Microbial fuel cell (MFC) is a novel technique can solve the waste water problem and generate electricity. This study focuses on the anode material by using co-axial electrospinning method to fabricate polylactic acid (PLLA) microtube array membrane (PLLA MTAM). These MTAM not only have high specific area but also enhance the microbial adhesion ability. The addictive, multi-walled carbon nanotubes (MWCNT), increase the conductivity that can improve the efficiency of MFC. Furthermore, the effects of different numbers of electro spun fiber membrane placed in the anode of dual-chamber MFC on electrical performances are analyzed. The scanning electron microscopy (SEM) images show that MTAM have thin walls and contain pores on its surface which is beneficial for fast mass transport. During acclimation, bacteria regularly accumulate on the surface and formed a lamellar structure. However the bacteria in the inner structure tend to group into individual block. The growth of biofilm obviously affected the electrical performance of the cell. In the acclimation process, more electro spun fiber membranes in the anode raise voltage rapidly due to better electron transfer ability. The use of MTAM as the anode in MFC is beneficial for attaching bacteria in shorter acclimation time. The MFC of 11 MTAMs has a maximum power density of 2.94 mW/m² and a minimum resistance of 833 Ω. Compared to sealing MTAM, hollow MTAM demonstrate better electrical performance.