A biocatalytic cascade reaction sensitive to the gas-liquid interface: Modeling and upscaling in a dynamic membrane aeration reactor

Abstract The oxidation of lactose to lactobionic acid by a bi-enzymatic system in a membrane-aerated reactor was scaled up 100-fold from a miniaturized membrane contactor to a 20-L dynamic membrane aeration reactor. The conversion was catalyzed by an enzyme cascade consisting of cellobiose dehydrogenase as synthesizing enzyme and laccase as regenerating enzyme coupled by a redox mediator. A model of the process, combining mass-transfer and enzyme kinetics, was developed to predict optimal conversion conditions. The dynamic membrane aeration reactor was successfully operated in discontinuous and CSTR mode to achieve maximum productivity at very low power input (27.7 W m−3) and also greatly reduced enzyme inactivation by eliminating the high gas/liquid interfacial area of conventionally aerated stirred reactors or bubble columns. The reaction product, lactobionic acid was obtained with a space–time yield of 74.4 g L−1 d−1 and a degree of conversion higher than 97%. The dynamic membrane aeration reactor is well suited for the bubbleless oxygenation of laccase-regenerated dehydrogenase reactions and other oxidase-catalyzed reactions on large scale to perform sustainable enzymatic oxidation reactions employing enzymes sensitive to shear or the gas/liquid interface.

[1]  D. Haltrich,et al.  Optimisation of cellobiose dehydrogenase production by the fungus Sclerotium (Athelia) rolfsii , 2003, Applied Microbiology and Biotechnology.

[2]  Dietmar Haltrich,et al.  Continuous enzymatic regeneration of redox mediators used in biotransformation reactions employing flavoproteins , 2001 .

[3]  Andreas Schmid,et al.  The production of fine chemicals by biotransformations. , 2002, Current opinion in biotechnology.

[4]  Dietmar Haltrich,et al.  Kinetic modeling of a bi‐enzymatic system for efficient conversion of lactose to lactobionic acid , 2009, Biotechnology and bioengineering.

[5]  H. J. Henzler,et al.  Oxygenation of cell cultures , 1993 .

[6]  D. Haltrich,et al.  Continuous Enzymatic Regeneration of Electron Acceptors Used by Flavoenzymes: Cellobiose Dehydrogenase-Catalyzed Production of Lactobionic Acid as an Example , 2004 .

[7]  U. Langer,et al.  Improving bioreactor cultivation conditions for sensitive cell lines by dynamic membrane aeration , 2009, Cytotechnology.

[8]  Udo Kragl,et al.  Technology transfer in biotechnology : from lab to industry to production , 2005 .

[9]  D. Haltrich,et al.  Characterization of the major laccase isoenzyme from Trametes pubescens and regulation of its synthesis by metal ions. , 2002, Microbiology.

[10]  D. Haltrich,et al.  Purification and Characterization of Cellobiose Dehydrogenase from the Plant Pathogen Sclerotium(Athelia) rolfsii , 2001, Applied and Environmental Microbiology.

[11]  John Villadsen,et al.  Scale‐up of enzymatic production of lactobionic acid using the rotary jet head system , 2007, Biotechnology and bioengineering.

[12]  D. Haltrich,et al.  Enhanced formation of laccase activity by the white-rot fungus Trametes pubescens in the presence of copper , 2001, Applied Microbiology and Biotechnology.

[13]  D. Haltrich,et al.  Bubble‐free oxygenation of a bi‐enzymatic system: effect on biocatalyst stability , 2009, Biotechnology and bioengineering.

[14]  U. Kragl,et al.  Synthesis of natural product precursors by Baeyer-Villiger oxidation with cyclohexanone monooxygenase from Acinetobacter , 2001 .

[15]  Z. Draelos,et al.  Cosmetic Formulation of Skin Care Products , 2005 .

[16]  U. Kragl,et al.  Synthesis of chiral ε-lactones in a two-enzyme system of cyclohexanone mono-oxygenase and formate dehydrogenase with integrated bubble-free aeration , 1997 .

[17]  Leon S. Lasdon,et al.  Design and Testing of a Generalized Reduced Gradient Code for Nonlinear Programming , 1978, TOMS.

[18]  D. Haltrich,et al.  A simple assay for measuring cellobiose dehydrogenase activity in the presence of laccase. , 1999, Journal of microbiological methods.