Biological interfacing of graphene has become crucial to improve its biocompatibility, dispersability, and selectivity. However, biofunctionalization of graphene without yielding defects in its sp(2)-carbon lattice is a major challenge. Here, a process is set out for biofunctionalized defect-free graphene synthesis through the liquid phase ultrasonic exfoliation of raw graphitic material assisted by the self-assembling fungal hydrophobin Vmh2. This protein ( extracted from the edible fungus Pleurotus ostreatus) is endowed with peculiar physicochemical properties, exceptional stability, and versatility. The unique properties of Vmh2 and, above all, its superior hydrophobicity, and stability allow to obtain a highly concentrated (approximate to 440-510 mu g mL(-1)) and stable exfoliated material (zeta-potential, + 40/+ 70 mV). In addition controlled centrifugation enables the selection of biofunctionalized few-layer defect-free micrographene flakes, as assessed by Raman spectroscopy, atomic force microscopy, scanning electron microscopy, and electrophoretic mobility. This biofunctionalized product represents a high value added material for the emerging applications of graphene in the biotechnological field such as sensing, nanomedicine, and bioelectronics technologies.