The interaction of materials with their surroundings proceeds mainly through their outer functional groups. Coating of solid materials with thin organic films using the Layer-by-Layer (LbL) assembly strategy is one of the most versatile approaches to fabricate functional surfaces for a wide range of applications. In this contribution, the functionalization of silica microparticles with LbL films containing poly(N,N-dimethylaminoethyl methacrylate) and polysaccharides (sodium alginate, carboxymethyl cellulose and chitosan) is investigated. All polymers used in this work are biocompatible, while the deposition strategy is environmentally benign and allows a precise control of the outer layer properties (chemical nature, charge, roughness, etc.). The application of the obtained films in immobilization of proteins/enzymes (i.e. pepsin, bovine serum albumin, hemoglobin and lysozyme), sorption/release of organic dyes (i.e. methylene blue, methyl orange, bromocresol green and Congo red), as well as their use as “green” catalysts for the synthesis of silver nanoparticles was investigated as a function of films’ composition, number of deposited layers and/or assembly conditions.