Different yeast species in distinct clades within Saccharomycetaceae family are interesting model organisms for inter-species comparative proteomics, in regard to different metabolic properties and genomic architectures. The aim in this analysis is revealing conserved and diverse expression profile of proteome under different culture conditions, including fermentative and non-fermentative growth. Our label-free quantitative proteomics analysis not only uncovered species-specific characters of metabolic proteins expression pattern and also provide intriguing insight into optimized protein cost allocation and evolutionary role for duplicated genes.