135 research outputs found

    High yield 1,3-propanediol production by rational engineering of the 3-hydroxypropionaldehyde bottleneck in Citrobacter werkmanii

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    Background: Imbalance in cofactors causing the accumulation of intermediates in biosynthesis pathways is a frequently occurring problem in metabolic engineering when optimizing a production pathway in a microorganism. In our previous study, a single knock-out Citrobacter werkmanii Delta dhaD was constructed for improved 1,3-propanediol (PDO) production. Instead of an enhanced PDO concentration on this strain, the gene knock-out led to the accumulation of the toxic intermediate 3-hydroxypropionaldehyde (3-HPA). The hypothesis was emerged that the accumulation of this toxic intermediate, 3-HPA, is due to a cofactor imbalance, i.e. to the limited supply of reducing equivalents (NADH). Here, this bottleneck is alleviated by rationally engineering cell metabolism to balance the cofactor supply. Results: By eliminating non-essential NADH consuming enzymes (such as lactate dehydrogenase coded by ldhA, and ethanol dehydrogenase coded by adhE) or by increasing NADH producing enzymes, the accumulation of 3-HPA is minimized. Combining the above modifications in C. werkmanii Delta dhaD resulted in the strain C. werkmanii Delta dhaD Delta ldhA.adhE::ChlFRT which provided the maximum theoretical yield of 1.00 +/- 0.03 mol PDO/mol glycerol when grown on glucose/glycerol (0.33 molar ratio) on flask scale under anaerobic conditions. On bioreactor scale, the yield decreased to 0.73 +/- 0.01 mol PDO/mol glycerol although no 3-HPA could be measured, which indicates the existence of a sink of glycerol by a putative glycerol dehydrogenase, channeling glycerol to the central metabolism. Conclusions: In this study, a multiple knock-out was created in Citrobacter species for the first time. As a result, the concentration of the toxic intermediate 3-HPA was reduced to below the detection limit and the maximal theoretical PDO yield on glycerol was reached

    PBDEs in marine and freshwater sediments from Belgium: levels, profiles and relations with biota

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    Sediments from the Belgian North Sea (BNS), the Western Scheldt Estuary (SE) and freshwater watercourses from the Scheldt basin were analysed for eight PBDE congeners, namely BDEs 28, 47, 99, 100, 153, 154, 183 and 209. Previously analysed biological samples from the same locations in the BNS and the SE have been shown to contain large amounts of PBDEs. Surprisingly, PBDE concentrations in the sediments were below the LOQ for samples from the BNS (except BDE 209), while in those from the SE the sum of PBDEs (not including BDE 209) were higher and ranged from 0.20 to 0.41 ng g-1 dw. BDE 209 could be detected in 83% of the samples from the BNS and in all the samples from the SE. Concentrations up to 1200 ng g-1 were hereby measured in the SE. Compared to the marine and estuarine locations, the sediments from the freshwater watercourses were relatively more polluted with the lower brominated PBDEs (-1 dw). BDE 209 concentrations up to 320 ng g-1 dw were measured in those sediments. However, the contribution of BDE 209 to the total amount of PBDEs varied much more at the freshwater locations than in the SE, which suggests a different input of pollutants. PBDE profiles observed in biological samples do not match the profiles of the sediments. BDE 183 and 209 could not be quantified in biota, although these congeners were undoubtedly present in the sediments. This raises questions about the bioavailability of these congeners in the environment

    A reassessment of the nomenclature of polychlorinated biphenyl (PCB) metabolites.

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    Polychlorinated biphenyls (PCBs) are a widespread class of persistent organic chemicals that accumulate in the environment and humans and are associated with a broad spectrum of health effects. PCB biotransformation has been shown to lead to two classes of PCB metabolites that are present as contaminant residues in the tissues of selected biota: hydroxylated (HO) and methyl sulfone (MeSO2) PCBs. Although these two types of metabolites are related structures, different rules for abbreviation of both classes have emerged. It is important that a standardized nomenclature for the notation of PCB metabolites be universally agreed upon. We suggest that the full chemical name of the PCB metabolite and a shorthand notation should be adopted using the International Union of Pure and Applied Chemistry's chemical name/original Ballschmiter and Zell number of the parent congener, followed by the assignment of the phenyl ring position number of the MeSO2- or HO-substituent. This nomenclature provides a clear, unequivocal set of rules in naming and abbreviating the PCB metabolite structure. Furthermore, this unified PCB metabolite nomenclature approach can be extended to the naming and abbreviation of potential metabolites of structurally analogous contaminants such as HO-polybrominated biphenyls and HO-polybrominated diphenyl ethers

    Microbial and dietary factors associated with the 8-prenylnaringenin producer phenotype: a dietary intervention trial with fifty healthy post-menopausal Caucasian women

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    Hop-derived food supplements and beers contain the prenylflavonoids xanthohumol (X), isoxanthohumol (IX) and the very potent phyto-oestrogen (plant-derived oestrogen mimic) 8-prenylnaringenin (8-PN). The weakly oestrogenic IX can be bioactivated via O-demethylation to 8-PN. Since IX usually predominates over 8-PN, human subjects may be exposed to increased doses of 8-PN. A dietary intervention trial with fifty healthy postmenopausal Caucasian women was under-taken. After a 4 d washout period, participants delivered faeces, blank urine and breath samples. Next, they started a 5 d treatment with hop-based supplements that were administered three times per d and on the last day, a 24 h urine sample was collected. A semi-quantitative FFQ was used to estimate fat, fibre, alcohol, caffeine and theobromine intakes. The recoveries of IX, 8-PN and X in the urine were low and considerable inter-individual variations were observed. A five-fold increase in the dosage of IX without change in 8-PN concentration resulted in a significant lower IX recovery and a higher 8-PN recovery. Classification of the subjects into poor (60 %), moderate (25 %) and strong (15 %) 8-PN producers based on either urinary excretion or microbial bioactivation capacity gave comparable results. Recent antibiotic therapy seemed to affect the 8-PN production negatively. A positive trend between methane excretion and 8-PN production was observed. Strong 8-PN producers consumed less alcohol and had a higher theobromine intake. From this study we conclude that in vivo O-demethylation of IX increases the oestrogenic potency of hop-derived products

    Pelagic seabirds as biomonitors of persistent organic pollutants in the Southwestern Atlantic

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    Persistent Organic Pollutants (POPs) are accumulated through time and can exert different effect on ecosystems. POPs and Chlorpyrifos, a current use pesticide, were assessed in body feathers of males and females of Black-browed albatross (Thalassarche melanophris, BBA) and Cape petrels (Daption capense, CAP) during their non-breeding seasons at the Patagonian Shelf, Argentina. Chlorpyrifos showed the highest values among all pollutants in both species (49.56–84.88 ng g−1), resulting from current agricultural practices. The pattern OCPs > PCBs > PBDEs was observed in both species, and CAP showed higher concentrations than BBA probably as a consequence of higher lipid mobilization and pollutants availability during dispersion. Non-significant differences between sexes about POPs levels were found; however a slight tendency was observed, females>males in CAP, and males>females in BBA. More attention and further studies are needed to understand seabirds' physiology and its relationship with the pollutants distribution in their tissues and considering breeding season.Fil: Quadri Adrogué, Agustina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Ciencias Marinas. Laboratorio de Ecotoxicología; ArgentinaFil: Miglioranza, Karina Silvia Beatriz. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Ciencias Marinas. Laboratorio de Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Copello, Sofía. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Favero, Marco. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Seco Pon, Juan Pablo. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentin
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