Pengaruh Konsentrasi Substrat terhadap Produksi Urease oleh Bakteri dari Lahan Pertanian Musiman

Haqq, Annida (2023) Pengaruh Konsentrasi Substrat terhadap Produksi Urease oleh Bakteri dari Lahan Pertanian Musiman. Other thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Pemanfaatan enzim urease yang dihasilkan oleh bakteri ureolitik saat ini banyak digunakan antara lain untuk menutupi keretakan beton dan mendegradasi limbah logam berat. Bakteri ureolitik ini dapat ditemukan di tanah lahan pertanian musiman. Hal ini disebabkan pada umumnya petani menggunakan pupuk urea yang dapat dimanfaatkan oleh bakteri ureolitik sebagai sumber energi. Salah satu penelitian yang penting dilakukan adalah untuk menemukan kondisi paling optimum untuk menghasilkan aktivitas urease paling besar sehingga pemanfaatan urease di berbagai bidang dapat lebih optimal. Aktivitas urease oleh bakteri ureolitik dapat diketahui dengan uji aktivitas menggunakan substrat berupa urea komersial. Namun biaya untuk memperoleh urea komersial cukup tinggi, sehingga pada penelitian ini digunakan urin sapi sebagai pengganti substrat urea komersial. Penelitian ini bertujuan untuk mengetahui hasil isolasi bakteri ureolitik dari tanah lahan pertanian dan mengetahui pengaruh kondisi lingkungan khususnya konsentrasi urin sapi sebagai substrat reaksi terhadap aktivitas urease. Skrining bakteri ureolitik dilakukan dengan menumbuhkan bakteri pada media selektif ureolitik (Christensen’s). Bakteri dengan kemampuan produksi urease tertinggi digunakan sebagai kultur starter produksi enzim. Variasi konsentrasi urin sapi yang digunakan saat fermentasi ialah 0,5%, 2,5%, 5%, 7,5%, 10%, dan 12,5% (v/v). Lalu dilakukan pemurnian parsial menggunakan Amonium Sulfat untuk memperoleh urease murni serta dilakukan Uji Aktivitas, Uji Bradford (kadar protein), dan Uji Titik Isoelektrik. Analisis data menggunakan One Way Anova untuk melihat signifikansi pengaruh konsentrasi urin sapi terhadap aktivitas urease yang dihasilkan. Hasil penelitian menunjukkan terdapat 17 jenis isolat ureolitik yang ditemukan di lahan pertanian. Sedangkan konsentrasi urin sapi berpengaruh signifikan (p value>0,05) terhadap aktivitas urease. Aktivitas urease tertinggi diperoleh pada konsentrasi urin sapi 5% yaitu sebesar 11.877,12 U/ml dengan kadar protein 0,046 mg/ml. Aktivitas spesifik terbesar diperoleh pada fraksi pengendapan Amonium Sulfat 45-60% yaitu sebesar 665.167,4 U/mg, dengan aktivitas enzim 7.587,46 U/ml dan kadar protein 0,0153 mg/ml.
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Utilization of the enzyme urease produced by ureolytic bacteria is currently widely used, among others, to cover concrete cracks and degrade heavy metal waste. This ureolytic bacteria can be found in seasonal agricultural land. This is because in general farmers use urea fertilizer which can be utilized by ureolytic bacteria as an energy source. One of the most important studies to be carried out is to find the most optimum conditions to produce the greatest urease activity so that the utilization of urease in various fields can be more optimal. Urease activity by ureolytic bacteria can be determined by an activity test using a substrate in the form of commercial urea. However, the cost of obtaining commercial urea is quite high, so that in this study cow urine was used as a substitute for commercial urea substrate. This study aims to determine the results of isolating ureolytic bacteria from agricultural soil and to determine the effect of environmental conditions, especially the concentration of cow urine as a reaction substrate on urease activity. Screening for ureolytic bacteria was carried out by growing the bacteria on selective ureolytic media (Christensen's). Bacteria with the highest ability to produce urease were used as starter cultures for enzyme production. Variations in the concentration of cow urine used during fermentation were 0.5%, 2.5%, 5%, 7.5%, 10% and 12.5% (v/v). Then, partial purification was carried out using Ammonium Sulfate to obtain pure urease and an Activity Test, Bradford Test (protein content), and Isoelectric Point Test were carried out. Data analysis used One Way Anova to see the significant effect of cow urine concentration on the resulting urease activity. The results showed that there were 17 types of ureolytic isolates found on agricultural land. While the concentration of cow urine has a significant effect (p value> 0.05) on urease activity. The highest urease activity was obtained at 5% cow urine concentration, which was 11,877.12 U/ml with a protein content of 0.046 mg/ml. The greatest specific activity was obtained in the 45-60% Ammonium Sulphate precipitation fraction, which was 65,167.4 U/mg, with an enzyme activity of 7,587.46 U/ml and a protein content of 0.0153 mg/ml.

Item Type:	Thesis (Other)
Uncontrolled Keywords:	Bakteri ureolitik, lahan pertanian, urease, urin sapi; Ureolytic bacteria, agricultural land, urease, cattle urine
Subjects:	Q Science > QR Microbiology > QR74.8 Bacteria
Divisions:	Faculty of Science and Data Analytics (SCIENTICS) > Biology > 46201-(S1) Undergraduate Thesis
Depositing User:	Annida Haqq
Date Deposited:	24 Aug 2023 04:18
Last Modified:	24 Aug 2023 04:18
URI:	http://repository.its.ac.id/id/eprint/102093

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