scholarly journals Kinetic studies of partially purified lipase from marine actinomycete streptomyces acrimycini ngp 1

2020 ◽  
Vol 7 (1) ◽  
pp. 63-68
Author(s):  
Vishnupriya B ◽  
Anbarasi G
Keyword(s):  
Specific Activity ◽  
Coastal Region ◽  
Kinetic Studies ◽  
Biodiesel Production ◽  
Partial Purification ◽  
Native Page ◽  
Ammonium Sulphate Precipitation ◽  
South Indian ◽  
Percent Recovery ◽  
Marine Actinomycete

This study was focused on partial purification and characterization of lipase from Streptomyces acrimyciniNGP 1, isolated from marine sediment of south Indian coastal region. In purification steps, 4.53 fold purification was achieved after 85% ammonium sulphate precipitation with 0.97 percent recovery. In further purification steps, 1.33 fold purification was achieved by Sephadex G-100 chromatography with 1.61 percent of recovery. The specific activity of purified enzyme was 1525 U/mg. Zymogram of crude enzyme on native-PAGE presented bands with lipase activity of molecular weight and Isoelectric point were 50 kDa and 7.4 respectively. These features render this lipase of interest as a biocatalyst for applications such as biodiesel production and detergent formulations.

scholarly journals New Lipase for Biodiesel Production: Partial Purification and Characterization of LipSB 25-4

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Aysel Ugur ◽  
Nurdan Sarac ◽  
Rukiye Boran ◽  
Berk Ayaz ◽  
Ozgur Ceylan ◽  
...  
Keyword(s):  
Specific Activity ◽  
Gel Filtration ◽  
Nitrogen Sources ◽  
Biodiesel Production ◽  
Partial Purification ◽  
Period Effect ◽  
Carbon And Nitrogen ◽  
Ammonium Sulphate Precipitation ◽  
Thermophilic Conditions

The lipolytic activities of 300 Streptomyces isolates were determined in Tributyrin and Rhodamine-B Agar. Lipase activities were also measured with p-nitrophenyl palmitate (p-NPP) as a substrate. The strain of Streptomyces bambergiensis OC 25-4 used in this study was selected among 300 strains of Streptomyces from MUCC as the best lipase producer. The incubation conditions were optimized and the inoculum amount, incubation period, effect of carbon and nitrogen sources, and rates of MgSO4 and CaCO3 were investigated. LipSB 25-4 (the lipase produced by S. bambergiensis OC 25-4 strain) was partially purified with ammonium sulphate precipitation, dialysis, and gel filtration chromatography 2.73-fold and with 92.12 U/mg specific activity. The optimal pH and temperature for LipSB 25-4 were determined as 8.0 and 50°C, respectively. The lipase has high stability in all pH and temperature values used in this study. While LipSB 25-4 was slightly activated in the presence of β-mercaptoethanol, it was slightly reduced by PMSF. The enzyme conserved approximately 75% of its activity at the end of 60 h, in the presence of methanol and ethanol. Since LipSB 25-4 displays high activity in the thermophilic conditions and stability in the presence of organic solvents, this lipase can catalyse the biodiesel production from olive oil by the transesterification reactions.

Partial purification and properties of apoplastic β-1,3 glucanases of groundnut leaves treated with glucan isolated from a biocontrol agent, Acremonium obclavatum

2000 ◽  
Vol 78 (2) ◽  
pp. 168-174 ◽  
Author(s):  
M Sathiyabama ◽  
R Balasubramanian
Keyword(s):  
Arachis Hypogaea ◽  
Partial Purification ◽  
Ph Optimum ◽  
Native Page ◽  
Puccinia Arachidis ◽  
Ammonium Sulphate Precipitation ◽  
Arachis Hypogaea L ◽  
Purification And Properties ◽  
Molecular Masses ◽  
Electrophoretic Techniques

Apoplastic β-1,3 glucanases (G1, G2) of Arachis hypogaea L. (peanut) leaves treated with glucan have been partially purified by ammonium sulphate precipitation, sephadex G-100, CM-Sephadex, DEAE-Sephadex chromatography, and preparative native PAGE electrophoretic techniques. The pI values of purified enzymes G1 and G2 were near 8 and 4, respectively. The apparent molecular masses of purified glucanases G1 and G2 from glucan-treated peanut leaves were 36 and 34 kDa, respectively. Both isoforms (G1 and G2) showed their pH optimum of 5.0 and temperature optimum of 40°C. The partially purified enzymes hydrolysed laminarin better than other substrates and inhibited uredospore germination of Puccinia arachidis. Both isoforms (G1 and G2) inhibited spore germination of some biocontrol agents such as Acremonium obclavatum W. Gams, Myrothecium verrucaria (Alb. Schw.) Ditmer, Fusarium solani (Mart.) Sacc., and Trichoderma harzianum Rifai.Key words: Acremonium obclavatum, Arachis hypogaea, β-1,3 glucanase, glucan, inhibition, Puccinia arachidis.

