FERMENTASI SUBSTRAT PADAT LIMBAH AMPAS KELAPA DENGAN KAPANG LIPOLITIK UNTUK PRODUKSI LIPASE

EKO SUYANTO, Prof. Dra. Endang Sutariningsih, M.Sc., Ph.D.

2015 | Tesis | S2 Biologi

Lipase (triacyl glyserol acyl hydrolases, E.C 3.1.1.3) yang dihasilkan oleh kapang lipolitik mempunyai nilai ekonomi tinggi dibidang industri dan termasuk kelompok enzim hidrolase yaitu meng-hidrolisis senyawa triasilgliserol menjadi gliserol dan asam lemak. Tujuan penelitian adalah mendapatkan kapang lipolitik yang memiliki aktivitas hidrolisis tinggi dan menguji pengaruh manipulasi faktor lingkungan pertumbuhan kapang lipolitik terpilih terhadap produksi dan aktivitas lipase. Penelitian diawali dengan purifikasi, skrining dan seleksi isolat kapang. Kapang yang menunjukkan aktivitas lipase tertinggi digunakan untuk percobaan selanjutnya. Peningkatan produksi dan aktivitas lipase kapang lipolitik dilakukan melalui percobaan manipulasi lingkungan tumbuh (pH substrat dan suhu inkubasi) pada ampas kelapa menggunakan metode solid state fermentation (SSF). Adanya pengaruh dianalisis melalui penentuan kadar protein Bradford, aktivitas hidrolisis lipase, perubahan pH, biomassa spora dan volume supernatan ekstrak kasar lipase. Karakterisasi lipase dilakukan pada lipase yang diperoleh saat kondisi optimum produksi lipase. Kapang lipolitik dikarakterisasi melalui pengamatan fenotipik kapang sesuai metode standart dan diidentifikasi secara profile matching. Hasil penelitian menunjukkan bahwa isolat kapang yang menghasilkan aktivitas hidrolisis terbesar adalah isolat KLC-33 sebesar 13,33 U/ml dengan aktivitas protease terkecil yaitu 0,145 U/ml. Kondisi optimum produksi lipase diperoleh pada penambahan jumlah spora kapang 1x106 spora/ml, volume larutan pengekstrak 75 ml, pH medium 4 dan suhu inkubasi 30oC dengan aktivitas hidrolisis sebesar 18,33 U/ml serta volume enzim 72,5 ml. Aktivitas lipase bekerja optimum pada pH 3, suhu inkubasi 30oC dan penambahan 0,01 M ion logam Co2+ sebesar 25,83 U/ml, 20,83 U/ml dan 48,33 U/ml secara berurutan. Isolat KLC-33 diidentifikasi sebagai Aspergillus niger. Dari hasil penelitian dapat disimpulkan bahwa limbah ampas kelapa merupakan medium efektif pertumbuhan kapang lipolitik. Produksi dan aktivitas lipase mengalami peningkatan yang dipengaruhi oleh faktor lingkungan diantaranya kondisi pH medium dan suhu inkubasi fermentasi.

Lipases (triacyl glyserol acyl hydrolases, E.C 3.1.1.3) derived from lipolytic fungi has high economical value and are class of hydrolase enzyme which able to hydrolyze of triglycerides and release glycerol and free fatty acids. The research aims were to obtain lipolytic fungi (molds) having the highest hydrolysis activity and to elucidate the effects of growth environmental factors only from selected lipolytic fungi for lipase production and its activity. This research was started with purification, screening and selection of fungi isolates. The fungi isolates shown the highest of lipase activity was selected for further experiment. Improve of its activity especially on lipase production using manipulation of environmental factors (pH substrate and temperature of incubation) in coconut oil cake with solid state fermentation (SSF). The effects of treatments was analyze with determine of protein Bradford concentration, hydrolysis activity, pH reaction, spore biomass and volume of enzyme had produced. Characterize was done on optimum condition for producing lipase. Morphology of fungi observed their characters and were identified using profile matching according standard methods. The results showed that coconut oil-cake was degraded by fungal isolate of KLC-33 during reaction so that produce highest hydrolysis activity that is 13.33 U/ml and smallest proteolytic activity that is 0.145 U/ml. Optimum condition for improving production of lipase and their activity has obtained on 1x106 spore/ml inoculums, 75 ml enzyme extracted solution, pH medium 4, and temperature incubation is 30oC with 72.5 ml of enzyme supernatant and the hydrolysis activity is 18.33 U/ml. The optimum pH, temperature and effects of 0.01 M ion metals for enzymatic activity were pH 3, temperature incubation 30oC and ion Co2+ that 25.83 U/ml, 20.83 U/ml and 48.33 U/ml, respectively. Isolate of KLC-33 has identified as Aspergillus niger. It could be conclude that the coconut oil cake is effectively substrate for growing up the lipolytic fungi. Improving of production of lipase and hydrolysis activity depend on environmental factors such pH and temperature of reaction.

Kata Kunci : Lipase, kapang, ampas kelapa, SSF, hidrolisis