Genetically Modifying the Protein Matrix of Macroscopic Living Materials to Control Their Structure and Rheological Properties

The field of engineering living materials (ELMs) seeks to engineer cells to form macroscopic materials with tailorable structures and properties. While the rheological properties of ELMs have been altered using synthetic biology methodology, the relationships connecting their sequence, structural, and rheological properties remain to be elucidated. Recently, our lab created centimeter-scale ELMs from <i>Caulobacter crescentus</i> that offer a platform to investigate this paradigm. Here, we explore how changing the elastin-like polypeptide (ELP) length within the protein matrix of this ELM impacts its microstructure and viscoelastic behavior. We demonstrate that shortening ELP produces fibers almost 2× thicker than other variants, resulting in a stiffer material at rest. Interestingly, the midlength ELP forms a complex structure with globules and multidirectional fibers with increased yield stress under flow conditions. Lengthening ELP creates thinner strands between cells with similar storage and loss moduli to those of the midlength ELP. This study begins to elucidate sequence-structure-property relationships in these ELMs and shows that they are complex with few parallels to other biocomposite models. Furthermore, it highlights that fine-tuning genetic sequences can create significant differences in rheological properties, uncovering new design principles of ELMs.