A comparative analysis of the amount and composition of the proteins adsorbed onto the surface of silver-sulfide (NpAg2S) nanoparticles obtained by biosynthesis using bacteria was carried out for the first time. These were the gram-negative bacteria Shewanella oneidensis MR-1 and Escherichia coli K12, as well as the gram-positive bacterium Bacillus subtilis 168. The biosynthesis of NpAg2S was carried out in nutrient broth with 1 mM of AgNO3 and Na2S2O3 · 5H2O salts in the presence of bacterial cells under aerobic conditions. Analysis of NpAg2S by transmission electron microscopy showed that the particles were spherical and had an average diameter of 8 ± 2 nm for S. oneidensis MR-1 and E. coli K12 and 10 ± 3 nm for B. subtilis 168. It was found that the highest amount of protein was sorbed on NpAg2S, when the strain B. subtilis 168 was used, and the smallest amount was sorbed by E. coli. The main proteins adsorbed on NpAg2S were determined by the MALDI TOF/TOF method, and the heterogeneity of the protein coating was revealed. The lowest heterogeneity of proteins on the surface of nanoparticles is observed in the case of B. subtilis (only one protein, flagellin, predominates); the highest heterogeneity of proteins was found on nanoparticles obtained using S. oneidensis MR-1. All proteins covering the surface of NpAg2S were shown to be outer-membrane or cytoplasmic-membrane proteins of the studied bacteria. The composition of the protein coating of nanoparticles is individual and constant for each bacterial strain used. The values of the ζ-potential and the effective diameter of nanoparticles were shown to differ depending on the “protein corona” of the strain that was used to obtain NpAg2S. The characteristic of the “protein corona” of biologically obtained nanoparticles is an important and necessary condition for their practical application. © 2017, Allerton Press, Inc.