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Development of a new method for d-xylose detection and quantification in urine, based on the use of recombinant xylose dehydrogenase from Caulobacter crescentus.

J Biotechnol.. 2016-09; 
Sánchez-Moreno I,García-Junceda E,Hermida C,Fernández-Mayoralas A.
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Codon Optimization ... The sequence of the gene xylB from C. crescentus strain NA1000 was codon optimized for expression in E. coli (OptimumGene? algorithm), chemically synthesized and cloned into pUC57 vector by GenScript Corporation (Piscataway, NJ, USA). ... Get A Quote

摘要

The gene xylB from Caulobacter crescentus has been cloned and expressed in Escherichia coli providing a high yield of xylose dehydrogenase (XylB) production and excellent purity (97%). Purified recombinant XylB showed an absolute dependence on the cofactor NAD(+) and a strong preference for d-xylose against other assayed mono and disaccharides. Additionally, XylB showed strong stability when stored as freeze-dried powder at least 250days both at 4°C and room temperature. In addition, more than 80% of the initial activity of rehydrated freeze-dried enzyme remained after 150days of incubation at 4°C. Based on these characteristics, the capability of XylB in d-xylose detection and quantification was studied. The... More

關鍵詞

Enzymatic detection; Gaxilose; Hypolactasia; Intestinal lactase activity; Xylose dehydrogenase; Xylose quantification
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