Aquaporin (AQP) is a water channel protein from the family of transmembrane proteins which facilitates the movement of water across the cell membrane. It is ubiquitous in nature, however the understanding of the water transport mechanism, especially for AQPs in microbes adapted to low temperatures, remains limited. AQP also has been recognized for its ability to be used for water filtration, but knowledge of the biochemical features necessary for its potential applications in industrial processes has been lacking. Therefore, this research was conducted to express, extract, solubilize, purify, and study the functional adaptations of the aquaporin Z family from Pseudomonas sp. AMS3 via molecular approaches. In this study, AqpZ1 AMS3 was successfully subcloned and expressed in E. coli BL21 (DE3) as a recombinant protein. The AqpZ1 AMS3 gene was expressed under optimized conditions and the best optimized condition for the AQP was in 0.5 mM IPTG incubated at 25°C for 20 h induction time. A zwitterionic mild detergent [(3‐cholamidopropyl) dimethylammonio]‐1‐propanesulfonate was the suitable surfactant for the protein solubilization. The protein was then purified via affinity chromatography. Liposome and proteoliposome was reconstituted to determine the particle size using dynamic light scattering. This information obtained from this psychrophilic AQP identified provides new insights into the structural adaptation of this protein at low temperatures and could be useful for low temperature application and molecular engineering purposes in the future.

中文翻译：

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南极假单胞菌重组水通道蛋白 Z 的表达和功能分析。AMS3

水通道蛋白 (AQP) 是跨膜蛋白家族的一种水通道蛋白，可促进水穿过细胞膜的运动。它在自然界中无处不在，然而对水传输机制的理解，尤其是适应低温的微生物中的 AQP，仍然有限。AQP 还因其用于水过滤的能力而得到认可，但对其在工业过程中潜在应用所需的生化特征的了解一直缺乏。因此，本研究旨在表达、提取、溶解、纯化水通道蛋白Z家族，并研究其功能适应性。假单胞菌属sp。AMS3 通过分子方法。本研究成功亚克隆AqpZ1 AMS3并在其中表达大肠杆菌BL21 (DE3) 作为重组蛋白。AqpZ1 AMS3 基因在优化条件下表达，AQP 的最佳优化条件是在 0.5 mM IPTG 中于 25°C 孵育 20 小时诱导时间。两性离子温和去污剂[(3-胆酰胺丙基)二甲基铵]-1-丙磺酸盐是适合溶解蛋白质的表面活性剂。然后通过亲和层析纯化蛋白质。使用动态光散射重构脂质体和蛋白脂质体以确定粒径。从这种嗜冷 AQP 中获得的信息为该蛋白质在低温下的结构适应提供了新的见解，并且可能对未来的低温应用和分子工程目的有用。