ARC Centre of Excellence in Innovations in Peptide and Protein Science, Australian National University, Research School of Chemistry, Canberra, ACT 2601, Australia.
ARC Centre of Excellence in Innovations in Peptide and Protein Science, Australian National University, Research School of Chemistry, Canberra, ACT 2601, Australia.
J Biotechnol. 2021 Jan 10;325:145-151. doi: 10.1016/j.jbiotec.2020.11.005. Epub 2020 Nov 6.
The human rhinovirus 14 3C protease (HRV3C protease), in fusion with glutathione S-transferase also referred to as PreScission™ protease, is a cysteine protease of particular interest for affinity tag removal from fusion proteins due to its stringent recognition sequence specificity (LEVLFQ/GX) and superior activity at low temperature. Here we report the expression, purification and use of a fusion construct of HRV3C protease, NT*-HRV3CP, that affords high expression yield in E. coli (over 300 mg/L cell culture), facile single-step purification, high solubility (>10 mg/mL) and excellent storage properties. NT*-HRV3CP cleaves affinity tags at 4 °C in minutes, making it an attractive tool for the production of recombinant proteins for biotechnological, industrial and pharmaceutical applications.
人鼻病毒 14 3C 蛋白酶（HRV3C 蛋白酶）与谷胱甘肽 S-转移酶融合，也称为 PreScissionTM 蛋白酶，是一种半胱氨酸蛋白酶，由于其严格的识别序列特异性（LEVLFQ/GX）和在低温下的优异活性，特别适合用于从融合蛋白中去除亲和标签。在这里，我们报告了 HRV3C 蛋白酶融合构建体 NT*-HRV3CP 的表达、纯化和用途，该构建体在大肠杆菌中的表达产量高（超过 300mg/L 细胞培养物），易于进行单步纯化，高溶解度（>10mg/mL）和出色的储存性能。NT*-HRV3CP 在 4°C 下几分钟内即可切割亲和标签，使其成为生物技术、工业和制药应用中生产重组蛋白的有吸引力的工具。
