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Immobilized lipase and preparation method thereof

A technology of immobilized lipase and lipase, which is applied in the direction of biochemical equipment and methods, fixed on/in organic carriers, enzymes, etc., can solve the problems of low reuse rate of immobilized enzymes, leakage of enzyme molecules, condensation Solve the problems of poor mechanical strength of rubber beads, and achieve good thermal stability and reusability

Active Publication Date: 2018-06-08
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sodium alginate is a commonly used immobilized enzyme carrier. The calcium alginate gel formed with calcium chloride has the advantages of high water content, easy molding, clean and non-toxic, etc., but due to the large size of its network pores , the mechanical strength of the gel beads is poor, the enzyme molecules are easy to leak out of the gel pores, and the reusability of the immobilized enzymes prepared by them is not high

Method used

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  • Immobilized lipase and preparation method thereof
  • Immobilized lipase and preparation method thereof
  • Immobilized lipase and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: Obtaining of immobilized lipase

[0021] 1.1 Screening the carrier compounded with sodium alginate

[0022] Set the concentration of the compound carrier in the compound carrier solution to be 2.5% by mass fraction, choose sodium alginate-gum arabic (mass ratio 3:2), sodium alginate-polyvinyl alcohol (mass ratio 3:2), seaweed Sodium alginate-xanthan gum (mass ratio 3:2), sodium alginate-gelatin (mass ratio 3:2), sodium alginate-carrageenan (mass ratio 3:2), sodium alginate-carrageenan ( Mass ratio 3:2), sodium alginate-sodium carboxymethyl cellulose (CMC) (mass ratio 3:2) was used as embedding carrier to study the immobilization of lipase.

[0023] The specific steps are:

[0024] S1. Weigh 1.5g of sodium alginate and 1g of gum arabic, add it to 97.5g of pH7.0 phosphate buffer, heat to dissolve, and stir and mix evenly to obtain a sodium alginate-gum arabic compound carrier solution; weigh 1.5g Add 97.5g of sodium alginate and 1g of polyvinyl alcohol in...

Embodiment 2

[0070] Example 2: Cross-linking of immobilized lipase

[0071] 2.1 The effect of cross-linking agent concentration on the activity of immobilized lipase

[0072] According to the content of sodium alginate in the composite carrier solution is 2.5% by mass fraction, the content of CMC is 1.5% by mass fraction, the concentration of calcium chloride solution is 5% by mass fraction and the fixation time is 50min, the effect of crosslinking agent concentration on immobilized lipase is studied. effect on enzyme activity.

[0073] The specific steps are:

[0074] S1. Weigh 2.5g sodium alginate and 1.5g CMC and add them to 96g pH7.0 phosphate buffer solution, heat to dissolve, and stir and mix evenly to obtain sodium alginate-CMC composite carrier solution P (sodium alginate content is 2.5% by mass fraction, and the content of CMC is 1.5% by mass fraction); lipase (Shenzhen Hengsheng Biotechnology Co., Ltd., article number: 9001-62-1) is dissolved in the phosphate buffer solution of...

Embodiment 3

[0086] Example 3: Comparison of enzymatic properties between immobilized lipase and free lipase after crosslinking

[0087] 3.1 The optimal reaction pH of immobilized lipase and free lipase

[0088] Set the pH of the enzymatic reaction system to 6-10, measure the enzymatic activity of immobilized lipase and free lipase at 40°C, the results are shown in Figure 4 a. Depend on Figure 4 It can be seen that the optimal reaction of the immobilized lipase (preparation method is the same as embodiment 2 step 2.3) of EGDE crosslinking, glutaraldehyde crosslinking immobilized lipase (preparation method is the same as embodiment 2 step 2.3) and free lipase The pHs were 8.5, 8.0 and 8.0, respectively. Compared with free lipase and glutaraldehyde-crosslinked immobilized lipase, the optimal pH of the reaction system of EGDE-crosslinked immobilized lipase shifted to the alkaline direction, and when the pH was between 8.0 and 9.0, it showed Higher enzyme activity. 3.2 Optimum reaction ...

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Abstract

The invention discloses immobilized lipase and a preparation method thereof. The preparation method comprises the following steps of respectively dissolving a compound carrier and lipase into a buffersolution with pH (potential of hydrogen) value of 6.0 to 8.0, so as to obtain a compound carrier solution and a lipase solution; uniformly mixing the compound carrier solution and the lipase solutionaccording to a ratio of compound carrier to lipase as (26 to 47)mg:(400 to 1600)U, so as to obtain a compound carrier / lipase mixed solution, wherein the content of the compound carrier in the compound carrier / lipase mixed solution is 12.8 to 23.6mg / mL; dripping the compound carrier / lipase mixed solution into a mixed solution containing a crosslinking agent and calcium chloride, forming gel microspheres, crosslinking and immobilizing, washing, suction filtering, so as to obtain the immobilized lipase. The prepared immobilized lipase has the advantages that the enzyme activity reaches 380U / g, the yield rate of enzyme activity is about 50.09%, the heat stability and repeated utilization rate are higher, and the requirement of industrialized application is met.

Description

Technical field: [0001] The invention belongs to the fields of biochemical industry and biotechnology, and in particular relates to an immobilized lipase and a preparation method thereof. Background technique: [0002] Lipase (Lipase, EC3.1.1.3) is a general term for a class of enzymes that catalyze the hydrolysis of glycerides. The catalytic reactions usually have substrate specificity and stereoisomerism, and the reaction conditions are relatively mild, and the catalytic efficiency is high. High, with few by-products, it is widely used in industrial fields such as food, tanning, washing, pharmaceuticals and environmental governance. As people pay more and more attention to food safety, lipase, as a green bio-efficient product, is gradually replacing the use of some chemical additives. For example, lipase and glucose oxidase can replace the chemical gluten-enhancing agent potassium bromate , Lipase replaces the chemical catalyst used in traditional oil hydrolysis, enzyme p...

Claims

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Application Information

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IPC IPC(8): C12N9/20C12N11/08C12N11/10C12N11/12
CPCC12N9/20C12N11/08C12N11/10C12N11/12C12Y301/01003
Inventor 胡云峰徐珊李任强张继福张云孙爱君
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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