Monorhamnolipids and 3-(3-hydroxyalkanoyloxy)alkanoic acids (HAAs) production using Escherichia coli as a heterologous host.
Pseudomonas aeruginosa produces the biosurfactants rhamnolipids and 3-(3-hydroxyalkanoyloxy)alkanoic acids (HAAs). In this study, we report the production of one family of rhamnolipids, specifically the monorhamnolipids, and of HAAs in a recombinant Escherichia coli strain expressing P. aeruginosa rhlAB operon. We found that the availability in E. coli of dTDP-L: -rhamnose, a substrate of RhlB, restricts the production of monorhamnolipids in E. coli. We present evidence showing that HAAs and the fatty acid dimer moiety of rhamnolipids are the product of RhlA enzymatic activity. Furthermore, we found that in the recombinant E. coli, these compounds have the same chain length of the fatty acid dimer moiety as those produced by P. aeruginosa. These data suggest that it is RhlAB specificity, and not the hydroxyfatty acid relative abundance in the bacterium, that determines the profile of the fatty acid moiety of rhamnolipids and HAAs. The rhamnolipids level produced in recombinant E. coli expressing rhlAB is lower than the P. aeruginosa level and much higher than those reported by others in E. coli, showing that this metabolic engineering strategy lead to an increased rhamnolipids production in this heterologous host.
Cabrera-Valladares
Natividad
N
Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México (UNAM), Mexico, Mexico.
Richardson
Anne-Pascale
AP
Olvera
Clarita
C
Treviño
Luis Gerardo
LG
Déziel
Eric
E
Lépine
François
F
Soberón-Chávez
Gloria
G
D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002264:Carboxylic Acids / Q000378:metabolism*; D003001:Cloning, Molecular; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D017930:Genes, Reporter; D006017:Glycolipids / Q000096:biosynthesis* / Q000235:genetics; D006602:Hexosyltransferases / Q000235:genetics / Q000378:metabolism*; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism; D009876:Operon / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics; D013501:Surface-Active Agents / Q000378:metabolism*; D013942:Thymine Nucleotides / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D002264:Carboxylic Acids; D003470:Culture Media; D006017:Glycolipids; D009702:Nucleoside Diphosphate Sugars; D013501:Surface-Active Agents; D013942:Thymine Nucleotides; C418382:rhamnolipid; C073293:thymidine diphosphate rhamnose; D006602:Hexosyltransferases; C088697:rhamnosyltransferase 1; D001616:beta-Galactosidase