scholarly journals Extraction and Characterization of Cathepsin Inhibitor from Milkfish

2015 ◽  
Vol 18 (1) ◽  
Author(s):  
Tati Nurhayati ◽  
Ruddy Suwandi ◽  
Syefri Rusyadi
Keyword(s):  
Protease Inhibitor ◽  
Metal Ion ◽  
Proteolytic Enzyme ◽  
Cysteine Proteases ◽  
Partial Purification ◽  
Optimum Temperature ◽  
Endogenous Inhibitors ◽  
Protease Enzyme ◽  
Myofibril Protein

Proteolytic enzyme is distributed acros all organism including fish. Cysteine proteases are the largest group of proteolytic enzyme. Lysosomal cathepsin, one of cysteine protease enzyme, cause softening and degradation of myofibril protein and it’s activity is regulated by endogenous inhibitors. The purposes of this study were to optimize the extraction cathepsin inhibitors from the skin, muscles, and viscera of fish, to partially purify the cathepsin inhibitors of selected sources, and to study the characteristics of the cathepsin inhibitor. The cathepsin inhibitor could be extracted from muscle fish and partially purified using ammonium sulfate of 70%. The purified cathepsin inhibitor had optimum temperature at 40°C and the optimum at pH 8. Metal ions decreased the activity of the protease inhibitor, except 1 mM of metal ion Mn2+ and Na+. Keywords: Cathepsin, characterization, partial purification, protease inhibitor

scholarly journals BIOCHEMICAL CHARACTERIZATION OF RECOMBINANT PHYTASE ENZYME (phyK) FROM Klebsiella sp. ASR1 ENCAPSULATED WITH ALGINATE

2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Muhammad Eka Hidayatullah ◽  
. Sajidan ◽  
Ari Susilowati ◽  
Baraka Stewart Mkumbe ◽  
Ralf Greiner
Keyword(s):  
Metal Ion ◽  
Biochemical Characterization ◽  
Extreme Conditions ◽  
Optimum Temperature ◽  
High Ph ◽  
Recombinant Phytase ◽  
Ion Effects ◽  
Metal Ion Effects

Karakterisasi Biokimia Enzim Fitase Rekombinan (phyK) dari Klebsiella sp. ASR1 Yang Dienkapsulasi Dengan AlginatEnzim fitase melepas molekul fosfor pada atom C dari benzena Inositol fitat. Tetapi fitase memiliki kelemahan tidak mampu bertahan terhadap kondisi ekstrim dalam lambung nonruminansia. Solusi dalam penelitian ini yaitu fitase dienkapsulasi menggunakan alginat. Penelitian ini bertujuan mengkarakterisasi fitase setelah dienkapsulasi menggunakan alginate. Hasil penelitian ini yaitu fitase yang dienkapsulasi memiliki aktivitas tertinggi pada pH 6,0, sedangkan fitase tanpa enkapsulasi pada pH 5,0. Suhu optimum untuk aktivitas tertinggi fitase yang dienkapsulasi yaitu 70ºC, sedangkan fitase tanpa enkapsulasi 37ºC. Untuk perlakuan penambahan ion logam, aktivitas tertinggi fitase yang dienkapsulasi terjadi dengan penambahan 0,1 mM Fe2+ dan 1,0 mM Ca2+, sedangkan fitase tanpa enkapsulasi dengan penambahan 0,1 mM Fe2+. Berdasarkan hasil penelitian ini, fitase yang dienkapsulasi memiliki keunggulan lebih banyak dibandingkan dengan fitase tanpa enkapsulasi, karena mampu bertahan pada pH dan suhu tinggi, dan beberapa efek ion logam.Kata Kunci: alginat, asam fitat, enkapsulasi, fitase, fitase rekombinanABSTRACTPhytase enzymes release phosphorus molecules on the C atom from benzene inositol phytate. But phytase has the disadvantage of being unable to withstand extreme conditions in the non-ruminant stomach. The solution in this research was phytase encapsulated using alginate. This study aims to characterize phytase after being encapsulated using alginate. The results of this study were the encapsulated phytase had the highest activity at pH 6.0, while the unencapsulated phytase at pH 5.0. The optimum temperature for the highest activity of the encapsulated phytase was 70ºC, while the unencapsulated phytase 37ºC. For treatment of metal ion addition, the highest activity of the encapsulated phytase occurred with the addition of 0.1 mM Fe2+ and 1.0 mM Ca2+, while the unencapsulated phytase with the addition of 0.1 mM Fe2+. Based on the results of this study, the encapsulated phytase had more advantages compared to the unencapsulated phytase, as the former withstand high pH and temperature, and some metal ion effects.

