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Lactose enzyme and common immobilization method of glucose isomerase

A technology of glucose isomerase and lactase, which is applied in the field of embedding-crosslinking and co-immobilization of lactase and glucose isomerase, can solve the problems of deep color, high cost and many by-products, and achieve Improve reaction efficiency and product quality, enhance market competitiveness, and save economic costs

Inactive Publication Date: 2008-05-21
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the industrial production of lactulose mainly adopts the chemical catalytic method, but the reaction conditions of this method are severe, the product obtained is dark in color, there are many by-products, and the separation of the catalyst is difficult, which brings difficulties to the further purification of the syrup and the preparation of crystallization.
However, the preparation of lactulose by enzymatic method has strong specificity, less by-products and light color of the product, but the cost is higher

Method used

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  • Lactose enzyme and common immobilization method of glucose isomerase
  • Lactose enzyme and common immobilization method of glucose isomerase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: prepare 20 g of pH 8.6 27% (w / w) gelatin solution, dissolve the gelatin completely in a water bath at 30° C., dissolve lactase and glucose isomerase in 2 mL of deionized water according to the activity ratio of 1:1, and dissolve Add the enzyme solution to the gelatin so that the amount of the two enzymes added is 3 U / g (gelatin solution), stir and mix well, then add 2 mL of glutaraldehyde with a volume fraction of 0.15%, pour it into a petri dish immediately after stirring, and It was cured overnight at 4°C, and the next day it was immersed in 0.15% glutaraldehyde for crosslinking at 4°C for 10 minutes, then washed with deionized water several times, the surface water was blotted with filter paper, and finally cut into small squares of 2mm square. The recovery rate of lactase in the co-immobilized enzymes was measured to be 30.85%, and the recovery rate of glucose isomerase was 83.48%.

[0030] Figure 1 shows the thermal stability changes of free lactase, fre...

Embodiment 2

[0033] Example 2: Prepare 20 g of mixed glue solution containing gelatin and sodium alginate with a mass concentration of 20% and 4% respectively, pH 7, dissolve completely in a water bath at 30°C, and isomerize lactase and glucose according to the activity ratio of 1:2 Dissolve the enzyme in 2mL of deionized water, add the enzyme solution to the mixed glue, make the addition amount of lactase and glucose isomerase respectively 9U / g and 18U / g (mixed glue), stir and mix well, then add 2mL of glutaraldehyde with a volume fraction of 0.25% was injected into the Petri dish, and after condensation, it was condensed with 4% CaCl 2 Soak overnight. The next day, it was washed with deionized water, and after clearing, it was cross-linked with 0.25% glutaraldehyde at 15°C for 30 minutes, then washed with deionized water several times, the surface water was blotted with filter paper, and cut into small squares of 2mm square.

Embodiment 3

[0034] Example 3: prepare 20 mL of carrageenan solution with pH 7.5 20%, completely dissolve the carrageenan in a water bath at 30°C, dissolve lactase and glucose isomerase in 2 mL of deionized water according to the activity ratio of 1:5, and dissolve the enzyme solution Add it to the carrageenan solution so that the amount of lactase and glucose isomerase added is 6U / g and 30U / g (carrageenan solution) respectively, stir and mix well, then add 2mL of glutaraldehyde with a volume fraction of 1%, Immediately after stirring, it was poured into a Petri dish, and after solidification, it was immersed in 0.2mol / L KCl to harden overnight at 4°C. The next day, cross-link with 1% glutaraldehyde at 30°C for 40 min, then wash with deionized water several times, blot the surface water with filter paper, and cut into small squares of 2 mm square.

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Abstract

A co-immobilization method for lactase and glucose isomerase belongs to the food biology technology area. The invention uses gelatine, sodium polymannuronate and / or carrageenan as the carrier; the first embedding and after cross linking process is applied; or the nylon net is used as the carrier; the first cross linking and after covalent combination method is used for the co-immobilization of lactase and glucose isomerase. The distinct characteristics of the invention are that the co-immobilization of lactase and glucose isomerase can not only realize recycle utilization of the enzyme which saves the economic cost and simplifies the downstream separation steps, but also exert the cooperation function between two enzymes which improves the reaction efficiency. The invention has a novel idea, a simple process and a strong practicality. The obtained lactase and glucose isomerase can be used for the transformation of lactose for the preparation of lactulose. The invention has good application prospect.

Description

technical field [0001] The invention relates to a co-immobilization method of lactase and glucose isomerase, in particular to an embedding-crosslinking method using gelatin, sodium alginate or / and carrageenan as a carrier; or a crosslinking method using nylon mesh as a carrier -Covalent method, co-immobilization of lactase and glucose isomerase, belonging to the field of food biotechnology. Background technique [0002] The use of free enzymes in large-scale industrial production will not only increase the production cost, but also cause the quality of the product to be low due to the addition of extra protein. After the enzyme is immobilized, it can not only realize continuous operation and repeated recycling, but also simplify the subsequent separation steps. The physical and chemical properties of the immobilized enzyme will change to a certain extent, and the stability to temperature and pH will be enhanced, which has great advantages. In 1970, Mosbach established the ...

Claims

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Application Information

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IPC IPC(8): C12N11/04C12N11/06
Inventor 杨瑞金刘芳
Owner JIANGNAN UNIV
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