Optimasi Ekstraksi Antimicrobial Peptide dari Daun Gandarusa Menggunakan Box-Behnken Design

Abstract

Antimicrobial Peptide (AMP) memiliki aktivitas spektrum luas dan dapat membunuh bakteri yang resistan terhadap berbagai obat. Daun gandarusa (Justicia gendarussa Burm. F.) memiliki aktivitas antibakteri namun kajian terkait antimicrobial peptide pada tanaman tersebut belum dilakukan. Penelitian ini bertujuan menentukan kondisi optimum ekstraksi serta mengidentifikasi AMP daun gandarusa yang berpotensi memiliki aktivitas antibakteri. Rancangan optimasi ekstraksi dibuat menggunakan Box-Behnken Design. Kadar total protein diukur dengan metode Bradford, identifikasi AMP dilakukan dengan metode SDS-PAGE, dan pengujian aktivitas antibakteri dilakukan dengan metode difusi sumuran. Hasil penelitian menunjukkan kondisi optimum ekstraksi diperoleh dari kombinasi konsentrasi pelarut 14,997%, waktu ekstraksi 2,998 jam, dan rasio antara sampel dengan pelarut yaitu 1:3. Identifikasi dengan SDS-PAGE menunjukkan AMP terduga dari daun gandarusa memiliki berat molekul sebesar 11 kDa, pengujian aktivitas antibakteri ekstrak kasar terhadap Escherichia coli dan Staphylococcus aureus tidak menunjukkan adanya daya hambat.

Antimicrobial Peptide (AMP) has broad-spectrum activity and can kill multidrug-resistant bacteria. Gandarusa leaves (Justicia gendarussa Burm. F.) have antibacterial activity, but studies regarding antimicrobial peptides in this plant have not been carried out. This study aimed to determine the optimum conditions of extraction and identify AMPs of gandarusa leaves that have potential antibacterial activity. Extraction optimization design was made using Box-Behnken Design. Total protein levels were measured by the Bradford method, AMP identification was carried out by the SDS-PAGE method, and antibacterial activity testing was carried out by the well diffusion method. The results showed that the optimum extraction conditions were obtained from a combination of 14,997% solvent concentration, 2,998 hours extraction time, and the ratio between sample and solvent of 1:3. Identification with SDS-PAGE showed that AMP from gandarusa leaves has a molecular weight of 11 kDa, testing the antibacterial activity of crude extracts against Escherichia coli and Staphylococcus aureus showed no inhibition.