scholarly journals Studies on Anti-Cancer Activity of Lysyl Oxidase from Trichoderma Viride MTCC 167

2016 ◽  
Vol 4 (1) ◽  
pp. 57-63
Author(s):  
Shipra Kalra ◽  
Kanav Midha ◽  
Monika ◽  
Manpreet Kaur
Keyword(s):  
Enzyme Activity ◽  
Protein Content ◽  
Ammonium Sulphate ◽  
Specific Activity ◽  
Lysyl Oxidase ◽  
Trichoderma Viride ◽  
Effect Of Temperature ◽  
Ovarian Cancer Cells ◽  
Acetone Precipitation ◽  
Ammonium Sulphate Precipitation

Lysyl oxidase produces H2O2 i.e. ROS by acting on L-Lysyine. In the present study, ROS thus produced has been used by in vitro cytotoxicity assay on ovarian cancer cells thereby achieving 250 percent inhibition. Lysyl oxidase activity was determined spectrophotometrically by Dintrophenylhydrazine (DNPH) reagent. For isolation of lysyl oxidase produced by Trichodermaviride, acetone precipitation and ammonium sulphate precipitation was carried out. The effect of temperature on lysyl oxidase production was determined by incubating the media with 1.5 % inoculum at different temperature ranging from  10 , 20, 30,40,50,60,70 , 80 0C for 72 hr . Anti-tumor properties of Lysyl oxidase was checked using Rhodamine assay and NBT Assay.  Comparative results for Acetone and Ammonium Sulphate precipitation showed that Enzyme activity(U/ml)  of acetone precipitation is 124.6 and 144.3 IU/ml and Protein Content is 1.8 and 2.2 mg/ml with the specific activity 68.4IU/mg and 64.1IU/mg . The optimum enzyme production was found to be at pH 8 and optimum temperature for lysyl oxidase production was 500C with maximum enzyme activity of 0.38 (U/ml). 7th fraction contained highest enzyme activity so the retention time of lysyl oxidase was found to be 7 min. The protein content was also calculated by using Bradford method and it was highest in the 1st fraction and in 6th, 7th and 8th fractions. The specific activity has been improved from 75.1 IU/mg to 86.5 IU/mg. 10 units of lysyl oxidase inhibits 3x104 cells in each well to 82.5% and inhibition achieved at same cell count at 200 units which was 250% as observed with NBT and Rhodamine assay.Int J Appl Sci Biotechnol, Vol 4(1): 57-63

scholarly journals Bromelain Activity of Waste Parts of Two Pineapple Varieties

2018 ◽  
Vol 2 ◽  
pp. 21-28 ◽  
Author(s):  
Edmund Ofosu Benefo ◽  
Isaac Williams Ofosu
Keyword(s):  
Enzyme Activity ◽  
Ammonium Sulphate ◽  
Crude Extract ◽  
Specific Activity ◽  
High Demand ◽  
Digestion Method ◽  
Waste Products ◽  
Food Industries ◽  
Ethanol Precipitation ◽  
Ammonium Sulphate Precipitation

Bromelain, a protease found in pineapples, is of high demand in the pharmaceutical, cosmetic and food industries. Along the pineapple processing chain, waste products such as peels, crowns, stems and cores result. These parts are usually discarded, though they contain significant amounts of the enzyme bromelain. This study sought to determine the bromelain activity of the crowns and peels of two pineapple varieties grown in Ghana;MD2andSugarloaf. The crude extract was obtained by homogenising the peels and crowns in a cold phosphate buffer and centrifuging at 3000 rpm for 15 min. Ethanol and ammonium sulphate precipitation were carried out on the extract between 30% – 80% precipitation levels. The enzyme activity was determined using the casein digestion method. Results showed that bromelain was precipitated mainly in the 30% – 60% precipitation range.Sugarloafcrowns yielded the highest enzyme activity of 20.82 U/ml and a specific activity of 194.58 U/mg at the 40% ammonium sulphate precipitation level. This was followed by theSugarloafpeels with an enzyme activity of 19.98 U/ml at 50% ethanol precipitation level. Ethanol precipitation resulted in fractions with lower bromelain activity. Enzyme activity was higher in theSugarloafvariety and also in the crowns of both varieties. The two pineapple varieties have significant levels of bromelain activity and could be exploited for commercialisation.

