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
View PDF0 Cites 9 Cited by
  • 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 la

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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

Example Embodiment

[0020] Example 1: Obtainment 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 2.5% by mass, and select sodium alginate-arabic gum (mass ratio 3:2), sodium alginate-polyvinyl alcohol (mass ratio 3:2), and seaweed. Sodium-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) Mass ratio 3:2), sodium alginate-sodium carboxymethyl cellulose (CMC) (mass ratio 3:2) as the embedding carrier for the immobilization of lipase.

[0023] The specific steps are:

[0024] S1. Weigh 1.5g of sodium alginate and 1g of gum arabic, add them to 97.5g of pH 7.0 phosphate buffer, heat to dissolve, and stir and mix well to obtain a sodium alginate-acacia compound carrier solution; weigh 1.5 g sodium alginate and 1g polyvinyl alcohol were added to 97.5g pH 7.0 phosphate b...

Example Embodiment

[0070] Example 2: Crosslinking of immobilized lipase

[0071] 2.1 The effect of crosslinker concentration on the activity of immobilized lipase

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

[0073] The specific steps are:

[0074] S1. Weigh 2.5g of sodium alginate and 1.5g of CMC into 96g of pH 7.0 phosphate buffer, heat to dissolve, and stir and mix uniformly to obtain sodium alginate-CMC composite carrier solution P (sodium alginate The content is 2.5% by mass, and the content of CMC is 1.5% by mass); lipase (Shenzhen Hengsheng Biotechnology Co., Ltd., article number: 9001-62-1) is dissolved in pH 7.0 phosphate buffer to prepare 800 U / mL of lipase solution;

[0075] S2. Stir and m...

Example Embodiment

[0086] Example 3: Comparison of enzymatic properties of immobilized lipase and free lipase after cross-linking

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

[0088] Set the pH of the enzymatic reaction system to 6-10, and measure the enzymatic activity of immobilized lipase and free lipase at 40°C. The results are shown in Figure 4 a. by Figure 4 a It can be seen that the optimal reaction of the immobilized lipase cross-linked by EGDE (the preparation method is the same as that in Example 2 step 2.3), the immobilized lipase cross-linked by glutaraldehyde (the preparation method is the same as that in Example 2 step 2.3) and free lipase The pH is 8.5, 8.0 and 8.0, respectively. Compared with the immobilized lipase cross-linked by free lipase and glutaraldehyde, the optimal pH of the reaction system of immobilized lipase cross-linked by EGDE shifts to the alkaline direction. When the pH is between 8.0 and 9.0, it shows Higher enzyme activity. ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
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
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap