scholarly journals SEMI PURIFIKASI DAN KARAKTERISASI ENZIM PROTEASE Bacillus sp.

2007 ◽  
Vol 13 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Elidar Naiola ◽  
Nunuk Widhyastuti
Keyword(s):  
Metal Ions ◽  
Gel Filtration ◽  
Liquid Waste ◽  
Enzyme Reaction ◽  
Inhibitory Effect ◽  
Crude Enzyme ◽  
Bacillus Sp ◽  
Optimum Temperature ◽  
Optimum Ph ◽  
Specific Activities

The aim of the research was to find the partial purified of enzyme protease from Bacillus sp. The crude enzyme of protease was produce in rice brand medium (100 gram of rice brand in a liter tofu liquid waste). The enzyme was semi-purified by the procedure of precipitation using ethanol in different percentages of saturation, gel filtration using Sephadex G 100 and Ion Exchanged Chromatography using DEAE Sephadex A50. Specific activities of the enzyme during purification were 5.71 U/mg (crude enzyme); 6.75 U/mg (ethanol precipitations); 37.16 U/mg (gel filtration) and 43.02 U/mg (Ion Exchanged Chromatography). The optimum temperature for enzyme reaction was 45–50 °C, while the optimum pH was 7.0–8.0. Protease was relatively stable after heating until 37–50 °C for 60 minutes. Metal ions had different effects to the enzyme. CaCl2, FeCl3, MnCl2, ZnCl2 and MgCl2 increased enzyme activity, CdCl2 and HgCl2 gave an inhibitory effect, and another of metal ions had no effects to the enzyme.

scholarly journals Characterization of Biosynthetic Enzymes for Ectoine as a Compatible Solute in a Moderately Halophilic Eubacterium, Halomonas elongata

1999 ◽  
Vol 181 (1) ◽  
pp. 91-99 ◽  
Author(s):  
Hisayo Ono ◽  
Kazuhisa Sawada ◽  
Nonpanga Khunajakr ◽  
Tao Tao ◽  
Mihoko Yamamoto ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Apparent Molecular Mass ◽  
Optimum Temperature ◽  
Sds Page ◽  
Halomonas Elongata ◽  
Optimum Ph

ABSTRACT 1,4,5,6-Tetrahydro-2-methyl-4-pyrimidinecarboxylic acid (ectoine) is an excellent osmoprotectant. The biosynthetic pathway of ectoine from aspartic β-semialdehyde (ASA), in Halomonas elongata, was elucidated by purification and characterization of each enzyme involved. 2,4-Diaminobutyrate (DABA) aminotransferase catalyzed reversively the first step of the pathway, conversion of ASA to DABA by transamination with l-glutamate. This enzyme required pyridoxal 5′-phosphate and potassium ions for its activity and stability. The gel filtration estimated an apparent molecular mass of 260 kDa, whereas molecular mass measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 44 kDa. This enzyme exhibited an optimum pH of 8.6 and an optimum temperature of 25°C and had Km s of 9.1 mM forl-glutamate and 4.5 mM for dl-ASA. DABA acetyltransferase catalyzed acetylation of DABA to γ-N-acetyl-α,γ-diaminobutyric acid (ADABA) with acetyl coenzyme A and exhibited an optimum pH of 8.2 and an optimum temperature of 20°C in the presence of 0.4 M NaCl. The molecular mass was 45 kDa by gel filtration. Ectoine synthase catalyzed circularization of ADABA to ectoine and exhibited an optimum pH of 8.5 to 9.0 and an optimum temperature of 15°C in the presence of 0.5 M NaCl. This enzyme had an apparent molecular mass of 19 kDa by SDS-PAGE and a Km of 8.4 mM in the presence of 0.77 M NaCl. DABA acetyltransferase and ectoine synthase were stabilized in the presence of NaCl (>2 M) and DABA (100 mM) at temperatures below 30°C.

scholarly journals Production and Characterization of Collagenase From Bacillus sp. 6-2 Isolated From Fish Liquid Waste

2019 ◽  
Vol 12 (1) ◽  
pp. 58
Author(s):  
Sartika Danial ◽  
Hasnah Natsir ◽  
Seniwati Dali ◽  
Leliani Leliani
Keyword(s):  
Liquid Waste ◽  
Industrial Applications ◽  
Selective Medium ◽  
Bacillus Sp ◽  
Fermentation Time ◽  
Collagen Peptides ◽  
Hydrolysis Products ◽  
Native Collagen ◽  
Optimum Ph

