Baker’s yeast (Saccharomyces cerevisiae) and other lignocellulosic materials use as energy producing biomass have improved over time. These to date have continued to play a massive and quality role in the developmental needs of energy for sustainable future. Baker’s yeast dominance in this energy trends has shown to be very productive as part realization of the set goal ahead. This through current research has shown to be positively supportive in the tackling of the rising greenhouse gas emissions (GHG) and global warming. In this study, the mechanical pre-treatment of yeast substrate was conducted through its homogenization in the high-pressure homogenizer (HPH). With the operating conditions of temperature, pressure and number of passes (cycles) ranging from (30 - 50 °C), (30 – 90 MPa) and (1-5) respectively, this was ensured that the HPH machine maximum pressure of 100MPa was not exceeded. The homogenized yeast was diluted in three levels of dilutions with buffer solutions; denoted as solution C in the ratio of; 10:90, 20:80; and 30:70 respectively. The resultant protein concentration yield so obtained was analyzed and optimized through using the Design Expert V.8 in the comparison of the three samples in the ratios given with the 3 input factors of pressure, temperature and number of cycles estimated within the considered limits.