This study involves the application of new phase separated biological pretreatment (PSBP) strategy on microalgal biomass using the nickel nanoparticle induced cellulase secreting bacterial disintegration. Particularly, interest was focussed on cell wall weakening (CWW) of microalgae biomass besides the cell disintegration (CD) and release of organics. During CWW, protein, carbohydrate, cellulose, hemicellulose and DNA were used as evaluation indexes. Similarly, during CD, soluble chemical oxygen demand was used as evaluation index to assess the disintegration effect. A higher CWW was achieved at nickel nanoparticle (Np) dosage of 0.004 g/g SS. During CD, a clear demarcation in biomass solubilisation was achieved by PSBP (36%) than the sole biological pretreatment -BP (24%). The biomethanogenesis test results showed that enhanced methane production of 411 mL/g COD was achieved by PSBP than BP. Energy analysis showed that a higher net energy production of 6.467 GJ/d was achieved by PSBP.