2024-03-29T14:22:49Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1369742021-06-10T12:05:51Zcom_10261_33com_10261_5col_10261_286
2016-09-19T12:58:47Z
urn:hdl:10261/136974
Development of a new method for D-xylose detection and quantification in urine, based on the use of recombinant xylose dehydrogenase from Caulobacter crescentus.
Sánchez-Moreno, Israel
García-Junceda, Eduardo
Hermida, Carmen
Fernández-Mayoralas, Alfonso
VenterPharma
European Commission
Ministerio de Economía y Competitividad (España)
Enzymatic detection
Gaxilose
Hypolactasia
Xylose quantification
Xylose dehydrogenase
Intestinal lactase activity
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 250 days both at 4 °C and room temperature. In addition, more than 80% of the initial activity of rehydrated freeze-dried enzyme remained after 150 days of incubation at 4 °C. Based on these characteristics, the capability of XylB in d-xylose detection and quantification was studied. The linearity of the method was maintained up to concentrations of d-xylose of 10 mg/dL and the calculated limits of detection (LoD) and quantification (LoQ) of xylose in buffer were 0.568 mg/dL and 1.89 mg/dL respectively. Thus, enzymatic detection was found to be an excellent method for quantification of d-xylose in both buffer and urine samples. This method can easily be incorporated in a new test for the diagnosis of hypolactasia through the measurement of intestinal lactase activity.
2016-09-19T12:58:47Z
2016-09-19T12:58:47Z
2016-09-20
artículo
Journal of Biotechnology 234: 50-57 (2016)
0168-1656
http://hdl.handle.net/10261/136974
10.1016/j.jbiotec.2016.07.019
http://dx.doi.org/10.13039/501100000780
http://dx.doi.org/10.13039/501100003329
eng
Postprint
http://dx.doi.org/10.1016/j.jbiotec.2016.07.019
Sí
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/MAT2015-65184-C2-2-R
http://creativecommons.org/licenses/by-nc-nd/4.0/
openAccess
Elsevier