The popular Saccharomyces cerevisiae laboratory strain CEN.PK113-7D is regularly grown in chemically defined also referred to synthetic media (SM) that contain a set of organic compounds known as class B vitamins to support fast growth. While previous work showed how specific vitamin requirements could be eliminated through directed evolution, this approach is still limited to the biotin requirement. In this study, the CEN.PK113-7D vitamin requirements were evaluated and showed that growth was suboptimal upon omission of thiamine, pyridoxine, para-aminobenzoic acid, pantothenic acid, inositol, or nicotinic acid. The S. cerevisiae strain was then evolved for fast growth in the absence of one of the vitamins. In all evolution lines, strains reached growth rates comparable to those of the strain grown in the presence of the vitamin. After few generations on medium without myo-inositol, nicotinic acid or pABA, CEN.PK113-7D exhibited fast prototrophic growth, conversely evolution for thiamine, pyridoxine and pantothenate had to