INTEGRAL PROCESS OF PRODUCTION AND RECOVERY OF LIPOPEPTIDES FROM PSEUDOMONAS SYRINGAE PV. TABACI USING A FOAM FRACTIONATOR

CARLA N. HAIDAR; MATHEUS M. PEREIRA; ALVARO S. LIMA; BIBIANA B. NERLI; LUCIANA P. MALPIEDI

Campos Ville - Araraquara, Sao Paulo

Conferencia; BPP 2019 Biopartitioning and Purification Conference; 2019

Faculdade de Ciências Farmacêuticas - Câmpus de Araraquara, UNESP

The bio-surfactants (BS) are amphipathic molecules that present diverse industrial applications. They emerged as an alternative to synthetics, for presenting biodegradability and several advantages. Most BS can be easily recovered from the culture medium using a combination of conventional techniques such as precipitation and solvent extraction. However, these methodologies are not recommended because they are expensive and harmful for the environment. Currently, a technique of low environmental impact and economical is used, the foam fractionation. This allows the surfactant compounds to adsorb at the air-liquid interface, with the consequent formation of foam. In this context, this work aimed to employ a foam fractionator to purify and concentrate BS from a culture broth of Pseudomonas syringae pv. tabaci. To achieve this, a 4 L bioreactor was integrated into a foam fractionation column in order to reduce the purification steps of the desired product. Foam production was controlled by fixed aeration and constant stirring for 12 hours. After the fermentation, the BS extract (BE) obtained was evaluated and partially characterized. These tests consisted of i) determination of the emulsifying capacity (EI) by using toluene and various commercial oils ii) thin layer chromatography (TLC) using different development methods iii) determination of the critical micelle concentration (CMC) and iv) determination of the critical micellar dilution (CMD). Regarding to the production and foaming processes, 80 mg of BE could be recovered in 280 mL of collected foam. The highest proportion of BS showed to be present in the BE but not in the culture medium. The BE presented EI between 65 and 80 % with stability greater than 90 % after 24 hours of incubation. Additionally, EI % in toluene did not show significant variation when the BE was previously subjected to high temperatures. With respect to the TLC, three separate main bands were observed. Ninhydrin staining and water spraying demonstrated the chemical lipopeptide nature of the samples. The estimated CMC for BE extract was 76 mg/mL. In the CMD tests, was observed two-step decay. This was attributed to the presence of different emulsifiers in the analyzed sample, agreeing well with CMC and TLC results. Finally, it can be concluded that the integrated process described is a simple and practical method for the concentration of lipopeptides using P. syringae pv. tabaci as a producer microorganism.