Experimental benchmarking remains crucial to the interpretation of sequence annotation data.

Determining the distribution and origin of bacterial synthesis of metabolites of interest relies on computational prediction of protein function. In their recent study, Lawrence and colleagues employed a rigorous pipeline for benchmarking their annotation methods (E. C. Lawrence, H. Pan, N. Ollikainen, and B. J. Belin, mSystems 11:e00619-26, 2026, https://doi.org/10.1128/msystems.00619-26). This allowed them to distinguish between related yet functionally distinct proteins, thereby refining their understanding of bacterial hopanoid production. They find that the ability to synthesize these compounds likely arose in marine alphaproteobacteria and is related to osmotic stress tolerance. This study additionally provides new insights into the extracellular transport of hopanoids. The combination of large-scale computational analysis with experimentally based benchmarking that they describe provides a useful model for any researcher interested in understanding bacterial biosynthetic capability.