This bio-chemical study focuses on obtaining high-quality hemp fiber. The effects of the structures and properties of hemp fibers in different treatment periods were studied. Moreover, the changes of the surface morphology, chemical composition, and breaking tenacity of hemp fibers were researched by scanning-electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), fluorescence microscopy, and fiber tensile testing. The results showed that by virtue of the enzyme scouring process, alkali refining process, and bleaching process, the pectin, lignin, and hemicellulose and other impurities were removed. Through the single factor experiment, the optimal process conditions for the bio-chemical combination of the degumming process were obtained. These conditions included 10 g of dried hemp fibers, 15% (v/v) pectinase solution, a temperature of 50 °C, a duration of 120 min, pH 8.0 (phosphate buffer), a liquor ratio (w/v) of 1:10, and 0.0625 mol/L NaOH. In these conditions, the residual gum content and breaking tenacity were 4.8% and 49.8 cN/tex, respectively, indicating that the treated hemp fibers met the requirements of the spinning process.