Phytate is an abundant organic phosphorus compound produced by plants and microbes and represents a major reservoir of organic phosphorus in soils. Phytate-utilising bacteria (PUB) degrade phytate into more bioavailable forms of phosphorus that can be used by plants, reducing reliance on rock-phosphate fertilisers. However, cultured representatives of PUB remain limited and often not publicly available and a systematic investigation of microbial phytate degradation mechanisms in soil is missing. Therefore, we aim to investigate phytate utilisation across a diverse range of bacterial species systematically and comparably, enabling a comprehensive assessment of bacterial phytate utilisation mechanisms and their regulation for the first time. Here we present a qualitative and quantitative approach to measure phytate degradation by a diverse collection of pure bacterial isolates which were selectively recovered from Irish soils and rhizospheres using a custom PUB-selecting medium developed by our lab group. This growing biobank includes new strains of known phytate-utilising genera, such as Pseudomonas, Bacillus, Priestia and Pantoea, and genera not previously known to degraded phytate, such as Herbiconiux, Variovorax, Ensifer, Peribacillus and Xanthomonas. Phytase activity was quantified using a high-throughput 96-well plate malachite green assay and normalised to total protein concentration. Pseudomonas and Bacillus strains exhibited the highest specific extracellular activity (~0.95 and ~0.60 U mg⁻¹ protein, respectively), followed by Priestia (~0.2 U mg⁻¹ protein), highlighting substantial inter-species variation. These findings contribute to the establishment of a diverse and functionally characterised PUB biobank.