A new bio-based polyamide 56/512 (PA56/512) has been synthesized with a higher bio-based composition compared to industrialized bio-based PA56, which is considered a lower carbon emission bio-based nylon. In this paper, the one-step approach of copolymerizing PA56 units with PA512 units using melt polymerization has been investigated. The structure of the copolymer PA56/512 was characterized using Fourier-transform infrared spectroscopy (FTIR) and Proton nuclear magnetic resonance (1H NMR). Other measurement methods, including relative viscosity tests, amine end group measurement, thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), were used to analyze the physical and thermal properties of the PA56/512. Furthermore, the non-isothermal crystallization behaviors of PA56/512 have been investigated with the analytical model of Mo’s method and the Kissinger method. The melting point of copolymer PA56/512 exhibited a eutectic point at 60 mol% of 512 corresponding to the typical isodimorphism behavior, and the crystallization ability of PA56/512 also displayed a similar tendency.