Karakterisasi Enzim Hidrolase Intraseluler dari Bacillus amyloliquefaciens-SLBD Asal Feses Kuda

Abstract

Bacillus amyloliquefacins SLBD (Ba-SLBD) berpotensi sebagai bakteri penghasil enzim hidrolitik untuk aplikasi bidang bioteknologi. Penelitian ini bertujuan mengevaluasi dinamika pertumbuhan dan mengkarakterisasi aktivitas enzim protease, lipase, dan fosfatase pada fraksi intraseluler Ba-SLBD. Kultur bakteri dipanen pada fase stasioner menggunakan media Luria Bertani (LB) broth, kemudian disentrifugasi, dilisis, dan diultrasentrifugasi untuk memperoleh fraksi SAL (solution after lysis) dan PAL (pellet after lysis). Total protein yang diperoleh pada fraksi SAL dan PAL masing-masing sebesar 3,98±0,11 mg dan 1,53±0,20 mg. Aktivitas enzim dianalisis berdasarkan indeks aktivitas enzim yang terbentuk pada media spesifik. Hasil indeks aktivitas enzim protease, lipase dan fosfatase tertinggi pada fraksi PAL, meskipun aktivitas ketiga enzim tersebut juga terdeteksi pada fraksi SAL. Hasil ini menunjukkan bahwa fraksi intraseluler Ba-SLBD berpotensi sebagai sumber enzim hidrolitik, terutama protease, lipase dan fosfatase.

Bacillus amyloliquefaciens SLBD (Ba-SLBD) has the potential to be a bacterium that produces hydrolytic enzymes for use in biotechnology applications. The aim of this study is to evaluate the growth dynamics and characterise the protease, lipase and phosphatase activities in the intracellular fraction of Ba-SLBD. The bacterial culture was harvested in the stationary phase using Luria Bertani (LB) broth, then centrifuged, lysed and ultrasentrifugated to obtain the SAL (Solution After Lysis) and PAL (Pellet After Lysis) fractions. The total protein obtained in the SAL and PAL fractions was 3.98±0.11 mg and 1.53±0.20 mg, respectively. The enzyme activity was analysed based on the enzyme activity index formed in the specific medium. The highest enzyme activity indices for protease, lipase and phosphatase were found in the PAL fraction, although activity of the three enzymes was also detected in the SAL fraction. These results indicate that the intracellular fractions of Ba-SLBD have potential as a source of hydrolytic enzymes, particularly proteases, lipases, and phosphatases.