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Abundant sulphur is present in the Late Miocene evaporitic sequence of the lacustrine Hellín basin in SE Spain. Weathering of Triassic evaporites controlled the chemical composition of the Miocene lake. The lacustrine deposits comprise gypsum, marlstones, diatomites and carbonate beds. Sulphur-bearing carbonate deposits predominantly consist of early diagenetic dolomite. Abundant dolomite crystals with a spheroidal habit are in accordance with an early formation and point to a microbial origin. The carbon isotopic composition of the dolomite (δ 13 C values between −10 and −4 ‰) indicates mixing of lake water carbonate and carbonate derived from the remineralization of organic matter by heterotrophic bacteria. Dolomite precipitated syngenetically under evaporitic conditions as indicated by high oxygen isotope values (δ 18 O between +6 and +11 ‰). Nodules of native sulphur are found in gypsum, carbonate beds and marlstone layers. Sulphur formed in the course of microbial sulphate reduction, as reflected by its strong depletion in 34 S (δ 34 S values as low as −17 ‰). Near to the surface many of the sulphur nodules were in part or completely substituted by secondary gypsum, which still reflects the sulphur isotopic composition of native sulphur (−18 to −10 ‰). This study exemplifies the role of bacterial sulphate reduction in the formation of dolomite and native sulphur in a semi-enclosed lacustrine basin during Late Miocene time.