Carboxyl groups in polyamide (PA) reverse osmosis (RO) membrane contribute significantly to fouling and scaling, hindering the sustainable operation of RO in practical applications. Herein, we developed a novel interfacial polymerization (IP) strategy to finely engineer the molecular structure of PA with no carboxyl groups, and to significantly enhance RO membrane fouling/scaling-resistance. During IP, trimesoyl chloride (TMC) at the interface was consumed completely by the diffused m-phenylenediamine (MPD) and glycerol (GLY) under the assistance of benzalkonium chloride (BAC) surfactant. The fabricated RO membrane with no carboxyl groups exhibits sustainable anti-fouling performance with low flux decline ratios and high flux recovery ratios during the five cycles of fouling and cleaning when treating real coke wastewater, surpassing the reported anti-fouling membranes and the renowned commercial fouling-resistant RO membrane (DuPont FilmTec™ CR100). This work provides some insights to precisely tailor the molecular structure of PA RO membrane with sustainable anti-fouling performance.