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The Fed-Batch Production of a Thermophilic 2-Deoxyribose-5-Phosphate Aldolase (DERA) in Escherichia coli by Exponential Feeding Strategy Control

Pei, Xiaolin, Wang, Qiuyan, Qiu, Xiaofeng, Ying, Longbin, Tao, Junhua, Xie, Tian
Applied biochemistry and biotechnology 2010 v.162 no.5 pp. 1423-1434
Escherichia coli, acetates, cell growth, chemistry, fructose-bisphosphate aldolase, protein synthesis, specific growth rate
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 (µ set) was evaluated at 0.20, 0.15, and 0.10 h⁻¹, respectively. The DERA concentration and volumetric productivity were associated with µ set. The cells synthesized the enzyme most efficiently at µ 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.