Wardani, KartikaKusuma (2014) Karakterisasi Enzim Organofosfat Hidrolase (OPH) dari Pseudomonas Putida yang Diproduksi Menggunakan Substrat Klorpirifos dan Profenofos. Sarjana thesis, Universitas Brawijaya.
Abstract
Organofosfat hidrolase (OPH) merupakan enzim intraseluler yang diperoleh dengan cara isolasi dari bakteri Pseudomonas putida. Enzim ini mampu mendegradasi pestisida golongan organofosfat. Penelitian ini dilakukan untuk mengetahui kemampuan enzim OPH dalam mendegradasi pestisida golongan organofosfat klorpirifos dan profenofos, serta mengetahui kondisi kerja optimum dan kinetika kimia reaksi enzimastisnya. Enzim OPH dimurnikan melalui fraksinasi bertingkat menggunakan ammonium sulfat dengan fraksi pengendapan 0-45% dan 45%-65%. Uji aktivitas enzim OPH dilakukan dengan mereaksikan enzim dan substrat profenofos dan klorpirifos pada pH optimumnya. Kinetika reaksi enzimatis enzim OPH dengan substrat klorpirifos dan profenofos ditentukan dengan cara mereaksikan enzim OPH dengan substrat profenofos dan klorpirifos dengan variasi konsentrasi 2-10 ppm, kemudian ditentukan Vmaks dan KM. Hasil penelitian menunjukkan aktivitas enzim spesifik untuk substrat klorpirifos adalah 0,017 unit/mg sedangkan untuk substrat profenofos 0,024 unit/mg. pH optimum OPH untuk substrat klorpirifos yaitu pH 7,5 sedangkan pH optimum untuk substrat profenofos yaitu pH 8,5. Parameter kinetika reaksi enzimatis enzim OPH untuk substrat klorpirifos Vmax= 1,34 x 10-3 μg. mL-1. menit-1, KM = 2,17 mg/L dan profenofos Vmax = 1,87 x 10-3 μg. mL-1. menit-1, KM = 3,08 mg/L. Hasil analisa dengan menggunakan pola rancangan acak lengkap (RAL) diperoleh nilai Fhitung>Ftabel maka H0 ditolak yang artinya pH berpengaruh nyata terhadap aktivitas OPH (p<0,05).
English Abstract
Organophosphate hydrolase (OPH) is an intracellular enzyme which can be obtained from isolation of Pseudomonas putida bacteria. This enzyme can degrade organophosphate group pesticides. This reasearch was conducted to examine the ability of OPH in degrading the organophosphate pesticides chlorpyrifos and profenofos, and to determine the optimum conditions of the chemical kinetics of the enzyme. The obtained OPH were purified through a graded fractioning by using centrifugation filtrate methods using ammonium sulphate, with fraction of precipitation of 0-45% and 45-65%. Enzyme activity tests were conducted by reacting the enzyme and substrate group organophosphate pesticides chlorpyrifos and profenofos, at the optimum pH conditions for enzyme. The chemical kinetics enzymatic reactions were conducted by reacting the enzyme and substrate at various concentration of 2-10 ppm, from which the values of Vmaks and KM are determined. The results showed a specific activity of the enzyme with chlorpiryfos substrate which was 0.017 units/mg, and 0.024 units/mg for profenofos substrate, with the optimum pH for this enzyme with chlorpiryfos substrate was 7.5, while for the profenofos substrate was 8.5, with fraction of precipitation of 0-45%. The results of the chemical kinetics of enzimatic reactions are Vmax= 1.34 x 10-3 μg. mL-1. min-1, KM = 2.17 mg/L for chlorpiryfos substrate; and Vmax = 1.87 x 10-3 μg. mL-1. min-1, KM = 3.08 mg/L for the profenofos substrate. The results of analysis by employing the complete random design (RAL) method showed Fcalculated>Ftable so that the H0 is denied, which means that pH has real influence over OPH activity (p<0,05).
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | SKR/MIPA/2014/303/051405301 |
| Subjects: | 500 Natural sciences and mathematics > 540 Chemistry and allied sciences |
| Divisions: | Fakultas Matematika dan Ilmu Pengetahuan Alam > Kimia |
| Depositing User: | Budi Wahyono Wahyono |
| Date Deposited: | 02 Sep 2014 10:12 |
| Last Modified: | 21 Oct 2021 05:01 |
| URI: | http://repository.ub.ac.id/id/eprint/153934 |
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