scholarly journals Produksi Enzim Selulase Kasar dari Isolat Bakteri B2S8 menggunakan Substrat Brangkasan Jagung dengan Perlakuan Konsentrasi Inokulum dan Komposisi Media yang berbeda

2020 ◽  
Vol 8 (2) ◽  
pp. 267
Author(s):  
Nursatria Purba ◽  
Ida Bagus Wayan Gunam ◽  
I Made Mahaputra Wijaya
Keyword(s):  
Enzyme Activity ◽  
Protein Content ◽  
Corn Stover ◽  
Bacterial Isolate ◽  
Specific Activity ◽  
Block Design ◽  
Cellulolytic Bacteria ◽  
Endoglucanase Activity ◽  
Cellulase Enzyme ◽  
The Media

The purpose of this study was determined the media and concentration of cellulolytic bacterial isolates to produce high cellulase enzyme activity. Production of crude cellulase enzyme in media and concentration of different bacterial isolate used a factorial Randomized Block Design (RBD) which consist of two factors. The first factor was the media production of different cellulase enzyme consisting of 3 levels, namely media 1, 2 and 3. The second factor was the concentration of bacterial isolate consisting of 5 levels namely 1, 2, 3, 4 and 5%. This study used a B2S8 cellulolytic bacterial isolate that has the highest value of cellulase enzyme activity and the highest degradation rate of cellulose in previous studied and determined the ability of exoglucanase enzyme activity, endoglucanase enzyme and dissolved protein content produced from cellulolytic bacterial isolate. This study used Carboxymethyl Cellulose (CMC) for enzyme activity test and 1% corn stover as a substrate on the media to produce crude cellulase enzyme. The result showed that the highest cellulase enzyme activity in the third media and 5% cellulolytic bacterial inoculum concentration resulted in endoglucanase activity of 0.0332 IU/mL, exoglucanases enzyme activity of 0.0060 IU/mL, dissolved protein content in the amount of 0.5670 mg/mL, the specific endoglucanase activity of 0.0807 IU/mg and the specific activity of exoglucanase of 0.0123 IU/mg. Keywords: Cellulolytic bacteria, Cellulase enzymes, Enzyme activity, Corn stover

scholarly journals Aspergillus niveus Blochwitz 4128URM: new source for inulinase production

2005 ◽  
Vol 48 (3) ◽  
pp. 343-350 ◽  
Author(s):  
Cristina Maria de Souza-Motta ◽  
Maria Auxiliadora de Queiroz Cavalcanti ◽  
Ana Lúcia Figueiredo Porto ◽  
Keila Aparecida Moreira ◽  
José Luiz de Lima Filho
Keyword(s):  
Enzyme Activity ◽  
Ammonium Sulphate ◽  
Crude Extract ◽  
Culture Medium ◽  
Food Industries ◽  
Ammonium Sulphate Precipitation ◽  
Aspergillus Niveus ◽  
Optimal Ph

Aspergillus niveus Blochwitz 4128 URM isolated from sunflower rhizosphere demonstrated a new source of inulinase. The enzyme was produced in culture medium containing inulin as substrate in the concentrations: 10, 15 and 20g L-1. Maximum enzyme activity was obtained in medium containing 20g L-1 inulin. The enzyme was partially purified using ammonium sulphate precipitation, followed by ion charge (DE-32) and molecular exclusion (Sephadex) chromatography. The results showed the optimal pH and temperature of inulinase from crude extract were 4.0 and 4.8 and 45ºC, respectively. The enzyme was purified 34.65 fold with yield of 53.63%. A. niveus 4128URM can be used in the inulinase production with use in the food industries.

