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Production of 7-methylxanthine from Theobromine by Metabolically Engineered E. coli
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In this work, a novel biocatalytic process for the production of 7-methylxanthines from theobromine, an economic feedstock has been developed. Bench scale production of 7-methlxanthine has been demonstrated. The biocatalytic process used in this work operates at 30 OC and atmospheric pressure, and is environmentally friendly. The biocatalyst was E. coli BL21(DE3) engineered with ndmB/D genes combinations. These modifications enabled specific N7- demethylation of theobromine to 7-methylxanthine. This production process consists of uniform fermentation conditions with a specific metabolically engineered strain, uniform induction of specific enzymes for 7-methylxanthine production, uniform recovery and preparation of biocatalyst for reaction and uniform recovery of pure 7-methylxanthine.

   Many E. coli BL21(DE3) strains metabolically engineered with single and/or multiple ndmB/D genes were tested for catalytic activity, and the best strains which had the higher activity were chosen to carry out the N-demethylation reaction of theobromine. Strain pBD2dDB had the highest activity for the production of 7-methylxanthine from theobromine. That strain was used to find the optimum amount of cells required to achieve complete conversion of theobromine to 7-methylxanthine within two hours. It was found that the optimum concentration of pBD2dDB strain to achieve 100% conversion of 0.5 mM theobromine to 7-methylxanthine was 5 mg/mL. The cell growth of pBD2dDB strain was studied using two different growth media, (Luria-Bertani Broth and Super Broth). Super broth was found to be the best medium to produce the highest amount of cell paste (1.5 g). Subsequently, the process was scaled up in which 2 L reaction volume was used to produce 7-methylxanthine (100% conversion) from 0.5 mM theobromine catalyzed by pBD2dDB strain. The reactions was carried out at 30 oC and 250 rpm shaker speed, and the reaction medium was 50 mM potassium phosphate buffer (pH=7). 7-methylxanthines was separated by preparative chromatography with high recovery, and the product solution was collected, purified by drying at 120-140 oC for 4 hours and, recovered (127 mg). Purity of the isolated 7-methylxanthine was comparable to authentic standards with no contaminant peaks, as observed by HPLC, LC-MS, and NMR. 

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Publication Date
Sun May 01 2016
Journal Name
Journal Of Engineering
Production of Methyl Ester (Biodiesel) from Used Cooking Oils via Trans-esterification process
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Used cooking oil was undergoing trans-esterification reaction to produce biodiesel fuel. Method of production consisted of pretreatment steps, trans-esterification, separation, washing and drying. Trans-esterification of treated oils was studied at different operation conditions, the methanol to oil mole ratio were 6:1, 8:1, 10:1, and 12:1, at different temperature 30, 40, 50, and 60 º C, reaction time 40, 60, 80, and 120 minutes, amount of catalyst 0.5, 1, 1.5, and 2 wt.% based on oil and mixing speed 400 rpm. The maximum yield of biodiesel was 91.68 wt.% for treated oils obtained by trans-esterification reaction with 10:1 methanol to oil mole ratio, 60 º C reaction temperature, 80 minute reactio

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Publication Date
Thu Mar 06 2014
Journal Name
Isolation, Screening, Identification And Improvement The Production Of Cellulase Produced From Iraqi Soil
Isolation, screening, identification and improvement the production of cellulase produced from Iraqi soil
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Publication Date
Wed Jan 01 2025
Journal Name
Medical Journal Of Babylon
Optimum Conditions for Pyomelanin Production and Characterization from Local Isolates of Pseudomonas aeruginosa
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Nowadays, there is increased interest in the biosynthesis of microbial melanin related to their numerous biological functions and applications in many fields, especially in medical fields, including immune-modulating, antimicrobial antibiotic, antiviral antivenin, anticancer, antitumor activity, and anti-biofilm activity. Pyomelanin is a hydrophobic macromolecule that is typically dark brown or black in color, formed by the oxidative polymerization of phenolic or indolic compounds. Pyomelanin is reported to be safe for consumption, thus providing a crucial strategy for biocontrol of biofilm. Furthermore, natural pyomelanin is known as a potent antioxidant, photoprotective, and free radical scavenging. Objective: This study focuses on the

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Publication Date
Wed Dec 24 2025
Journal Name
Iraqi Journal Of Agricultural Sciences
ISOLATION, SCREENING AND PRODUCTION OF PHYTATE DEGRADING ENZYME (PHYTASE) FROM LOCAL FUNGI ISOLATE
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Publication Date
Sun Sep 04 2011
Journal Name
Baghdad Science Journal
Effect of Lactobacilli sources on Escherichia coli and Staphylococcus aureus adherence to uroepithelial cells
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The inhibitory action of four lactobacilli isolates Lactobacillus bulgaricus, L. acidophilus, L. plantarum and L. fermentum, isolated from four different samples; yoghurt, vinegar, saliva and vagina respectively, on Escherichia coli and Staphylococcus aureus adhesion to uroepithelial cells were investigated. Results showed that all Lactobacillus isolates or their supernatant were able to reduce the number of the uropathogens attached to uroepithelial cells. However, inhibition level of lactobacilli cells was higher than their supernatant. Nevertheless, the human indigenous lactobacilli (L. fermentum and L. plantarum) were more competitive than food lactobacilli (L. acidophilus and L. bulgaricus).

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Publication Date
Wed Dec 01 2021
Journal Name
Gene Reports
The molecular study for evaluation the antibiotic resistance of Escherichia coli and Klebsiella pneumoniae bacteria isolated from urinary tract infection patients
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Urinary tract infection is a bacterial infection that often affects the bladder and thus the urinary system. E. coli is one of the leading uropathogenic bacteria that cause urinary tract infections. Uropathogenic E. coli is highly effective and successful in causing urinary tract infections through biofilm formation and urothelial cell invasion mechanisms. Other organisms that cause urinary tract infections include members of the Enterobacteriaceae family, streptococci and staphylococci species and perch. In addition, K.penumoniae is another important gram-negative bacterium that causes urinary tract infections. With the PCR technique, unseen bacterial species can be detected using standard clinical microbiology methods. In this study, the

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Publication Date
Thu Nov 02 2023
Journal Name
Missouri Journal Of Mathematical Sciences
e*_lifting midule
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Publication Date
Sat Jan 01 2022
Journal Name
European Journal Of Pure And Applied Mathematics
E*_essential submodule
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Publication Date
Sat Dec 01 2012
Journal Name
Iraqi Journal Of Science,
Monitoring Vegetation Growth of Spectrally Landsat Satellite Imagery ETM+ 7 & TM 5 for Western Region of Iraq by Using Remote Sensing Techniques.
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Landsat-5 Thematic Mapper (TM) has been imaging the Earth since March 1984 and Landsat-7 Enhanced Thematic Mapper Plus (ETM+) was added to the series of Landsat instruments in April 1999. In this paper the two sensors are used to monitoring the agriculture condition and detection the changing in the area of plant covers, the stability and calibration of the ETM+ has been monitored extensively since launch although it is not monitored for many years, TM now has a similar system in place to monitor stability and calibration. By referring to statistical values for the classification process, the results indicated that the state of vegetation in 1990 was in the proportion of 42.8%, while this percentage rose to 52.5% for the same study area in

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Publication Date
Sun Apr 14 2024
Journal Name
Kufa Journal For Agricultural Sciences
Growth and production of three potato cultivars as affected by organic foliar nutrition
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