Integration of whole-cell reaction and product isolation: Highlyhydrophobic solvents promote in situ substrate supply and simplify extractive product isolation
Leis Pfennig Nidetzky Kratzer et al. 2016 Integration of whole-cell reaction and product isolation - Highly hydrophobic solvents promote in situ substrate supply and simplify extractive product isolation.pdf
Whole-cell bioreduction; E. coli stabilized emulsion; Emulsion separation
Abstract :
[en] Product isolation from aqueous-organic reaction mixtures that contain high concentrations of whole cells constitutes a challenging task in bioprocessing. Stirring of the biphasic reaction media leads to the formation of solvent droplets coated by cells and other surface active components and an emulsion forms. We used an early focus on phase separation to simplify a whole-cell bioreduction. Octanol, heptanol, hexanol, hexane and dipropylether were tested as co-solvents in the E. coli catalyzed reduction of ochloroacetophenone. All solvents showed very similar performance in bioreductions and highest yields were obtained with low organic-to-aqueous phase ratios. Reaction mixtures were directly investigated for organic-phase recovery. Phase separation was optimized in small-scale settling experiments and confirmed by the isolation of 20.4 g (S)-1-(2-chlorophenyl)ethanol from a 0.5 L batch reduction containing 40 gCDW/L whole-cell catalyst. Solvent consumption during product isolation could be halved by the simple addition of sodium hydroxide prior to product extraction. Basification to pH 13.5 and three extraction steps with a total of 1.2 v/v hexane led to an isolated yield of 87% (97% reduction yield). A general emulsion destabilizing effect under harsh conditions, as extreme pH values and presence of toxic reactants, was observed.
Disciplines :
Chemical engineering
Author, co-author :
Leis, Dorothea
Lauß, Bernhard
Macher-Ambrosch, Robert
Pfennig, Andreas ; Université de Liège - ULiège > Department of Chemical Engineering > PEPs - Products, Environment, and Processes
Nidetzky, Bernd
Kratzer, Regina
Language :
English
Title :
Integration of whole-cell reaction and product isolation: Highlyhydrophobic solvents promote in situ substrate supply and simplify extractive product isolation
Bräutigam, S., Dennewald, D., Schürmann, M., Lutje-Spelberg, J., Pitner, W.-R., Weuster-Botz, D., Whole-cell biocatalysis: evaluation of new hydrophobic ionic liquids for efficient asymmetric reduction of prochiral ketones. Enzyme Microb. Technol. 45 (2009), 310–316.
Chen, G., Tao, D., An experimental study of stability of oil-water emulsion. Fuel Process. Technol. 86 (2005), 499–508.
Collins, J., Grund, M., Brandenbusch, C., Sadowski, G., Schmid, A., Bühler, B., The dynamic influence of cells on the formation of stable emulsions in organic-aqueous biotransformations. J. Ind. Microbiol. Biotechnol. 42 (2015), 1011–1026.
Cuellar, M.C., van der Wielen, L.A.M., Recent advances in the microbial production and recovery of apolar molecules. Curr. Opin. Biotechnol. 33 (2015), 39–45.
Dennewald, D., Pitner, W.-R., Weuster-Botz, D., Recycling of the ionic liquid phase in process integrated biphasic whole-cell biocatalysis. Process Biochem. 46 (2011), 1132–1137.
Eixelsberger, T., Woodley, J.M., Nidetzky, B., Kratzer, R., Scale-up and intensification of (S)-1-(2-chlorophenyl)ethanol bioproduction: economic evaluation of whole cell-catalyzed reduction of o-chloroacetophenone. Biotechnol. Bioeng. 110 (2013), 2311–2315.
Furtado, G.F., Picone, C.S.F., Cuellar, M.C., Cunha, R.L., Breaking oil-in-water emulsions stabilized by yeast. Colloids Surf. B 128 (2015), 568–576.
Gröger, H., Chamouleau, F., Orologas, N., Rollmann, C., Drauz, K., Hummel, W., Weckbecker, A., May, O., Enantioselective reduction of ketons with ‘designer cells’ at high substrate concentrations: highly efficient access to functionalized optically active alcohols. Angew. Chem. Int. Ed. 45 (2006), 5677–5681.
Heeres, A.S., Schroën, K., Heijnen, J.J., van der Wielen, L.A.M., Cuellar, M.C., Fermentation broth components influence droplet coalescence and hinder advanced biofuel recovery during fermentation. Biotechnol. J. 10 (2015), 1206–1215.
