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UJI AKTIVITAS ANTIBAKTERI ISOLAT DAN HIDROLISAT PROTEIN KERANG DARAH (Anadara Granosa L)

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UJI AKTIVITAS ANTIBAKTERI ISOLAT DAN HIDROLISAT PROTEIN KERANG DARAH (Anadara Granosa L) UJI AKTIVITAS ANTIBAKTERI ISOLAT DAN HIDROLISAT PROTEIN KERANG DARAH (Anadara granosa L) SKRIPSI HALAMAN JUDUL OLEH : VIVIENNE WIJAYA NIM 171501050 PROGRAM SARJANA FARMASI FAKULTAS FARMASI UNIVERSITAS SUMATERA UTARA MEDAN 2021 i UJI AKTIVITAS ANTIBAKTERI ISOLAT DAN HIDROLISAT PROTEIN KERANG DARAH (Anadara granosa L) SKRIPSI Diajukan sebagai salah satu syarat untuk memperoleh gelar Sarjana Farmasi pada Fakultas Farmasi Universitas Sumatera Utara OLEH : VIVIENNE WIJAYA NIM 171501050 PROGRAM SARJANA FARMASI FAKULTAS FARMASI UNIVERSITAS SUMATERA UTARA MEDAN 2021 ii PENGESABAN SKRIPSI un AK11VIT AS ANTIBAKTERI ISOLAT DAN BIDROLISAT PROTEIN KERANG DARAH (A • JNII.. L) OLI.II: VIVIENNE WUA YA NIM 171!11158 Dipertahanbn di Hadapln Penguji Skripli Fakultas Fanwi Uui\'asibd Sumlten Utara pada TIIIIPI: OI Juni 2021 Disetujui oleh: b"ig) Panitia "'-JL :· ~ ~Sri Yul" i. S.Fann.,M.Si., Apt Prof.Dr.rer.nat.Effi:ady De Lux Pun, SU., Apt NIP. 198207032001122002 NIP. 195306191913031001 Pembimbing II. Sri ~.Fam.,M.Si, Apt NIP. l 98207032008122002 YodeW:.::.-:.Farm, M.Si, Apt. NIP. 191704212017042001 Dr. SitOnls, M.Si., Apt. Ketua Pqnm Studi Sarjana Farmasi, NIP. 195310301980031002 Dr. Sumaiyah,- .Si., Apt. ftnlna1a, S.f11111., M.Si .. Apt. NIP. l 977 l 2262008122002 7042120 17042001 ., ~ Pa.I)., Apt. l S2008122001 iii KATA PENGANTAR Puji syukur kehadirat Tuhan Yang Maha Esa yang telah memberikan rahmat dan karunia-Nya sehingga penulis dapat menjalani penelitian hingga akhirnya menyelesaikan penyusunan skripsi yang berjudul “Uji Aktivitas Antibakteri Isolat dan Hidrolisat Protein Kerang Darah (Anadara granosa L.)”. Skripsi ini diajukan sebagai salah satu syarat untuk memperoleh gelar Sarjana Farmasi pada Fakultas Farmasi Universitas Sumatera Utara. Penulis menyampaikan ucapan terimakasih kepada Ibu Sri Yuliasmi, S.Farm., M.Si., Apt. selaku dosen pembimbing I dan Ibu Yade Metri Permata, S.Farm., M.Si., Apt. selaku dosen pembimbing II , yang telah membimbing dengan penuh kesabaran, tulus dan ikhlas selama penelitian dan penulisan skripsi ini berlangsung. Penulis juga berterimaksih kepada Bapak Prof. Dr.rer.nat. Effendy De Lux Putra, SU., Apt. dan Bapak Dr. Panal Sitorus., M.Si., Apt. selaku dosen penguji yang telah memberikan kritik dan saran yang membangun demi kelengkapan skripsi ini. Pada kesempatan ini, penulis juga mengucapkan terimakasih kepada Dekan Fakultas Farmasi Universitas Sumatera Utara, Ibu Khairunnisa, S.Si., M.Pharm., Ph.D., Apt., yang telah memberikan fasilitas selama masa pendidikan. Penulis juga menyampaikan rasa terimakasih dan penghargaan yang tulus kepada kedua orang tua, Ayahanda Ng Kian Tjai dan Ibunda Ong Kui Bie, kepada abang saya David, kakak yaitu Fiona dan Intan serta adik saya Vonny, atas doa, dukungan dan pengorbanan baik moril maupun materil selama perkuliahan hingga penyelesaian skripsi ini. Penulis juga ingin mengucapkan terimakasih kepada rekan penelitian, sahabat dan teman-teman yang telah memberikan dukungan selama masa perkuliahan, penelitian dan penulisan skripsi