STUDIES ON THE PURIFICATION OF BOVINE PLASMINOGEN (PROFIBRINOLYSIN)

1960 ◽  
Vol 38 (1) ◽  
pp. 1029-1034 ◽  
Author(s):  
Arthur Dalby ◽  
Edmond R. Cole ◽  
Edwin T. Mertz
Keyword(s):  
Calcium Phosphate ◽  
Ammonium Sulphate ◽  
Bovine Serum ◽  
Specific Activity ◽  
Gel Chromatography ◽  
Isoelectric Precipitation ◽  
Ammonium Sulphate Precipitation ◽  
Ph Range

A crude bovine plasminogen product obtained from bovine serum by 30% ammonium sulphate precipitation has been purified 20-fold by means of isoelectric precipitation and calcium phosphate gel chromatography. A threefold purification was achieved by isoelectric precipitation. Plasminogen was precipitated in greatest yield and highest specific activity at 40-fold dilution in the pH range of 5.25–5.50. The conditions under which plasminogen is eluted from calcium phosphate gel columns have been investigated. Plasminogen fractions possessing specific activity seven times that of the isoelectric-precipitated materials have been obtained by elution with phosphate buffers over the range of 0.05 to 0.1 M, pH 6.8.

Purification and Partial Characterization of Cellulase Produced by Aspergillus niger Cultured on Vitellaria paradoxa shells

2017 ◽  
Vol 6 (2) ◽  
Author(s):  
Abdulhakeem Olarewaju Sulyman ◽  
Yusuf A Iyanda ◽  
Afolabi Olaniyi Opasola ◽  
OtunOla Adedayo ◽  
Raliat Abimbola Aladodo
Keyword(s):  
Aspergillus Niger ◽  
Specific Activity ◽  
Gel Filtration ◽  
Inoculum Size ◽  
Partial Characterization ◽  
Effect Of Temperature ◽  
Vitellaria Paradoxa ◽  
Endoglucanase Activity ◽  
Ammonium Sulphate Precipitation

This research investigated the purification and partial characterization of cellulase produced by Aspergillus niger cultured on Vitellaria paradoxa shells. Cellulase (endoglucanase) from A. niger was produced under optimum fermentation conditions at 35 °C, pH 4.7, V. paradoxa, 4 g/L, inoculum size of 10 mm and the fermentation media incubated for 120 hours. The crude endoglucanase obtained were partially purified by subjecting to ammonium sulphate precipitation, dialysis and gel filtration chromatography for further purification. The effect of temperature and pH on the activity of purified endoglucanase was determined. Cellulase was purified to 734.33 folds by Sephadex G-100 column chromatography with a specific activity and yield of 4.406 U/mg and 63.03% respectively. Fractions 4 and 7 contained the highest endoglucanase activity out of 18 fractions collected and the two fractions were pooled for further analysis. The activity of purified endoglucanase was optimum at a temperature of 40 °C and pH 5. Therefore, the purified endoglucanase produced may be explored in detergent industry.

THE DEVELOPMENT OF A RADIOIMMUNOASSAY FOR OXYTOCIN: RADIO-IODINATION, ANTIBODY PRODUCTION AND SEPARATION TECHNIQUES

1970 ◽  
Vol 46 (2) ◽  
pp. 269-278 ◽  
Author(s):  
T. CHARD ◽  
M. J. KITAU ◽  
J. LANDON
Keyword(s):  
Lymph Node ◽  
Human Plasma ◽  
Rapid Method ◽  
Ammonium Sulphate ◽  
Antibody Production ◽  
Specific Activity ◽  
High Specific Activity ◽  
Separation Techniques ◽  
Ammonium Sulphate Precipitation

SUMMARY A simple and rapid method is described for labelling oxytocin with 131I at a high specific activity. This method is compared with those of previous workers. A satisfactory antiserum has been raised by direct intra-lymph node injection of oxytocin adsorbed to carbon microparticles. A number of methods for separating antibody-bound from free oxytocin are described, and reasons given for preferring a procedure using ammonium sulphate precipitation. These data form the basis for developing a radioimmunoassay intended for the determination of oxytocin in human plasma.

Purification of a Fibrinolytic Enzyme from Bacillus Sp. Isolated from Budu

2012 ◽  
Vol 59 (1) ◽  
Author(s):  
Nurulhanis Ahmad Sanusi ◽  
Haryati Jamaluddin
Keyword(s):  
Ammonium Sulphate ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Culture Broth ◽  
Fibrinolytic Enzyme ◽  
Total Protein Content ◽  
Bacillus Sp ◽  
Ammonium Sulphate Precipitation ◽  
Sds Page

