Cultivation Conditions for Phytase Production from Recombinant Escherichia coli DH5α

Abstract

Response surface methodology (RSM) was used to optimize the cultivation conditions for the production of phytase by recombinant Escherichia coli DH5α. The optimum predicted cultivation conditions for phytase production were at 3 hours seed age, a 2.5% inoculum level, an L-arabinose concentration of 0.20%, a cell concentration of 0.3 (as measured at 600 nm) and 17 hours post-induction time with a predicted phytase activity of 4194.45 U/mL. The model was validated and the results showed no significant difference between the experimental and the predicted phytase activity (P = 0.305). Under optimum cultivation conditions, the phytase activity of the recombinant E. coli DH5α was 364 times higher compared to the phytase activity of the wild-type producer, Enterobacter sakazakii ASUIA279. Hence, optimization of the cultivation conditions using RSM positively increased phytase production from recombinant E. coli DH5α.

Safety evaluation of a novel variant of consensus bacterial phytase

“…Advancements in science and technology over the last 30 years have facilitated the production and development of increasingly more efficacious phytases. Recombinant DNA technologies have enabled the use of host microbial organisms as vehicles (‘production strains’) for producing large quantities of phytases identified as having desirable characteristics, by insertion of the candidate phytase gene into the host genome followed by expression using an appropriate promoter system [ 31 ]. Thus, the safety of both the enzyme itself and the production strain must be considered when assessing new phytases for commercial applications.…”

Microtiter miniature shaken bioreactor system as a scale-down model for process development of production of therapeutic alpha-interferon2b by recombinant Escherichia coli

“…Results from this study have demonstrated that the inoculum size plays an important role in the optimization of recombinant α-IFN2b production. The effect of inoculum size was suggested to be related to the length of the lag phase in the cultivation, which resulted in different cell densities of bacterial expression [ 10 ]. Small inoculum size may cause slow adaptation of E. coli culture when inoculated into a new fermentation medium [ 11 ] resulted in a long lag phase with insufficient number of cells that would reduce the production of α-IFN2b.…”

Improved expression and immobilization of Geobacillus thermoleovorans CCR11 thermostable recombinant lipase

“…Numerous studies have been conducted in order to improve recombinant heterologous gene expression using Escherichia coli as a host [1,[6][7][8][9]. Induction conditions for such systems are considered important parameters in the production of recombinant proteins, and they need to be carefully adjusted for every new expressed protein [10,11]. Therefore, different strategies based on response surface methodology have been applied to assess significant factors and achieve maximum metabolite production, reporting increments as high as 364fold when compared with nonoptimal conditions in native microorganisms [11].…”