Nannochloropsis oceanica is a microalgal species that has high potential in both lipid and protein accumulation capacity. For this reason, it has been receiving attention as a microalgal candidate for biofuel production. This study evaluated the effect of six different protein extraction treatments on defatted Nannochloropsis, whole soybean, and soybean meal samples (25mL, 10% dry weight), by performing nutrients assays that measure soluble protein extraction rate (%), inorganic phosphorus, soluble glucose, and free iron in the extracted supernatant solution. Alkaline boiling, protease digestion, and lipase digestion are found to be effective in releasing 44.6%, 31.48% and 25.6% total protein, respectively. Three combination treatments are then designed based on the extraction efficiency single treatments. Although combination treatment has increased the soluble protein yields in WSB and SBM samples (up to 66.17% and 80.06%, respective), no further improvement in DN soluble protein yield is found (at max 44.69% released, resembling the yield in alkaline single treatment). This indicates that the three combination treatments did not further disintegrate microalgal structure for nutrient release. Two-way ANOVA analysis shows that both sample type and treatment method have statistically significant effect on protein extraction rates. Soluble forms of inorganic phosphorus, glucose, and free iron were extracted, yet were only detected in relatively low values (less than 10% of total predicted), regardless of treatment method applied.