Coconut presscake is a byproduct of processing VCO which has a relatively high protein content of 24.22%. Coconut presscake protein not fully utilized. Coconut presscake protein have functional properties that can affect the characteristics of food products. This study aims to determine the separation method of VCO which can produce protein isolate withthe best functional properties. Three methods of separation are compared: chemical, physical and enzymatic method. In the chemical method with the precipitation of protein in coconut milk at the isoelectric point using acid. In the physical method by solving coconut milk emulsion system with the addition of coconut oil to increase the interfacial tensionof protein-water. Enzymatic method using a protease that can break the chains of peptides of proteins into simpler molecules. Parameters were measured protein content and functional properties of protein isolates of coconut presscake of VCO covering Emulsion Activity Index (EAI), Emulsion Stability Index (ESI), Water Holding Capacity (WHC), Oil Holding Capacity (OHC), Hidrophylic Lipophylic Balance (HLB). Data were analyzed by analysis of variance and Honestly Significant Difference test at 5% level. The results showed that the protein isolate of coconut presscakeof VCO made by physical processed yealded the highest protein content of 95.12 ± 2.9% db and the best functional properties EAI (37.87 ± 6.6 m2/g) and HLB (42.87 ± 1.2%) value is highest as compared to chemical and enzymatic processed. ESI,WHC and OHC showed no significant differences between methods (p≤ 0.05). The results of this studyconcluded that the physical method produces the best functional properties.Keywords: Functional properties, protein isolate,coconut presscake, VCO ABSTRAKBlondo merupakan hasil samping pengolahan VCO yang memiliki kadar protein cukup tinggi sebesar 24,22%. Protein blondo belum dimanfaatkan secara maksimal. Protein blondo memiliki sifat fungsional yang dapat mempengaruhi karakteristik produk pangan. Penelitian ini bertujuan untuk menentukan metode pemisahan VCO yang menghasilkanisolat protein dengan sifat fungsional terbaik. Tiga metode pemisahan yang dibandingkan: metode kimia, fisik dan enzimatis. Pada metode kimia dengan pengendapan protein dalam santan pada titik isoelektris dengan menggunakan asam. Pada metode fisik dengan pemecahan sistem emulsi santan dengan penambahan minyak kelapa untuk memperbesar tegangan antar muka protein-air. Metode enzimatis dengan menggunakan protease yang dapat memutus rantai peptidadari protein menjadi molekul yang lebih sederhana. Paramater yang diamati adalah kadar protein, dan sifat fungsional isolat protein blondo VCO yang meliputi Indeks Aktivitas Emulsi (IAE), Indeks Stabilitas Emulsi (ISE), Water Holding Capacity (WHC), Oil Holding Capacity (OHC), Hidrophylic Lipophylic Balance (HLB). Data hasil penelitian dianalisisdengan analisis keragaman dan uji beda nyata jujur pada taraf 5%. Hasil penelitian menunjukkan bahwa isolat protein blondo VCO yang dibuat dengan metode fisik memberikan kadar protein (95,12 ± 2,9 %db), nilai IAE (37,87 ± 6,6 m2/g) dan nilai HLB (42,87 ± 1,2%) tertinggi dibandingkan dengan metode kimia dan enzimatis. ISE,WHC danOHC tidak menunjukkan perbedaan yang signifikan antar metode (p≤ 0,05). Hasil penelitian ini menyimpulkan bahwa metode fisik menghasilkan sifat fungsional terbaik.Kata kunci: Sifat fungsional, isolat protein, blondo, VCO