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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
Fri May 30 2025
Journal Name
University Of Anbar Sport And Physical Education Science Journal
That the child is aged 7 years, surrounded by information, knowledge and skillsvaried,
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Abstract That the child is aged 7 years, surrounded by information, knowledge and skillsvaried, which constitute the raw material of experience teaching and is in the rule of inputlearning, if received by the student of these data are positive, these data require research and audit, the style becomes more positive, effective, and then becomes the explorer, butnot done exploration efficiency without the visual, auditory and sensory owned by thechildren. So study aimed to identify the values of the optical track and explore mathematical and find the relationship between them. And use the descriptive approach in a manner the linkon the children of the first year of primary school age (6-7) years in my school Waziriya and Karkh, during the peri

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Publication Date
Thu Oct 01 2015
Journal Name
Iraqi Journal Of Science
optimization of pectinase production from pesudomonas sp. isolated from iraqi soi
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Publication Date
Wed Jan 01 2014
Journal Name
Infection And Drug Resistance
Virulence and antimicrobial resistance of Escherichia coli isolated from Tigris River and children diarrhea
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Publication Date
Wed May 17 2017
Journal Name
Ibn Al-haitham Journal For Pure And Applied Sciences
Sensitivity of Bacteria Escherichia coli Iisolated from Urinary Tract Infection toward Ciprofloxacin and Lomefloxacin.
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   Four hundred and fifty urine samples were collected from patients suffering from urinary tract infection from the General Azadi hospital in Kurkuk province ,during the period of october 2007 till march 2008 .       Results of bacteriological culture revealed that (168) out of (450) studied samples (37.3%) gave positive culture using blood agar and macConkey agar ; different species of bacterial isolates were detected using morphological and biochemical tests ,from these isolates the highest percentage of the isolates were from Escherichia coli when it was (100) isolates out of (190) isolate  (52.63%)  .  one hundred isolates were distributed between (77) from females and (23) from

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Publication Date
Wed May 01 2024
Journal Name
Biology
Lysophosphatidylcholine Acetyltransferase 2 (LPCAT2) Influences the Gene Expression of the Lipopolysaccharide Receptor Complex in Infected RAW264.7 Macrophages, Depending on the E. coli Lipopolysaccharide Serotype
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Escherichia coli (E. coli) is a frequent gram-negative bacterium that causes nosocomial infections, affecting more than 100 million patients annually worldwide. Bacterial lipopolysaccharide (LPS) from E. coli binds to toll-like receptor 4 (TLR4) and its co-receptor’s cluster of differentiation protein 14 (CD14) and myeloid differentiation factor 2 (MD2), collectively known as the LPS receptor complex. LPCAT2 participates in lipid-raft assembly by phospholipid remodelling. Previous research has proven that LPCAT2 co-localises in lipid rafts with TLR4 and regulates macrophage inflammatory response. However, no published evidence exists of the influence of LPCAT2 on the gene expression of the LPS receptor complex induced by smooth or rough b

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Publication Date
Thu Sep 30 2004
Journal Name
Iraqi Journal Of Chemical And Petroleum Engineering
Production of Formaldehyde by Catalytic Conversion of Methanol
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Publication Date
Sat Oct 01 2011
Journal Name
Journal Of Engineering
Production of Biodiesel Fuel from Used vegetable Oil
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Used vegetable oil was introduced to transesterfication reaction to produce Biodiesel fuel suitable for diesel engines. Method of production was consisted of filtration, transesterfication, separation and washing. Transesterfication was studied extensively with different operating conditions, temperature range (35-80oC), catalyst concentration (0.5-2 wt. % based on oil), mixing time (30-120 min.) with constant oil/methanol weight ratio 5:1 and mixing speed 1300 rpm. The concentration of Fatty acid methyl esters (Biodiesel) was determined for the transesterficated oil samples, besides of some important physical properties such as specific gravity, viscosity, pour point and flash point. The behavior of methyl esters production and the physica

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Publication Date
Wed Mar 01 2006
Journal Name
Journal Of Engineering
Production of Graphite Electrodes Binder from Iraqi Asphalt
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Basrah crude oil Vacuum residue 773+ K with specific gravity 1.107 and 4.87wt. % sulfur, was treated with hexane commercial fraction provided from Al-Taji Gas Company for preparing deasphaltened oil(DAO)suitable for hydrotreating process. Deasphaltening was carried out with 1h mixing time, 10ml:1g solvent to oil ratio and at room temperature. Hexane deasphaltened oil was hydrotreated on presulfied commercial Co-Mo/γ-Al2O3 catalyst in a trickle bed reactor. The hydrotreating process was carried out at temperature 660 K,LHSV 1.3 h^ –1, H2/oil ratio 300 l/l and constant pressure of 4MPa. The hydrotreated product was distillated under vacuum distillation unit. It is found that the mixture of 75% of vacuum residue with 25% anthracene satisfie

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Publication Date
Sun Oct 30 2011
Journal Name
Journal Of Engineering
Production of Biodiesel Fuel from Used vegetable Oil
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Used vegetable oil was introduced to transesterfication reaction to produce Biodiesel fuel suitable for diesel engines. Method of production was consisted of filtration, transesterfication, separation and washing. Transesterfication was studied extensively with different operating conditions, temperature range (35-80o C), catalyst concentration (0.5-2 wt. % based on oil), mixing time (30-120 min.) with constant oil/methanol weight ratio 5:1 and mixing speed 1300 rpm. The concentration of Fatty acid methyl esters (Biodiesel) was determined for the transesterficated oil samples, besides of some important physical properties such as specific gravity, viscosity, pour point and flash point. The behavior of methyl esters production and the

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Publication Date
Mon Jul 31 2017
Journal Name
Journal Of Engineering
Production of Biofuels from Selected Cellulosic Waste materials
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