Enhanced extracellular expression of gene-optimized Thermobifida fusca cutinase in Escherichia coli by optimization of induction strategy

Parallel steady state studies on a milliliter scale accelerate fed‐batch bioprocess design for recombinant protein production with Escherichia coli

“…The performance of this bioprocess is in the range of the most efficient recombinant protein production process with E. coli ( 5 , 7.0–9.7 g protein L −1 , 50–190 mg protein ). The beneficial effect of feeding the inducer on the metabolic activity of cells and consequently on the protein yield has already been shown for a few fed‐batch studies . In summary, suitable induction strategies identified in a time‐saving cost‐reducing manner making use of parallel chemostats on a milliliter‐scale were successfully used for the improvement of a high performance protein production process in fed‐batch mode.…”

Extracellular expression of natural cytosolic arginine deiminase from Pseudomonas putida and its application in the production of l-citrulline

“…The gene of cutinase has a high GC content of 67.3%, together with complicated RNA secondary structure, which probably made the expression of cutinase at an unfavorable position in the co-expression system (Su et al, 2015), and low expression level as well. Except for that, it has been reported that foreign protein expression generally causes a metabolic burden on the cell (Donovan et al, 1996).…”

“…E. coli strains JM109, BL21(DE3), and the expression vector pETDuet-1, were obtained from Novagen (Madison, USA). The plasmid pETDuet-cut and pET24a-cut opt , which harbor the native and optimized genes encoding T. fusca cutinase respectively, were constructed and stocked in our previous work (Su et al, 2015(Su et al, , 2013b. The pMD18-T simple vector, DNA polymerase, restriction enzymes, alkaline phosphatase, and T 4 DNA ligase were obtained from Takara (Dalian, China).…”

“…Thus, the preparation of recombinant ADI by high density fermentation of the engineered E. coli in a 3-L fermenter was further carried out, and two factors, induction temperature and inducer concentration, which have important effect on cell growth and enzyme production, were optimized in the current study. In addition, in the previous study for extracellular expression of the recombinant cutinase in a 3-L fermenter, an IPTG/lactose combined induction strategy was proposed, which offers advantages of using less inducer lactose, and increased extracellular expression of the target protein (Su et al, 2015). Thus, similar induction approach was also used in this study.…”

Efficient androst-1,4-diene-3,17-dione production by co-expressing 3-ketosteroid-Δ1-dehydrogenase and catalase inBacillus subtilis

“…It has been reported that gene codon optimization could significantly increase the expression level of the target enzyme (Su et al . ,b). Therefore, codon optimization of the ksdd gene based on the codon preference of B. subtilis is necessary for KSDD enzyme activity improvement.…”