Reagents and equipment:
1. ammonium sulfate (containing 10mmol / L Tris-HCl, (pH 7.4) and 0.2mmol / L MgCl 2 of 1mol / L and 0.2mol / L ammonium sulfate solution);
2. Magnesium chloride (1 mol/L stock solution);
3. Phenylmethylsulfonyl fluoride (PMSF) (1 mol/L stock solution prepared from absolute ethanol);
4. The purified nuclei are stored in a solution containing 0.25 mol/L sucrose, 10 mmol/L Tris-HCl, (pH 7.4), 5 mmol/L MgCl 2 ;
5. DNase I;
6. Tris-HCl (1 mol/L stock solution, pH 7.4);
7. Suspension buffer (SB): 5 mmol/L MgCl2, 10 mmol/L Tris-HCl, pH 7.4 (containing 0.1 mmol/L phenylmethylsulfonyl fluoride);
8. Triton X-100;
9. Low speed centrifuge with fixed angle rotor and centrifuge tube;
experimental method:
All reagents were placed at 0-4 °C and all operations were done at 0-4 °C.
1. Add Triton X-100 to the nuclear suspension to a concentration of 1% (volume fraction), incubate for 30 minutes while carefully mixing with a pipette to avoid air bubbles;
2. Centrifuge at 1000g for 10min and discard the supernatant. Resuspend the extracted nuclear pellet (eg 5 ml) in small volume with suspension buffer;
3. Digest the nucleus with DNase I (30 U/mg DNA) for 30 min;
4. Increasing the salt concentration by slowly adding 1 mol/L ammonium sulfate solution to achieve a final ionic strength of 0.6 (0.2 mol/L ammonium sulfate), adding 0.2 mol/L ammonium sulfate to a volume of 40 ml, and then incubating for 30 min;
5. Centrifuge the core substrate at 1000g for 15min, discard the supernatant;
6. Resuspend the pellet in suspension buffer and examine the obtained product using phase contrast microscopy or thin-section electron microscopy;
Besides improving the baking quality of wheat flour, it enhances the rheological properties of the dough by stabilizing pore size and elasticity. Other benefits of TG-ase include its positive impact on the sensory properties of bread, texture and loaf volume.
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Transglutaminase For Plant Based Meat
TGase applied in plant based meat, Transglutaminase for vegan meat, TG enzyme for plant based meat
Guangdong Kelong Biotechnology Co., Ltd. , https://www.kelongfood.com