Heterologeous production of rhamnolipids in Pseudomonas putida is characterized by advantages of a non-pathogenic host and avoidance of the native quorum sensing regulation in Pseudomonas aeruginosa. Yet, downstream processing is a major problem in rhamnolipid production and increases in complexity at low rhamnolipid titers and when using chemical foam control. This leaves the necessity of a simple concentrating and purification method. Foam fractionation is an elegant method for in situ product removal when producing microbial surfactants. However, up to now in situ foam fractionation is nearly exclusively reported for the production of surfactin with Bacillus subtilis. So far no cultivation integrated foam fractionation process for rhamnolipid production has been reported. This is probably due to excessive bacterial foam enrichment in that system. In this article a simple integrated foam fractionation process is reported for heterologous rhamnolipid production in a bioreactor with easily manageable bacterial foam enrichments. Rhamnolipids were highly concentrated in the foam during the cultivation process with enrichment factors up to 200. The described process was evaluated at different pH, media compositions and temperatures. Foam fractionation processes were characterized by calculating procedural parameter including rhamnolipid and bacterial enrichment, rhamnolipid recovery, YX/S, YP/X, and specific as well as volumetric productivities. Comparing foam fractionation parameters of the rhamnolipid process with the surfactin process a high effectiveness of the integrated foam fractionation for rhamnolipid production was demonstrated.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-016-0183-2) contains supplementary material, which is available to authorized users.
Heterologous mono-rhamnolipid production by Pseudomonas putida KT2440 pSynPro8oT_rhlAB using glucose as the single carbon source was characterized in fed-batch bioreactor cultivations. For the described experiments, a defined mineral salt medium was used, and a two phase glucose feeding profile was applied, which yielded a final rhamnolipid concentration of 14.9 g/L. Applying the feeding profile, glucose stayed almost constant until 28 h of cultivation and decreased afterwards to limiting levels. Until the end of cultivation 253.0 ± 0.1 g glucose was added to the bioreactor of which a total of 252.0 ± 0.6 g glucose was metabolized. By modeling the fed-batch bioreactor cultivations the time courses of generated biomass, rhamnolipid and consumed glucose were described. The model was furthermore used to derive key process parameters from the collected data. The obtained values for the specific product formation rates (qRL) reached 18 mg/(g h) and yield coefficients (YRL/S) 10 mg/g respectively.
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