During the last few decades, increasing interest in biological surfactants led to an intensification of research for the cost-efficient production of biosurfactants compared with traditional petrochemical surface-active components. The quest for alternative production strains also is associated with new demands on biosurfactant analysis. The present paper gives an overview of existing analytical methods, based on the example of rhamnolipids. The methods reviewed range from simple colorimetric testing to sophisticated chromatographic separation coupled with detection systems like mass spectrometry, by means of which detailed structural information is obtained. High-performance liquid chromatography (HPLC) coupled with mass spectrometry currently presents the most precise method for rhamnolipid identification and quantification. Suitable approaches to accelerate rhamnolipid quantification for better control of biosurfactant production are HPLC analysis directly from culture broth by adding an internal standard or Fourier transform infrared attenuated total reflectance spectroscopy measurements of culture broth as a possible quasi-online quantification method in the future. The search for alternative rhamnolipid-producing strains makes a structure analysis and constant adaptation of the existing quantification methods necessary. Therefore, simple colorimetric tests based on whole rhamnolipid content can be useful for strain and medium screening. Furthermore, rhamnolipid purification from a fermentation broth will be considered depending on the following application.

Increasing interest in biological surfactants has led to intensified research directed at more cost-efficient production of biosurfactants, relative to traditional surface-active components based on petrochemical feedstocks. This publication will focus on a new integrated process for continuous rhamnolipid (RL) production. RL was synthesized by Pseudomonas aeruginosa DSM 2874 and was continuously removed in situ by foam fractionation. To prevent loss of the biocatalyst through foaming, bacteria were entrapped in magnetic alginate beads. Immobilizates were retained from the foam by high-gradient magnetic separation and back-flushed in the bioreactor at constant intervals. It was demonstrated that continuous RL production in a 10-L bioreactor over several cycles with intermediate growth periods is feasible. Complete separation of RLs from the production medium with an average enrichment ratio of 15 in the collapsed foam was demonstrated, yielding a final RL amount of 70 g after four production cycles.

During the last decades, whole-cell immobilization has been used successfully in many bioprocesses. In particular, it is aimed at implementing continuous production processes, reaching higher production rates, and reusing the biocatalyst. In some cases, effective retention of immobilizates in the bioprocess is not feasible by membranes or sieves due to pore plugging or undesired losses of immobilizates. In the present publication, it is reported about the investigation of magnetic immobilizates of Pseudomonas aeruginosa for application in continuous biosurfactant production of rhamnolipids by foam fractionation and retention of entrained immobilizates by high-gradient magnetic separation from foam. Different materials and methods were tested with respect to important parameters, such as stability, diffusion properties or magnetic separation. Good magnetic separation of immobilizates was achieved at 5% (w/w) magnetite loading. Best results in terms of homogeneous embedding, good diffusion properties, and stability enhancement vis-à-vis pure alginate beads was achieved with alginate beads with embedded Bayoxide magnetite or MagPrep silica particles. Although polyurethane immobilizates showed higher stabilities compared with alginate beads, rhamnolipid diffusion in immobilizates was superior in magnetic alginate beads. Regarding bead production, smaller immobilizates were achieved with suspension polymerization compared to droplet extrusion by the JetCutting technology. In total, magnetic immobilizates are a promising tool for an easier handling of biocatalysts in a continuous biological production process, but they have to be adapted to the current production task.

Gegenstand der Arbeit ist die kostengünstige Produktion von Rhamnolipiden (Biotensid) durch magnetische Ganzellimmobilisate von Pseudomonas aeruginosa DSM 2874 und deren Separation mittels Schaumfraktionierung in einem integrierten Reaktorsystem. Als Kohlenstoffquelle/Substrat wird Glycerin verwendet, das als nachwachsender Rohstoff beispielsweise in größeren Mengen während der Biodieselherstellung anfällt.