scholarly journals Indigeneous Streptomyces spp. isolated from Cyperus rotundus rhizosphere indicate high mercuric reductase activity as a pontential bioremediation agent

2021 ◽  
Vol 22 (3) ◽  
Author(s):  
Hanum Mukti Rahayu ◽  
Wahyu Aristyaning Putri ◽  
Anis Uswatun Khasanah ◽  
LANGKAH SEMBIRING ◽  
Yekti Asih Purwestri
Keyword(s):  
Specific Activity ◽  
Reductase Activity ◽  
Cyperus Rotundus ◽  
Cell Free Extract ◽  
Mercuric Reductase ◽  
Streptomyces Spp ◽  
Sds Page ◽  
Optimum Ph ◽  
Bioremediation Agent ◽  
Optimum Ph And Temperature

Abstract. Rahayu HM, Putri WA, Khasanah AU, Sembiring L, Purwestri YA. 2021. Indigenous Streptomyces spp. isolated from Cyperus rotundus rhizosphere indicate high mercuric reductase activity as a potential bioremediation agent. Biodiversitas 22: 1519-1526. The purification and characterization of mercuric reductase of four indigenous Streptomyces spp. from Cyperus rotundus L. rhizosphere in mercury-contaminated area have been investigated. Cell-free extract was obtained by disrupting cells using sea sand at 4 °C followed by centrifugation. Mercuric reductase was purified by ammonium sulfate precipitation, dialysis, and chromatography column (DEAE Sepharose anion column chromatography). The determination of optimum pH and temperature of mercuric reductase activity was measured based on the number of NADPH2 oxidized to NADP per mg protein per minute using a spectrophotometer. The molecular weight of mercuric reductase was determined using SDS-PAGE. Result showed that the highest specific activity of mercuric reductase was recorded from Streptomyces spp. BR28. The optimum pH and temperature of cell-free extract enzyme mercuric reductase were 7.5 and 80 °C, respectively. The enzyme was purified to 431.87-fold with specific activity 21918.95 U/mg protein. SDS PAGE showed that the molecular weight of mercuric reductase in Streptomyces spp. BR 28  ranged from 50 kDa to 75 kDa. It can be concluded that Streptomyces isolates contain mercuric reductase and have potential as mercury bioremediation agent to overcome mercury contamination in the environment.

scholarly journals Thermostable glucose isomerase from psychrotolerant Streptomyces species

1970 ◽  
Vol 1 ◽  
pp. 6-10 ◽  
Author(s):  
Bidur Dhungel ◽  
Manoj Subedi ◽  
Kiran Babu Tiwari ◽  
Upendra Thapa Shrestha ◽  
Subarna Pokhrel ◽  
...  
Keyword(s):  
Specific Activity ◽  
Fold Increase ◽  
Glucose Isomerase ◽  
Enzyme Concentration ◽  
Maximal Velocity ◽  
Ph Optimum ◽  
Streptomyces Spp ◽  
Optimum Ph ◽  
Optimum Reaction ◽  
Sepharose 4B

Glucose isomerase (EC 5.3.1.5) was extracted from Streptomyces spp., isolated from Mt. Everest soil sample, and purified by ammonium sulfate fractionation and Sepharose-4B chromatography. A 7.1 fold increase in specific activity of the purified enzyme over crude was observed. Using glucose as substrate, the Michaelis constant (KM<) and maximal velocity (Vmax) were found to be 0.45M and 0.18U/mg. respectively. The optimum substrate (glucose) concentration, optimum enzyme concentration, optimum pH, optimum temperature, and optimum reaction time were 0.6M, 62.14μg/100μl, 6.9, 70ºC, and 30 minutes, respectively. Optimum concentrations of Mg2+ and Co2+ were 5mM and 0.5mM, respectively. The enzyme was thermostable with half-life 30 minutes at 100ºC.DOI: 10.3126/ijls.v1i0.2300 Int J Life Sci 1 : 6-10

scholarly journals Characterization of amylase and protease activity in the digestive tract of two teleosts (Labeo rohita and Anabas testudineus) with different feeding habits

2021 ◽  
Vol 64 (2) ◽  
pp. 173-179
Author(s):  
Sanjeet Debnath ◽  
Surjya Kumar Saikia
Keyword(s):  
Digestive Tract ◽  
Protease Activity ◽  
Specific Activity ◽  
Labeo Rohita ◽  
Feeding Habits ◽  
Anabas Testudineus ◽  
Pyloric Caeca ◽  
Optimum Ph ◽  
Optimum Ph And Temperature

