Method for enriching D-psicose 3-epimerase by using membrane and application of D-psicose 3-epimerase
A technology of epimerase and psicose, which is applied in the field of using membrane to enrich D-psicose 3-epimerase, and can solve the problem of D-psicose 3-epimerase The problems of low enzyme utilization rate and high preparation cost of psicose can achieve the effect of low production cost, large social benefit and broad economic benefit.
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Embodiment 1
[0024] (1) Bacteria removal: the saccharification solution with a sugar concentration of 42% without enzymes is used for sterilization operation with a ceramic membrane with a pore size of 0.1um. During the operation, the temperature of the material inside the ceramic membrane is controlled at 18°C, and the pressure is controlled at 0.2MPa After the filtration is completed, rinse the ceramic membrane with 0.3 times water (V / V) until the sugar concentration of the permeated phase is zero;
[0025] (2) Ultrafiltration Enrichment Enzyme: Step (1) The permeate after ceramic membrane treatment is ultrafiltered with an ultrafiltration membrane. The molecular weight cut-off of the ultrafiltration membrane is 10,000 Daltons. , the pressure is controlled at 0.4Mpa, and the ultrafiltration membrane is washed with 0.4 times of water (V / V) until the sugar concentration of the permeated phase is zero, and the concentrated phase is collected, which is the recovered D-psicose 3-epimide Const...
Embodiment 2
[0028] (1) Bacteria removal: the saccharification solution with a sugar concentration of 50% without enzymes is used for sterilization operation with a ceramic membrane with a pore size of 0.1um. During the operation, the temperature of the material inside the ceramic membrane is controlled at 20°C, and the pressure is controlled at 0.4MPa After the filtration is completed, wash the ceramic membrane with 0.5 times water (V / V) until the sugar concentration of the permeated phase is zero;
[0029] (2) Ultrafiltration Enrichment Enzyme: Step (1) The permeate after ceramic membrane treatment is ultrafiltered by ultrafiltration membrane, the molecular weight cut-off of ultrafiltration membrane is 10000 Daltons, and the temperature of the material inside the ultrafiltration membrane is controlled at 18°C , the pressure is controlled at 0.6Mpa, and the ultrafiltration membrane is washed with 0.4 times of water (V / V) until the sugar concentration of the permeated phase is zero, and the...
Embodiment 3
[0032] Reuse of the D-psicose 3-epimerase recovered in Example 1:
[0033] Prepare a fructose solution with a sugar concentration of 45%, adjust the pH value to 6.2, add the D-psicose 3-epimerase recovered in Example 1, and add the enzyme in an amount of 35 U / g fructose dry basis, After reacting at 60°C for 20 h, the components of the saccharification solution were determined to be: 0.27% polysaccharide, 0.73% disaccharide, 0.99% glucose, 1.21% mannose, 70.45% fructose, and 26.18% allulose; 1) Compared with the composition of the saccharification solution in the medium, the conversion rate of fructose into allulose is roughly the same (the amount of enzyme added is equal), so the D-psicose 3-epimerase recovered by the present invention Still has a strong transformation ability.
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Abstract
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