Ngoc Trang Nhu Ngo - Lund, Sverige Professionell profil

Patrick Adlercreutz - Research Outputs - Lund University

One unit of activity was defined as the amount of enzyme which decreases 10% of amylose-iodine complex optical density per min 2005-2-16 · Enzyme assay CGTase activity was assayed by the Lejeune et al. (1989) method slightly modified by Gawande and Patkar (1999). A 1.0 mL amount of 1% soluble starch prepared in 50 mmol/L phosphate buffer, pH 7.0, was added with 0.1 mL of properly diluted enzyme and incubated at 40 ºC for 10 min. Enzyme reaction was stopped by immediately cooling the enzyme was determined by incubating the CGTase assay at di V erent temperatures, ranging from 30–90°C for 10 min. Then the subsequent steps were do ne according to the Using this assay, it was shown that the crude enzyme from an alkalophilic Bacillus sp. (ATCC 21783) showed cyclization activity with three distinct pH optima, at pH 4.5, 6.0 and 8.5.

Cgtase enzyme assay

with slight modiWcation. The reaction mixture containing 40 mg of soluble starch in 1.0 mL of 0.1 M phosphate buVer (pH 6.0) and 0.1 mL of enzyme solution was incubated at 60°C for 10 min. Cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) is a unique member of glycoside hydrolase family 13. Its defining property is the ability to form cyclodextrins from starch, through an intramolecular transglycosylation reaction (cyclization). CGTase enzyme production levels. Physiological Characterization of CGTase Enzyme Enzyme assay results at different pH conditions ranging from 6.5 to 7.0 are depicted in Fig. 6. Both enzymes (from different sources) showed similar pattern of enzyme activity and indicated parabolic nature.

The amount of β-cyclodextrin produced was estimated from the standard graph of 0-500μg/mL βCD concentration against absorbance. One unit of CGTase was defined as the amount of enzyme required to produce 1μmol 2020-02-14 2016-10-22 This type of capture assay is called a “sandwich” assay because the analyte to be measured is bound between two primary antibodies, each detecting a different epitope of the antigen–the capture antibody and the detection antibody.

Ngoc Trang Nhu Ngo - Lund, Sverige Professionell profil

Anal. Biochem.

Patrick Adlercreutz - Research Outputs - Lund University

These include the choice of buffer and its composition, the type of enzyme and its concentration, as well as the type of substrate and concentrations, the reaction conditions, and the appropriate assay technology. Enzyme assay of N-acetylglucosaminyltransferase III (GnT-III) Authors Korekane, Hiroaki Department of Disease Glycomics (Seikagaku Corporation), The Institute of Scientific and Industrial Research, Osaka University ATP synthase Enzyme Activity Microplate Assay Kit ab109714 is used to determine the activity of ATP synthase (Complex V) in a human or rat sample. The ATP synthase enzyme is immunocaptured within the wells of the microplate and the enzyme activity is measured by monitoring the decrease in absorbance at 340 nm. 2013-08-26 · The luciferase reporter assay is commonly used as a tool to study gene expression at the transcriptional level. It is widely used because it is convenient, relatively inexpensive, and gives quantitative measurements instantaneously. It has broad applications across various fields of cell and molecular biology - wherever you want to measure or track expression of a cloned gene. As with many Prior to use, transfer 10 µl of the supplied enzyme to another vial and dilute with 190 µl of diluted Sample Buffer and keep on ice.

Synonyms. cyclodextrin glycosyltransferase, cyclodextrin glucanotransferase, cyclomaltodextrin glucanotransferase, alpha-cgtase, beta-cgtase, toruzyme, 10 May 2013 glycosyltransferase (CGTase), produced by bacteria of the genera. Bacillus In the pH and thermo stability assays, 50% of enzyme activity was CGTase is an extracellular enzyme identified in a number of bacteria, mainly Bacillus [4]. Assay of outer and inner membrane permeability.
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The extracellular enzyme cyclodextrin glucanotransferase (CGTase) synthesizes cyclic malto-oligosaccharides called cyclodextrins (CDs) from starch and related α-1,4-glucans. CGTases are produced by a variety of bacteria, mainly Bacillus species, by submerged culture in complex medium. CGTases differ in the amount and types of CDs produced. function of the quantity of cellulase enzyme in the assay mixture.

Determination of CGTase activity and protein content: CGTase assay was carried out as per slightly modified phenolphthalein method (13), 4% soluble starch in 0.1M phosphate buffer, pH 7.0 was prepared by heating it in boiling water bath for 3 min. To 650 μL of substrate, 250 μL of 0.1 M phosphate buffer used for further assays.
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GLYCOSIDE HYDROLASE FAMILY 16 - Avhandlingar.se

Determination of CD concentrationThe α-CD concentration was assayed by the decrease in absorbance at 507 nm caused by a methyl orange α-CD complex formation . CGTase enzyme production levels. Physiological Characterization of CGTase Enzyme Enzyme assay results at different pH conditions ranging from 6.5 to 7.0 are depicted in Fig. 6. Both enzymes (from different sources) showed similar pattern of enzyme activity and indicated parabolic nature. The maximum activity waqs observed at pH 6.7. The hydrolysis and cyclization properties of γ-CGTase were detected under the standard assay conditions with buffers of various pHs and different reaction temperatures.

GLYCOSIDE HYDROLASE FAMILY 16 - Avhandlingar.se

All bacterial CGTases studied convert starch into a mixture of cyclodextrins mostly consisting of 6, 7 or 8 α(1,4)‐linked glucose residues (α‐, β‐ or γ‐cyclodextrins, respectively) [ [ 2 ] , [ 3 ] ]. In the present study, this method was used to investigate the actions of three different CGTases: CGTase from B. macerans (Bmac CGTase), which produces mainly CD6 ; CGTase from alkalophilicBacillus sp.

Cyclodextrin glycosyltransferase (CGTase) is an important industrial enzyme used The β-CD produced in the assay was determined based on colour fading at Analysis of the maltose-dependent CGTase crystal structure revealed that each enzyme molecule contained three maltose molecules, situated at contact points Enzyme assay. CGTase activity was assayed with soluble starch as the substrate by measurement of the decrease in iodine-staining power; 0.5 ml of 1.5% One unit of CGTase activity was defined as the amount of enzyme releasing one µmol of β-CD per min under the defined assay conditions.