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首页> 外文期刊>Bioprocess and Biosystems Engineering >Enhancement of human granulocyte-colony stimulating factor production in recombinant E. coli using batch cultivation
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Enhancement of human granulocyte-colony stimulating factor production in recombinant E. coli using batch cultivation

机译：分批培养提高重组大肠杆菌中人粒细胞集落刺激因子的产生

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Development of inexpensive and simple culture media is always favorable for recombinant protein over-expression in E. coli. The effects of medium composition on the production of recombinant human granulocyte-colony stimulating factor (rh-GCSF) were investigated in batch culture of E. coli BL21 (DE3) [pET23a-hgcsf]. First, the optimum medium for production of rh-GCSF was determined; and, then it was shown that mixture of amino acid addition at induction time, which was determined on the basis of amino acids frequency in the recombinant protein, increases recombinant protein expression level significantly. Furthermore, the effect of glucose concentration on productivity of rh-GCSF was investigated; 20 g/l of glucose will result in maximum attainable biomass and rh-GCSF in this process. At optimum conditions, a cell dry weight of 10.5 g/l, an expression level of about 35% of total cellular protein, rh-GCSF concentration of 1.75 ± 0.1 g/l, and overall rh-GCSF yield of 165 ± 5 mg/g were obtained.
机译：开发廉价和简单的培养基总是有利于重组蛋白在大肠杆菌中的过表达。在大肠杆菌BL21（DE3）[pET23a-hgcsf]的分批培养中，研究了培养基组成对重组人粒细胞集落刺激因子（rh-GCSF）产生的影响。首先，确定生产rh-GCSF的最佳培养基；并且，显示了根据重组蛋白中氨基酸频率确定的诱导时添加氨基酸的混合物显着提高了重组蛋白表达水平。此外，研究了葡萄糖浓度对rh-GCSF生产力的影响。 20 g / l的葡萄糖将在此过程中获得最大的生物量和rh-GCSF。在最佳条件下，细胞干重为10.5 g / l，表达水平约为总细胞蛋白的35％，rh-GCSF浓度为1.75±0.1 g / l，rh-GCSF总体产量为165±5 mg / l获得g。

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