Immobilized lipase and preparation method thereof

A technology of immobilized lipase and lipase, which is applied in the direction of immobilization on/in organic carriers, can solve the problems of unsuitability for large-scale application, high purity requirements, serious loss of enzyme activity, etc., and achieve the convenience of industrial continuous Sexual production, strong binding mode, and less loss of lipase activity

Inactive Publication Date: 2009-03-11
BEIJING CTA NEW CENTURY BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This immobilized lipase has high activity, large enzyme load and long service life, but the process is complicated, the cost is high, the enzyme activity loss is serious (generally more than 60%) in the immobilization process, and the process has high requirements for the purity of the enzyme itself. high
The price of this kind of immobilized enzyme is very high, generally more than 1,000 yuan / kg, and some even exceed 10,000 yuan, which is not suitable for large-scale application in the market

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

[0024] Embodiment 1, the preparation of immobilized lipase of the present invention and its effect verification

[0025] 1, the preparation of immobilized lipase of the present invention

[0026] Candida sp 99-125 was expanded and cultured in a 1000L fermenter, and the medium included soybean meal 50g / L, soybean oil 40g / L, K 2 HPO 4 1g / L, KH 2 PO 4 1g / L, defoamer 3g / L, at 28°C, the stirring radius is 0.2m, and the stirring frequency is 175r / min. Get fermented liquid, detect and show that the lipase hydrolysis activity of fermented liquid is 4000U / g fermented liquid with olive oil hydrolysis method (industry standard QB / T 1803-1993).

[0027] Take the Candida sp 99-125 fermentation broth obtained above, centrifuge at 2200g to take the supernatant, add PEG6000 to make the mass percentage concentration 2‰, after completely dissolving, add 5% polyacrylic acid of the supernatant of the fermentation broth Ester emulsion (solid content 35%, viscosity 305cP, the polyacrylate in t...

Embodiment 2

[0033] Embodiment 2, the preparation of immobilized lipase of the present invention and its effect verification

[0034] 1, the preparation of immobilized lipase of the present invention

[0035] Precipitate lipase from the fermentation broth obtained in step 1 of Example 1 by organic solvent precipitation, and dry the lipase to obtain enzyme powder.

[0036]After the enzyme powder is dissolved in an appropriate amount of water, the obtained lipase powder solution is tested with an olive oil hydrolysis method (industry standard QB / T 1803-1993) to show that the lipase hydrolysis activity of the enzyme liquid is 5000U / g enzyme liquid.

[0037] Take the enzyme powder dissolving solution obtained above, add PEG6000 to make its mass percent concentration 4‰, after completely dissolving, add 8% polyacrylate emulsion (viscosity is 200cP, solid content is 46%; Polyacrylate is a copolymer obtained by the polymerization reaction of butyl acrylate, ethyl acrylate, isooctyl acrylate and ...

Embodiment 3

[0043] Embodiment 3, the preparation of immobilized lipase of the present invention and its effect verification

[0044] 1, the preparation of immobilized lipase of the present invention

[0045] The fermented liquid comes from a 5-ton fermenter, and the bacterial strain and medium are the same as step 1 of Example 1, with a stirring radius of 0.35 m and a stirring frequency of 136 r / min.

[0046] Get new tank fermented liquid, the lipase hydrolysis activity of this fermented liquid is 8000U / g fermented liquid, filter to remove impurities such as thalline, add gelatin, make its mass percentage concentration be 3 ‰, fully dissolve, add 12% of fermented liquid quality % of polymer emulsion (this emulsion is a mixed emulsion of polyacrylate and polyurethane with a mass ratio of 2:1 as a solute, a viscosity of 280cP, and a total solid content of 36%, wherein polyacrylate is butyl acrylate, ethyl acrylate It is a copolymer obtained by polymerization reaction with methyl methacryla...

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Abstract

The invention discloses immobilized lipase and a method for preparing the same. The method for preparing the immobilized lipase comprises the following steps: evenly mixing lipase solution with polymer emulsion the weight of which is between 3 and 60 percent against that of the lipase solution, immersing a non-woven fabric by the obtained mixed liquid, and drying the mixed liquid to obtain the immobilized lipase, wherein the polymer emulsion is polyacrylate emulsion or the mixed emulsion containing polyacrylate and polyurethane as solutes. Experiments prove that the immobilized lipase has high catalysis performance and long service life, can still keep the catalysis efficiency above 90 percent after various continuous catalytic esterification reactions and transesterification reactions, and the required amount of the immobilized lipase for catalytically producing each ton of bio-diesel is less than or equal to 4 kilograms.

Description

technical field [0001] The invention relates to an immobilized lipase and a preparation method thereof. Background technique [0002] The immobilization technology of biological enzymes is one of the core contents of enzyme engineering. It successfully overcomes the shortcomings of free enzymes in industrial applications, such as poor stability, poor heat resistance, non-recyclability and difficult separation from products. The stability and catalytic activity of the immobilized enzyme are greatly improved, and its reusability can greatly reduce the production cost. Enzyme immobilization technology can improve the microenvironment of enzyme molecular catalysis, facilitate the contact between enzyme and substrate and the diffusion of products. In addition, the immobilized carrier endows the immobilized enzyme with certain physical properties and mechanical properties, so that it can be conveniently applied to various bioreactors, and facilitates the continuous and automatic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08
Inventor 张增伟张文波周浩顾燕松
Owner BEIJING CTA NEW CENTURY BIOTECH
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