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| Zapata-perez, Ruben; Garcia-saura, Antonio G.; Jebbar, Mohamed; Golyshin, Peter N.; Sanchez-ferrer, Alvaro. |
| Nicotinamidases catalyze the hydrolysis of the amide bond in nicotinamide (NAM) to produce ammonia and nicotinic acid (NA). These enzymes are an essential component of the NAD(+) salvage pathway and are implicated in the viability of several pathogenic organisms. Its absence in humans makes them a promising drug target. In addition, although they are key analytical biocatalysts for screening modulators in relevant biomedical enzymes, such as sirtuins and poly-ADP-ribosyltransferases, no commercial sources are available. Surprisingly, the finding of an affordable source of nicotinamidase from metagenomic libraries is hindered by the absence of a suitable and fast screening method. In this manuscript, we describe the development of two new whole-cell methods... |
| Tipo: Text |
Palavras-chave: Amidohydrolases; Nicotinamidase; Functional screening; Metagenome; Library; High-throughput screening assays. |
| Ano: 2016 |
URL: http://archimer.ifremer.fr/doc/00362/47269/47232.pdf |
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| El Kheir, Sara M.; Cherrat, Lamia; Awussi, Ahoefa A.; Ramia, Nancy E.; Taha, Samir; Rahman, Abdur; Passerini, Delphine; Leroi, Francoise; Petit, Jeremy; Mangavel, Cecile; Revol-junelles, Anne-marie; Borges, Frederic. |
| This article describes a method for high-throughput competition assays using a bioluminescent strain of L. monocytogenes. This method is based on the use of the luminescent indicator strain L. monocytogenes EGDelux. The luminescence of this strain is correlated to growth, which make it suitable to monitor the growth of L. monocytogenes in mixed cultures. To this aim, luminescence kinetics were converted into a single numerical value, called the Luminescence Disturbance Indicator (LDI), which takes into account growth inhibition phenomena resulting in latency increase, decrease in the luminescence rate, or reduction of the maximum luminescence. The LDI allows to automatically and simultaneously handle multiple competition assays which are required for... |
| Tipo: Text |
Palavras-chave: Biopreservation; Listeria monocytogenes; High-throughput screening assays; Bioluminescence; Competition; Mixed culture; Bioprotection; Antibacterial activities. |
| Ano: 2018 |
URL: https://archimer.ifremer.fr/doc/00454/56534/58236.pdf |
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