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一种用于嗜冷栖假交替单胞菌TAC125亚零生长和重组蛋白生产的新型合成培养基及表达系统。

A novel synthetic medium and expression system for subzero growth and recombinant protein production in Pseudoalteromonas haloplanktis TAC125.

作者信息

Sannino F, Giuliani M, Salvatore U, Apuzzo G A, de Pascale D, Fani R, Fondi M, Marino G, Tutino M L, Parrilli E

机构信息

Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario Monte Sant'Angelo, via Cinthia, I-80126, Naples, Italy.

GSK Vaccines, Via Fiorentina, 1, 53100, Siena, Italy.

出版信息

Appl Microbiol Biotechnol. 2017 Jan;101(2):725-734. doi: 10.1007/s00253-016-7942-5. Epub 2016 Oct 27.

Abstract

The Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 is a model organism of cold-adapted bacteria. The interest in the study of this psychrophilic bacterium stems from its capability either as a non-conventional system for production of recombinant protein and as a rich source of bioactive compounds. To further explore the biotechnological ability of P. haloplanktis TAC125, we have developed a synthetic medium, containing D-gluconate and L-glutamate (GG), which allows the bacterium to grow even at subzero temperatures. P. haloplanktis TAC125 growing in GG medium at low temperature displays growth kinetic parameters which confirm its spectacular adaptation to cold environment and subzero lifestyle, paving the way to the definition of the underlying molecular strategies. Moreover, in this paper, we report the setup of a finely regulated gene expression system inducible by D-galactose to produce recombinant protein in GG synthetic medium at temperatures as low as -2.5 °C. Thanks to the combination of the novel medium and the new expression system, we obtained for the first time the production of a recombinant protein at subzero temperature, thus providing an innovative strategy for the recombinant production of "difficult" proteins.

摘要

南极细菌嗜冷栖假交替单胞菌TAC125是冷适应细菌的模式生物。对这种嗜冷细菌的研究兴趣源于它既可以作为生产重组蛋白的非传统系统,又可以作为生物活性化合物的丰富来源。为了进一步探索嗜冷栖假交替单胞菌TAC125的生物技术能力,我们开发了一种含有D-葡萄糖酸盐和L-谷氨酸(GG)的合成培养基,该培养基能使这种细菌即使在零下温度也能生长。嗜冷栖假交替单胞菌TAC125在低温下于GG培养基中生长时,其生长动力学参数证实了它对寒冷环境和零下生存方式的显著适应性,为确定潜在的分子策略铺平了道路。此外,在本文中,我们报告了一种由D-半乳糖诱导的精细调控基因表达系统的建立,该系统可在低至-2.5°C的温度下于GG合成培养基中生产重组蛋白。由于新型培养基和新表达系统的结合,我们首次在零下温度生产出了重组蛋白,从而为“难”蛋白的重组生产提供了一种创新策略。

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