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中试规模优化毕赤酵母表达人血清白蛋白及其高效纯化。

Medium optimization for high yield production of human serum albumin in Pichia pastoris and its efficient purification.

机构信息

China State Institute of Pharmaceutical Industry, Zhangjiang Institute, Shanghai, 201203, PR China.

China State Institute of Pharmaceutical Industry, Zhangjiang Institute, Shanghai, 201203, PR China.

出版信息

Protein Expr Purif. 2021 May;181:105831. doi: 10.1016/j.pep.2021.105831. Epub 2021 Jan 26.

DOI:10.1016/j.pep.2021.105831
PMID:33508474
Abstract

OBJECTIVE

To improve the yield of recombinant human serum albumin (HSA) in Pichia pastoris by medium optimization and establish the related purification scheme.

RESULTS

A simplified version of the generally used buffered glycerol complex medium (BMGY), which contained yeast extract, glycerol and potassium salts, was found to be applicable. By decreasing the salt concentration of basal salt medium (BSM) to half of the original formula further, we achieved a high yield of 17.47 g/L HSA in the supernatant within a 192 h induction, which is the highest rHSA yield ever reported as far as we know. Accompanied with a three-step purification procedure which recovered two thirds of the desired protein at high purity, our work lays a solid foundation for large-scale industrial production of HSA.

CONCLUSION

Medium optimization plays a significant role in improving the yield of desired protein, lowering the production cost and helping to explore the producing strain's character.

摘要

目的

通过优化培养基提高毕赤酵母表达的重组人血清白蛋白（HSA）的产量，并建立相关的纯化方案。

结果

发现一种简化的缓冲甘油复合培养基（BMGY）可替代常用的培养基，该培养基含有酵母提取物、甘油和钾盐。进一步将基础盐培养基（BSM）的盐浓度降低至原始配方的一半，我们在 192 小时的诱导期内实现了上清液中 17.47g/L 的 HSA 高产，这是迄今为止我们所知的最高的 rHSA 产量。伴随着三步纯化程序，我们以高纯度回收了三分之二的所需蛋白，我们的工作为 HSA 的大规模工业化生产奠定了坚实的基础。

结论

培养基优化在提高目的蛋白的产量、降低生产成本和帮助探索生产菌株的特性方面起着重要作用。

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