A liposome-based artificial cell (LBAC) consists of a liposome encapsulating a cell-free protein expression system (CFPES) and protein-encoding DNA. It is surrounded by a lipid bilayer membrane and synthesizes proteins that resemble actual cells. Hence, they have been one of the most studied artificial cells. According to recent studies, they have been able to sense bio-functional molecules by synthesizing fluorescent proteins in response to target molecules. Therefore, they are expected to be used as biosensors. However, previously reported LBACs encapsulated the CFPES derived from Escherichia coli, resulting in the most productive protein expression at 20–40 °C. To broaden the range of their working temperatures to lower temperatures, in this study, we constructed LBACs using a CFPES derived from Pseudomonas fluorescens that grows at a temperature range of 4 °C to 30 °C. We then demonstrated that the constructed LBAC expressed proteins at 8 °C and that, the protein expression capability of the LBAC derived from P. fluorescens was four-fold higher than that derived from E. coli at 8 °C. This study will pave the way for the development of artificial cell-based biosensors that work in cold environments or for the synthesis of heat-labile proteins in LBACs.

中文翻译：

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利用荧光假单胞菌衍生的细胞提取物构建能够低温表达蛋白质的人工细胞

基于脂质体的人造细胞（LBAC）由包裹无细胞蛋白质表达系统（CFPES）和蛋白质编码DNA的脂质体组成。它被脂质双层膜包围，并合成类似于实际细胞的蛋白质。因此，它们已经成为研究最多的人造细胞之一。根据最近的研究，他们已经能够通过响应目标分子而合成荧光蛋白来感知生物功能分子。因此，期望它们被用作生物传感器。但是，先前报道的LBAC包封了衍生自大肠杆菌的CFPES ，导致在20–40°C时蛋白质产量最高。为了将它们的工作温度范围扩展到更低的温度，在这项研究中，我们使用了衍生自CFPES的LBAC荧光假单胞菌在4°C至30°C的温度范围内生长。然后，我们证明了构建的LBAC在8°C时表达蛋白质，并且源自荧光假单胞菌的LBAC的蛋白质表达能力比在8°C时源自大肠杆菌的LBAC高四倍。这项研究将为开发在寒冷环境中工作的基于人工细胞的生物传感器或在LBAC中合成热不稳定蛋白铺平道路。