Optimasi Aktivitas Protease oleh Bacillus sp. MD24 menggunakan  Plackett Burman Design (PBD) dan Response Surface Methodology (RSM)

Suharti Suharti; Mieke Alvionita; Puji Putri Isnaini

doi:10.17977/um0260v8i22024p024

Authors

Suharti Suharti Universitas Negeri Malang
Mieke Alvionita Universitas Negeri Malang
Puji Putri Isnaini Universitas Negeri Malang

DOI:

https://doi.org/10.17977/um0260v8i22024p024

Keywords:

Optimasi, Aktivitas protease, PBD, CCD-RSM

Abstract

Protease merupakan kelompok enzim yang memiliki fungsi katalitik untuk menghidrolisis ikatan peptida pada protein. Penelitian pendahuluan telah berhasil mengisolasi bakteri Bacillus sp. MD24 yang dapat menghasilkan protease pada fermentasi menggunakan metode Sub-merge Fermentation (SmF) dengan aktivitas 16 U/mL yang diukur menggunakan substrat kasein. Namun demikian, aktivitas protease yang dihasilkan tergolong rendah sehingga pada penelitian ini dilakukan optimasi kondisi aktivitas protease dari Bacillus sp. MD24. Faktor optimasi yang digunakan meliputi faktor: temperatur, pH dan penambahan ion logam. Penelitian dilakukan dalam beberapa tahapan yaitu regenerasi Bacillus sp. MD24, fermentasi protease dari Bacillus sp. MD24, optimasi aktivitas protease menggunakan metode OFAT (One Factor at A Time), PBD (Plackett Burman Design) dan CCD-RSM (Central Composite Design-Response Surface Methodology). Hasil penelitian Metode OFAT menghasilkan aktivitas optimum pada keadaan temperatur 37℃ sebesar 544,7±10,48 U/mL; pH 8 sebesar 640,42±6,47 U/mL; CaCl₂ sebesar 609,04±4,6 U/mL. Metode PBD menghasilkan faktor-faktor yang berpengaruh secara signifikan terhadap aktivitas protease yaitu temperatur, pH, dan penambahan ion Ca²⁺. Metode CCD-RSM memberikan hasil optimasi faktor-faktor yang berpengaruh signifikan terhadap aktivitas protease yaitu pada temperatur 53,81℃, pH 7,677 dan Ca²⁺ 0,568 mM menghasilkan aktivitas protease 715,96 U/mL yang lebih tinggi dibandingkan dengan aktivitas pada studi pendahuluan maupun yang diperoleh melalui optimasi menggunakan metode OFAT.

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