Judul: Production and Immobilization of Pectinase from Bacillus sp. 2P11 Using Alginate Beads
Abstrak: Enzymes are biocatalysts with high catalytic power that have the potential to be used in the industrial field. Pectinase is an enzyme that degrades plant cell walls and is commonly used in the food industry to accelerate the extraction of fruit juices. Microbes are a source of enzymes that are often used because they grow on various media and can easily be increased by adjusting the growth conditions. Immobilization is the stage of developing a sustainable bioprocess by reusing enzymes bound to a matrix. Therefore, this study aimed to immobilize the pectinase enzyme and the efficiency of repeated use of the immobilized pectinase. This research begins with the rejuvenation of Bacillus sp. 2P11, which has been isolated from previous studies. The cocoa husk contains pectin, which can be used as a substrate to produce pectinase. Pectinase production was carried out by growing Bacillus sp. 2P11 on liquid production media with a cocoa liquid substrate with variations (1-15%) was tested for enzyme activity. Crude extract enzyme activity was obtained by optimizing microbial growth on the media by making an enzyme production curve. The enzyme production curve makes with an incubation time of 96 hours and observations every 12 hours. Enzyme activity was measured using Dinitrosalicylic Acid (DNS) reagent with a standard solution of galacturonic acid. Precipitation was carried out with ammonium sulfate salt and then dialyzed. The enzyme was precipitated gradually from 0% to the ammonium sulfate saturation level of 80%. Dialysis was carried out by inserting a cellophane tube containing the enzyme into a 0,05 M pH 5 acetate buffer. The purified pectinase was then immobilized by trapping using sodium alginate and CaCl2. Immobilized pectinase stored for 14 days was measured for its enzyme activity repeatedly. The activity of the crude extract of the pectinase enzyme obtained was 0,41 U/mL. Precipitation with ammonium sulfate and dialysis increased pectinase activity by 0,45 U/mL and 0,58 U/mL, respectively. Enzymes can be used repeatedly with enzyme immobilization. In this study, the immobilization of the enzyme entrapment method with sodium alginate and CaCl2 was designed with the response surface method. The optimum concentration of sodium alginate was 0,5%, and CaCl2 0,3 M, with the percentage of an immobilized enzyme was 40,7%, and pectinase activity was 0,23 U/mL. Stability and repeated use of immobilized pectinase can be done up to 5 times., Enzim adalah biokatalis dengan daya katalik tinggi yang memiliki potensi untuk digunakan pada bidang industri. Pektinase merupakan enzim pendegradasi dinding sel tanaman yang umum digunakan untuk industri pangan seperti mempercepat ekstraksi sari buah. Mikroba merupakan sumber penghasil enzim yang sering digunakan karena tumbuh pada berbagai media dan mudah ditingkatkan hasilnya dengan mengatur kondisi pertumbuhan. Imobilisasi merupakan tahapan pengembangan bioproses berkelanjutan dengan menggunakan kembali enzim yang terikat pada suatu matriks. Oleh karena itu, tujuan penelitian ini yaitu melakukan imobilisasi enzim pektinase dan mengukur efisiensi penggunaan berulang pektinase imobil tersebut. Penelitian ini diawali dengan peremajaan isolat Bacillus sp. 2P11 yang telah diisolasi dari penelitian terdahulu. Kulit kakao mengandung pektin sehingga dapat digunakan sebagai substrat untuk menghasilkan pektinase. Produksi pektinase dilakukan dengan menumbuhkan Bacillus sp. 2P11 pada media
Keyword: Alginate beads, Bacillus sp. 2P11, cacao, pectinase, immobilization