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Cloning, Expression and Purification of Putative Isovaleryl-CoA Dehydrogenase from Paracoccus denitrificans Pd1222

In Paracoccus denitrificans Pd1222 bacterium, Pden_3633 encoding gene has been nominated to encode for Isovaleryl CoA dehydrogenase (IVDH) [1], the enzyme which involve in leucine catabolism pathway. In this study, this putative IVDH was investigated. IVDH encoding gene from P. denitrificans Pd1222 in addition to desired features for cloning, expression and purification have been designed and synthesized. The synthetic coding sequence was expressed in Escherichia coli. The enzyme was purified as a Strep-Tagged protein with a total protein 220.5 mg. An apparent molecular weight of 42.9 kDa was determined on SDS gel. Amino acid alignment showed a very high similarity (91-96%) with corresponding IVDH from several other Paracoccus species. As for genera other than Paracoccus; Roseovarius mucosus, Catellibacterium nectariphilum and Oceanicola nanhaiensis recorded the highest similarity (85-86%), Suggesting that these mentioned species all have similar IVDH.

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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
Sun Apr 26 2020
Journal Name
Iraqi Journal Of Science
Extraction and Partial Purification of Exo-Polysaccharide from Lactobacillus Rhamnosus Isolated from Vaginal Specimens

Lactobacillus spp. is one of the most important strains used worldwide in different applications that range from medical to industrial uses. Therefore, this study aimed to determine the potential capability of the putative probiotic L. rhamnosus isolated from clinical vaginal specimens to produce exopolysaccharide (EPS). From a total of 100 clinical samples, only 13 (13%) samples were represented as Lactobacillus spp, as characterized by the use of the API 50CHL system. The results revealed that the number of L. rhamnosus isolates constituted 4/13 (30.8%), with a confident percentage of more than 80%. In addition, characterization by 16S rRNA sequencing showed 100% similarity to the characterized species of L. rhamnosus. Also, the result

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Publication Date
Thu May 25 2017
Journal Name
Ibn Al-haitham Journal For Pure And Applied Sciences
Purification, Characterization and Antifungal Activity of Chitinase from Serratia marcescens Isolated from Fresh Vegetables

  Seven [35%] and five [25%] Serratia marcescens  isolates were obtained out of 20 samples of lettuce and 20 samples of spinach, respectively, taken from different locations in a farm in Baghdad city. The isolate that produced chitinase in higher level was chosen to purify chitinase through several stages of purification including: ammonium sulfate precipitation, DEAE- sephadex ion exchange chromatograpgy and sephadex G-200 gel filtration with 89.5- fold purification and 30% recovery.       The purified chitinase was characterized and the molecular weight of enzyme was 59000 daltons by using gel filtration chromatography. The optimum pH and temperature of the purified chitinase were 6.0 and 50°

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Publication Date
Fri Jun 24 2022
Journal Name
Iraqi Journal Of Science
Production, Purification and characterization of bioactive compounds from locally Streptomycesrochei M78

The aim of the current study was to optimize different cultural and environmental conditions for production the antibacterial bioactive metabolites by Streptomyces rochei M78 isolated from agriculture soil, in Baghdad, Iraq. The effect of various parameters such as, culture media, incubation time, pH, carbon and nitrogen sources, C: N ratios and inducers on antibacterial metabolite production was studied by varying single parameter at a time. It was found from the results that higher metabolite production by isolate observed using starch casein broth (SCB) as the best production medium, at initial pH 7.0.Starch andcasein +yeast extract + peptone appeared to be the best carbon and nitrogen sources respectively and C: N ratio of 4: 1 after

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Publication Date
Thu Mar 30 2023
Journal Name
Iraqi Journal Of Science
Extraction ،Purification and Characterization Of Peroxidase From cabbage (Brassica oleracea Var)

The activity of peroxidase (POD) in cabbage was evaluated using
spectrophotometric method. The enzyme was extracted from the cabbage leaves
with 0.1 M phosphate buffer solution pH 7. 0 . POD activity was determined using
(O-dianisidine) as a substrate. The effects of the amounts of enzyme extract,
substrate concentration, pH and temperature were investigated. The highest activity
of POD was recored at 2 mg/ml. The highest activity of POD was optimized with
16 mM O-dianisidine, The optimum pH was 7.0 for POD , The optimum
temperature was 30°C for POD. These optimum conditions were used to
determined the enzyme activities in cabbage sample. Acetone fractionated
peroxidase from crude extract of Brassica oleracea

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Publication Date
Sun Apr 30 2023
Journal Name
Iraqi Journal Of Science
Production, Purification and Characterization of Cellulose from Local Isolate of Pantoea spp

Soil samples from fields cultivated with barley and wheat in addition to samples
from spoiled orange and apple fruits and carrot roots were collected with the aim to
isolate cellulase producing bacterial strains. Bacterial isolates obtained from these
samples were grown on a selective medium containing carboxymethyl cellulose
(CMC) as a sole source for carbon and energy. Results showed that nine isolates out
of fifty were able to produce cellulase.The specific activity of cellulase in culture
filtrate of the most efficient isolate was 1.601 u/mg protein.This isolate was
identified according to its morphological characteristics and biochemical tests, and
then by using Api 20-E and VITEK-II identification systems an

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Publication Date
Tue Sep 29 2020
Journal Name
Iraqi Journal Of Science
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 spe

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Publication Date
Sat Sep 30 2023
Journal Name
Iraqi Geological Journal
Purification of Aqueous Solutions from Nickel Using Ceramic Waste

This study aims to test ceramic waste's capacity to remove nickel from aqueous solutions through adsorption. Ceramic wastes were collected from the Refractories Manufacturing Plant in Ramadi. Through a series of lab tests, the reaction time (5, 10, 15, 20, 25, 30, 35, 40, 45, and 50 minutes, and Ni concentrations (20, 40, 60, and 80) were tested using ceramic wastes with a solid to liquid ratio of 2g/30ml. At a temperature of 30ºC, the pH, total dissolved solids (TDS), and electrical conductivity (EC) were all measured. The equilibrium time was set at 30 min. Thereafter, the sorption (%) somewhat increased positively with the Ni concentration. Freundlich's equation showed that the adsorption intensity is 1.1827 and the Freundlich c

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Publication Date
Fri Jan 26 2024
Journal Name
Iraqi Journal Of Science
The Extraction and Partial Purification of Wedelolactone from Local Eclipta alba Plant

Abstract:
Eclipta alba is a weed growing in damp, moist puddles distributed in the tropical and subtropical regions, so the most weight of the plant is water, which reached in to 90%. The extraction method by using different solvents (Methanol, Ethanol, Hexane and Aqueous) showed, the best yield with methanol reached to 76%, and the yield decreased with ethanol and hexane reached 55% and 53% respectively, while the minimum yield observed with aqueous hot and cold extraction reached 11% and 5% respectively. The phytochemical compound characterization showed the compounds (Coumarines , Flavones ,Volatile Oil ,Tannins , Saponines , Glycosides ,Carbohydrates ,Alkaloids , Resins) with different percentage. The thin layer chromatography det

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