Pseudomonas aeruginosa ATCC 9027 is a non-virulent strain suitable for mono-rhamnolipids production
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Rhamnolipids produced by Pseudomonas aeruginosa are biosurfactants with a high biotechnological potential, but their extensive commercialization is limited by the potential virulence of P. aeruginosa and by restrictions in producing these surfactants in heterologous hosts. In this work, we report the characterization of P. aeruginosa strain ATCC 9027 in terms of its genome-sequence, virulence, antibiotic resistance, and its ability to produce mono-rhamnolipids when carrying plasmids with different cloned genes from the type strain PAO1. The genes that were expressed from the plasmids are those coding for enzymes involved in the synthesis of this biosurfactant (rhlA and rhlB), as well as the gene that codes for the RhlR transcriptional regulator. We confirm that strain ATCC 9027 forms part of the PA7 clade, but contrary to strain PA7, it is sensitive to antibiotics and is completely avirulent in a mouse model. We also report that strain ATCC 9027 mono-rhamnolipid synthesis is limited by the expression of the rhlAB-R operon. Thus, this strain carrying the rhlAB-R operon produces similar rhamnolipids levels as PAO1 strain. We determined that strain ATCC 9027 with rhlAB-R operon was not virulent to mice. These results show that strain ATCC 9027, expressing PAO1 rhlAB-R operon, has a high biotechnological potential for industrial mono-rhamnolipid production.
KeywordsMetabolic engineering Biosurfactant production Pseudomonas aeruginosa virulence Quorum-sensing response
We acknowledge the support in providing the radioactive material and the use of laboratory facilities of Guadalupe Espín of the Instituto de Biotecnología, Universidad Nacional Autónoma de México.
Compliance with ethical standards
This work was supported in part by grant PAPIIT IN200416 (DGAPA-UNAM) and from a grant from Fundación Miguel Alemán for the project “Análisis genómico de cepas de Pseudomonas aeruginosa que presentan una respuesta de detección de quórum atípica.”
Conflict of interest
The authors declare that they have no conflict of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. In particular, all mouse studies were conducted in accordance with the Guide for the Care and Use of Laboratory Animals (Committee for the Update of the Guide for the Care and Use of Laboratory Animals and Institute for Laboratory Animal Research, Washington, DC, 2011) and the Comité para el Cuidado y Uso de Animales de Laboratorio (CCUAL) and were approved by the ethics committee of Instituto de Investigaciones Biomédicas—UNAM (approval No. ID201 09/02-2010).
- Abdel-Maugoud AM, Hausmann R, Lépine F, Muller MM, Déziel E (2011) Rhamnolipids: detection, analysis, biosynthesis, genetic regulation and bioengineering of production. In: Soberón-Chávez G (ed) Biosurfactants: from genes to applications, Microbiol Monographs, vol 20. Springer, Berlin Heilderberg, pp. 13–55CrossRefGoogle Scholar
- Aguirre-Ramírez M, Medina G, González-Valdez A, Grosso-Becerra V, Soberón-Chávez G (2012) Pseudomonas aeruginosa rmlBDAC operon, encoding dTDP-L-rhamnose biosynthetic enzymes, is regulated by the quorum-sensing transcriptional regulator RhlR and the alternative sigma S factor. Microbiol-UK 158:908–916CrossRefGoogle Scholar
- Aziz RK, Bartels D, Best AA, DeJongh M, Disz T, Edwards RA, Formsma K, Gerdes S, Glass EM, Kubal M, Meyer F, Olsen GJ, Olson R, Osterman AL, Overbeek RA, McNeil LK, Paarmann D, Paczian T, Parrello B, Pusch GD, Reich C, Stevens R, Vassieva O, Vonstein V, Wilke A, Zagnitko O (2008) The RAST server: rapid annotations using subsystems technology. BMC-Genomics 9:75CrossRefPubMedPubMedCentralGoogle Scholar
- Boukerb AM, Decor A, Ribun S, Tabaroni R, Rousset A, Commin L, Buff S, Doléans-Jordheim A, Vidal S, Varrot A, Inverty A, Cournoyer B (2016) Genomic rearrangements and functional diversification of lecA and lecB lectin-coding regions impacting the efficacy of glycomimetics directed against Pseudomonas aeruginosa. Front Microbiol 7:811. doi: 10.3389/fmicb.2016.00811 CrossRefPubMedPubMedCentralGoogle Scholar
- Chandrasekaran EV, Bemiller JN (1980) Constituent analyses of glycosaminoglycans. Methods Carbohydr Chem 8:89–96Google Scholar
- Grosso-Becerra MV, Santos-Medellín C, González-Valdez A, Méndez JL, Delgado G, Morales-Espinosa R, Servín-González L, Alcaraz LD, Soberón-Chávez G (2014a) Pseudomonas aeruginosa clinical and environmental isolates constitute a single population with high phenotypic diversity. BMC-Genomics 15:318CrossRefPubMedPubMedCentralGoogle Scholar
- Laemmli UK (1990) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277(5259):680–685Google Scholar
- Lee DG, Urbach JM, Wu G, Liberati NT, Feinbaum RL, Miyata S, Diggins LT, He J, Déziel E, Friedman L, Li L, Grills G, Montgomery K, Kucherlapati R, Rahme LG, Ausubel FM (2006) Genomic analysis reveals that Pseudomonas aeruginosa virulence is combinatorial. Genome Biol 7:R90CrossRefPubMedPubMedCentralGoogle Scholar
- Miller J (1972) In: Experiments in molecular genetics. Published by Cold Spring Harbor Laboratory (Cold Spring Harbor Laboratory, NY), pp 352–355Google Scholar
- Morales-Espinosa R, Soberón-Chávez G, Delgado-Sapién G, Sandner-Miranda L, Mendez J, Gonzalez-Valencia G, Cravioto A (2012) Genetic and phenotypic characterization of a Pseudomonas aeruginosa population with high frequency of genomic islands. PLoS One 7(5):e37459CrossRefPubMedPubMedCentralGoogle Scholar
- Rahim R, Ochsner U, Olvera C, Graninger M, Messner P, Lam JS, Soberón-Chávez G (2001) Cloning and functional characterization of the Pseudomonas aeruginosa rhlC gene that encodes rhamnosyltransferase 2, an enzyme responsible for di-rhamnolipid biosynthesis. Mol Microbiol 40:708–718CrossRefPubMedGoogle Scholar
- Rahman PK, Randhawa KK (2015) Editorial: Microbiotechnology based surfactants and their applications. Frontiers Microbiol 1 6:1344Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Lab Press, Cold Spring Harbor, NYGoogle Scholar
- Stover CK, Pham XQ, Erwin AL, Mizoguchi SD, Warrener P, Hickey MJ, Brinkman FS, Hufnagle WO, Kowalik DJ, Lagrou M, Garber RL, Goltry L, Tolentino E, Westbrock-Wadman S, Yuan Y, Brody LL, Coulter SN, Folger KR, Kas A, Larbig K, Lim R, Smith K, Spencer D, Wong GK, Wu Z, Paulsen IT, Reizer J, Saier MH, Hancock RE, Lory S, Olson MV (2000) Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 406:959–964CrossRefPubMedGoogle Scholar
- Wittgens A, Tiso T, Arndt TT, Wenk P, Hemmerick J, Müller C, Wichmann R, Küpper B, Zwick M, Wilhelm S, Hausmann R, Syldatk C, Rosenau F, Blank LM (2011) Growth independent rhamnolipid production from glucose using the non-pathogenic Pseudomonas putida KT2440. Microb Cell Factories 10:80CrossRefGoogle Scholar