
- Pseudomonas aeruginosa
- Gene knockout