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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
Tue Jan 01 2019
Journal Name
Biochemical And Cellular Archives
Phenotypic and molecular detection of Escherichia coli efflux pumps from UTI patients
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Publication Date
Mon Mar 05 1990
Journal Name
وقائع المؤتمر العلمي الخامس لمجلس البحث العلمي في المجلة العراقية
INACTIVATION OF SELECTED ANTIBIOTICS AGAINST ESCHERICHIA COLI BY VAMIN NUTRITIO- NAL SUPPLEMENTATION
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Posible interference of vamin with the activity of several antibiotics against E. coli was evaluated in vitro. In MBS- glucose medium, significant growth delay was induced by 8 ug/ml of terramycin (oxytetracycline- polymyxin B) and bactrim (trimethoprim-sulphamethoxazole), and by 16 ug/ml of refocin, lincomycin, and chloramphenicol. Rapid growth inhibition was induced by 32 ug/ml of all an- tibiotic tested separately. Significant inactivation of up to 64 ug/ml of licomycin and bactrim was in- duced by the addition of vamin at a concentration of 1:20 v/v of the medium. This effect was found to be due to the presence of specific amino acids in vamin. Among them is valine, leucine, isoleucine tyrosine, tryptophan, phenylalanine, cysteine, meth

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Publication Date
Mon Sep 01 2014
Journal Name
Al-khwarizmi Engineering Journal
Production of Bioethanol from Waste Potatoes
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In this research, production of ethanol from waste potatoes fermentation was studied using Saccharmyses cerevisiae. Potato Flour was prepared from potato tubers after cooking and drying at 85°C. Homogenous slurry of potato flour was prepared in water at solid liquid ratio 1:10. Liquefaction of potato flour slurry with α-amylase at 80°C for 40 min followed by saccharification with glucoamylase at 65°C for 2 hr .Fermentation of hydrolysate with Saccharomyces cerevisiae at 35°C for two days resulted in production of 33 g/l ethanol.

      The parameters studied were; temperature, time of fermentation and pH. It was found that Saccharification process is affected by enzyme Amylo 300 conc

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Publication Date
Fri Apr 04 2014
Journal Name
International Journal Of Sciences: Basic And Applied Research
Production of bioethanol from reed (Phragmitesaustralis)
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Publication Date
Tue Sep 01 2020
Journal Name
Baghdad Science Journal
Molecular Characterization of Plasmid-Mediated Non-O157 Verotoxigenic Escherichia coli Isolated from Infants and Children with Diarrhea
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A significant increase in the incidence of non-O157 verotoxigenic Escherichia coli (VTEC) infections have become a serious health issues, and this situation is worsening due to the dissemination of plasmid mediated multidrug-resistant microorganisms worldwide. This study aims to investigate the presence of plasmid-mediated verotoxin gene in non-O157 E. coli. Standard microbiological techniques identified a total of 137 E. coli isolates. The plasmid was detected by Perfectprep Plasmid Mini preparation kit. These isolates were subjected to disk diffusion assay, and plasmid curing with ethidium bromide treatment. The plasmid containing isolates were subjected to a polymerase chain reaction (PCR) for investigating

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Publication Date
Sun Jun 01 2025
Journal Name
Al-khwarizmi Engineering Journal
Biodiesel Production from Castor Oil
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Biodiesel is becoming one of the most attractive alternative biofuels for petroleum-based diesel fuels. The castor plant is one of the abundant non-edible oils found in many countries. This paper aims to study Libyan castor oil and its potential for diesel conversion. Experiments were carried out in the laboratories of the Specific Center for Training in the Oil Industries in Al-Zawiya. The oil was extracted using a Soxhlet extractor and n-hexane solvent at 60 °C. Transesterification reactions were conducted in a batch reactor (a three-neck flask was used, where the middle opening carries a reflux condensation unit) at 65 °C. The methanol-to-castor oil molar ratio was 6:1, with a catalyst concentration of 1 wt.% relative to the ca

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Publication Date
Sun Jul 23 2023
Journal Name
Al-rafidain Journal Of Medical Sciences ( Issn 2789-3219 )
Future of Engineered Phage Therapy for Clostridium difficile Infections
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Bacteriophages have the potential to eliminate both antibiotic-resistant and sensitive bacteria; as a result, they have become a major focus of such research. In contrast to antibiotics, which assault the entire bacterial population without discrimination, bacteriophages have a limited set of characteristics that allow them to target infectious microbes while avoiding friendly species (commensal microbiota). Nevertheless, large groups of naturally occurring bacteriophages that are well-differentiated and selective for the most clinically recognized pathogenic bacterial strains are required. Utilizing genetic engineering techniques that modify the target phage genome to synthesize phages with known characteristics in a brief period o

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Publication Date
Sun Jun 05 2016
Journal Name
Baghdad Science Journal
Synthesis and Characterization of New Mannich Bases Derived from 7-hydroxy-4-methyl Coumarin
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Coumarin is a natural substance isolated from different plants. It belonges to a group of benzobyrones which consists of a benzene ring joined to a pyrone nucleus. In the present research, a new series of coumarin derivatives were formed. Compound (1) (7-hydroxy-4-methyl Coumarin) was converted into 4-methylquinolin-2(H) derivative (2) by reaction with acetamide, and then reaction of (2) with thiosemicarbazide in ethanol leads to the synthesize of hydrazincarbothioamide derivative (3).The reaction of (3) with ethylchloroacetate in presence of sodium acetate leads to closure ring to get [(1-(5-oxo-2-thioxoimidazolidin-1-ylimino) ethyl)]quinolin-2(1H)-one (4). Mannich bases were prepared through the reaction of (4) with primary

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Crossref (1)
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Publication Date
Thu Apr 01 2021
Journal Name
Biochemical And Cellular Archives
Evaluation the Effect of Allium Sativum (garlic) oil on The Expression of Maz E and Maz F Genes in Escherichia Coli Clinical Isolates
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Scopus (4)
Scopus
Publication Date
Sun Jan 19 2014
Journal Name
Romanian Biotechnological Letters
Optimization conditions for bioemulsifier production by local Streptomyces sp. SS20 isolated from hydrocarbon contaminated soil
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