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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 Nov 05 2017
Journal Name
International Journal Of Science And Research
Production of Biodiesel from Waste Cooking Oil using Cao-Egg Shell Waste Derived Heterogeneous Catalyst
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Biodiesel is an environmentally friendly fuel and a good substitution for the fossil fuel. However, the purity of this fuel is a major concern that challenges researchers. In this study, a calcium oxide based catalyst has been prepared from local waste eggshells by the calcination method and tested in production biodiesel. The eggshells were powdered and calcined at different temperatures (700, 750, 800, 850 and 900 °C) and periods of time (1, 2, 3, 4 and 5 hr.). The effect of calcination temperature and calcination time on the structure and activity of the solid catalyst were examined by X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM), and Brunaure-Emmett-Teller (BET). The optimum catalyst performance was obtained at 900 °C

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Publication Date
Sun Dec 07 2008
Journal Name
Baghdad Science Journal
Optium Conditions for the Production of Neutral Protease from local strain Aspergillus niger var cabonarius
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The optimum conditions for the production of neutral protease from local strain Aspergillus niger var carbonarius by solid – state fermentation system (Wheat bran) moisted with 0.2 M phosphate buffer (PH7.0) . the hydration ratio was 1:5 (V:W) . the concentration of inoculum was 1×106 spores per 10 gram of solid materials , initial P H 6.5 and 96 hours of incubation period at 30? C .the enzyme activity was 1300 unit / ml and specific activity was 1550 unit / mg protein .

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Publication Date
Thu Nov 07 2024
Journal Name
Journal Of Ecological Engineering
Microwave assisted production of biodiesel using CaO nano-catalyst produced from mango fallen leaves extract
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Publication Date
Mon Nov 14 2022
Journal Name
Biomedicine
Cytotoxic effect of cloned EGFP gene on NCI-H727 cell line via genetically engineered gene transfer system
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Introduction and Aim: Cancers are a complex group of genetic illnesses that develop through multistep, mutagenic processes which can invade or spread throughout the body. Recent advances in cancer treatment involve oncolytic viruses to infect and destroy cancer cells. The Newcastle disease virus (NDV), an oncolytic virus has shown to have anti-cancer effects either directly by lysing cancer cells or indirectly by activating the immune system. The green fluorescent protein (GFP) has been widely used in studying the anti-tumor activity of oncolytic viruses. This study aimed to study the anticancer effect of a recombinant rNDV-GFP clone on NCI-H727 lung carcinoma cell line in vitro.   Materials and Methods: The GFP gene was inserted t

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Publication Date
Mon Nov 27 2023
Journal Name
Korean Journal Of Orthodontics
Effectiveness of laser-engineered copper-nickel titanium versus superelastic nickel-titanium aligning archwires: A randomized clinical trial
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Publication Date
Mon Nov 14 2022
Journal Name
Biomedicine
Cytotoxic effect of cloned EGFP gene on NCI-H727 cell line via genetically engineered gene transfer system
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Introduction and Aim: Cancers are a complex group of genetic illnesses that develop through multistep, mutagenic processes which can invade or spread throughout the body. Recent advances in cancer treatment involve oncolytic viruses to infect and destroy cancer cells. The Newcastle disease virus (NDV), an oncolytic virus has shown to have anti-cancer effects either directly by lysing cancer cells or indirectly by activating the immune system. The green fluorescent protein (GFP) has been widely used in studying the anti-tumor activity of oncolytic viruses. This study aimed to study the anticancer effect of a recombinant rNDV-GFP clone on NCI-H727 lung carcinoma cell line in vitro.   Materials and Methods: The GFP gene was inserted t

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Publication Date
Sat Jan 09 2010
Journal Name
Journal Of Al-nahrain University
DETECTIONOF GAMMARADIATIONEFFECT INDUCEDBY COBELT-60 ONESCHERICHIA COLI CELLS
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Publication Date
Tue Mar 24 2009
Journal Name
كتاب الوقائع /المؤتمر العلمي الثالث لكلية العلوم جامعة بغداد
Using antibiotics as mutagenic and curing agents for Prodigiosin Production By Serratia marcescens
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The effect of different antibiotics on growth pigment and plasmid curing of Serratia marcescens were studied, S. marcescens was cultured in media containing(16_500)µg/ml of antibiotics, curing mutants unable to produce prodigiosin and lost one plasmid band were obtained of of ampicillin, amoxillin, antibiotics concentrations (64 500) µg/ml metheprim, ultracloxam, azithromycin, cephalexin and erythromycin treated with (350 500) µg/ml of The mutant cells rose- light color and and refampicin revealed S.marcescens inhibited ciprodar and tetracyclin, lincomycin did not lost the plasmid band chlaforan

Publication Date
Mon Jun 30 2014
Journal Name
Al-kindy College Medical Journal
The prevalence and antimicrobial sensitivity of Esbl Escherichia Coli. in clinical isolates
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Background: The antimicrobial resistance is one of the most serious and expanding health problems world -wide in the last decades. The esbl escherichia coli. (extended – spectrum beta-lactamase e.coli) represents an important aspect of it .Objectives: To get an overview on the esbl e.coli prevalence profile in general. Also to assess the antibiotic sensitivity of esbl e. coli trying to specify the most effective antibiotics in combating this micro-organism.Methods: this study tries to focus on this problem in Iraq which through a prospective study approach by taking 35 clinical samples from various sources (urine, blood, abscess, eye ,vagina ,stool and others),and after confirming the presence of e.coli, the presence of esbl e.coli and

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Publication Date
Fri Dec 29 2023
Journal Name
Iraqi Journal Of Pharmaceutical Sciences ( P-issn 1683 - 3597 E-issn 2521 - 3512)
Protective Effect of Omega-7 against Doxorubicin-Induced Cardiotoxicity in Male Rats
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Background: Doxorubicin is considered one of the most effective anticancer drugs, yet it is use is limited by its side effect mediated by the generation of reactive oxygen species. Omega-7, an antioxidant has shown to have a cardioprotective effect.

Aim of the study: evaluate a possible protective effect of omega-7 against doxorubicin-induced cardiotoxicity in male rats.

Methods: twenty-eight male rats were divided into 4 groups (7 for each group).  Group 1 (Negative control): healthy animals received normal saline orally as the vehicle for eight successive days and were sacrificed on day 9. Group 2 (positive control): animals that r

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