High-level cell-free expression and functional characterization of a novel aquaporin from Photobactetrium profundum SS9

•The bioactivity of aquaporin in E.coli was demonstrated by in-vivo expression of AqpSS9 from P. profundum SS9.•A high productivity of soluble AqpSS9 (571mg/l) was achieved by using in-vitro E. coli cell-free expression system (D-CF mode).•High water permeability (Pf 310.7±3.2μm/s) was determined with AqpSS9 incorporated proteoliposome.•The construction of AqpSS9 embedded layer-by-layer (LbL) membrane with improved water permeability and salt rejection.