The cytoplasm is a dynamic, self-organizing system spanning scales from organelles to biomolecular condensates and multienzyme assemblies. While enzymatic regulation has traditionally been attributed to transcriptional control and allostery, emerging evidence highlights the cytoplasm's physical properties─macromolecular crowding, viscosity, and diffusion─as a crucial yet underappreciated regulatory layer. These biophysical factors actively modulate enzymatic activity and metabolic flux, maintaining homeostasis and enabling adaptive cellular responses. This Perspective explores the biochemical interplay between the cytosolic environment and enzymatic function, emphasizing how cytoplasmic organization contributes to metabolic regulation.