Betterle Nico, Hidalgo Martinez Diego, Melis Anastasios
Department of Plant and Microbial Biology, University of California, Berkeley, Berkeley, CA, United States.
Front Plant Sci. 2020 Mar 3;11:237. doi: 10.3389/fpls.2020.00237. eCollection 2020.
Efforts to express human therapeutic proteins in photosynthetic organisms have been described in the literature. Regarding microalgae, most of the research entailed a heterologous transformation of the chloroplast, but transformant cells failed to accumulate the desired recombinant proteins in high quantity. The present work provides methods and DNA construct formulations for over-expressing in photosynthetic cyanobacteria, at the protein level, human-origin bio-pharmaceutical and bio-therapeutic proteins. Proof-of-concept evidence is provided for the design and reduction to practice of "" for the generation of the bio-therapeutic protein interferon alpha-2 (IFN). IFN is a member of the Type I interferon cytokine family, well-known for its antiviral and anti-proliferative functions. Fusion construct formulations enabled accumulation of IFN up to 12% of total cellular protein in soluble form. In addition, the work reports on the isolation and purification of the fusion IFN protein and preliminary verification of its antiviral activity. Combining the expression and purification protocols developed here, it is possible to produce fairly large quantities of interferon in these photosynthetic microorganisms, generated from sunlight, CO, and HO.
文献中已描述了在光合生物中表达人类治疗性蛋白质的相关研究。就微藻而言,大多数研究涉及叶绿体的异源转化,但转化细胞未能大量积累所需的重组蛋白。本研究提供了在光合蓝细菌中过表达人类来源的生物制药和生物治疗蛋白的方法及DNA构建体制剂。为“生物治疗蛋白干扰素α - 2(IFN)的产生”的设计及实际应用提供了概念验证证据。IFN是I型干扰素细胞因子家族的成员,以其抗病毒和抗增殖功能而闻名。融合构建体制剂使IFN以可溶性形式积累至总细胞蛋白的12%。此外,该研究报告了融合IFN蛋白的分离和纯化及其抗病毒活性的初步验证。结合此处开发的表达和纯化方案,有可能在这些利用阳光、二氧化碳和水产生的光合微生物中大量生产干扰素。