Development of in situ product removal strategies in biocatalysis applying scaled-down unit operations.
An experimental platform based on scaled-down unit operations combined in a plug-and-play manner enables easy and highly flexible testing of advanced biocatalytic process options such as in situ product removal (ISPR) process strategies. In such a platform, it is possible to compartmentalize different process steps while operating it as a combined system, giving the possibility to test and characterize the performance of novel process concepts and biocatalysts with minimal influence of inhibitory products. Here the capabilities of performing process development by applying scaled-down unit operations are highlighted through a case study investigating the asymmetric synthesis of 1-methyl-3-phenylpropylamine (MPPA) using ω-transaminase, an enzyme in the sub-family of amino transferases (ATAs). An on-line HPLC system was applied to avoid manual sample handling and to semi-automatically characterize ω-transaminases in a scaled-down packed-bed reactor (PBR) module, showing MPPA as a strong inhibitor. To overcome the inhibition, a two-step liquid-liquid extraction (LLE) ISPR concept was tested using scaled-down unit operations combined in a plug-and-play manner. Through the tested ISPR concept, it was possible to continuously feed the main substrate benzylacetone (BA) and extract the main product MPPA throughout the reaction, thereby overcoming the challenges of low substrate solubility and product inhibition. The tested ISPR concept achieved a product concentration of 26.5 gMPPA· L-1, a purity up to 70% gMPPA· gtot-1and a recovery in the range of 80% mol · mol-1of MPPA in 20 h, with the possibility to increase the concentration, purity, and recovery further. Biotechnol. Bioeng. 2017;114: 600-609. © 2016 Wiley Periodicals, Inc.
Department of Chemical and Biochemical Engineering, CAPEC-PROCESS Research Center, The Technical University of Denmark (DTU), Building 229, Lyngby 2800 Kgs., Denmark.
Søren
S
Heintz
Department of Biotechnology, Lund University, Lund, Sweden.
Tim
T
Börner
Department of Chemical and Biochemical Engineering, CAPEC-PROCESS Research Center, The Technical University of Denmark (DTU), Building 229, Lyngby 2800 Kgs., Denmark.
Rolf H
RH
Ringborg
Department of Chemical and Biochemical Engineering, CAPEC-PROCESS Research Center, The Technical University of Denmark (DTU), Building 229, Lyngby 2800 Kgs., Denmark.
Gustav
G
Rehn
Department of Biotechnology, Lund University, Lund, Sweden.
Carl
C
Grey
Department of Chemical and Biochemical Engineering, CAPEC-PROCESS Research Center, The Technical University of Denmark (DTU), Building 229, Lyngby 2800 Kgs., Denmark.
Mathias
M
Nordblad
Department of Chemical and Biochemical Engineering, CAPEC-PROCESS Research Center, The Technical University of Denmark (DTU), Building 229, Lyngby 2800 Kgs., Denmark.
Ulrich
U
Krühne
Department of Chemical and Biochemical Engineering, CAPEC-PROCESS Research Center, The Technical University of Denmark (DTU), Building 229, Lyngby 2800 Kgs., Denmark.
Krist V
KV
Gernaey
Department of Biotechnology, Lund University, Lund, Sweden.
Patrick
P
Adlercreutz
Department of Chemical and Biochemical Engineering, CAPEC-PROCESS Research Center, The Technical University of Denmark (DTU), Building 229, Lyngby 2800 Kgs., Denmark.
John M
JM
Woodley
D061249:Batch Cell Culture Techniques; D055162:Biocatalysis; D001688:Biological Products / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D019149:Bioreactors; D001709:Biotechnology; D004800:Enzymes, Immobilized / Q000378:metabolism; D007218:Industrial Microbiology; D008954:Models, Biological; D011437:Propylamines / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013237:Stereoisomerism; D000637:Transaminases / Q000378:metabolism
D001688:Biological Products; D004800:Enzymes, Immobilized; D011437:Propylamines; D000637:Transaminases
amino transferase (ATAs); biocatalysis; chiral amines; in situ product removal (ISPR); process intensification; scale-down unit operations; substrate supply