Purification and characterization of a biosurfactant produced by Pseudomonas sp. G11 by asymmetrical flow field-flow fractionation (AsFlFFF)

Sung Kwang Cho; So Hee Shim; Kyeong Ryang Park; Seong-Ho Choi; Seungho Lee
December 2006
Analytical & Bioanalytical Chemistry;Dec2006, Vol. 386 Issue 7/8, p2027
Academic Journal
Three hundred and thirty two bacterial colonies were isolated from soil contaminated by an oil spill. All the bacteria were cultured in a liquid medium individually, and the surface tensions of the media were compared. The bacterium whose culture medium had the lowest surface tension was identified as Pseudomonas sp. G11. A biosurfactant was produced by cultivation of the Pseudomonas sp. G11 in the LB media. For extraction of the biosurfactant, two solvent systems were used ( n-hexane and a 2:1 (v/v) mixture of chloroform/MeOH), and the results were compared. Various experimental conditions (solvent composition, flow rate, etc.) were tested to optimize the analysis of the biosurfactant by asymmetrical flow field-flow fractionation (AsFlFFF). The biosurfactant was successfully separated from the culture medium by AsFlFFF when pure water was used as the carrier. From the retention data, the hydrodynamic diameter ( d H) and molecular weight ( M) of the biosurfactant were determined by AsFlFFF. The molecular weight was determined by using pullulans as the calibration standards. The d H and M were 49 nm and 2.3 × 105 Da when extracted with n-hexane, and 39 nm and 1.13 × 105 Da when extracted with the 2:1 mixture of chloroform/MeOH, respectively. [Figure not available: see fulltext.]


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