To probe the effect of kelp waste extracts (KWE) combined with acetate on biochemical composition of Chlorella sorokiniana, the cultures were performed under independent/combined treatment of KWE and acetate. The results showed that high cell density and biomass were obtained by KWE combined with acetate treatments, whose biomass productivity increased by 79.69-102.57% and 20.04-35.32% compared with 3.0gL-1 acetate and KWE treatments respectively. The maximal neutral lipid per cell and lipid productivity were gained in KWE combined with 3.0gL-1 acetate treatment, which increased by 16.32% and 129.03% compared with 3.0gL-1 acetate, and 253.35% and 70.74% compared with KWE treatment. Meanwhile, C18:3n3 and C18:2n6c contents were reduced to 4.90% and 11.88%, whereas C16:0 and C18:1n9c were improved to 28.71% and 37.76%. Hence, supplementing appropriate acetate in KWE cultures is supposed to be a great potential method for large-scale cultivation of C. sorokiniana to generate biofuel.