<P><B>Abstract</B></P> <P>Microalgae have been considered as one of the most promising biomass for the generation of biofuels. The anaerobic digestion (AD) has been proved to be a promising technique to transfer the microalgal biomass into biofuels. Previous study demonstrated that anaerobic pretreatment of microalgae biomass by <I>Bacillus licheniformis</I> could improve methane production. In this study micro-aerobic bio-pretreatment of microalgal biomass by the facultative anaerobic bacteria <I>Bacillus licheniformis</I> was invested with different loads of oxygen supplied. The bio-hydrogen and biomethane productions were tested to calculate total energy outcomes. The transmission electron microscope (TEM) photographs suggested that the novel micro-aerobic bio-pretreatment (MBP) could effectively damage the firm cell wall of algal cells. The processing time of the novel method (24h) was less than the previous anaerobic pretreatment (60h). Results showed that the group with 5mL oxygen/g VS<SUB>fed</SUB> had the highest total energy outcomes, which was 17.6% higher than that of the anaerobic pretreatment.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The micro-aerobic pretreatment was used to improve energy yield of <I>Chlorella</I> sp. </LI> <LI> The <I>Bacillus licheniformis</I> was confirmed to damage the cell wall of microalgae. </LI> <LI> Obtained energy from <I>Chlorella</I> sp. was improved by 12.3%. </LI> <LI> Pretreatment time was decreased from 60h to 24h. </LI> <LI> The VS degradation efficiency was increased from 75.7% to 82.1%. </LI> </UL> </P>