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Method for preparing biodiesel by catalyzing vector-free immobilized lipase in ionic liquid system

A carrier-free immobilization and ionic liquid technology, which is applied in biofuels, hydrolytic enzymes, fermentation, etc., can solve the problems of high lipase immobilization cost, low oil conversion rate, and easy decline in enzyme activity, which is beneficial to industrial applications. Good stability and high activity per unit volume

Inactive Publication Date: 2015-09-16
GUANGZHOU INST OF ENERGY CONVERSION - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problems of high fixed cost of lipase existing in the current biodiesel production process, easy decrease or even inactivation of enzyme activity, low oil conversion rate, etc. A method for preparing biodiesel catalyzed by lipase, which has the advantages of high enzyme activity, good stability, recyclable utilization, and cost saving.

Method used

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  • Method for preparing biodiesel by catalyzing vector-free immobilized lipase in ionic liquid system

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Preparation of carrier-free immobilized lipase:

[0030] ① Dissolve the free Rhizomucor miehei lipase and bovine serum albumin (BSA) protective agent with a mass ratio of 5:1 in a phosphate buffer solution with a pH of 5.8 and a molar concentration of 0.1mol / L to make 10mg / mL For enzyme solution, take 50mL of enzyme solution, add 100mL of 0.4mol / L calcium chloride aqueous solution, mix well, quickly add an equal amount of equimolar sodium carbonate solution of 100mL, stir magnetically at 4-25°C for 1min, and stand still for 2h , through co-precipitation, the free enzyme molecules are evenly dispersed in CaCO 3Inside the particulate template agent, stable spherical templated calcium carbonate particles are formed; then add 150mL of saturated ammonium sulfate solution precipitant dropwise, shake slowly at 4-25°C for 1-2min, let stand for 2h, and carry out lipase coagulation precipitation, 3000rpm Centrifuge and wash for 3 minutes to obtain coprecipitated particles;

[0...

Embodiment 2

[0036] Preparation of carrier-free immobilized lipase:

[0037] ①Dissolve free Candida antarctica lipase and bovine serum albumin (BSA) protective agent with a mass ratio of 6:1 in pH=7, molar concentration of 0.2mol / L phosphate buffer to make 11mg / mL enzyme liquid. Take 50mL of enzyme solution, add 100mL of 0.4mol / L calcium chloride aqueous solution, mix evenly, quickly add the same amount of equimolar sodium carbonate solution 100mL, stir magnetically for 1min at 4~25℃, let it stand for 2h, pass Co-precipitation allows free enzyme molecules to be evenly dispersed inside the CaCO3 particle template to form stable spherical templated calcium carbonate particles; then add 180mL of saturated ammonium sulfate solution precipitant dropwise, shake slowly at 4-25°C for 1-2min, and then statically Set aside for 2 hours, carry out coagulation and precipitation of lipase, centrifuge and wash at 3000rpm for 3 minutes to obtain coprecipitated particles;

[0038] ② Add 1ml of 50mmol / L d...

Embodiment 3

[0043] Preparation of carrier-free immobilized lipase:

[0044] ①Dissolve free Penicillium extensa lipase and bovine serum albumin (BSA) protective agent with a mass ratio of 10:1 in pH=8, molar concentration of 1mol / L phosphate buffer to prepare 15 mg / mL enzyme solution. Take 50mL of enzyme solution, add 100mL of 0.5mol / L calcium chloride aqueous solution, mix evenly, quickly add the same amount of equimolar sodium carbonate solution of 100mL, stir magnetically for 1min at 4-25℃, let stand for 2h, pass Co-precipitation allows the free enzyme molecules to be evenly dispersed inside the CaCO3 particle template to form stable spherical templated calcium carbonate particles; then add 200mL of saturated ammonium sulfate solution precipitant dropwise, shake slowly at 4-25°C for 1-2min, and then statically Set aside for 2 hours, carry out coagulation and precipitation of lipase, centrifuge and wash at 3000rpm for 3 minutes to obtain coprecipitated particles;

[0045] ②Add 1.2ml of ...

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Abstract

The invention discloses a method for preparing biodiesel by catalyzing vector-free immobilized lipase in an ionic liquid system. The method comprises the following two steps: (1) preparing vector-free immobilized lipase; and (2) preparing the biodiesel by catalyzing the prepared vector-free immobilized lipase in the ionic liquid system. The prepared vector-free immobilized lipase is high in activity and free of high-cost vectors; an ionic liquid medium is used as an immobilized enzyme for preparing the biodiesel; the stability and the stereoselectivity of substrates of the enzyme can be obviously improved; the problem of the enzyme poisoning caused by methyl alcohol and glycerol can also be avoided; the efficient catalysis of the enzyme is ensured; the yield of the biodiesel is greatly increased; the immobilized enzyme and the ionic liquid used in the method can be recycled and reused; the waste of resources is avoided; the production cost is reduced; the method has a good industrial application prospect.

Description

technical field [0001] The invention relates to the technical field of biochemical industry, in particular to a method for preparing biodiesel by catalyzing carrier-free immobilized lipase in an ionic liquid system. technical background [0002] With the dwindling of petroleum resources and the enhancement of people's awareness of environmental protection, it is imminent to find and develop clean and renewable energy. Biodiesel is a renewable energy source made from vegetable oil or animal oil. It does not contain sulfur and aromatics, has a high cetane number, good lubricity, and has the advantages of environmental friendliness and renewable resources. It is an ideal fuel to replace petroleum diesel. It has attracted much attention in recent years. The traditional process of preparing biodiesel uses a homogeneous inorganic acid-base as a catalyst, which has the disadvantages of large alcohol consumption, difficult product recovery, and large environmental pollution. Theref...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/64C12N9/20
CPCC12N9/20C12P7/6454C12P7/649Y02E50/10
Inventor 袁振宏苗长林吕鹏梅庄新姝王忠铭罗文李惠文杨玲梅刘姝娜
Owner GUANGZHOU INST OF ENERGY CONVERSION - CHINESE ACAD OF SCI
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