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.
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Acknowledgments
We are grateful to Dr. Anming Wang for helpful suggestions on the revised manuscript. The work was supported by the Major Science and Technology Project of Zhejiang Province (2007C01004-2), Technology Research and Development Program of Hangzhou (20090331 N03), and National Natural Science Foundation of China (20906016).
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Pei, X., Wang, Q., Qiu, X. et al. The Fed-Batch Production of a Thermophilic 2-Deoxyribose-5-Phosphate Aldolase (DERA) in Escherichia coli by Exponential Feeding Strategy Control. Appl Biochem Biotechnol 162, 1423–1434 (2010). https://doi.org/10.1007/s12010-010-8924-1
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DOI: https://doi.org/10.1007/s12010-010-8924-1