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Extraction and Purification of Lipases Enzyme from Germinating Seeds of Four Crops

Lipase enzyme has attracted a lot of attention in recent years because of its diverse biotechnological applications. The present study was conducted to screen germinated seeds of four crops, namely sunflower (Helianthus annuus), flaxor linseed (Linum usitatissimum ), peanut (Arachis hypogaea  ) and castor bean (Ricinus communis), for the activity of their lipases. to the study also included the extraction and purification of lipase from the seeds of  the most promising crop using different solvents.

The results indicated that the maximum enzymatic activity (0.669 U/ml) was  obtained when 0.1 M Tris-HCl buffer extract was used after 3 days of seed germination of all the tested species, as compared to the other test solvents (acetone and water). Sunflower germinated seeds showed the highest lipase activity, which was higher by 159.67, 185.32, and 285.90 % over the activities of castor bean, flax, and peanut seeds, respectively. Among the used ranges of saturation of ammonium sulfate, the  ratio of 70% was the best in precipitating the crude enzyme, showing a highest specific activity of 2.576 U/ mg protein.

The first stage of gel filtration chromatography column by Sephadex G-200  indicated the presence of two non-identical peaks, one for protein and another for lipase activity, between the fractions of 18 to 23. The active fractions were pooled and loaded again in the Sephadex G-200 column and the eluted fractions showed two identical peaks, one for protein and another for lipase activity, between the fractions of 19 to 23. The final purification step by gel filtration showed a specific activity of 6.482 U/mg proteins with a yield of 38.75 % and 11.33 folds of time of purification. The study revealed that sunflower seeds are a better source of lipase as compared to  the other used plant seeds, which can be used in different industries.

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Publication Date
Tue Dec 05 2023
Journal Name
Baghdad Science Journal
Purification and characterization of L-asparaginase produced from Bacillus sp.

The objective of this study was to isolate and identify the asparaginase-producing bacteria, then purify and characterize the enzyme in order to investigate their properties in the future. Fifteen local bacterial isolates were isolated from various sites in the city of Baghdad, identified by conventional morphological and biochemical procedures, and confirmed using vitek 2 methods, and submitted to primary screening processes for asparaginase production. For secondary screening, eight isolates with the greatest yellow zone ability on a specific solid medium were chosen. Bacillus sp. was reported to have the highest enzyme production (7.5 U/mg proteins). After 24 hours of incubation, submerged fermentation yielded optimal conditi

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Publication Date
Sun Jun 01 2014
Journal Name
Baghdad Science Journal
Partial purification and characterization of xylanase from Bacillus cereus X3

Three strain of Bacillus cereus were obtained from soil sours Laboratories of Biology Department/ College of Science/ University of Baghdad. The bacteria secreted extracellular xylanase in liquid cultur the test ability of xylanase production from these isolates was studied semi quantitative and quantitative screening appeared that Bacillus cereus X3 was the highest xylanase producer. The enzyme was partial purification 191 fold from cultur by reached step by 4 U/mg proteins by ammonium sulfat precipitation 80%, Ion exchang DEAE-cellulos chromatography Characterization study of the partial purifation enzyme revealed that the enzyme had a optimum activity pH8 and activity was stable in the pH rang (8-10) for 30min. maximal activity was attai

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Publication Date
Sun Mar 13 2011
Journal Name
Baghdad Science Journal
Evaluation the efficiency of different techniques for extraction and purification of Tomato yellow leaf curl virus (TYLCV)

This study was conducted to evaluate the efficacy of different techniques for extraction and purification of Tomato yellow leaf curl virus (TYLCV). An isolate of the virus free of possible contamination with other viruses infecting the same host and transmitted by the same vector Bemisia tabaci Genn. was obtained. This was realized by indicator plants and incubation period in the vector. Results obtained revealed that the virus infect Nicotiana glutinosa without visible symptoms, while Nicotiana tabaccum var. White Burley was not susceptible to the virus. The incubation period of the virus in the vector was found to be 21 hrs. These results indicate that the virus is TYLCV. Results showed that Butanol was more effective in clarification the

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Publication Date
Mon Nov 05 2018
Journal Name
Iraqi National Journal Of Nursing Specialties
Evaluation of Aqueous and Ethanolic Extraction for Coriander Seeds, Leaves and Stems and Studying their Antibacterial Activity

Objective: To evaluate two kinds of extraction (aqueous and ethanolic) for coriander using seeds, leaves and stems and
studying their antibacterial activity against nine different microorganisms.
Methodology: Coriander was selected to carry out this study. Seeds, leaves and stems were collected from local markets in
Baghdad then dried in shade for at least 10 days and grinded to fine powder. Aqueous hot extracts for 1hr. at (50
c) and
cold extracts for 24 hrs at (4
c) were performed by using seeds, leaves and stems then studied antibacterial effect against
nine different microorganisms by using well diffusion technique. Cold aqueous extracts of coriander seeds for 48 hrs. and
72 hrs and ethanolic extraction

