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

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 (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.

Influence of culture medium on the production of eif antigen from Leishmania chagasi in recombinant Escherichia coli

“…The gene encoding the eIF antigen was expressed as a fusion protein containing histidine tag at the N-terminal end of the peptide using pQE-40 cloned in E. coli M15 cells (Qiagen, U.S.A). The strain was maintained on Luria-Bertani (LB) medium in the presence of ampicillin and kanamycin, as described elsewhere (16).…”

“…Sen. Salgado Filho, 3000 -Campus Universitário, 59072-970, Natal, RN, Brazil. ; Fax: +55 84 3215-3756.; E-mail: rossana@eq.ufrn.br Production of eIF antigen from L. chagasi expression in cytoplasm often leads to inadequate protein structure and agglutination in insoluble inclusion bodies (16).…”

Efficient Production of a Thermophilic 2-Deoxyribose-5-Phosphate Aldolase in Glucose-Limited Fed-Batch Cultivations of Escherichia coli by Continuous Lactose Induction Strategy

“…A semi-synthetic medium (SM-B) with glucose as the sole carbon source was employed in all fermenter cultures [15]. For the fed-batch growth phase, the feeding solution contained (per liter): 600 g glucose, 15 …”

“…When the optical density (OD 600 ) reached 0.8, induction was done by addition of 0.5 mM IPTG or the different concentration of lactose under the same conditions. Subsequent fed-batch studies were conducted in controlled fermentations of initial 5.0 L SM-B medium in 13 L fermenter (BIOENGINEERING L1523, Bioengineering AG, Switzerland) [15]. All fed-batch fermentations were performed using an exponential feeding profile to keep a constant specific growth rate, 0.15 h −1 .…”

“…The enzyme exhibited higher thermal stability and remarkable resistance to acetaldehyde [14]. In addition, the effects of cell growth rate in fed-batch cultures on the production of the enzyme was also investigated [15]. The current work was undertaken to investigate a continuous lactose induction strategy in glucose-limited fed-batch cultivations of E. coli for the production of the thermophilic DERA.…”