ini berlangsung. iv Skripsi ini masih jauh dari kata sempurna, oleh karena itu penulis meminta maaf atas kesalahan dan kekhilafan dalam penulisan skripsi. Penulis bersedia menerima kritik dan saran yang membangun dari pembaca. Semoga skripsi inii bisa memberikan sumbangsih untuk menambah pengetahuan para pembaca dan berguna untuk ilmu pengetahuan. Medan, 08 Juni 2021 Vivienne Wijaya NIM 171501050 v SURAT PERNYATAAN ORISINILITAS Saya yang bertanda tangan dibawah ini: Nama : Vivienne Wijaya Nomor Induk Mahasiswa : 171501050 Program Studi : Sarjana Farmasi Judul Skripsi : Uji Aktivitas Antibakteri Isolat dan Hidrolisat Protein Kerang Darah (Anadara granosa L.) Dengan ini menyatakan bahwa skripsi yang saya buat adalah asli karya sendiri dan bukan plagiat. Apabila dikemudian hari diketahui bahwa skripsi saya tersebut terbukti plagiat karena kesalahan sendiri, maka saya bersedia diberi sanksi apapun oleh Program Studi Sarjana Farmasi Fakultas Farmasi Universitas Sumatera Utara. Saya tidak akan menuntut pihak manapun atas perbuatan saya tersebut. Demikian surat pernyataan ini saya perbuat dengan sebenarnya dan dalam keadaan sehat. Medan, 08 Juni 2021 Vivienne Wijaya NIM 171501050 vi UJI AKTIVITAS ANTIBAKTERI ISOLAT DAN HIDROLISAT PROTEIN KERANG DARAH (Anadara granosa L) ABSTRAK Latar Belakang : Selama dekade terakhir beberapa tipe antibiotika yang dikembangkan untuk keperluan klinis belum cukup untuk mengatasi laju kecepatan resistensi bakteri. Oleh karena itu, dicari alternatif pengganti antibiotika yang aman dan tidak menimbulkan efek samping yang merugikan, salah satunya adalah peptida antimikroba (Antimicrobial peptides). Kerang darah (Anadara granosa L) adalah salah satu jenis bivalvia, dan merupakan sumber protein hewani yang tergolong dalam Complete Protein, karena kadar asam amino essensialnya yang tinggi. Tujuan : Tujuan dari penelitian ini yaitu untuk mengetahui kandungan protein terlarut serta aktivitas antibakteri dalam isolat dan hidrolisat protein kerang darah (Anadara granosa L) terhadap bakteri Escherichia coli dan Staphylococcus aureus. Metode : Pada penelitian ini dilakukan proses isolasi protein kerang darah (Anadara granosa L) menggunakan larutan buffer A (Tris-HCl 0,1 M pH 8,3; NaCl 2 M; CaCl2 0,01 M; β-mercaptoetanol 1%, Triton X-100 0,5%) dan dilakukan proses hidrolisis dengan enzim tripsin. Pengukuran kadar protein ditentukan berdasarkan metode Lowry dengan menggunakan BSA (Bovine Serum Albumin) sebagai larutan standar. Pengujian aktivitas antibakteri dilakukan dengan metode difusi agar terhadap Escherichia coli dan Staphylococcus aureus dengan menentukan Konsentrasi Hambat Minimum. Hasil : Penelitian ini menunjukkan bahwa kerang darah mengandung protein dengan konsentrasi protein pada isolat yaitu 32,445 mg/ml, hidrolisat 1 jam yaitu 34,65 mg/ml, hidrolisat 3 jam yaitu 36,435 mg/ml, dan pada hidrolisat 5 jam yaitu 39,48 mg/ml. Pada hasil pengujian, aktivitas antibakteri tertinggi terdapat pada hidrolisat 5 jam terhadap Escherichia coli dan Staphylococcus aureus masing – masing sebesar 12,23 ± 0,12 mm dan 14,03 ± 0,15 mm pada konsentrasi 100%. Dengan konsentrasi hambat minimum pada isolat yaitu 50%, pada hidrolisat 1 jam yaitu 50%, hidrolisat 3 jam yaitu 25% dan pada hidrolisat 5 jam yaitu 25% terhadap Escherichia coli dan Staphylococcus