OPTIMASI PRODUKSI KITINASE Bacillus cereus SMG 1.1 MENGGUNAKAN RESPONSE SURFACE METHODOLOGY

ADNAN WIDODO, Indun Dewi Puspita, S.P., M.Sc., Ph.D.; Prof. Dr. Ir. Ustadi, M.P.

2022 | Tesis | MAGISTER ILMU PERIKANAN

Limbah cangkang udang berpotensi memiliki nilai tambah apabila diolah menjadi kitin dan turunannya. Pengolahan limbah dengan pendekatan biologi lebih disukai dan ramah lingkungan dibandingkan kimiawi. Kitinase merupakan enzim yang berperan penting dalam pengolahan limbah cangkang udang. Bacillus cereus SMG 1.1 telah diisolasi dari produk terasi dan menunjukkan kemampuan produksi kitinase yang baik. Penelitian ini bertujuan untuk mengetahui komposisi optimum media produksi kitinase B. cereus SMG 1.1 melalui Response Surface Method (RSM) menggunakan desain faktorial dengan 15 perlakuan. Optimasi media dilakukan dengan menentukan faktor yang berpengaruh terhadap produksi kitinase melalui desain Plackett-Burman dilanjutkan dengan optimasi konsentrasi media melalui desain Box-Behnken. Pengujian dilakukan untuk mengetahui pengaruh variabel bebas terpilih terhadap aktivitas kitinase. Hasil analisis Plackett-Burman menunjukkan bahwa koloidal kitin, fruktosa, dan MgSO4.5H2O merupakan komponen medium yang paling berpengaruh terhadap produksi kitinase B. cereus SMG 1.1. Analisis Box-Behnken menghasilkan model linier yang dapat digunakan untuk memprediksi respon. Nilai respon maksimal didapatkan pada konsentrasi 1,5% koloidal kitin, 0,75% fruktosa, dan 0,075% MgSO4.5H2O dengan aktivitas kitinase optimal sebesar 0,0016 U/ml.

Shrimp shell waste are potential to be processed further into value-added products, such as N-acetylglucosamine (GlcNAc). In the production of GlcNAc, biological approach is preferred and environmentally friendly to chemical treatment. Chitinase is an enzyme that plays a vital role in bioprocessing of shrimp shell waste into GlcNAc. Previously, Bacillus cereus SMG 1.1 was isolated from fermented shrimp paste (terasi) and showed the ability to produce chitinase. This study was designed to determine the optimum medium for the production of chitinase from B. cereus SMG 1.1 through the Response Surface Method (RSM) using a factorial design with 15 treatments. The optimization of the media was carried out by determining the factors that influence the production of chitinase through the Plackett-Burman design followed by optimization of the concentration of the media through the Box-Behnken design. The test was designed to assess the effect of the independent variables on chitinase activity. Placket Burman analysis shows that colloidal chitin, fructose, and MgSO4.5H2O were the significant components in the medium affecting the production of chitinase by B. cereus SMG 1.1. Box-Behnken analysis developed a linear model capable of predicting the response. The highest response value was achieved at a concentration of 0.75% fructose, 1.5% colloidal chitin, and 0.075% MgSO4.5H2O resulted in optimum chitinase activity of 0.0016 U/ml.

Kata Kunci : Kitinase, Kitin, Fruktosa, MgSO4.5H2O, RSM