Methanogens, thought to be present on early Earth, have a high requirement for Ni, suggesting that Ni utilization could be a potential biosignature for methanogens if enhanced Ni extraction from surrounding minerals accompanies methanogenic growth. To test the potential for such Ni extraction from minerals by methanogens, Ni release from Ni-containing silicate glass was measured in Ni-free growth medium in the presence of the methanogen Methanothermobacter thermoautotrophicus (average pH ∼7.0) and observed to be higher than an abiotic control (average pH ∼6.8). However, batch dissolution experiments and a siderophore assay indicate that cell exudates such as siderophores, low molecular weight organic acids, or lysates accompanying cell death are not responsible for the observed increase in Ni release rate. In addition, scanning electron microscopy (SEM) shows little to no evidence of direct microbe–mineral interactions such as biofilms or pitting. Instead, comparison with abiotic experiments suggests that changes in pH due to CO2 uptake may be responsible for enhanced dissolution in the presence of metabolizing cells. These results document that methanogens may not preferentially extract Ni from surrounding minerals although they may indirectly affect mineral reaction rates that are pH sensitive. Thus identifiable Ni biosignatures may not exist in the rock record to document the presence of methanogens on early Earth or Mars.