aureus. Kesimpulan : Kerang darah (Anadara granosa L) memiliki kandungan protein yang berbeda pada masing – masing perlakuan dan memiliki aktivitas antibakteri terhadap Escherichia coli dan Staphylococcus aureus. Kata kunci : Kerang darah, Protein, Hidrolisat Protein, Antibakteri, Escherichia coli, Staphylococcus aureus. vii ANTIBACTERIAL ACTIVITY OF ISOLATE AND HYDROLYSATE PROTEIN FROM BLOOD COCKLES (Anadara granosa L) ABSTRACT ABSTRACT Background: Over the last decade several types of antibiotics were developed for clinical purposes has not been enough to overcome bacterial resistance rate of speed. Therefore, alternatives to antibiotics that are safe and do not cause adverse side effects are sought, one of which is antimicrobial peptides (AMPs). Blood cockle (Anadara granosa L) is a type of bivalve and is a source of animal protein which is classified as Complete Protein, because of its high essential amino acid levels. Objective: The purpose of this study was to determine the dissolved protein content and antibacterial activity in blood cockle (Anadara granosa L) protein isolates and hydrolyzates against Escherichia coli and Staphylococcus aureus bacteria. Method: In this study, the isolation process of blood cockle protein (Anadara granosa L) was carried out using buffer A (Tris-HCl 0,1 M pH 8,3; NaCl 2 M; CaCl2 0,01 M; β-mercaptoetanol 1%, Triton X-100 0,5%) solution and hydrolysis was carried out with trypsin enzyme. Measurement of protein content was determined based on the Lowry method using BSA (Bovine Serum Albumin) as the standard solution. Antibacterial activity was carried out by the agar diffusion method against Escherichia coli and Staphylococcus aureus by determining the Minimum Inhibitory Concentration. Results: This study showed that blood cockles contained a protein with a protein concentration in the isolate that was 32.445 mg/ml, 1-hour hydrolyzate was 34.65 mg/ml, 3 hours hydrolyzate was 36.435 mg/ml, and 5 hours hydrolyzate was 39.48 mg/ml. In the test results, the highest antibacterial activity was found in the 5-hour hydrolyzate against Escherichia coli and Staphylococcus aureus, respectively 12.23 ± 0.12 mm and 14.03 ± 0.15 mm at a concentration of 100%. The minimum inhibitory concentration of the isolate is 50%, the 1-hour hydrolyzate is 50%, the 3 hours hydrolyzate is 25%, and the 5 hours hydrolyzate is 25% against Escherichia coli and Staphylococcus aureus. Conclusion: Blood cockles (Anadara granosa L) contained different proteins in each treatment and had antibacterial activity against Escherichia coli and Staphylococcus aureus. Keywords: Blood cockle, Protein, Protein Hydrolysate, Antibacterial, Escherichia coli, Staphylococcus aureus. viii DAFTAR ISI HALAMAN JUDUL ................................................................................................ i HALAMAN PENGESAHAN ................................................................................ iii KATA PENGANTAR ........................................................................................... iii SURAT PERNYATAAN ORISINILITAS............................................................ vi ABSTRAK ...........................................................................................................
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