Exploring the effect of a peptide additive on struvite formation and morphology: a high-throughput method

Precipitation of struvite (MgNH₄PO₄·6H₂O), a slow-release fertilizer, provides a means of recycling phosphate from wastewater streams. In this work, a high-throughput struvite precipitation method is developed to investigate the effects of a peptide additive. The reactions occurred in small volumes (300 μL or less) in a 96-well plate for 45 minutes. The formation of struvite was monitored by fitting absorbance at 600 nm over time to a first order model with induction time, with the addition of peptide inducing significant changes to the yield parameter and formation constant in that model. The impact of struvite seed dosing was also investigated, highlighting the importance of optimization when peptide is present. The composition of the precipitate was confirmed through Fourier-transform infrared spectroscopy, while morphology and crystal size were analyzed through optical microscopy. Crystals had a higher aspect ratio when precipitated with the peptide. Finally, the utility of the high-throughput platform was demonstrated with a 2⁵ full factorial design to capture the effects and interactions of: magnesium dose, mixing time, seed dose, pH, and temperature. Overall, this study quantifies novel effects of a sequence-defined peptide on struvite formation and morphology via a newly developed high throughput platform.