Light-responsive hydrogels are highly valued for their dynamic mechanical properties and biocompatibility. In this study, we present a hydrogel system that can either soften or strengthen on green light exposure, or remain unresponsive to light, depending on the addition of adenosyl cobalamin (AdoCbl) and Co2+. These protein-based hydrogels were formed using genetically encoded SpyTag-SpyCatcher chemistry and included green light-sensitive CarHc protein domains. As previously reported, these hydrogels formed in the dark with the addition of AdoCbl, due to the tetramerization of the CarHc domains. Under green light exposure, the CarHc tetramers disassembled, leading to a rapid transition from gel to sol. Interestingly, we discovered that an excess of AdoCbl leads to photo-strengthening rather than photo-weakening. This occurred because light exposure induces interchain crosslinks between AdoCbl and poly-histidine tags (His6-tags) of the proteins. Furthermore, incorporating Co2+ ions enhanced hydrogel stiffness by coordinating to His6-tags. This not only suppressed photo-weakening but also promoted photo-strengthening behaviour. These findings highlight the role of His6-tags in photochemical crosslinking with excess AdoCbl and in coordination to Co2+ ions, providing a novel strategy for designing tuneable, light responsiveness in materials.
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