How can 70-18-8 extract glutathione from glutathione fermentation broth? Research level GSH
The method of extracting glutathione from glutathione fermentation broth is to prepare high-quality and high content glutathione using glutathione fermentation broth. The prepared glutathione is used as a product in the form of food additives rich in GSH.
The specific operation steps are as follows:
1. Yeast cell fluid from Lixinde:
Centrifuge the fermentation broth at 3000rpm for 10-15min to obtain wet yeast cell precipitate.
2. Adjusting pH value:
Adjust the pH value of yeast cell fluid to 1-2 using 2mol/l H2SO4.
3. Boiling water breaks walls:
Based on the mass of wet yeast cells, calculate the volume of distilled water according to the material ratio of 10-12ml/g. First, heat most of the distilled water to boiling, pour the yeast cell solution with the adjusted pH into the boiling water, stir quickly, rinse the beaker with the remaining small amount of distilled water, and pour it in together.
When the temperature is reheated to 95-100 ℃, start timing. After 12 minutes, immediately place it in an ice bath to cool to 4 ℃
4. Cation exchange:
Change the model to 001 × 7 cation exchange resin columns with a column volume of φ 2.6cm × 30cm, directly add yeast cell fluid containing bacterial cells to the column, with a pH value of 2-3 and a flow rate of 1ml/min during sample loading, leaving the unadsorbed yeast cell fluid to circulate until saturation;
Wash the resin column with distilled water adjusted to pH 3 with hydrochloric acid at 3-4 times the column volume, remove impurities such as bacterial and cell precipitates, and then wash with 1.5% HCl at 3-4 times the column volume. The elution rate is 0.5ml/min, and the eluent is concentrated to 5-6 times the concentration. Collect the concentrated solution rich in GSH.
5. Vacuum concentration:
Concentrate the glutathione concentrate under vacuum again, following the following conditions: vacuum degree of 0.9MPa, rotation speed of 130rpm, temperature of 55-60 ℃, concentrated to 8-10 times, to obtain the GSH super concentrate.
6. Freeze drying:
Freeze dry the concentrated solution of glutathione at -84 ℃ for 28 hours to obtain a white powdered glutathione product.
