Objective The ecosystem of alpine lakes in northwestern Yunnan Province is well-preserved, while the microbial communities and functional characteristics in the sediments remain unclear. This study aims to elucidate the vertical distribution patterns of microbial communities in alpine lake sediments within this region and their functional differentiation in carbon, nitrogen, and sulfur cycling. Methods Three adjacent alpine lakes (Taiji Lake, Tiancai Lake, and Rencai Lake) in Laojunshan National Park, Lijiang City, Yunnan Province were selected. Samples were collected from four depths (0-1, 10-11, 20-21, 30-31 cm) of sediment cores and subjected to metagenomic sequencing. Medium and high-quality metagenome-assembled genomes (MAGs) were recovered through binning analysis, with taxonomic annotation conducted against the GTDB database. Meanwhile, functional gene annotation was performed against the CAZymes and KEGG databases to characterize the vertical stratification of community structure and biogeochemical cycling functions. Results A total of 478 MAGs (belonging to 27 bacterial phyla and 9 archaeal phyla) were obtained. Approximately 95.0% of these MAGs could not be identified at the species level, indicating that there were a large number of uncultured microbial groups in the sediments. The bacterial communities exhibited distinct succession with depth. The surface layer was dominated by Cyanobacteriota and Bacteroidota, while the middle and lower layers were mainly occupied by Pseudomonadota and Chloroflexota. The archaeal community was mainly composed of Nanobdellota, Thermoproteota, and Halobacteriota, and exhibited increasing stability with sediment depth. Carbohydrate-active enzymes (CAZymes) in the surface layer were mainly enzymes (e.g., glycosyl transferases GT51 and glycoside hydrolases GH59) targeting readily degradable carbon sources, while those in deeper layers were mainly enzymes (e.g., carbohydrate-binding modules CBM38 and auxiliary activity family AA6) acting on recalcitrant carbon sources. Nitrogen and sulfur cycling functions also exhibited a distinct vertical hierarchical structure. The bacteria primarily participated in nitrogen and sulfur cycling processes in surface sediments, whereas archaea predominated in deeper sediment layers. Conclusion The microbial communities in the sediments of alpine lakes in northwestern Yunnan Province exhibited distinct vertical distribution patterns related to carbon, nitrogen, and sulfur cycling, reflecting the influences of the sediment redox gradient and the organic matter composition on microorganisms. This study provides a new perspective for understanding the microbial ecology and biogeochemical cycling process in alpine lakes.