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Bioprocess 2022
发酵条件对天冬氨酸转氨酶重组菌高效表达与质粒稳定性的影响
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Abstract:
通过对以pET-22b质粒为载体，大肠杆菌BL21为宿主构建的产天冬氨酸转氨酶的重组菌BL21/pET-aspC，质粒稳定性与重组菌产酶活性的发酵条件优化，对比了不同发酵条件对基因工程菌菌体生长、质粒稳定性和转氨酶酶活性的影响，得到较工程菌常用LB培养基更利于工程菌生长的培养基CSN。考察了该培养基主要成份玉米浆、酵母粉和柠檬酸钠对重组菌生长及质粒稳定性的影响。优化后，该发酵体系的发酵成本为LB培养基的10%，外源基因的质粒稳定性得到明显改善，重组菌生物量与天冬氨酸转氨酶活性显著提高。优化后培养体系成本低廉，发酵稳定，适于基因工程菌的大规模工业化生产发酵使用。
Using the recombinant strain Bl21/pet-AspC producing aspartate aminotransferase which was constructed with pET-22b as the vector and E. coli Bl21 as the host, the stability of plasmid and the most suitable fermentation conditions for the production of enzyme were studied. By comparing the relationship between cell growth, plasmid stability, transaminase activity and medium composition of genetically engineered bacteria, the CSN medi-um, which is more favorable for the growth of engineering bacteria than the LB medium commonly used for engineering bacteria, was obtained. The effects of corn syrup, yeast powder and sodium citrate on the growth of recombinant bacteria and the stability of plasmid were investigated. The fermentation cost of the new medium was 10% of that of LB medium. The stability of plasmid of for-eign gene, the biomass of recombinant bacteria and the activity of aspartate aminotransferase were significantly improved. The optimized culture system has low cost and stable fermentation, which is suitable for large-scale industrial production and fermentation of genetic engineering bacteria.
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