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Immobilized lactase with hydrophobic property and method for preparing lactulose by applying immobilized lactase

A lactase and hydrophobic technology, applied in the direction of immobilized enzymes, biochemical equipment and methods, hydrolytic enzymes, etc., can solve the problems of unfavorable lactulose production and high hydrolysis activity, and achieve easy recycling, simplified purification, and cost reduction Effect

Active Publication Date: 2021-03-12
苏州福赛思生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problem that the hydrolysis activity of lactase is too high in the reaction of catalyzing the preparation of lactulose from lactose, which is not conducive to the production of lactulose, the present invention immobilizes lactase on a hydrophobic carrier innovatively, and further surface lactase Agglomeration is carried out to endow the prepared immobilized lactase with certain hydrophobic characteristics, and then provide a hydrophobic microenvironment for lactase in the reaction system, thereby inhibiting its hydrolysis activity to a certain extent and promoting the formation of lactulose

Method used

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  • Immobilized lactase with hydrophobic property and method for preparing lactulose by applying immobilized lactase
  • Immobilized lactase with hydrophobic property and method for preparing lactulose by applying immobilized lactase
  • Immobilized lactase with hydrophobic property and method for preparing lactulose by applying immobilized lactase

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Embodiment 1

[0035] Example 1: Immobilized lactase

[0036] DSM's Maxilact ® Take lactase as an example, add it to the phosphate buffer solution with pH 6.8, the enzyme protein concentration is 2.5 mg / mL, and add commercially available amino resin (take Duolite A568 amino resin of DOW Company as an example), the amount of resin added to 0.5 mg / mL, react at 4°C for 12 h, then slowly add ground ammonium sulfate powder until its saturation reaches 70%, continue stirring at 4°C for 30 min, then add glutaraldehyde solution (commercially available product, 25 %, v / v) until the concentration of glutaraldehyde in the whole system was 0.5% (v / v), stirred slowly at 4°C for 4 h, then collected the immobilized lactase particles by filtration, and used phosphate buffer solution of pH 6.8 to The collected immobilized lactase particles were washed three times, and then freeze-dried. The schematic diagram of the operation of immobilized lactase is as follows: figure 1 As shown, the electron micrograph ...

Embodiment 2

[0037] Example 2: Preparation of Lactulose from Lactose Catalyzed by Immobilized Lactase

[0038]Add 250 g of lactose and 250 g of fructose to the pH 6.8 phosphate buffer, stir to dissolve (heating can be used to assist dissolution), and prepare 2 parts (A and B) each 1 L with a final lactose and fructose content of 250 g / L of the reaction substrate solution, add 2 mL of free lactase to A as a catalyst, add 5 g of immobilized lactase to B as a catalyst, stir and react for 6 h at 37°C, and take 1 mL of the reaction solution each , inactivate the enzyme in a boiling water bath for 5 min, dilute to an appropriate multiple, and then use high performance liquid chromatography to detect the composition and content of various sugars in the reaction solution system. The obtained HPLC chromatograms are shown in Figure 3 and Figure 4 below. It can be seen from the analysis that the final lactulose content of group A using free enzyme is 46.18 g / L, including 38.65 g / L 1-lactulose and 7....

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Abstract

The invention discloses immobilized lactase with a hydrophobic property and a method for preparing lactulose by using the immobilized lactase. The method comprises the following steps: firstly, immobilizing lactase to endow the lactase with a hydrophobic property; taking the lactulose as a bio-catalyst, and catalyzing lactose under mild conditions to prepare lactulose, wherein the yield of the lactulose can reach 33.6%, the content of the lactulose is 83.9 g / L, and the yield of the lactulose is nearly two times that of lactulose obtained by free lactase. The method is characterized in that through immobilization operation, the immobilized lactase is endowed with a certain hydrophobic property, so that the hydrolysis activity and transglycosylation activity of the lactase are regulated andcontrolled, and the lactase is enabled to be carried out in the transglycosylation direction beneficial to generation of lactulose. The method effectively solves the problem that the hydrolysis activity is too high when lactase is applied to lactulose production, the immobilized lactase with a hydrophobic characteristic is adopted, lactulose can be efficiently prepared without assistance of an organic phase, the practicability is high, and the method is a beneficial substitute for preparing lactulose through a chemical method and a free enzyme method.

Description

technical field [0001] The invention belongs to the field of food biotechnology, and in particular relates to an immobilized lactase with hydrophobic properties and a method for its application in preparing lactulose. Background technique [0002] Lactulose (C 12 h 22 o 11 , 4- O - β - D -Galactopyranosyl- D -Fructose) is a functional oligosaccharide formed by the combination of galactose and fructose by β-1,4-glycosidic bonds. Lactulose cannot be metabolized and decomposed by the human body, and cannot be absorbed in the small intestine of the human body. After entering the colon, it can be decomposed by the β-galactosidase produced by bifidobacteria and become a carbon source for the proliferation of bifidobacteria, thereby promoting beneficial bacteria in the human intestinal tract It can inhibit the growth of harmful bacteria, and has good curative effect on patients with intestinal flora imbalance such as constipation, diarrhea, enteritis, etc., so it is also cal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24C12N11/00C12P19/12C12P19/14
CPCC12N9/2402C12Y302/01108C12N11/00C12P19/12C12P19/14
Inventor 杨瑞金汪明明
Owner 苏州福赛思生物科技有限公司
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