Due to various production stages involved, textiles and clothing industry is known for causing carbon dioxide emissions, water pollution, soil erosion and huge waste generation. It is a need of the hour to seek for natural, renewable, and bio-degradable fabrication materials and environmentally friendly production methods. This study proposes an eco-friendly approach to prepare bacterial cellulose/electrospun nanofibers membrane-based hybrid non-woven fabrics using in-situ self-assembly method. This fabrication method enables bacterial cellulose cultivation on nanofibrous membrane support to create custom-made seamless tubular hybrid fabrics in desirable dimensions, to be used for various textile applications with minimized material wastage. As-prepared nano-composite fabric was characterized using SEM, X-ray diffraction, and FTIR. FTIR and X-ray diffraction results confirmed the presence of bacterial cellulose in the composite structure. SEM analysis showed as the bacterial cellulose cultivates, its nanofibrils penetrate and grow into empty voids of membrane's structure, which results in secure binding and interlocking of electrospun nonofibers. Sample thickness and weight gain measurements after the modification were found to be approx. 33.90% and 39.02%, respectively. Reduced surface hydrophobicity, water uptake, and increased tensile strength might contribute towards better fabric performance and comfort. Overall, this study suggests an eco-friendly approach to prepare nano-composite fabrics that might be used for bio-textiles and related applications.