Abstract：
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.
摘要：
水通道蛋白（AQP）是跨膜蛋白家族的水通道蛋白，可促进水穿过细胞膜的运动。它在自然界中无处不在，然而，对水运机制的理解，特别是对于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被认可，但是缺乏其在工业过程中的潜在应用所必需的生化特征的知识。因此,这项研究是为了表达，Extract,溶解，净化，并研究假单胞菌水通道蛋白Z家族的功能适应。AMS3通过分子方法。在这项研究中,AqpZ1AMS3被成功地亚克隆并作为重组蛋白在大肠杆菌BL21(DE3)中表达。AqpZ1AMS3基因在优化条件下表达，AQP的最佳优化条件是在25°C孵育的0.5mMIPTG中诱导20小时。两性离子温和洗涤剂[（3-胆碱丙基）二甲基铵]-1-丙磺酸盐是蛋白质溶解的合适表面活性剂。然后通过亲和层析纯化蛋白质。使用动态光散射重建脂质体和蛋白脂质体以确定粒度。从鉴定的嗜冷AQP获得的此信息为该蛋白质在低温下的结构适应提供了新的见解，并且将来可用于低温应用和分子工程目的。