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Publication Date
Sat Dec 01 2018
Journal Name
Al-khwarizmi Engineering Journal
Production of Protease Enzyme from Wheat Straw

      Protease enzyme production was studied and optimized as  a first step to collect information about solid state fermenter) to produce protease enzyme. A local isolated Aspergillus niger was used for this study with constant spores feeding in every experiment at (105/g). Experiments carried out in conical flasks with (250 ml) containing (10 g) of wheat straw as a substrate with different conditions included temperature, pH, hydration ratio, and fermentation time, the results comprised by measuring protease activity (u). The results showed that the best activity can be obtained at (T = 32°C, t= 100 hrs, pH= 2.5 and hydration ratio is 1:3). On the other hand the results is courage to p

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Publication Date
Sat Dec 02 2017
Journal Name
Al-khwarizmi Engineering Journal
Extraction of Chlorophyll from Alfalfa Plant

The extraction process of chlorophyll from dehydrated and pulverized alfalfa plant were studied by percolation method. Two  solvent systems were used for the extraction namely; Ethanol-water and Hexane-Toluene systems . The effect of circulation rate, solvent concentration, and solvent volume to solid weight ratio were studied. In  both ethanol water, and Hexane-Toluene systems it appears that solvent concentration is the most effective variable.

 

 

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Publication Date
Thu Mar 07 2013
Journal Name
International Journal Of Pharma Sciences
Separation and Purification of Hemolysin from Local Isolate of Serratia marcescens

Twenty isolates of Serratia marcescens were isolated from inflammation of the urinary tract (UTI)., These isolates were found to produce hemolysin as indicated by blood agar plates in which the hemolysis of red blood cell indicate a positive result. Isolates were selected according to their hemolysis activity by measuring absorbance of hemoglobin at 405 nm that released from red blood cell. Hemolysin was completely purified using 50-75% saturation of ammonium sulphate followed by ion exchange chromatography with DEAE-cellulose then gel filtration chromatography by sepharose 4B. Accordingly molecular weight for the purified toxin was estimated as 45 KD.

Publication Date
Sun Sep 01 2013
Journal Name
Baghdad Science Journal
Purification and Characterization of Endoglucanase from local isolate of Aspergillus flavus

Endoglucanase produced from Aspergillus flavus was purified by several steps including precipitation with 25 % ammonium sulphate followed by Ion –exchange chromatography, the obtained specific activity was 377.35 U/ mg protein, with a yield of 51.32 % .This step was followed by gel filtration chromatography (Sepharose -6B), when a value of specific activity was 400 U/ mg protein, with a yield of 48 %. Certain properties of this purified enzyme were investigated, the optimum pH of activity was 7 and the pH of its stability was 4.5, while the temperature stability was 40 °C for 60 min. The enzyme retained 100% of its original activity after incubation at 40 °C for 60 min; the optimum temperature for enzyme activity was 40 °C.

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Publication Date
Thu Aug 01 2019
Journal Name
Biochemical And Cellular Archives
PARTIAL PURIFICATION AND CHARACTERIZATION OF ACID PHOSPHATASE FROM SERA OF OBESE DIABETES MELLITUS PATIENTS

The study aimed to purification of acid phosphatase (ACP) from sera of obesetype 2 diabetes mellitus patients, this study included from thirty T2DM patients and thirty control, purification process was done with several steps included precipitation with inorganic salt (NH4 ) 2SO4 30%-80%, dialysis, ion exchange chromatography by DEAE sepharose anion column and size exclusion chromatography by Sepharose 6B.ACP, BMI, FBS, HbA1c, Lipid profile, Urea, Creatinie, Insuline, Homa-IR were determined. Results showed the precipitate and concentrated protein appeared four peaks in ion exchange column. ACP located in the first and second peak with purification fold (21.1), (37.2) yield of enzyme and specific activity (173.3) IU/ml, which obtained a si

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Publication Date
Sun Jul 03 2005
Journal Name
Journal Of The Faculty Of Medicine Baghdad
Purification and characterization of DNA from whole blood of children with ALL and normal children

Summary:
Background: The acute lymphocytic leukemias (All) make up about 76% of children leukemias . lymphoid leukemias occur more often than expected in patients with immunodeficiency , chromosomal abnormalities & ataxia telangiectasia . A number of chnges occur in preneoplastic & neoplastic cells as the progress towards a greater degree of malignancy . Nuclear DNA may be used as an aid in diagnosis , to predict prognosis & to determent of certain neoplasia.
Aim of the work: is to purify DNA from whole blood of predict & normal children & to characterize it by spectral studies.
Patients & Methods: Fifteen EDTA-treated blood samples of children with acute lymphoblastic leukemia (ALL) and the same number f

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