Collagenases are enzyme that are able to hydrolyze native collagen into fragment collagen peptides. Collagenases and its hydrolysis products have received tremendous attention in medical and industrial applications. The present study was conducted to isolate and identify new collagenase producing bacteria from fish liquid waste, then produce and characterize collagenase. A total of 7 isolate from fish liquid waste were screened on selective medium containg 2 % collagen and its activity was confirmed by the formation of clear zone. Isolat 6-2 was positif as collagenase producer and identified as Bacillus sp. 6-2 by morphological and biochemical characteristics. The optimum fermentation time of enzyme was investigated. Collagenase crude extract was characterized by the effect of pH, temperature, and metal ions. Isolat 62 optimally produced collagenase enzyme after 30 h of incubation with activity of   0.072 U/mL and protein content of 3.768 mg/mL. The optimum pH and temperature were 7.0 and 40 oC, respectively. The enzyme was activated by 1 mM Ca2+and  Mg2+, and inhibited by   1 mM  Zn2+ and Co2+. Collagenase from Bacillus sp. 6-2 may have potentials for medical and industrial applications.

Characterisation of a Neutral Protease Produced by Micrococcus luteus

1996 ◽  
Vol 51 (5-6) ◽  
pp. 429-431 ◽  
Author(s):  
M.O. Ilori ◽  
O.O. Amund ◽  
O. Omidiji
Keyword(s):  
Molecular Weight ◽  
Citric Acid ◽  
Proteolytic Enzyme ◽  
Micrococcus Luteus ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Optimum Temperature ◽  
Optimum Ph

Abstract A proteolytic enzyme produced by a cassava-ferment­ing strain of Micrococcus luteus was extracted and puri­fied 50-fold by gel filtration and ion exchange chromatography. The optimum pH for the enzyme was 7.0, the optimum temperature 25 °C, the apparent molecular weight 42 kDa and the Km value, 0.45 mg ml-1 with casein as substrate. The enzyme was stimulated by Ca2+ and Mg2+ but inhibited by Zn2+ and Co2+ ions. Other inhibitors were EDTA, KCN, citric acid and L-cysteine indicating the enzyme to be a metalloprotease.

Purification and properties of an exocellular β-glucosidase of Candida molischiana (Zikes) Meyer and Yarrow capable of hydrolyzing soluble cellodextrins

1985 ◽  
Vol 63 (11) ◽  
pp. 1160-1166 ◽  
Author(s):  
Pierre Gondé ◽  
Robert Ratomahenina ◽  
Alain Arnaud ◽  
Pierre Galzy
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Optimum Temperature ◽  
Purification And Properties ◽  
Optimum Ph

The exocellular enzyme β-glucosidase of Candida molischiana was studied. This strain is able to ferment soluble cellodextrins. The enzyme was partially purified by ion-exchange chromatography and gel filtration. The molecular weight of this enzyme was 120 000; its optimum pH was between 4 and 4.5 and its optimum temperature was 60 °C. This enzyme was active against different soluble glucosides and was inhibited by p-chloromercuribenzoate, gluconolactone, and glucose. A "glucosyltransferase" activity appeared in the presence of ethanol. The biosynthesis of the enzyme was constitutive but repressed by glucose.

scholarly journals Isolasi dan Karakterisasi Fitase dari Mikroba yang Terdapat pada Pupuk Kompos, Rumen Sapi, Ragi dan Tanah Sawah

2016 ◽  
Vol 7 (1) ◽  
pp. 14
Author(s):  
Prof. Dr.rer.nat. Sajidan
Keyword(s):  
Key Words ◽  
Complex Compound ◽  
Relative Activity ◽  
Luria Bertani ◽  
Crude Enzyme ◽  
Soy Sauce ◽  
Optimum Temperature ◽  
Ph Optimum ◽  
Optimum Ph

<div class="WordSection1"><p><em> </em></p><p><em>            This research was aimed to isolate and characterize  phytase from different source</em><em>s</em><em> of higger phosphat complex compound</em><em>s</em><em> (compost, cattle rumen, and yeast). Bacteria of phytase was isolated on Luria Bertani (LB) medium and it was incubated on 37 <sup>o</sup>C for 16 hours. Microbe from yeast was isolated on Pottato Dextrose Agar (PDA) medium. Crude enzyme from supernatant was extracted with centrifuge on 5.000 rpm for 5 minutes. Crude enzyme was characterized for optimum pH, temperature, and influence of matalo ion efector on relative activity of enzyme. B1 isolate from P88 compost had optimum relative activity on pH 5, temperature 50 <sup>o</sup>C and activator Zn<sup>2+</sup> (10<sup>-3</sup> and 10<sup>-4</sup> M). B2 Isolate from cattle rumen had optimum relative activity on pH 5, temperature 50 <sup>o</sup>C and activator Zn<sup>2+</sup> (10<sup>-3</sup> M) and Mg<sup>2+</sup> (10<sup>-4</sup> M). B3 isolated from soy sauce yeast had optimum relative activity on pH 5, temperature 60 <sup>o</sup>C, and  activators Mg<sup>2+</sup> (10<sup>-3 </sup>M) and, Fe<sup>2+</sup> (10<sup>-4</sup> M). B4 isolated from tempeh yeast had optimum relative activity on pH 5, temperature 50 <sup>o</sup>C and activators Mg<sup>2+</sup> (10<sup>-3 </sup>M) and Ca<sup>2+</sup> (10<sup>-4</sup> M).</em></p><p><strong><em> </em></strong></p><p><strong><em>Key words</em></strong><em>: phytase, optimum pH, optimum temperature, metalo ion efector, isolate.</em></p></div><strong><br clear="all" /> </strong>

