Pengaruh Ph Terhadap Aktivitas Enzim Kitinase dari Isolat Actinomycetes dengan Metode Somogyi-Nelson

Welly Anggraini

Abstract


The pH effect on chitinase enzyme activity of  actinomycetes isolate  by Somogyi-Nelson method has been done. Chitinase enzymes can be produced by chitinolytic micro organisms, actinomycetes. This actinomycetes is taken from Mangrove from Coastal Ringgung of Teluk Lampung Waters, and has been isolated in previous research. The actinomycetes isolates used were ANL-12, ANL-9, ANLd-2b-3, and ANL-4, with chitinolytic activity of 1.9 cm, 2.0 cm, 2.3 cm, and 5.0 cm.  The ANL-4 isolate, which has the largest chitinolytic activity, was selected for the next process, the solidstate fermentationprocess (SSF) by the Somogyi-Nelson method. For this SSF process is done in one place. Chitinase enzyme activity was measured using micro plate reader by the somogyi-Nelson method. enzyme activity is calculated by measuring the amount of glucose released in μg / ml rough enzyme / hour (U / mL) by substrate reaction under certain conditions. From the research data showed that the purification enzyme had optimum activity at pH 7.0 with unit activity of 11,166 U / mL, while on chitin substrate without washing with NaOH yield optimum activity at pH 6 with unit activity equal to 10,929 U / ml.

Telah dilakukan pengaruh pH terhadap aktivitas enzim kitinase dari isolat actinomycetes dengan metode Somogyi-Nelson. Enzim kitinase dapat diproduksi oleh mikroorganisme kitinolitik, yaitu actinomycetes. Actinomycetes ini diambil dari Lumpur Hutan Bakau asal Pantai  Ringgung Perairan Teluk Lampumg, dan telah diisolasi pada penelitian sebelumnya.  Isolat actinomycetes yang digunakan adalah ANL-12, ANL-9, ANLd-2b-3, dan ANL-4, dengan memiliki aktivitas kitinolitik berturut-turut 1,9 cm, 2,0 cm, 2,3 cm, dan 5,0 cm.Isolat ANL-4 yang memiliki aktivitas kitinolitik terbesar ini dipilih untuk proses selanjutnya, yaitu proses Solid State Fermentation (SSF) dengan metode Somogyi-Nelson. Untuk proses SSF ini dilakukan dalam satu tempat. Aktivitas enzim kitinase diukur menggunakan microplate reader dengan metode Somogyi-Nelson.  Aktivitas enzim dihitung dengan mengukur jumlah glukosa yang dilepaskan dalam µg/ml enzim kasar/jam (U/mL) oleh reaksi substrat dengan kondisi tertentu.  Dari data penelitian menunjukkan bahwa enzim hasil pemurnian mempunyai aktivitas optimum  pada pH 7,0 dengan aktivitas unit sebesar 11,166 U/mL, sedangkan  pada substrat kitin tanpa dicuci dengan NaOH  menghasilkan aktivitas optimum pada pH 6 dengan aktivitas unit sebesar 10,929 U/mL.



Keywords


actinomycetes; kitin; kitinase; solid state fermentation (SSF)

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DOI: http://dx.doi.org/10.24042/jipfalbiruni.v4i2.94

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