Large scale production of cyclohexanone monooxygenase from Escherichia coli TOP10 pQR239.
The cyclohexanone monooxygenase (CHMO) from Acinetobacter calcoaceticus NCIMB 9871 has been cloned into Escherichia coli in an L-arabinose inducible vector. The recombinant E. coli containing the L-arabinose inducible CHMO was grown at 1.5 litres under controlled conditions to determine the parameters for growth and induction. It was found that induction with 0.1% (w/v) L-arabinose at late logarithmic phase of growth and growth for a further 2.5 to 3 h gave the optimal CHMO titre ( approximately 3500 U.l(-1,) 630 U. g dry cell weight(-1)). High dissolved oxygen concentrations were shown to be deleterious to the CHMO titre. This influenced the strategy for growth and induction, and was optimal when the oxygen uptake rate was maximized but the dissolved oxygen concentration was zero. Finally, a 300 litre scale fermentation was carried out giving a total CHMO titre of >8 x 10(5) U.
Doig
S D.
SD
Department of Biochemistry and Molecular Biology, University College London, Gower Street, WC1E 6BT, London, United Kingdom
O'Sullivan
L M.
LM
Patel
S
S
Ward
J M.
JM
Woodley
J M.
JM