scholarly journals Isolasi dan Karakterisasi Fitase dari Mikroba yang Terdapat pada Pupuk Kompos, Rumen Sapi, Ragi dan Tanah Sawah

2016 ◽  
Vol 7 (1) ◽  
pp. 14
Author(s):  
Prof. Dr.rer.nat. Sajidan
Keyword(s):  
Key Words ◽  
Complex Compound ◽  
Relative Activity ◽  
Luria Bertani ◽  
Crude Enzyme ◽  
Soy Sauce ◽  
Optimum Temperature ◽  
Ph Optimum ◽  
Optimum Ph

<div class="WordSection1"><p><em> </em></p><p><em>            This research was aimed to isolate and characterize  phytase from different source</em><em>s</em><em> of higger phosphat complex compound</em><em>s</em><em> (compost, cattle rumen, and yeast). Bacteria of phytase was isolated on Luria Bertani (LB) medium and it was incubated on 37 <sup>o</sup>C for 16 hours. Microbe from yeast was isolated on Pottato Dextrose Agar (PDA) medium. Crude enzyme from supernatant was extracted with centrifuge on 5.000 rpm for 5 minutes. Crude enzyme was characterized for optimum pH, temperature, and influence of matalo ion efector on relative activity of enzyme. B1 isolate from P88 compost had optimum relative activity on pH 5, temperature 50 <sup>o</sup>C and activator Zn<sup>2+</sup> (10<sup>-3</sup> and 10<sup>-4</sup> M). B2 Isolate from cattle rumen had optimum relative activity on pH 5, temperature 50 <sup>o</sup>C and activator Zn<sup>2+</sup> (10<sup>-3</sup> M) and Mg<sup>2+</sup> (10<sup>-4</sup> M). B3 isolated from soy sauce yeast had optimum relative activity on pH 5, temperature 60 <sup>o</sup>C, and  activators Mg<sup>2+</sup> (10<sup>-3 </sup>M) and, Fe<sup>2+</sup> (10<sup>-4</sup> M). B4 isolated from tempeh yeast had optimum relative activity on pH 5, temperature 50 <sup>o</sup>C and activators Mg<sup>2+</sup> (10<sup>-3 </sup>M) and Ca<sup>2+</sup> (10<sup>-4</sup> M).</em></p><p><strong><em> </em></strong></p><p><strong><em>Key words</em></strong><em>: phytase, optimum pH, optimum temperature, metalo ion efector, isolate.</em></p></div><strong><br clear="all" /> </strong>

scholarly journals A highly inducible β-galactosidase from Enterobacter sp.

2020 ◽  
Vol 85 (5) ◽  
pp. 609-622
Author(s):  
Bestoon Shaikhan ◽  
Kemal Güven ◽  
Fatma Bekler ◽  
Ömer Acer ◽  
Reyhan Güven
Keyword(s):  
Gel Filtration ◽  
Gel Permeation Chromatography ◽  
Inhibitory Effect ◽  
Native Page ◽  
Environment And Health ◽  
Low Concentrations ◽  
Sds Page ◽  
Gel Permeation ◽  
Optimum Ph ◽  
Strong Inhibitory Effect

Enterobacter sp. 3TP2A isolated from a petroleum station was found to produce a novel, highly inducible mesophilic intracellular ?-galactosidase in the presence of lactose up to 76.5 U mg-1. The enzyme was purified to 17.3- -fold after gel permeation chromatography with a yield of approximately 11 %. The optimum pH and temperature values of the purified enzyme were found to be 8.0?9.0 and 35 ?C, respectively. The molecular weight of the enzyme was approx. 60 kDa with a single band by both SDS-PAGE and native-PAGE, and estimated by gel filtration chromatography. The enzyme was inhibited by Zn2+ and EDTA, while Cu2+ had strong inhibitory effect even at low concentrations. Activation by Mg2+ and inhibition by EDTA show that the enzyme is metal- -dependent or a metalloenzyme. The enzyme was slightly activated by 2-mercaptoethanol, while slightly inhibited by iodoacetamide. On the other hand, PCMB inhibited the enzymatic activity to a great extent, whereas it was completely inhibited by N-ethylmaleimide. The Vmax and Km values were calculated as 0.701 ?mol min-1 and 0.104 mM, respectively. The results indicated that the ?-galactosidase Enterobacter sp. 3TP2A might well be a good candidate for use in biotechnology, particularly in the area of environment and health.