Two teleosts (Rohu, Labeo rohita and Koi, Anabas testudineus), both with contrasting feeding habits (herbivorous versus carnivorous) were studied for amylase and protease activity concerning different regions of their digestive tracts. Significant differences in enzymatic activity across different regions of the digestive tracts were observed. Rohu, with three equal regions of the stomachless gut, showed the highest amylolytic activity at the posterior digestive tract but the highest proteolytic activity is limited to mid region. Contrary to such observation, Koi with three distinct regions of the digestive tract (stomach, pyloric caeca and intestine), the pyloric caeca exhibited the highest specific activity for both amylase and total protease. The optimum pH and temperature conditions were determined concerning the activity for both amylase and protease.

scholarly journals Expression and characterization of Trichoderma reesei endoglucanase II in Pichia pastoris under the regulation of the GAP promoter

2020 ◽  
Vol 25 (2) ◽  
pp. 127
Author(s):  
Kezia Abib Yerah Tjandra ◽  
Kartika Sari Dewi ◽  
Asrul Muhamad Fuad ◽  
Trisanti Anindyawati
Keyword(s):  
Pichia Pastoris ◽  
Trichoderma Reesei ◽  
Specific Activity ◽  
Catalytic Efficiency ◽  
Endoglucanase Activity ◽  
Gap Promoter ◽  
Sds Page ◽  
Optimum Ph ◽  
High Concentrations

Trichoderma reesei is known to be one of the organisms capable for producing various types of cellulase in high concentrations. Among these cellulases, the highest catalytic efficiency of endoglucanases II (EGII, EC 3.2.1.4) are considered important for industrial application. The characterization of the EGII is necessary since it is widely used in high-temperature reactions in the industries. In this study, the recombinant EGII protein was expressed in Pichia pastoris and it has a molecular mass of approximately 52 kDa. Recombinant EGII was purified using Ni-NTA affinity chromatography and characterized by SDS-PAGE and western blot analyses. The enzyme activity of recombinant EGII was measured using the Nelson Somogyi method to determine its optimum pH and temperature. The result showed that the maximum EGII expression was achieved after 72 h of culture incubation. The crude enzyme has optimum activity at pH 5.0, resulting in 16.3 U/mL and 14.6 U/mL activity at 40 °C and 50 °C, respectively. While the purified enzyme gave the specific activity of 115.7 U/mg under the optimum condition. Finally, our study demonstrated that recombinant EGII could retain the endoglucanase activity for 89% and 80% at 40 °C and 50 °C, respectively.

scholarly journals Peroxidase from Coleus Forskohlii: Purification and Biochemical Characterization

2020 ◽  
Vol 5 (1) ◽  
pp. 9-20
Author(s):  
Yaaser Q. Almulaiky ◽  
Yaaser Q. Almulaiky
Keyword(s):  
Ion Exchange ◽  
Peroxidase Activity ◽  
Biochemical Characterization ◽  
Specific Activity ◽  
Gel Filtration ◽  
Catalytic Efficiency ◽  
Industrial Applications ◽  
Coleus Forskohlii ◽  
Sds Page ◽  
Optimum Ph

In this study, a peroxidase from new source was purified using ion exchange and gel filtration techniques. The recovery for peroxidase activity was 19% with 11-fold purification and specific activity of 749 unit/mg protein. Purified peroxidase demonstrated a molecular mass of 39 kDa using gel filtration and was confirmed as a single band on SDS-PAGE. The purified peroxidase revealed a broad optimum pH activity at 6.0-6.5 and 50°C temperature. The kinetic parameters for purified peroxidase toward H2O2 and guaiacol as substrates were found to be Km = 3.355, 5.395 mM, Kcat = 99.52, 79.56 s-1 and Vmax =1.531, 1.242 µmole ml-1 min-1, respectively. The catalytic efficiency (kcat/Km) of the purified peroxidase was 14.75 and 29.66 s−1 mM−1 for guaiacol and H2O2, respectively. Peroxidase activity was observed to be enhanced by Cu2+, Co2+, Ni2+ and inhibited in the presence of Sn2+, Al3+, Hg2+, NaN3, EDTA and urea. Characterization showed that peroxidase purified from C. forskohlii has the ability to be used for food industrial applications.

scholarly journals Production and Characterization of Crude Protease from RS1 Isolate from silage of Floating Bladderwort (Utricularia gibba)

2020 ◽  
Vol 10 (3) ◽  
pp. 289-293
Author(s):  
Ace Baehaki ◽  
Arif Hidayat ◽  
Nuni Gofar ◽  
Rodiana Nopianti
Keyword(s):  
Molecular Weight ◽  
Production Time ◽  
Molecular Weights ◽  
Sds Page ◽  
Optimum Ph ◽  
Utricularia Gibba ◽  
Optimum Production ◽  
Optimum Ph And Temperature

The purpose of this research was to produce and characterizing crude protease from RS1 isolate of swamp plant silage. The optimum production time of RS1 isolate was 40 h. The optimum pH and temperature of protease from RS1 isolate were 10 and 45℃, respectively.  Ion Mg3+ increased RS1 protease whereas ion of Na+, K+, Fe2+, and Zn2+ inhibited protease from RS1 isolate. Study on the effect of metals ion indicated that protease from RS1 isolate was metaloenzyme. Based analysis on SDS-PAGE, the molecular weight of RS1 protease had 12 bands with molecular weights ranging from 34.75 kDa to 263.53 kDa.

