The fed-batch production of a thermophilic 2-deoxyribose-5-phosphate aldolase (DERA) in Escherichia coli by exponential feeding strategy control.
2-Deoxyribose-5-phosphate aldolase (DERA) catalyzes a sequential aldol reaction useful in synthetic chemistry. In this work, the effect of a feeding strategy on the production of a thermophilic DERA was investigated in fed-batch cultures of recombinant Escherichia coli BL21 (pET303-DERA008). The predetermined specific growth rate (micro(set)) was evaluated at 0.20, 0.15, and 0.10 h(-1), respectively. The DERA concentration and volumetric productivity were associated with micro(set). The cells synthesized the enzyme most efficiently at micro(set) = 0.15 h(-1). The maximum enzyme concentration (5.12 g/L) and total volumetric productivity (0.256 g L(-1) h(-1)) obtained were over 10 and five times higher than that from traditional batch cultures. Furthermore, the acetate concentration remained at a relatively low level, less than 0.4 g/L, under this condition which would not inhibit cell growth and target protein expression. Thus, a specific growth rate control strategy has been successfully applied to induce fed-batch cultures for the maximal production of the thermophilic 2-deoxyribose-5-phosphate aldolase.
Pei
Xiaolin
X
Center for Biomedicine and Health, Hangzhou Normal University, Hangzhou 310012, People's Republic of China.
Wang
Qiuyan
Q
Qiu
Xiaofeng
X
Ying
Longbin
L
Tao
Junhua
J
Xie
Tian
T
D000446:Aldehyde-Lyases / Q000096:biosynthesis*; D019149:Bioreactors / Q000382:microbiology; D001709:Biotechnology / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D005285:Fermentation / Q000502:physiology*; D013696:Temperature
D000446:Aldehyde-Lyases; C020019:deoxyribose-phosphate aldolase