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Determination of transglutaminase activity using fluorescence spectrophotometer

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dc.creator KARAHAN, Aynur Guel
dc.creator Sokullu, Esen
dc.creator Bas, Deniz
dc.creator Cakir, Ibrahim
dc.creator Cakmakci, Luetfue
dc.creator Oener, Zübeyde
dc.creator BOYACI, İSMAİL HAKKI
dc.date 2007-12-31T22:00:00Z
dc.date.accessioned 2020-10-06T09:48:15Z
dc.date.available 2020-10-06T09:48:15Z
dc.identifier 3ee8081f-a963-41f5-86eb-32d7536eb50b
dc.identifier 10.1080/08905430802265775
dc.identifier https://avesis.sdu.edu.tr/publication/details/3ee8081f-a963-41f5-86eb-32d7536eb50b/oai
dc.identifier.uri http://acikerisim.sdu.edu.tr/xmlui/handle/123456789/58159
dc.description An improved fluorometric assay for determining the activity of the microbial transglutaminase (TGase) in the culture medium samples has been developed. The assay procedure measures the fluorescence enhancement due to the incorporation of monodansyl cadaverine (Substrate A) into pentafluorophenylester of CBZ-Gln-Gly (Substrate Q) at exc. 260 nm and em 538 nm. The effect of the competitive inhibitors in the culture medium samples on TGase activity was determined. The assay was combined with HPLC method for determining enzyme activity as an international unit (IU). Enzymatic reaction was monitored by HPLC and the rate of product formation was measured via amine substrate consumption rate. A conversion factor was obtained using HPLC and fluorescence spectrophotometer data together. This was formulated for quantification of TGase activity as IU using fluorometric assay reported in this study. The detection limit of the assay was determined as 0.0014 IU (0.5 mg). TGase activity remained linear upto the enzyme concentraion of 20 mg. This technique dramatically decreases the incubation time of enzyme to a few minutes of activity measurement.
dc.language eng
dc.rights info:eu-repo/semantics/closedAccess
dc.title Determination of transglutaminase activity using fluorescence spectrophotometer
dc.type info:eu-repo/semantics/article


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