Bacillus sp. strain B1 producing wild type fibrinolytic enzyme was isolated from Budu. The fibrinolytic enzyme was collected from the supernatant of Bacillus sp. strain B1 culture broth and purified to electrophoretic homogeneity through a combination of various purification schemes, which include ammonium sulphate precipitation, followed by anion exchange chromatography using DEAE–Sepharose Fast Flow and gel filtration chromatography on Sephadex G–75 column. During ammonium sulphate precipitation screening, it was observed that the crude enzyme from Bacillus sp. strain B1 precipitated at 40% and 50% of ammonium sulphate saturation respectively. The fibrinolytic enzyme was purified 58.5–fold with a final yield of 0.51%. The specific activity was determined to be 1.17 Units/mg using plasmin as standard and the final total protein content was 8.58 mg/ml. After the successive purification steps, the estimated molecular mass of fibrinolytic enzymes from strain B1 was estimated via sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE). SDS–PAGE analysis showed a single band at 45 kDa corresponding to the purified fraction with fibrinolytic activity.

scholarly journals Biobleaching of Industrial Important Dyes with Peroxidase Partially Purified from Garlic

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Akudo Chigozirim Osuji ◽  
Sabinus Oscar O. Eze ◽  
Emmanuel Emeka Osayi ◽  
Ferdinand Chiemeka Chilaka
Keyword(s):  
Ammonium Sulphate ◽  
Specific Activity ◽  
Low Cost ◽  
Gel Filtration ◽  
Enzyme Concentration ◽  
Gel Filtration Chromatography ◽  
Vat Dyes ◽  
Ammonium Sulphate Precipitation ◽  
Ph Range ◽  
Better Than

An acidic peroxidase was extracted from garlic (Allium sativum) and was partially purified threefold by ammonium sulphate precipitation, dialysis, and gel filtration chromatography using sephadex G-200. The specific activity of the enzyme increased from 4.89 U/mg after ammonium sulphate precipitation to 25.26 U/mg after gel filtration chromatography. The optimum temperature and pH of the enzyme were 50°C and 5.0, respectively. The Km andVmaxfor H2O2and o-dianisidine were 0.026 mM and 0.8 U/min, and 25 mM and 0.75 U/min, respectively. Peroxidase from garlic was effective in decolourizing Vat Yellow 2, Vat Orange 11, and Vat Black 27 better than Vat Green 9 dye. For all the parameters monitored, the decolourization was more effective at a pH range, temperature, H2O2concentration, and enzyme concentration of 4.5–5.0, 50°C, 0.6 mM, and 0.20 U/mL, respectively. The observed properties of the enzyme together with its low cost of extraction (from local sources) show the potential of this enzyme for practical application in industrial wastewater treatment especially with hydrogen peroxide. These Vat dyes also exhibited potentials of acting as peroxidase inhibitors at alkaline pH range.

scholarly journals PURIFICATION OF β-GLUCOSIDASE PRODUCED FROM Trichoderma viride USING COW DUNG AS CARBON SOURCE

2020 ◽  
Vol 4 (2) ◽  
Author(s):  
S. T. Tyohemba ◽  
S. Aliyu ◽  
N. N. Ndukwe ◽  
G. G. Memi ◽  
U. O. Edem
Keyword(s):  
Specific Activity ◽  
Trichoderma Viride ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Broad Peak ◽  
Cow Dung ◽  
Elution Profile ◽  
Ammonium Sulphate Precipitation ◽  
Sds Page ◽  
Peak Fraction

β-glucosidases have characteristics of biotechnological interest and have thus become important industrial enzymes.In this study, β-glucosidase produced by Trichoderma viride from cow dung was subjected to a three step purification process involving ammonium sulphate precipitation, gel filtration by Sephadex G-100 and ion exchange chromatography by DEAE-Sephadex A-25. The elution profile on Sephadex G-100 resulted in a single broad peak (fractions 9-21) which had a yield of 3.7% and a purification fold of 4.29 with a specific activity of 25.70 µmol/min/mg proteins while the elution profile on DEAE-Sephadex A-25 resulted in a single broad peak (fraction 8-14) which had a yield of 2.76% and a purification fold of 22.14 with a specific activity of 132.41µmol/min/mg of protein. The purified enzyme was obtained as a single band and had a molecular mass of 51.8 kDa on SDS-PAGE. This results provide support for further studies of this enzyme towards revealing its potential biotechnological applications.

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