Production and biochemical and molecular characterization of a keratinolytic serine protease from chicken feather-degradingBacillus licheniformisRPk

2009 ◽  
Vol 55 (4) ◽  
pp. 427-436 ◽  
Author(s):  
Nahed Fakhfakh ◽  
Safia Kanoun ◽  
Laila Manni ◽  
Moncef Nasri
Keyword(s):  
Serine Protease ◽  
Gel Filtration ◽  
Chicken Feather ◽  
Feather Degradation ◽  
Subtilisin Carlsberg ◽  
Degrading Bacterium ◽  
Step Procedure ◽  
Sds Page ◽  
Optimum Ph ◽  
Optimum Ph And Temperature

A novel feather-degrading bacterium was isolated from a polluted river and identified as Bacillus licheniformis RPk. The isolate exhibited high proteinase production when grown in chicken-feather media. Complete feather degradation was achieved during cultivation. Maximum protease activity (4150 U/mL with casein as a substrate and 37.35 U/mL with keratin as a substrate) was obtained when the strain was grown in a medium containing 7.5 g/L chicken feathers, 2 g/L yeast extract, 0.5 g/L NaCl, 0.1 g/L MgSO4·7H2O, 0.7 g/L KH2PO4, and 1.4 g/L K2HPO4for 48 h with agitation of 200 rev/min at 37 °C. The major protease produced by B. licheniformis RPk was purified to homogeneity by a 3-step procedure. The molecular mass of the purified enzyme was estimated to be 32 kDa by SDS–PAGE and gel filtration. The optimum pH and temperature for the caseinolytic activity were around 11.0 and 65 °C, respectively. The optimum pH and temperature for the keratinolytic activity were 9.0 and 60 °C, respectively. The activity of the enzyme was totally lost in the presence of phenylmethylsulfonyl fluoride, which suggests that the purified enzyme is a serine protease. The thermostability of the enzyme was considerably enhanced in the presence of Ca2+at temperatures >50 °C. The kerRP gene, which encodes the keratinolytic protease, was isolated, and its DNA sequence was determined. The deduced amino acid sequence revealed that the keratinase KerRP differs from KerA of B. licheniformis PWD-1, subtilisin Carlsberg, and keratinase of B. licheniformis by 2, 4, and 62 amino acids, respectively.

scholarly journals Purification, crystallization and characterization of N-acetylneuraminate lyase from Escherichia coli

1991 ◽  
Vol 276 (2) ◽  
pp. 541-546 ◽  
Author(s):  
K Aisaka ◽  
A Igarashi ◽  
K Yamaguchi ◽  
T Uwajima
Keyword(s):  
Escherichia Coli ◽  
Gel Filtration ◽  
Genetically Engineered ◽  
E Coli ◽  
Sds Page ◽  
Optimum Ph ◽  
Molecular Masses ◽  
Related Compounds ◽  
Optimum Ph And Temperature

N-Acetylneuraminate lyase produced by Escherichia coli was purified and crystallized from a genetically engineered strain (E. coli SF8/pNAL1). The enzyme showed apparent molecular masses of 105,000 Da on gel filtration and 35,000 Da on SDS/PAGE, suggesting that the enzyme is a trimer. The apparent optimum pH and temperature were found to be 6.5-7.0 and 80 degrees C respectively. The Km values for N-acetylneuraminate and N-glycollylneuraminate were 3.3 and 3.3 mM respectively. The enzyme was inhibited by reduction with NaBH4 in the presence of the substrate, indicating that the enzyme belongs to the Schiff-base-forming Class I aldolases. The enzyme was strongly inhibited by Cu2+ ions, p-chloromercuribenzoate and N-bromosuccinimide, and also inhibited competitively by the reaction product, pyruvate, and its structurally related compounds, dihydroxyacetone and DL-glyceraldehyde.

scholarly journals Purification, Characterization and De-Staining Potentials of a Thermotolerant Protease Produced by Fusarium oxysporum

