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利用杆状病毒表达系统在低温下高效生产重组蛋白磷酸酶2A(PP2A)

Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system.

作者信息

Ikehara Tsuyoshi, Nakashima Shihoko, Nakashima Junichi, Kinoshita Tsubasa, Yasumoto Takeshi

机构信息

Department of Food Science and Technology, National Fisheries University, 2-7-1 Nagata-honmachi, Shimonoseki, Yamaguchi, 759-6595, Japan.

Faculty of Sports and Health Science, Fukuoka University, Fukuoka, 814-0180, Japan.

出版信息

Biotechnol Rep (Amst). 2016 Aug 5;11:86-89. doi: 10.1016/j.btre.2016.07.004. eCollection 2016 Sep.

Abstract

Protein phosphatase 2A (PP2A) is an enzyme useful for detecting several natural toxins represented by okadaic acid and microcystins. We found that the production of the recombinant human PP2A catalytic subunit (rhPP2Ac) in High Five insect cells could markedly increase when the cells were cultured at 19 °C instead of 27 °C used under conventional conditions. The yield and purity of the enzyme increased four- and three-folds, respectively. The benefit of the altered culturing temperature was observed with the recombinant human protein phosphatase 2B but not 2Cα. The different responses among the enzymes suggest the involvement of an enzyme-specific mechanism that leads to the catalytic subunit overexpression. This is the first report to produce rhPP2Ac at a temperature lower than that used under conventional culture conditions (27 °C) used in the baculovirus expression system with High Five insect cells.

摘要

蛋白磷酸酶2A(PP2A)是一种可用于检测以冈田酸和微囊藻毒素为代表的多种天然毒素的酶。我们发现,当在19°C而非传统条件下使用的27°C培养High Five昆虫细胞时,重组人PP2A催化亚基(rhPP2Ac)的产量会显著增加。该酶的产量和纯度分别提高了四倍和三倍。在重组人蛋白磷酸酶2B中观察到了改变培养温度的益处,但在2Cα中未观察到。这些酶之间的不同反应表明存在一种导致催化亚基过表达的酶特异性机制。这是首次报道在低于杆状病毒表达系统与High Five昆虫细胞常规培养条件(27°C)的温度下生产rhPP2Ac。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/79c3/5042294/e1b23c5b7771/gr1.jpg

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