scholarly journals Alfa-amilaz Enzimlerini Üreten Termofilik Bacillus Suşlarının İzolasyonu ve Enzimlerin Kısmi Karakterizasyonu

2015 ◽  
Vol 3 (6) ◽  
pp. 387 ◽  
Author(s):  
Celal Türker ◽  
Bahri Devrim Özcan
Keyword(s):  
Molecular Weight ◽  
Ph Value ◽  
Soil Samples ◽  
The Other ◽  
Bacillus Sp ◽  
Thermophilic Bacillus ◽  
Amylase Production ◽  
Optimum Ph ◽  
Specific Activities ◽  
Bacillus Strains

In the present study, we isolated three thermophilic Bacillus strains from the soil samples collected from the coast sediments of the Burnaz Stream located in Erzin. The isolates were entitled as Bacillus sp. CT1, CT2, and CT3, respectively. The maximum α-amylase production was revealed at 60°C for CT1 strain, and at 80°C for CT2 and CT3 strains, respectively. The optimum enzyme activity was observed at 90°C for CT1 α-amylase, whereas at 60°C for CT2 and CT3 α-amylases. On the other hand, optimum pH value for CT2 α-amylase was 7.0, whereas 8.0 for CT1 and CT3 α-amylases. The specific activities of CT1, CT2, and CT3 amylases were 317.6, 113.3 and 362.7 U/mg at 55°C, respectively. The estimated molecular weight of CT1 and CT3 α-amylase was 65 kDa, and for CT2 α-amylase was 38 kDa by zymogram analysis.

scholarly journals Encapsulation of HRP Enzyme onto a Magnetic Fe3O4 Np–PMMA Film via Casting with Sustainable Biocatalytic Activity

Catalysts ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 181 ◽  
Author(s):  
Wesam H. Abdulaal ◽  
Yaaser Q. Almulaiky ◽  
Reda M. El-Shishtawy
Keyword(s):  
Metal Ions ◽  
Immobilized Enzyme ◽  
Pmma Film ◽  
Optimum Temperature ◽  
Initial Activity ◽  
Casting Method ◽  
Ph Optimum ◽  
Optimum Ph ◽  
Triton X ◽  
Enzyme Changes

Horseradish peroxidase (HRP) enzyme was effectively encapsulated onto an Fe3O4 nanoparticle–polymethyl methacrylate (PMMA) film via the casting method. The HRP was immobilized on the 0.5% Fe3O4Np–PMMA film and characterized by Fourier transform infrared spectroscopy and field emission scanning electron microscopy. Moreover, the reusability, thermal stability, optimum pH, optimum temperature, the influence of metal ions, and the effects of detergent and organic solvent were investigated. After optimizing the immobilization conditions, the highest efficiency of the immobilized enzyme was 88.4% using 0.5% Fe3O4Np–PMMA. The reusability of the immobilized HRP activity was 78.5% of its initial activity after being repeatedly used for 10 cycles. When comparing the free and immobilized forms of the HRP enzyme, changes in the optimum temperature and optimum pH from 30 to 40 °C and 7.0 to 7.5, respectively, were observed. The Km and Vmax for the immobilized HRP were estimated to be 41 mM, 0.89 U/mL for guaiacol and 5.84 mM, 0.66 U/mL for H2O2, respectively. The high stability of the immobilized HRP enzyme was obtained using metal ions, a high urea concentration, isopropanol, and Triton X-100. In conclusion, the applicability of immobilized HRP involves the removal of phenol in the presence of hydrogen peroxide, therefore, it could be a potential catalyst for the removal of wastewater aromatic pollutants.

scholarly journals Pengaruh pH dan Suhu terhadap Produksi Antibiotika dari Isolat Bakteri Endofitik pada Tumbuhan Andalas (Morus macroura Miq.)

2019 ◽  
Vol 6 (1) ◽  
pp. 90
Author(s):  
Riski Budi Yani ◽  
Anthoni Agustien ◽  
Feskaharny Alamsjah
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Survey Method ◽  
Bacillus Sp ◽  
Optimum Temperature ◽  
Paper Disk ◽  
Optimum Ph ◽  
Disk Method ◽  
And Staphylococcus Aureus ◽  
Selection Of

The study used survey method and the data were analysed descriptively. The selection of the bacteria which produce antibiotic had been with paper disk method and used Escherichia coli and Staphylococcus aureus as the sample bacteria. This result showed pH 7,0 was succesful optimum pH antibiotic produced for Bacillus sp.1 and Bacillus sp.2. and 370C was the optimum temperature to antibiotic produced from Bacillus sp.1 and Bacillus sp.2.

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