Column flow reactor using acetohydroxyacid synthase I from Escherichia coli as catalyst in continuous synthesis of R-phenylacetyl carbinol.
We tested the possibility of utilizing acetohydroxyacid synthase I (AHAS I) from Escherichia coli in a continuous flow reactor for production of R-phenylacetyl carbinol (R-PAC). We constructed a fusion of the large, catalytic subunit of AHAS I with a cellulose binding domain (CBD). This allowed purification of the enzyme and its immobilization on cellulose in a single step. After immobilization, AHAS I is fully active and can be used as a catalyst in an R-PAC production unit, operating either in batch or continuous mode. We propose a simplified mechanistic model that can predict the product output of the AHAS I-catalyzed reaction. This model should be useful for optimization and scaling up of a R-PAC production unit, as demonstrated by a column flow reactor.
Engel
Stanislav
S
Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
Vyazmensky
Maria
M
Berkovich
Dvora
D
Barak
Ze'ev
Z
Merchuk
Jose
J
Chipman
David M
DM
D000095:Acetolactate Synthase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000096:Acetone / Q000031:analogs & derivatives* / Q000378:metabolism*; D019149:Bioreactors / Q000382:microbiology*; D002384:Catalysis; D002482:Cellulose / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000096:biosynthesis; D004926:Escherichia coli / Q000201:enzymology*; D017434:Protein Structure, Tertiary
D004800:Enzymes, Immobilized; D000096:Acetone; C035293:1-hydroxy-1-phenyl-2-propanone; D002482:Cellulose; D000095:Acetolactate Synthase