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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

Author:
Pei, Xiaolin, Wang, Qiuyan, Qiu, Xiaofeng, Ying, Longbin, Tao, Junhua, Xie, Tian
Source:
Applied biochemistry and biotechnology 2010 v.162 no.5 pp. 1423-1434
ISSN:
0273-2289
Subject:
Escherichia coli, acetates, cell growth, chemistry, fructose-bisphosphate aldolase, protein synthesis, specific growth rate
Abstract:
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.
Agid:
640855