scholarly journals Study on Isolation and Partial Purification of Lactase (β-galactosidase) Enzyme from Lactobacillus Bacteria Isolated from Yogurt

2011 ◽  
Vol 4 (1) ◽  
pp. 239 ◽  
Author(s):  
N. H. M. R. Mozumder ◽  
M. Akhtaruzzaman ◽  
M. A. Bakr ◽  
F. Tuj Zohra
Keyword(s):  
Specific Activity ◽  
Culture Media ◽  
Deae Cellulose ◽  
Chemical Properties ◽  
Total Activity ◽  
Partial Purification ◽  
Anion Exchange Column ◽  
Ammonium Sulphate Precipitation ◽  
Agar Media ◽  
Modified Media

Lactase has many applications in dairy industry including for the treatment of lactose intolerance. The present study was conducted to identify the activity of lactase enzyme produced by Lactobacillus bacteria isolated from yogurts available in Dhaka city. The strains were identified to be gram positive, catalase negative, fermentative and lactase producer when cultured on selective MRS agar media by using standard bacteriological procedures and techniques. The study revealed that enzymes produced by lactobacilli were capable to produce glucose from substrate lactose in lactose modified media using lactase assay Kit Glu IB and their highest protein concentration (17.25 mg/ml) was observed in the supernatant of culture media isolated from L. lactis. Highest total activity (850.69 U/l) and specific activity (50.04 U/mg) of lactase enzyme was observed in the strain of L. bulgaricus. The crude extract which showed highest activity was further purified by ammonium sulphate precipitation followed by anion exchange column chromatography (DEAE cellulose). Final specific activity and fold purification of lactase enzyme reached to 62.80 U/mg and 1.47 respectively. The highest physic-chemical properties (Effect of pH and temperature) of lactase enzyme were observed at PH 6.0 which was 43.98 U/mg of protein and at 70°c temperature which was 111.11 U/mg of protein.Keywords: β-Galactosidase; Specific activity; DEAE cellulose; Fold purification; Yogurt.© 2012 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved.doi: http://dx.doi.org/10.3329/jsr.v4i1.8478J. Sci. Res. 4 (1), 239-249 (2012)

Partial purification and characterization of a soluble acid phosphatase from the tapeworm, Hymenolepis diminuta

1991 ◽  
Vol 65 (2) ◽  
pp. 103-110
Author(s):  
Michael J. Bumbulis ◽  
Peter W. Pappas
Keyword(s):  
Acid Phosphatase ◽  
Cellular Localization ◽  
Specific Activity ◽  
Sodium Dodecyl ◽  
Hymenolepis Diminuta ◽  
Partial Purification ◽  
Transferase Activity ◽  
Crude Enzyme Preparation ◽  
Ammonium Sulphate Precipitation ◽  
And Function

ABSTRACTAn acid phosphatase activity (APA; EC 3.1.3.2) was demonstrated in homogenates of adult Hymenolepis diminuta. The APA was soluble based on the observation that it did not sediment at 130 000 g. APA was partially purified using a combination of differential centrifugation, ammonium sulphate precipitation, chloroform extraction, and gel and fast-protein-liquid-chromatography. This combination of techniques resulted in a preparation with a specific activity approximately 500 times greater than the crude enzyme preparation. The temperature and pH optima of the partially purified APA were 44°C and pH 5·0. The enzyme appeared to be a monomer with a molecular weight of approximately 62 000. APA had a higher affinity for a greater activity towards aromatic than aliphatic phosphoesters and phosphoryl transferase activity was demonstrable using 1-butanol and ethylene glycol as acceptors. APA was inhibited significantly by sodium dodecyl sulphate, fluoride, molybdate and tartrate, but CuSO4 and Fast Garnet GBC were poor inhibitors. The precise cellular localization and function of this enzyme remains unknown since it possesses characteristics of both cytoplasmic and lysosomal APA's of other organisms.

scholarly journals Partial Purification of Alkaline Protease as Thrombolytic Agent from Mutant Strain Bacillus licheniformis EMS250-O-1

2017 ◽  
Vol 15 (2) ◽  
pp. 135-141 ◽  
Author(s):  
Md Asad Uz Zaman ◽  
Md Arafat Al Mamun ◽  
Shakila Nargis Khan ◽  
Md Mozammel Hoq ◽  
Md Abdul Mazid
Keyword(s):  
Mutant Strain ◽  
Bacillus Licheniformis ◽  
Specific Activity ◽  
Cardiovascular Complications ◽  
Clot Lysis ◽  
Partial Purification ◽  
Thrombolytic Activity ◽  
Ammonium Sulphate Precipitation ◽  
Maximum Stability

