Fungal glucan oligomer chrome complex and preparation method thereof
A technology of chromium complex and oligoglucose, which is applied in the field of chromium oligosaccharide and its preparation, can solve the problems of large difference in ratio, harsh reaction conditions, and low bioavailability, and achieve mild synthesis conditions, The conditions are easy to control and the effect of low molecular weight
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Embodiment 1
[0053] Preparation of Sclerotinia β-glucooligosaccharide
[0054] Suspend 15g of Sclerotin glucan in 100mL of water, stir to fully swell, adjust the pH to 4-5, add 30mg of β-glucanase when heated to 50°C, and hydrolyze for 1-2 hours under stirring; then add glucose Polysaccharide 10g, keep stirring for 1-2h, the solution becomes thinner and gradually transparent; adjust the pH to 7-8, add 20mg of neutral protease, keep warm for 2-4h; raise the temperature to 90°C, inactivate the enzyme activity; centrifuge at 6000r / min for 20min, Ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of 10 kDa to collect the filtrate, and then ultrafiltration with a nanofiltration membrane with a molecular weight cut-off of 200 Dalton to collect the retentate; that is, oligoglucose with a weight average molecular weight of 2528 Dalton. The gluco-oligosaccharide solution contains 22.51% (w / v) of β-gluco-oligosaccharide.
Embodiment 2
[0056] Synthesis and purification of β-glucooligosaccharide chromium from Sclerotinia sp.
[0057] Sclerotinia β-glucooligosaccharide solution 100mL, containing 15.37% (w / v) of glucooligosaccharide with a weight average molecular weight of 2240Dalton, adjusted the pH value to 12 with 10mol / L NaOH, heated to 75°C, and added dropwise 2mol / L During this period, use 2mol / L NaOH to adjust the pH to stabilize the pH value at about 12. When green flocculent precipitates appear in the solution, stop adding chromium chloride and NaOH dropwise, and the reaction solution continues to be heated at 75°C. Keep warm for 1h. Ultrafiltration with a nanofiltration membrane with a molecular weight cut-off of 200 Daltons was used to collect the retentate; further purified by 200-300 mesh silica gel column chromatography, and then freeze-dried to obtain 8.6 g of Sclerotinia β-glucooligosaccharide chromium. The chromium yield of β-glucooligosaccharide is 56%, and the chromium content is 7.43%w / w. ...
Embodiment 3
[0066] Preparation of Aureobasidium pullulans α-gluco-oligosaccharide
[0067] Suspend 10g of pullulan in 100mL of water, stir to fully swell, adjust the pH to 4-5, heat at 55°C, add 10mg of α-1,6-glucanase, hydrolyze for 2 hours under stirring; then add 5g of dextran , 10 mg of α-amylase was continuously stirred for 1-2 hours, the solution became thinner and gradually became transparent; adjust the pH to 7-8, add 15 mg of alkaline protease, and keep warm for 2-4 hours; raise the temperature to 100°C to inactivate the enzyme; centrifuge at 6000r / min After 20 minutes, use ultrafiltration with a molecular weight cut-off of 6000 Dalton to collect the filtrate, and then use a nanofiltration membrane with a molecular weight cut-off of 300 Dalton to collect the retentate; that is, the gluco-oligosaccharide with a weight average molecular weight of 1650 Dalton. The gluco-oligosaccharide solution contains 12.6% (w / v) of α-gluco-oligosaccharide.
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