Eksplorasi Protein Larva Black Soldier Fly (Hermetia illucens) Menggunakan Trans-Proteomic Pipeline

Abstract

Kebutuhan akan protein kian meningkat dalam beberapa tahun terakhir. Larva lalat tentara hitam (BSF) telah banyak dipelajari terkait kemampuan bio-konversinya serta optimalisasi kultur biakannya; sementara itu, pengetahuan terkait informasi genetik serta proteomiknya masih terbatas. Studi ini bertujuan mengevaluasi metode hidrolisis berdasarkan jumlah protein bervariasi, identifikasi protein yang memiliki efektivitas di bidang biomedis, serta rekomendasi metode hidrolisis yang menghasilkan jumlah protein biomedis terbanyak. Larva BSF dihidrolisis dengan empat perlakuan berbeda menggunakan enzim tripsin dengan waktu inkubasi 3, 5, dan 7 jam dan enzim papain dengan durasi inkubasi 5 jam. Kemudian dianalisis menggunakan LC-MS/MS diikuti dengan analisis trans-proteomic pipeline. Metode hidrolisis yang menghasilkan jumlah protein terbanyak dan bervariasi adalah metode yang menggunakan enzim papain dengan waktu inkubasi selama 5 jam (150 protein), diikuti oleh metode dengan tripsin 7 jam (127 protein), tripsin 3 jam (95 protein), serta tripsin 5 jam (68 protein). Protein dalam hidrolisat larva BSF yang memiliki efektivitas di bidang biomedis meliputi protein Nazo, autophagy-related protein 13, coronin, protein yellow, cytochrome P450, dan kolagen. Rekomendasi metode hidrolisis untuk menghasilkan protein dengan efektivitas biomedis terbanyak adalah tripsin 3 jam.

The demand for protein has kept increasing in recent years. Black soldier fly (BSF) larvae have been studied for their bioconversion and optimization of medium culture; meanwhile, there is still limited knowledge of genetic and proteomic information from these larvae. This study aims to explore the proteomic information from BSF larvae and evaluate the hydrolysis methods based on the amount of protein produced and the number of unique proteins. The BSF larvae were hydrolyzed with four different treatments using trypsin incubated for 3, 5, and 7 hours and using papain incubated for 5 hours. Then, they were analyzed using LC-MS/MS, followed by trans-proteomic pipeline analysis. The hydrolysis method that has the highest number of identified proteins was papain incubated for 5 hours (150 proteins), followed by trypsin incubated for 7 hours (127 proteins), trypsin for 3 hours (95 proteins), and lastly, trypsin for 5 hours (68 proteins). Several proteins from BSF larval hydrolysate that have biomedical effectivity include Nazo protein, autophagy-related protein 13, coronin, yellow protein, cytochrome P450, and collagen. The recommended hydrolysis method to produce the highest number of proteins with biomedical effectivity was trypsin incubated for 3 hours.