Thrombosis leads to myocardial infarction, stroke and other cardiovascular complications. Microbial thrombolytic agents such as urokinase, streptokinase etc. are used to treat complications related to thrombosis. To search for new microbial enzymes as thrombolytics having better efficacy and specificity, Bacillus licheniformis EMS-O-1 mutant strain was cultured in modified urea-molasses media followed by purification using ammonium sulphate precipitation and ultrafiltration through centricon tube of 100 MWCO value. The yield of crude enzyme was 11129.14 U/mg and after purification 40180.46 U/mg. Purification process increased the specific activity of purified enzyme to 12.28 fold with a recovery of 17.79%. The purified enzyme was a serine protease with molecular weight of 25.5 kDa as confirmed by irreversible inhibition of activity with phenylmethylsulfonyl fluoride (PMSF) followed by SDS-PAGE gel image and by LC-MS analyses. In vitro clot lysis assay of the purified enzyme exhibited 38.30% thrombolytic activity. The crude enzymes from the mutant strain EMS-O-1 were found to be stable up to 50oC and showed maximum stability between pH range 7.5 to 8.5. These findings signify that proteases produced by B. licheniformis mutant have the potential to be developed as a viable thrombolytic agent.Dhaka Univ. J. Pharm. Sci. 15(2): 135-141, 2016 (December)

PARTIAL PURIFICATION OF BOVINE ANTIPLASMIN (ANTIFIBRINOLYSIN)

1960 ◽  
Vol 38 (9) ◽  
pp. 1021-1027
Author(s):  
Edmond R. Cole ◽  
Arthur Dalby ◽  
Edwin T. Mertz
Keyword(s):  
Calcium Phosphate ◽  
Ammonium Sulphate ◽  
Bovine Serum ◽  
Specific Activity ◽  
Large Diameter ◽  
Small Diameter ◽  
Partial Purification ◽  
Isoelectric Precipitation ◽  
Ammonium Sulphate Precipitation ◽  
Column Eluate

Bovine antiplasmin (antifibrinolysin) solutions with activities of 12–19 units/mg nitrogen have been obtained from bovine serum by 60% ammonium sulphate precipitation and isoelectric precipitation at pH 5.0 representing approximately a 6-fold purification of antiplasmin from serum. An additional 6.4-fold purification of antiplasmin was obtained by chromatography on a large diameter calcium phosphate gel column. Dialysis and lyophilization of the column eluate caused a 30% loss in activity. The dry antiplasmin was 25 times more potent than serum protein and was stable for 12 months when stored in a desiccator over P2O5 at −10°. Antiplasmin solutions with a specific activity twice that obtained with the large column could be eluted from a small diameter calcium phosphate column, but the activity decreased too rapidly to permit preparation of a lyophilized product.

PARTIAL PURIFICATION OF BOVINE ANTIPLASMIN (ANTIFIBRINOLYSIN)

1960 ◽  
Vol 38 (1) ◽  
pp. 1021-1027
Author(s):  
Edmond R. Cole ◽  
Arthur Dalby ◽  
Edwin T. Mertz
Keyword(s):  
Calcium Phosphate ◽  
Ammonium Sulphate ◽  
Bovine Serum ◽  
Specific Activity ◽  
Large Diameter ◽  
Small Diameter ◽  
Partial Purification ◽  
Isoelectric Precipitation ◽  
Ammonium Sulphate Precipitation ◽  
Column Eluate

Bovine antiplasmin (antifibrinolysin) solutions with activities of 12–19 units/mg nitrogen have been obtained from bovine serum by 60% ammonium sulphate precipitation and isoelectric precipitation at pH 5.0 representing approximately a 6-fold purification of antiplasmin from serum. An additional 6.4-fold purification of antiplasmin was obtained by chromatography on a large diameter calcium phosphate gel column. Dialysis and lyophilization of the column eluate caused a 30% loss in activity. The dry antiplasmin was 25 times more potent than serum protein and was stable for 12 months when stored in a desiccator over P2O5 at −10°. Antiplasmin solutions with a specific activity twice that obtained with the large column could be eluted from a small diameter calcium phosphate column, but the activity decreased too rapidly to permit preparation of a lyophilized product.

