Changes in stable chromium isotopes (denoted as δ
53Cr) in ancient carbonate sediments are increasingly used to reconstruct the oxygenation history in Earth’s atmosphere and oceans through time. As a significant proportion of marine carbonate older than the Cambrian is microbially-mediated, the utility of δ
53Cr values in ancient carbonates hinges on whether these sediments accurately capture the isotope composition of their environment. We report Cr concentrations (Cr) and δ
53Cr values of modern marginal marine and non-marine microbial carbonates. These data are supported by stable C and O isotope compositions, as well as rare earth elements and yttrium (REY) concentrations. In addition, we present data on ancient analogs from Precambrian strata. Microbial carbonates from Marion Lake (Australia, δ
53Cr ≈ 0.99‰) and Mono Lake (USA, ≈0.78‰) display significantly higher δ
53Cr values compared with ancient microbialites from the Andrée Land Group in Greenland (720 Ma, ≈0.36‰) and the Bitter Springs Formation in Australia (800 Ma, ≈−0.12‰). The δ
53Cr values are homogenous within microbialite specimens and within individual study sites. This indicates that biological parameters, such as vital effects, causing highly variable δ
53Cr values in skeletal carbonates, do not induce variability in δ
53Cr values in microbialites. Together with stable C and O isotope compositions and REY patterns, δ
53Cr values in microbialites seem to be driven by environmental parameters such as background lithology and salinity. In support, our Cr and δ
53Cr results of ancient microbial carbonates agree well with data of abiotically precipitated carbonates of the Proterozoic. If detrital contamination is carefully assessed, microbialites have the potential to record the δ
53Cr values of the waters from which they precipitated. However, it remains unclear if these δ
53Cr values record (paleo-) redox conditions or rather result from other physico-chemical parameters.
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