The study also investigated the behavior of furan-based copolyesters during crystallization using holographic imaging and profiling. The results showed that the spherulite diameters fall within the range of 4-7 µm and the phase change between the crystalline and amorphous phase was smaller for certain copolymers, indicating a higher degree of crystallinity or more homogeneous distribution of the amorphous phase. The diol structure was also found to have a significant impact on the homogeneity of the crystallization process, with increasing Sa values as the number of atoms in the diol structure increases. In summary, this research has successfully synthesized new bio-based copolymers and demonstrated the significant impact that the properties of starting materials have on the enzymatic synthesis process. The study sheds light on the relationship between diol chain length and the crystalline structure of the copolymers, providing valuable information for optimizing the properties of these materials for green packaging materials in the medical sector. The use of monomers and enzymes derived from biomass in the synthesis of bio-based copolymers is a significant step towards resource-efficiency optimization. By utilizing renewable resources, this process reduces dependence on non-renewable petroleum-based materials and supports sustainability efforts.