2019 ◽  
Vol 64 (4) ◽  
pp. 539-547
Author(s):  
Mohammed Inuwa Ja’afaru ◽  
Konjerimam Ishaku Chimbekujwo ◽  
Obinna Markraphael Ajunwa
Keyword(s):  
Enzyme Activity ◽  
Fusarium Oxysporum ◽  
Treatment Time ◽  
Specific Activity ◽  
Total Yield ◽  
Tween 20 ◽  
Optimum Temperature ◽  
Industrial Enzymes ◽  
Sds Page ◽  
Optimum Ph

Proteases are important industrial enzymes and fungi prove to be good sources of such enzymes. Purification techniques are however necessary for increased specificity in activity and better industrial value. Based on this, a protease produced by a Fusarium oxysporum was purified to homogeneity by Sephadex G-200 column and α–casein agarose chromatography. The enzyme had a molecular weight of 70 kDa in SDS-PAGE. Purified Fusarium oxysporum protease had a specific activity of 93.88 U/mg protein. The purification magnitude was 7.7 and the total yield was 20 %. Purified protease had an optimum pH of 5.0 while the optimum temperature was 40 °C. The enzyme was also thermotolerant (approximately 100 % at 40 °C for 2 h). The enzyme activity was stimulated by surfactants and metal ions like, Tween-20 and Mg2+. Enzyme activity was inhibited in presence of PMSF and EDTA. Casein was found to be the best substrate for protease activity of Fusarium oxysporum FWT1. Protease were tested upon blood stain for de-clotting of blood and was found to exhibit good de-clotting and de-staining activity after 15 minutes treatment time.

scholarly journals Biochemical Properties of Mercuric Reductase from Local Isolate of Bacillus sp for Bioremediation Agent

Molekul ◽  
2017 ◽  
Vol 12 (2) ◽  
pp. 182
Author(s):  
Purkan Purkan ◽  
Yuliana Firdausi Nuzulla ◽  
Sofijan Hadi ◽  
Endang Triwahyu Prasetyawati
Keyword(s):  
Reductase Activity ◽  
Biochemical Properties ◽  
Growth Time ◽  
Resistant Bacteria ◽  
Bacillus Sp ◽  
Resistance Level ◽  
Mercuric Reductase ◽  
Mercury Reductase ◽  
Important Enzyme ◽  
Bioremediation Agent

Mercuric reductase is the important enzyme which catalyzes a reduction of a toxic Hg2+ to non-toxic Hg0. The enzyme which has been potentially used as mercury bioremediation agent is produced by mercury resistant bacteria. These research aims are to determinate the resistance level of a local Bacillus sp to HgCl2 in media, to determine the mercuric reductase activity from the bacteria, and to determine the biochemical properties of the mercuric reductase. The Bacillus sp was grown in the Nutrient Broth media with various of  0; 20; 40; 60; 120; and 160 µM HgCl2 to know the response of the bacteria against mercury, The cell growth of Bacillus sp was measured by optical density (OD) method of at λ 600 nm. The mercuric reductase activity was assayed in the solution of MRA (Mercury Reductase Assay), then the oxidized NADPH was observed by the spectrophotometry method at λ340 nm. The result showed that the Bacillus sp has been resistant to media containing mercury at 120 µM, but the microbial growth was decreased by 50% in media containing mercury 80 µM. The Bacillus sp could produce highly the mercuric reductase enzyme at 16 hours of growth time with enzyme activity as 0.574 Unit/µg. The mercuric reductase from the bacteria has an  optimum activity at pH 6 and temperature 37 °C

scholarly journals Isolasi dan Karakterisasi Inulinase dari Aspergillus niger Gmn11.1 Galur Lokal

2012 ◽  
Vol 11 (1) ◽  
pp. 19 ◽  
Author(s):  
Saryono Saryono
Keyword(s):  
Molecular Weight ◽  
Aspergillus Niger ◽  
Incubation Time ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Salt Precipitation ◽  
Sds Page ◽  
Optimum Ph ◽  
Relative Molecular Weight ◽  
Optimum Ph And Temperature

Inulin is a naturally potential polysaccharide used to produced fructose and fructooligosaccharide. Inulinaseknown also as ß-fructosidase can hydrolise inulin to fructose or fructooligosaccharide. Inulinase production fromAspergillus niger Gmn11.1 isolated from dahlia tubers is conducted using medium containing 1% inulin and 0,2%yeast extract. The crude enzyme (filtrate culture) is purified by means of ammonium sulphate salt precipitation,followed by Sephadex G25 gel filtration column chromatography and DEAE cellulose anion exchanger columnchromatography. The result indicated that the enzyme had optimum pH and temperature of 4,6 and 450C, respectivelywith incubation time of 15 hours. The Km and Vmaxs values obtained from this experiment are 20 mg/ml and 0,769mg/ml/hours, respectively. Whereas the relative molecular weight of inulinase was monitored by SDS PAGE is 63KDa.

Sign in / Sign up

Export Citation Format

Share Document