In this paper, the mulberry fibers were successfully obtained by a new pretreatment named alkali-assisted microwave plus biological enzymatic technique (AMBET). The morphology, microstructure, physico-mechanical and antibacterial properties of the mulberry bast fibers were investigated by means of scanning electron microscope (SEM), Fourier Transform-Infrared (FTIR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD), instron tensile tester and antibacterial testing. The results showed that impurities of the bast fibers could be removed by AMBET treatment. AMBET treated mulberry fiber was even, smooth and fine, and typical cellulose I in the mulberry fibers was confirmed by FTIR and XRD analysis. The crystallinity of the AMBET treated fibers was higher than that of the raw mulberry and chemical treated mulberry fibers. Thermal analysis indicated that the mulberry fibers had a good thermal stability. Moreover, the AMBET treated mulberry fibers showed excellent antimicrobial activities against $S.aureus$. The physical properties of the mulberry fibers indicated the AMBET treated mulberry fibers were ideal candidates for new textile materials.