In a variety of applications, functionalization of gold nanoparticles is needed to ensure adequate surface charge and hydrophobicity for their biodistribution, interparticle interactions, or self-organization. In the present paper, we provide an economic way for the synthesis of hydrophobized poly(ethylenimine) (PEI) capped gold nanoparticles at room temperature using sodium dodecyl sulfate (SDS). The approach is based on the controlled competition between the nucleation of gold nanophases within the PEI molecules and the SDS binding onto their amine groups. This can be achieved via utilizing the strongly irreversible nature of the association between the oppositely charged polymer and that of the surfactant molecules. Specifically, by varying the order and timing of SDS addition during the process of gold nanoassembly formation, the size distribution, the morphology, and the local hydrophobic environment of the produced Au-PEI/SDS nanohybrids can be tuned even at one composition of the system. The results may be further exploited for the preparation of noble metal nanoassemblies with controlled hydrophobicity and charge.