scholarly journals Molecular Identification of Indigenous Lactic Acid Bacteria, Partial Purification and Characterization of β-Galactosidase Produced

2017 ◽  
Vol 3 (4) ◽  
pp. 247
Author(s):  
Tatik Khusniati ◽  
Sulistiania . ◽  
Bellen Nastitie Pamela Fury ◽  
Syamsul Falah ◽  
Abdul Choliq
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Ammonium Sulphate ◽  
Molecular Identification ◽  
Specific Activity ◽  
Partial Purification ◽  
Optimum Temperature ◽  
Purification And Characterization

<p>β-Galactosidase is enzyme which hydrolyze lactose to glucose and galactose, as lactose hydrolyzer. To know indigenous lactic acid bacteria (LAB) characteristics, indigenous LAB identification, partial purification and characterization of β-galactosidase produced was researched. LAB was molecularly identified, β-galactosidase partial purification was conducted by precipitation followed dialysis. β-Galactosidase  characterization was based on optimum activities of pH and temperature. The results show that LAB was identified as <em>Lactobacillus plantarum</em> B123. Optimum activity of precipited β-galactosidase was reached at 50 % ammonium sulphate. Activity and specific activity of 50 % precipited β-galactosidase were 95.675 U · mg<sup>–1</sup> and 32.268 U · mg<sup>–1</sup>, respectively. Precipited β-galactosidase resulted a purification level of 3.99 fold, a yield of 38.73 %, and a specific activity of 32.27 U · mg<sup>–1</sup> protein, while dialyzed β-galactosidase resulted 7.61 fold, 10.67 %, and 61.53 U· mg<sup>–1</sup> protein. Optimum temperature and pH for crude β-galactosidase were found at 55 °C and 7.0, respectively, while that dialyzed β-galactosidase were optimized at 50 °C and 7.0. Based on partial purification and characterization, <em>Lactobacillus plantarum</em> B123 is indigenous LAB which good for production of β-galactosidase.</p><div><p class="Els-keywords"><em> </em></p><p class="Els-keywords"><strong>Keywords:</strong> characterization; dialysis; lactic acid bacteria; <em>Lactobacillus plantarum</em> B 123; β-galactosidase.</p></div>

scholarly journals PURIFICATION AND CHARACTERIZATION OF THE NEWLY THERMOSTABLE PROTEASE PRODUCED BY Brevibacillus thermoruber LII ISOLATED FROM PADANG CERMIN HOTSPRING, INDONESIA

2014 ◽  
Vol 9 (1) ◽  
pp. 1 ◽  
Author(s):  
Dewi Zeswita Zilda ◽  
Eni Harmayani ◽  
Jaka Widada ◽  
Widya Asmara ◽  
Hari Eko Irianto ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Metal Ion ◽  
Ethylenediaminetetraacetic Acid ◽  
Chloride Salt ◽  
Optimum Temperature ◽  
Purification And Characterization ◽  
The Stability ◽  
Thermo Stability ◽  
Edta Ethylenediaminetetraacetic Acid

Thermo stability is among of the vital enzyme characteristics for industrial application. Brevibacillus thermoruber LII was obtained as a potential isolate from the previous researchwhich screened the potential thermostable protease producing bacteria from Indonesian hotspring.The newly thermostable protease produced by thermophilic Brevibacillus thermoruber LII hadbeen purified and characterized. It was predicted that the pure enzyme obtained from Brevibacillusthermoruber LII was homo hexameric, having molecular weight of 36 kDa unit protein and itsnative was 215 kDa. In addition, it was also a neutral metalo serine protease according tobiochemical tests that it was totaly inhibited by PMSF (Phenylmethanesulfonyl fluoride) and EDTA(Ethylenediaminetetraacetic acid). It showed optimum activity at pH of 8 and active in acidic buffer(up to pH of 4). All of metal ion in the form of chloride salt (2.5 mM) which were tested on theenzyme enhanced the enzyme activity but Li2+. Ca2+ion increased the activity and the stability ofenzyme against thermal. The enzyme also showed the stability against solvent. The protease LIIhad optimum temperature at 60oC without CaCl 2and 80 – 85oC with addition of 2.5 mM CaCl 2. TheK Mand V maxvalues for the purified protease LII were 27.2 mg/ml or 0.362 – 0.272 M for substrateHammersteinCasein (MM 75–100 kDa) and 261.1 µg/minute/ml, respectively.

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