Herein, we describe a yeast-laden hydrogel ink that can be printed using a direct-write 3D printer and used for the production of a peptide. A poly(alkyl glycidyl ether)-based triblock copolymer was synthesized and formulated as a hydrogel that was characterized via rheometry to evaluate the printability of the hydrogel ink. An engineered yeast strain with an upregulated α-factor production pathway was incorporated into the hydrogel ink and 3D printed. The immobilized yeast cells exhibited adequate viability of 87.5% within the hydrogel. The production of the up-regulated α- factor was detected using a detecting yeast strain and quantified at 268 nM (s = 34.6 nM) over 72 h. The reusability of these systems was demonstrated by immersion of the yeast-laden hydrogel lattice in fresh SC media and confirmed by the detection of similar amounts of up-regulated α-factor 259 nM (s = 45.1 nM). These yeast-laden materials represent an attractive opportunity for whole-cell catalysis of other high-value products in a sustainable and continuous manner.
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