Aktivitas Antioksidan dan Antibakteri Peptida Bioaktif Daging Ayam IPB D1 Hasil Hidrolisis dengan Enzim Kimotripsin

Abstract

Ayam IPB D1 merupakan galur ayam lokal unggul yang memiliki kandungan protein tinggi dan berpotensi dikembangkan sebagai sumber senyawa fungsional. Selama ini pemanfaatannya masih terbatas, padahal fraksi protein ototnya, khususnya fraksi sarkoplasmik yang larut dalam air, berpotensi menghasilkan peptida bioaktif melalui proses hidrolisis enzimatik. Penelitian ini bertujuan untuk mengembangkan peptida bioaktif dari fraksi sarkoplasmik ayam IPB D1 menggunakan enzim kimotripsin, serta mengevaluasi aktivitas antioksidan dan antibakterinya. Penelitian dilakukan melalui beberapa tahap, yaitu ekstraksi fraksi sarkoplasmik dari daging dada ayam IPB D1, hidrolisis menggunakan kimotripsin dengan variasi konsentrasi dan waktu inkubasi, analisis derajat hidrolisis, analisis total protein dan SDS-PAGE, serta pengujian aktivitas antioksidan dan antibakteri. Hidrolisat dengan derajat hidrolisis optimum kemudian difraksinasi menggunakan membran ultrafiltrasi =3 kDa untuk memperoleh peptida berukuran kecil. Aktivitas antioksidan diuji menggunakan metode DPPH, sedangkan aktivitas antibakteri diuji terhadap bakteri Gram-positif dan Gram-negatif menggunakan metode difusi cakram. Hasil menunjukkan bahwa kombinasi 5 U mL?¹ kimotripsin dan 6 jam inkubasi menghasilkan derajat hidrolisis tertinggi sebesar 17,93%, serta menurunkan total protein secara signifikan. SDS-PAGE menunjukkan bahwa protein utama dalam fraksi sarkoplasmik telah terdegradasi setelah hidrolisis, menghasilkan peptida kecil yang tidak terdeteksi dalam gel. Fraksi dengan berat molekul <3 kDa menunjukkan peningkatan aktivitas antioksidan dan antibakteri, terutama terhadap bakteri Gram negatif. Hal ini menunjukkan bahwa peptida kecil hasil hidrolisis berkontribusi besar terhadap bioaktivitas yang diamati. Penelitian ini menunjukkan bahwa fraksi sarkoplasmik ayam IPB D1 dapat dimanfaatkan sebagai sumber peptida bioaktif dengan aktivitas antioksidan dan antibakteri. Penggunaan enzim kimotripsin memungkinkan optimalisasi protein lokal menjadi bahan fungsional bernilai tambah tinggi, mendukung pengembangan pangan fungsional atau suplemen nutrasetikal dari hasil unggas lokal.

IPB D1 chicken is a superior local chicken strain with high protein content and potential to be developed as a source of functional compounds. Its utilization has been limited so far, despite the fact that its muscle protein fraction, particularly the water-soluble sarcoplasmic fraction, has the potential to produce bioactive peptides through enzymatic hydrolysis. This study aimed to develop bioactive peptides from the sarcoplasmic fraction of IPB D1 chicken using the enzyme chymotrypsin and to evaluate their antioxidant and antibacterial activities. The research was conducted in several stages, including extraction of the sarcoplasmic fraction from IPB D1 chicken breast, hydrolysis using chymotrypsin with variations in enzyme concentration and incubation time, analysis of the degree of hydrolysis, total protein content, SDS-PAGE profiling, and evaluation of antioxidant and antibacterial activities. The hydrolysate with the optimum degree of hydrolysis was further fractionated using an ultrafiltration membrane =3 kDa to obtain small-sized peptides. Antioxidant activity was tested using the DPPH method, while antibacterial activity was assessed against Gram-positive and Gram-negative bacteria using the disc diffusion method. The results showed that the combination of 5 U mL?¹ chymotrypsin and 6 hours of incubation produced the highest degree of hydrolysis at 17.93%, along with a significant reduction in total protein content. SDS-PAGE analysis confirmed that major proteins in the sarcoplasmic fraction were degraded after hydrolysis, resulting in small peptides that were not detectable on the gel. The <3 kDa peptide fraction demonstrated increased antioxidant and antibacterial activity, particularly against Gram-negative bacteria. These findings indicate that the small peptides produced through hydrolysis significantly contribute to the observed bioactivity. This study demonstrates that the sarcoplasmic fraction of IPB D1 chicken can be utilized as a source of bioactive peptides with antioxidant and antibacterial properties. The use of specific proteolytic enzymes such as chymotrypsin enables the optimization of local animal protein sources into high-value functional ingredients, supporting the development of functional foods or nutraceutical supplements from local poultry products.