Fructo-oligosaccharide producing method with fossilized endoinulinase in bacterial cellulose
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A technology for endo-inulinase and bacterial cellulose, which is applied in the field of fructooligosaccharide production to achieve the effects of high purity, high content and high reuse rate
Inactive Publication Date: 2015-09-23
WEIDE QINGHAI BIOTECH
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[0005] Bacterial cellulose has high mechanical strength, thermal stability, good biocompatibility and non-toxicity, so that it can be widely used in the technical fields of immobilized enzymes and immobilized cells. Production of fructo-oligosaccharides by carrier-immobilized endo-inulinase
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Embodiment 1
[0029] Example 1: Bacterial bacterial cellulose surface and treatment.
[0030] Add bacterial cellulose membrane into pyridine at 5g / L~10g / L, stir evenly, then add sulfuric acid, the ratio of bacterial cellulose to sulfuric acid is 0.5~2g:100ml, react at 110°C for 1h, and prepare the modified bacterial cellulose membrane.
Embodiment 2
[0031] Example 2: Effect of the content of endo-inulinase on the immobilization effect.
[0032] The modified bacterial cellulose film obtained in Example 1 is added in the endo-inulinase solution at 0.5g / L, and the content of the endo-inulinase in the endo-inulinase solution is set as: 1000U / L, 2000U / L , 3000U / L, 4000U / L, 5000U / L. The immobilization temperature is 4°C, and the reaction time is 12h. After immobilization, the bacterial cellulose membrane immobilized with endo-inulinase was placed in 20 g / L inulin solution, and reacted at 32° C. for 12 h. The results are shown in Table 1.
[0033] The impact of the content of endo-inulinase in table 1 on immobilization effect
[0034] Enzyme amount
Embodiment 3
[0035] Example 3: Effect of immobilization time on immobilization effect.
[0036] The experimental method is the same as that in Example 2, except that the content of endo-inulinase in the endo-inulinase solution is set at 3000 U / L. The immobilization time is set to 3h, 6h, 12h, 18h, 24h. With the prolongation of immobilization time, the yield of fructo-oligosaccharide reached the highest at 12h, and the yield of fructo-oligosaccharide began to decrease after 12h.
[0037] The impact of table 2 immobilization time on immobilization effect
[0038] time
[0039] When the immobilization time is 12h, in the 0.25L system, the total enzyme activity is 750U, the total activity of the immobilized enzyme is 500U, and the recovery rate of the enzyme activity is 67%.
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Abstract
The invention discloses a fructo-oligosaccharide producing method with fossilized endoinulinase in bacterial cellulose. The fructo-oligosaccharide producing method with the fossilized endoinulinase in bacterial cellulose is characterized in that the fructo-oligosaccharide producing method comprises the steps that 1 pretreatment is conducted on the surface of a bacterial cellulose film, the surface hole size of the bacterial cellulose film is enlarged, and modified bacterial cellulose film is acquired; 2 the modified bacterial cellulose film is added into the endoinulinase solution, and the endoinulinase is fixed to the surface of the bacterial cellulose film; 3 washing is conducted through 0.5 mol /L Tris-HCI buffer solution, and bacterial cellulose film which fossilizing the endoinulinase is obtained; 4 the bacterial cellulose film which fossilizing the endoinulinase is placed into inulin solution, the endoinulinase is used for resolving the inulin, and fructo-oligosaccharide solution is obtained; 5 purification is conducted on the obtained fructo-oligosaccharide solution, and fructo-oligosaccharide with the purity over 90% is obtained. According to the fructo-oligosaccharide producing method with fossilized endoinulinase in bacterial cellulose, the prepared fructo-oligosaccharide is high in purity and little in impurity, the content of the fructo-oligosaccharide is over 99%, and significant industrial application prospect is possessed.
Description
technical field [0001] The invention belongs to the technical field of fructo-oligosaccharide synthesis, and in particular relates to a method for producing fructo-oligosaccharide by using bacterial cellulose to immobilize endo-inulinase. Background technique [0002] Fructose-oligosaccharides, also known as fructo-oligosaccharides, are kestose, kestose and kestrose formed by combining 1 to 3 fructosyl groups with fructosyl groups in sucrose through β(2-1) glycosidic bonds. A mixture of sugar and the like; about 60-70 grams of inulin in 100 grams of dry weight chicory, inulin is a fructan connected by a linear β-2,1-glycosidic chain, and its end is a sucrose group. Therefore, chicory powder is used as raw material to hydrolyze with endo-inulinase, and after refining, fructo-oligosaccharides can be finally obtained. [0003] Fructose-oligosaccharides have good health effects, are non-toxic, and are suitable for diabetic patients. They can prevent dental caries, regulate lipi...
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