Protein-based hydrogels have a variety of biomedical and nanotechnology applications including drug delivery, tissue engineering and development of biosensors and nanowires. Proteins as polymeric hydrogel building blocks are uniquely adept at simultaneously playing structural and bioactive roles. Alternative approaches to modifying hydrogels to facilitate their interaction with biological systems requires time consuming and expensive synthesis and purification steps. Here, we demonstrate a novel method for the rapid biosynthesis of tailored bioactive hydrogels in a single step, directly from bacterial culture, using a protocol that requires no protein purification. The hydrogel scaffold is based on the engineered extracellular matrix protein CsgA, which self-assembles into a fibrous mesh-like network. Subsequent addition of the ionic surfactant sodium dodecyl sulfate leads to rapid gelation. This unique method provides quick and cost effective generation of amyloid-based hydrogels that possess high stability and mechanical strength. Moreover, by fusing functional proteins to the amyloid fibers we can biofabricate a variety of customized hydrogel for several applications.<br/>This basic hydrogel fabrication protocol works well with a range of domains fused to CsgA, including mucin-binding and anti-inflammatory trefoil factors (TFFs), and a panel of fluorescent proteins, which results in optically active hydrogels. By displaying functional proteins on curli fibers, not only can we reprogram the function of the hydrogels, we can also tune the mechanical properties to make these customized hydrogels suitable as injectable materials.

Culturing customized hydrogels from engineered biofilm matrix proteins

254th National Meeting (2017)

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