Biological soil crusts are integral to Arctic ecosystems, playing a crucial role in primary production, nitrogen fixation and nutrient cycling, as well as maintaining soil stability. However, the composition and complex relationships between the diverse organisms within these biocrusts are not well studied. This study investigates how the microbial community composition within Arctic biocrusts is influenced by environmental factors along an altitudinal gradient (101 m to 314 m). Metagenomic analyses were used to provide insights into the community composition, revealing that temperature, pH, and nutrient availability significantly shaped the community. In contrast, altitude did not directly influence the microbial composition significantly. Eukaryotic communities were dominated by Chloroplastida and fungi, while Proteobacteria and Actinobacteria prevailed among prokaryotes. Cyanobacteria, particularly orders such as Pseudoanabaenales, Pleurocapsales, and Nostocales, emerged as the most abundant photoautotrophic organisms. Our findings highlight the impact of environmental gradients on microbial diversity and the functional dynamics of biocrusts, emphasizing their critical role in Arctic tundra ecosystems. Arctic biocrusts are intricate micro-ecosystems, whose structure is strongly shaped by local physicochemical parameters, likely affecting essential ecological functions.