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A technology for immobilizing lipase and lipase enzyme, which is applied in biochemical equipment and methods, immobilized on/in organic carriers, enzymes, etc., to achieve good storage stability, good pH tolerance, and high reuse. rate effect
Inactive Publication Date: 2018-09-04
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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However, there are few reports on immobilized lipase based on epoxy resin carrier at present.
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Embodiment 1
[0036] Embodiment 1: Obtaining of immobilized lipase
[0037] 1.1 Effect of different epoxy resins on lipase immobilization
[0038] Take 1g of different epoxy resins (epoxy resins LXEP-120, LX-1000EP, ES-1, ES-3, ES-5, ES-7), add 4mL enzyme solution (50mmol / L phosphoric acid at pH=7.0 Dissolve in sodium buffer solution, enzyme solution concentration 2mg / mL, centrifuge at 10000r / min for 12min), place in a shaker at 25°C, fix at 150rpm for 12h. It can be seen from Table 1 that the fixation effect of epoxy resin LXEP-120 is the best, and the recovery rate of enzyme activity reaches 12.21%.
[0039] Table 1 Immobilization effect of different epoxy resins
[0040]
[0041] 1.2 Effect of different immobilization buffers on lipase immobilization
[0042]Potassium phosphate buffer solution, sodium phosphate buffer solution, citric acid-disodium hydrogen phosphate buffer solution, Tris-HCl buffer solution, glycine-sodium hydroxide buffer solution with a pH of 8.0 were respective...
Embodiment 2
[0053] Embodiment 2: the enzymatic property comparison of epoxy resin immobilized lipase and free enzyme
[0054] In this embodiment, epoxy resin immobilized lipase is prepared by the following method:
[0055] Use 1.0mol / L of potassium phosphate buffer solution with a pH of 6.0 to dissolve lipase enzyme powder to prepare an enzyme solution with a concentration of 2.0mg / mL, centrifuge at 10000r / min to obtain the supernatant as the enzyme solution, and accurately weigh 5.0g of the model as The epoxy resin of LXEP-120 was placed in 4mL enzyme solution, immobilized at 150rpm and 22°C for 12h, then washed three times with 1.0mol / L potassium phosphate buffer solution with a pH of 6.0, filtered with suction, and placed at 40°C. ℃ drying to obtain immobilized lipase. The enzyme activity of the prepared immobilized lipase is about 400U / g, and the recovery rate of the enzyme activity reaches 92.79%.
[0056] 2.1 The optimal reaction pH of immobilized lipase and free lipase
[0057] ...
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Abstract
The invention discloses immobilized lipase and a preparation method thereof. The preparation method comprises the following steps: dissolving lipase enzyme powder by utilizing a potassium phosphate buffering solution with the concentration of 1.0mol/L and the pH (Potential of Hydrogen) of 6.0, so as to prepare an enzyme solution with the concentration of 2.0mg/mL; after centrifuging at the speed of 10000r/min, obtaining a clarified enzyme solution; putting epoxy resin with the model of LXEP-120 into the enzyme solution according to the ratio of 5.0g to 4mL; immobilizing for 12h under the conditions of 150rpm and 22 DEG C; then washing for 3 times by utilizing the potassium phosphate buffering solution with the concentration of 1.0mol/L and the pH of 6.0; filtering and drying at 40 DEG C toobtain the immobilized lipase, wherein the enzyme activity is 400U/g and the enzyme activity recovery rate reaches 92.79 percent. The immobilized lipase has good pH tolerance, temperature tolerance,operation stability and storage stability and meets the requirements of industrial application.
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 very important biocatalyst that can decompose substrates to produce various natural oils. As a biocatalyst, lipase has the characteristics of high catalytic efficiency, strong specificity, few by-products, and mild reaction conditions, and is widely used in the resolution of chiral compounds, the preparation of biodiesel, food processing and daily chemical industry and other industries. However, due to the poor stability of the free enzyme itself, the difficulty of separation and recovery, and the difficulty in realizing repeated and continuous utilization, its practical application in industrial production is largely limited. In order to solve the above shortcomings of free enzymes and really apply enzyme preparations to indust...
Claims
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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08
CPCC12N9/20C12N11/08C12Y301/01003
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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