Production and characterisation of a biosurfactant isolated from Pseudomonas aeruginosa UW-1

Abstract

Pseudomonas aeruginosa UW-1 produced 17-24 g L-1 rhamnolipid in vegetable oil-containing media in shake flask cultures in 13 days. In time course studies of growth and rhamnolipid production in a salts medium containing 6% canola oil, total bacterial count reached 2.6 x 10(10) CFU ml-1 after 48 h and a maximum rhamnolipid yield of 24.3 g L-1 was obtained after 9 days. Rhamnolipid components were purified and separated by chloroform-methanol extraction and TLC chromatography. The major rhamnolipid components were characterised as L-rhamnosyl-beta-hydroxydecanoyl-beta-hydroxydecanoate and L-rhamnosyl-L-rhamnosyl-beta-hydroxydecanoyl-beta-hydr oxydecanoate by nuclear magnetic resonance and mass spectrometry. The components were separated preparatively by silica gel column chromatography. The recovered monorhamnosyl fraction contained no dirhamnosyl moiety while the recovered dirhamnosyl fraction contained 5% of the monorhamnosyl moiety when analyzed by HPLC. The ratio of mono- to dirhamnosyl components produced by P. aeruginosa UW-1 was determined by HPLC to be 4:1 by weight. Purified mono- and dirhamnosyl components had the same CMC value of 40 micrograms ml-1 and decreased the surface tension of water to 27.7 and 30.4 dynes cm-1, respectively.