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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

关键词

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