PURIFICATION AND CHARACTERIZATION OFESTERASE ENZYME ISOLATED FROM CONTAMINATED SOIL SAMPLE

2019 ◽  
pp. 1-3
Author(s):  
S. Chithra ◽  
K. Silambarasi ◽  
I. Yasmin
Keyword(s):  
Soil Sample ◽  
Ammonium Sulphate ◽  
Contaminated Soil ◽  
Specific Activity ◽  
Nitrogen Sources ◽  
Partial Purification ◽  
Carbon And Nitrogen ◽  
Crude Enzyme Preparation ◽  
Ammonium Sulphate Precipitation ◽  
Highly Active

Isolation and partial purification of esterase from contaminated soil sample was the main aim of this study. The production medium for organism was optimized by using different pH,Temperature,Carbon and Nitrogen sources.The esterase enzyme was highly active and stable from pH 5.0 to 9.0 with an optimum at pH 9.Its optimum temperature was 35°C. The best carbon and nitrogen sources were mannitol and yeast extract. Esterase was partially purified by ammonium sulphate precipitation,dialysis.The specific activity of partially purified esterase obtained from ammonium sulphate fractionation is found to be 8.6485U/mg and the fractionation is 5.4 fold purified from the crude enzyme preparation yielding 17.5U/mg from the crude protein. This result showed that Bacillus subtilis under study is a good producer of extra cellular esterase,which can be beneficial for industries

scholarly journals The Screening, Characterization and Identification of Chitinolytic Actinomycetes Streptomyces sp. ANU5a.

2021 ◽  
Author(s):  
Piyush Kumar Tiwari ◽  
Shubhjeet Mandal ◽  
Anchal Anchal
Keyword(s):  
Ammonium Sulphate ◽  
Casein Hydrolysate ◽  
Plant Growth Promoting Rhizobacteria ◽  
Chitinase Activity ◽  
Precipitation Method ◽  
Partial Purification ◽  
Native Page ◽  
Streptomyces Sp ◽  
Mangrove Soil ◽  
Ammonium Sulphate Precipitation

Abstract Streptomyces sp. ANU5a is a potential plant growth-promoting Rhizobacteria and a highly effective biocontrol agent. It produces Chitinases in broth media containing Chitin. Earlier reports on the chitinase production revealed the key importance of casein hydrolysate and yeast extract as additional sources of carbon and nitrogen to increase the yield. 16S rRNA-based method was used to identification of a Chitinolytic Actinomycete a Streptomyces sp. ANU5a was completed from Mangrove soil samples. Ammonium sulphate precipitation method was used for partial purification of Chitinases. The Chitinases have optimum activity at a temperature of 50-55°C and pH 5-6. Three protein bands of 20kDa, 67kDa, and 240kDa are resolved by Native-PAGE of Ammonium Sulphate fractions; the smallest band of 20kDa belongs to Chitinase. Enzyme activity for chitinase was recorded by zymogram analysis. Streptomyces sp. ANU5a was found to have notable antifungal activity on culture plates. We correlate the antimicrobial activity of Streptomycetes sp. ANU5a with Chitinase activity and other PGPR traits.

scholarly journals Extraction, Characterization and Partial Purification of L-Asparaginase from the Leaves of Arachis Hypogaea L.

2019 ◽  
Vol 16 (3) ◽  
pp. 681-691
Author(s):  
Niranjana. J Niranjana ◽  
Kandasamy Arun Gandhi ◽  
D. Sunmathi ◽  
P. Nanthavanan
Keyword(s):  
Metal Ions ◽  
Arachis Hypogaea ◽  
Specific Activity ◽  
Lymphoblastic Leukemia ◽  
Gel Filtration ◽  
Size Exclusion ◽  
Partial Purification ◽  
Crude Enzyme Extract ◽  
Ammonium Sulphate Precipitation ◽  
Arachis Hypogaea L

L-asparaginase has been a promising therapeutic agent in the treatment of acute lymphoblastic leukemia and lymphoma. In recent times, due to the side effects of commercially available bacterial L-asparaginase and its unavoidable importance, plants are being explored as the source of L-asparaginase. The enzyme L-asparaginase was partially purified from Arachis hypogaea L. The crude enzyme extract was subjected to different purification steps including ammonium sulphate precipitation, dialysis followed by separation on Sephadex G-100 gel filtration (size exclusion chromatography) to obtain partially pure form of L - asparaginase. The enzyme was partially purified to 118 folds and contained specific activity of 4686.86 U/mg with 9.85% yield. SDS-PAGE electrophoresis of the partially purified enzyme revealed that it was a single protein with molecular weight of 70 kDa. The study on physiochemical properties showed that L - asparaginase from Arachis hypogaea L. was potassium-dependent in nature, where its optimum pH of enzyme activity was found to be 8.0 and temperature as 40°/50°C with reaction time of 15 - 20 minutes. Also it was observed that the L-asparaginase activity increased with the presence of metal ions such as Na+, Mg++, making it an enzyme dependent on metal ions for its reaction. In addition to this, it was revealed that the enzyme was partially inhibited in presence of certain chelators. The specificity of L-asparaginase obtained from Arachis hypogaea L. with lack of urease activity and minimal glutaminase activity along with less cytotoxicity on human blood indicated it as an efficient chemotherapeutic agent that could be investigated further in future studies.

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