Rhubarb horsetails Erwinia sp. and its application in preparation of isomaltulose
A technology of isomaltulose and Erwinia, applied in the fields of fermentation engineering and enzyme engineering
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Embodiment 1
[0036] Slant medium: peptone 10g / L, beef extract 3g / L, NaCl 5g / L, agar 20g / L, pH 7.0.
[0037] Shake flask medium: sucrose 50g / L, yeast extract 10g / L, Na 3 PO 4 ·12H 2 O 5g / L, pH7.0.
[0038] Erwinia rhubarb NX-5 CGMCC No.2222 was cultured on a slant medium at 30°C for 24h, then a loop of this bacteria was placed in a shake flask medium, cultured at 30°C for 10h, and the shake flask speed was 200r / min. The bacterial content in the solution is 20g / L, the enzyme production reaches 2.5-5.0U / ml, and it is transformed with free cells. Weigh 10g of cells and add them to 100ml of 450g / L sucrose solution (the amount of cells added is 1L55% per 100g of cells). sucrose solution), under the condition of 30 ℃, shake and transform to produce isomaltulose in a shaker flask, the reaction time is 8h, and the substrate conversion rate and product concentration are measured after the reaction is terminated. The conversion rate of sucrose is 99.5%, and the conversion rate of isomaltulose is ...
Embodiment 2
[0040] With 50g / L maltose as carbon source, 5g / L bean cake flour as nitrogen source, other components of medium and bacterial culture conditions are the same as in Example 1, prepare 1.5% sodium alginate colloidal solution, and the bacterial suspension prepared with physiological saline After mixing evenly, add 8% diatomaceous earth for adsorption, drop 2% CaCl with a syringe 2 It was solidified in the solution to make spherical immobilized cells with a diameter of about 3-4 mm, which were placed in a refrigerator at 4 °C for several hours, then the supernatant was poured out, washed twice with distilled water, and 10 g of wet cells were wrapped in sodium alginate. 20 g of immobilized cells were obtained by burying, and 100 ml of 550 g / L sucrose solution was transformed with this in a shaker flask at 30 °C. The transformation time of each batch was 10 h, and 40 batches were transformed. The average conversion rate of sucrose was 99.5%. The average conversion of maltulose was 8...
Embodiment 3
[0042] 225g of glucose, 45g of corn steep liquor, 22.5g of NaCl, pH 6.0 were made up to 4.5L with water to form a culture medium, put into a 7.5L glass stirring fermenter, and steam sterilized at 121°C for 15min. Preparation of seed medium: sucrose 50g / L, yeast extract 10g / L, Na 2 HPO 4 ·12H 2 O 5g / L, pH 6.0. Connect Erwinia rhubarb NX-5 CGMCC No.2222 to the seed medium, cultivate at 30°C for 8 hours to obtain the seed liquid, and insert the seed liquid into the cooled fermentation medium according to the inoculum of 3% of the volume of the fermentation liquid. , 30 ℃ culture (ventilation volume 1vvm, stirring speed of 600r / min) 12h. The fermentation broth is filtered bacteria in an ultrafilter, and the ultrafiltration membrane cut-off molecular weight is 10 6 Dalton, the operating pressure is 0.2MPa, the control temperature is 50°C, and the membrane surface speed is 4m / s. After the 90g cells obtained by the interception are immobilized according to the method of Example 2...
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