Phosphoglucoisomerase

Manufactured by Merck Group

Sourced in Switzerland, United States, Japan, Germany

Phosphoglucoisomerase is an enzyme that catalyzes the isomerization of glucose-6-phosphate to fructose-6-phosphate, a key step in glycolysis. It is an important component in the energy production pathway of cells.

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Lab products found in correlation

Hexokinase, by Merck Group (2 mentions) Glucose 6 phosphate dehydrogenase, by Roche (2 mentions) Glucose 6 phosphate dehydrogenase, by Merck Group (1 mentions) Fructose 1 6 bisphosphate, by Merck Group (1 mentions) Invertase, by Merck Group (1 mentions)

4 protocols using phosphoglucoisomerase

Quantitative Determination of Soluble Sugars

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Determination of soluble sugars was performed as described in Stitt et al.^[⁵⁴
^] by sequential enzymatic degradation of glucose, fructose, and sucrose; using one unit each of glucose‐6‐phosphate dehydrogenase (Hoffmann‐La Roche, Basel, Switzerland) for baseline generation, and then hexokinase, phosphoglucoisomerase, and invertase (β‐fructofuranosidase, all from Sigma‐Aldrich), respectively. Quantities were calculated based on reference curves for authentic standards in the range from 0.1 to 1.0 mM.

Pahlke G., Ahlberg K., Oertel A., Janson‐Schaffer T., Grabher S., Mock H., Matros A, & Marko D. (2021). Antioxidant Effects of Elderberry Anthocyanins in Human Colon Carcinoma Cells: A Study on Structure–Activity Relationships. Molecular Nutrition & Food Research, 65(17), 2100229.

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Hexokinase Activity Assay Protocol

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Hexokinase activity with fructose or glucose as substrate was measured at saturating conditions as described (Lowry and Passoneau, 1972 ). The reaction mixture was (in mmol/L) Tris buffer, 25 (pH 8.1), NADP, 0.5, MgCl₂, 2, fructose or glucose, 10, glucose-6-phosphate dehydrogenase, 0.2 U/mL, and phosphoglucoisomerase 0.4 U/mL (Sigma, St Louis, MO, USA). Homogenates, 5% (weight/volume) in sucrose, 0.32 mol/L, were made from frontal cortex (without the underlying white matter). Triton X-100 was added to a final concentration of 0.3 % (vol/vol); the samples were left on ice for 10 minutes and centrifuged at 5 000 × g. Two μL of supernatant were added to 100 μL of reaction mixture. The reaction was started by adding 5 μL ATP, 10 mmol/L. Formation of NADPH was measured spectrophotometrically at 340 nm and 37°C. The reaction was linear with time and added homogenate.

Hassel B., Elsais A., Frøland A.S., Taubøll E., Gjerstad L., Quan Y., Dingledine R, & Rise F. (2015). Uptake and metabolism of fructose by rat neocortical cells in vivo and by isolated nerve terminals in vitro. Journal of neurochemistry, 133(4), 572-581.

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Determination of Cytosolic Fructose-1,6-bisphosphatase Activity in Rice

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Cytosolic fructose-1,6-bisphosphatase activity was determined according to the method of Sharkey et al. (1991 (link)) with a slight modification. The basal portions of rice shoots harvested at the four-leaf stage were frozen at − 80 °C then milled with a mortar and pestle in 20 mM HEPES (4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid)-NaOH (pH 7.5), 100 mM KCl, and 0.5 mM EDTA. Each extraction solution was centrifuged at 13,000×g and 4 °C for 10 min. The supernatant was added to a reaction mixture consisting of 100 mM HEPES-NaOH (pH 7.5), 100 mM KCl, 5 mM MgCl₂, 0.5 mM EDTA, 0.5 mM NADP, one unit of phosphoglucoisomerase (Sigma-Aldrich Corp., Tokyo, Japan), and two units of glucose-6-phosphate dehydrogenase. The reaction was initiated by adding 40 μM fructose-1,6-bisphosphate (Sigma-Aldrich Corp., Tokyo, Japan). It was then spectrophotometrically monitored at 340 nm for increases in NADPH concentration for 5 min after the start of the enzymatic reaction. The amount of product formed was calculated from the increase in absorbance using the NADPH extinction coefficient of 6220 L mol^− 1 cm^− 1.

Ohashi M., Ishiyama K., Kusano M., Fukushima A., Kojima S., Hayakawa T, & Yamaya T. (2018). Reduction in sucrose contents by downregulation of fructose-1,6-bisphosphatase 2 causes tiller outgrowth cessation in rice mutants lacking glutamine synthetase1;2. Rice, 11, 65.

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Enzymatic Determination of Sugars in Blackberry

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Determination of soluble sugars in each fraction (anthocyanin A1–A4, polyphenols and the cleared blackberry raw juice) was performed as described previously through sequential enzymatic degradation of glucose, fructose and sucrose; using one unit each of glucose-6-phosphate dehydrogenase (Hoffmann-La Roche, Basel, Switzerland) for baseline generation, and then hexokinase (Sigma-Aldrich, Munich, Germany), phosphogluco-isomerase (Sigma-Aldrich) and invertase (β-fructofuranosidase, Sigma-Aldrich), respectively [26 (link),55 (link)]. Quantities were calculated based on reference curves for authentic standards in the range from 0.1 mM to 1.0 mM.

Cenk E., Schmutz C., Pahlke G., Oertel A., Kollarova J., Mock H.P., Matros A, & Marko D. (2021). Immunomodulatory Properties of Blackberry Anthocyanins in THP-1 Derived Macrophages. International Journal of Molecular Sciences, 22(19), 10483.

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