Carvajal Felipe, Vargas-Torres Valentina, Becerra Daniela, González-Quezada Nicolás, Egaña José Tomás
Institute for Biological and Medical Engineering, Pontificia Universidad Católica de Chile, Vicuña Mackenna 4860, Santiago, 7820436, Chile.
J Biol Eng. 2025 Jun 4;19(1):54. doi: 10.1186/s13036-025-00525-9.
Genetically engineered photosynthetic microorganisms have been proposed as a therapeutic approach for the localized delivery of oxygen and recombinant proteins to tissues in various pathological conditions. However, the effect of recombinant protein production and secretion on microalgal fitness, as well as the impact of key environmental conditions on their potential therapeutic performance, has not yet been described. Therefore, in this study, the microalga was genetically engineered to produce and release the reporter protein mVenus and was then challenged by exposure to different media, temperatures, and substrates.
The genetically modified microalgae were able to produce and release the mVenus protein under standard culture conditions without affecting overall fitness, including cell size and shape, growth potential, and oxygen metabolism, compared to the wild-type strain. Under mammalian cell culture conditions, the strains continued to produce and secrete mVenus protein for up to four days at 22 °C, 30 °C, and 37 °C. Additionally, photosynthetic biomaterials containing the engineered microalgae showed continuous recombinant protein release at 30 °C and 37 °C for up to four days.
The microalga can be genetically engineered to produce and release recombinant proteins without detrimental effects on its fitness, showing therapeutic potential under mammalian culture conditions and within biomaterials designed to promote tissue regeneration. Overall, these findings support the use of genetically engineered photosynthetic microalgae for the localized and controlled release of oxygen and recombinant proteins for several therapeutic applications.
基因工程改造的光合微生物已被提议作为一种治疗方法,用于在各种病理条件下向组织局部输送氧气和重组蛋白。然而,重组蛋白的产生和分泌对微藻适应性的影响,以及关键环境条件对其潜在治疗性能的影响,尚未见报道。因此,在本研究中,对微藻进行基因工程改造以产生并释放报告蛋白mVenus,然后使其暴露于不同的培养基、温度和底物中进行挑战。
与野生型菌株相比,基因改造后的微藻在标准培养条件下能够产生并释放mVenus蛋白,且不影响其整体适应性,包括细胞大小和形状、生长潜力以及氧代谢。在哺乳动物细胞培养条件下,这些菌株在22℃、30℃和37℃下持续产生并分泌mVenus蛋白长达四天。此外,含有工程微藻的光合生物材料在30℃和37℃下持续释放重组蛋白长达四天。
微藻可以通过基因工程改造来产生并释放重组蛋白,而不会对其适应性产生不利影响,在哺乳动物培养条件下以及在旨在促进组织再生的生物材料中显示出治疗潜力。总体而言,这些发现支持使用基因工程改造的光合微藻进行氧气和重组蛋白的局部可控释放,用于多种治疗应用。