Heeres, A.S., Heijnen, J.J., van der Wielen, L.A.M., Cuellar, M.C., Gas bubble induced oil recovery from emulsions stabilised by yeast components. Chem. Eng. Sci. 145 (2016), 31–44.
Henschke, M., Schlieper, L.H., Pfennig, A., Determination of a coalescence parameter from batch-settling experiments. Chem. Eng. J. 85 (2002), 369–378.
Horozov, T.S., Binks, B.P., Gottschalk-Gaudig, T., Effect of electrolyte in silicone oil-in-water emulsions stabilised by fumed silica particles. Biophys. J. 102 (2007), 2641–2648.
Kratzer, R., Pukl, M., Egger, S., Vogl, M., Brecker, L., Nidetzky, B., Enzyme identification and development of a whole-cell biotransformation for asymmetric reduction of o-chloroacetophenone. Biotechnol. Bioeng. 101 (2011), 1094–1101.
Lin, D.-Q., Zhong, L.-N., Yao, S.-J., Zeta potential as a diagnostic tool to evaluate the biomass electrostatic adhesion during ion-exchange expanded bed application. Biotechnol. Bioeng. 95 (2006), 185–191.
Lindahl, M., Faris, A., Wadström, T., Hjertén, S., A new test based on 'salting out’ to measure relative hydrophobicity of bacterial cells. Biochim. Biophys. Acta 677 (1981), 471–476.
Möller, J., Schroer, M.A., Erlkamp, M., Grobelny, S., Paulus, M., Tiemeyer, S., Wirkert, F.J., Tolan, M., Winter, R.r., The effect of ionic strength, temperature, and pressure on the interaction potential of dense protein solutions: from nonlinear pressure response to protein crystallization. Biophys. J. 102 (2012), 2641–2648.
Moreira, T.C.P., da Silva, V.M., Gombert, A.K., da Cunha, R.L., Stabilization mechanisms of oil-in-water emulsions by Saccharomyces cerevisiae. Colloids Surf. B 143 (2016), 399–405.
Park, S., Kazlauskas, R.J., Biocatalysis in ionic liquids – advantages beyond green technology. Curr. Opin. Biotech. 14 (2003), 432–437.
Pollard, D.J., Woodley, J.M., Biocatalysis for pharmaceutical intermediates: the future is now. Trends Biotechnol. 25 (2007), 66–73.
Qiao, X., Wang, L., Shao, Z., Sun, K., Miller, R., Stability and rheological behaviors of different oil/water emulsions stabilized by natural silk fibroin. Colloids Surf. A 475 (2015), 84–93.
Rayner, M., Marku, D., Eriksson, M., Sjöö, M., Dejmek, P., Wahlgren, M., Biomass-based particles for the formulation of Pickering type emulsions in food and topical applications. Colloids Surf. A 458 (2014), 48–62.
Saini, G., Nasholm, N., Dolan, M.E., Wood, B.D., Application of salting-out agent to enhance the hydrophobicity of weakly hydrophobic bacterial strains. J. Adhes. Sci. Technol. 25 (2011), 2169–2182.
Schmölzer, K., Mädje, K., Kratzer, R., Nidetzky, B., Bioprocess design guided by in situ substrate supply and product removal: process intensification for synthesis of (S)-1-(2-chlorophenyl) ethanol. Bioresour. Technol. 108 (2012), 216–223.
Schmid, A., Kollmer, B., Witholt, B., Effects of biosurfactant and emulsification on two-liquid phase Pseudomonas oleovorans cultures and cell-free emulsions containing n-decane. Enzyme Microb. Technol. 22 (1998), 487–493.
Tadros, T.F., Emulsion formation, stability, and rheology. Emulsion Formation and Stability, 2006, Wiley-VCH Weinheim, Germany, 1–67.
Weber, F.J., de Bont, J.A.M., Adaptation mechanisms of microorganisms to the toxic effects of organic solvents on membranes. Biochim. Biophys. Acta 1286 (1996), 225–245.
Wriessnegger, T., Augustin, P., Engleder, M., Leitner, E., Müller, M., Kaluzna, I., Schürmann, M., Mink, D., Zellnig, G., Schwab, H., Pichler, H., Production of the sesquiterpenoid (+)-nootkatone by metabolic engineering of Pichia pastoris. Metab. Eng. 24 (2014), 18–29.
Zhang, F., Qian, X., Si, H., Xu, G., Han, R., Ni, Y., Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering. Microb. Cell Fact. 14 (2015), 1–11.
Zita, A., Hermansson, M., Effects of bacterial cell surface structures and hydrophobicity on attachment to activated sludge flocs. Appl. Environ. Microbiol. 63 (1997), 1168–1170.
