Preparation method of magnetic response composite lipase and application in biodiesel synthesis

A composite lipase, magnetic responsive technology, applied in biofuels, immobilized on or in inorganic carriers, immobilized on/in organic carriers, etc., can solve the needs of industrial production, enzyme loss of enzymes, etc. Activity, poor stability and other problems, to achieve the effect of improving enzyme transesterification efficiency, reducing enzyme dosage, and improving transesterification efficiency

Inactive Publication Date: 2015-01-28
TAIYUAN UNIV OF TECH
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  • Abstract
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Problems solved by technology

[0005]However, the compound enzyme technology currently used in biodiesel production is based on the existing immobilized enzyme, and the price is relatively high. For example, the most widely used one is Novi Shin's Novozym 435, priced at $2,270 per kilogram
On the other hand, since the lipid substrate is hydrophobic, lipase must be combined with the substrate to catalyze the reaction, so the catalytic reaction of lipase on esters can only occur at the oil-water interface.
Therefore, during the reaction process, constant stirring...

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  • Preparation method of magnetic response composite lipase and application in biodiesel synthesis

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Embodiment 1

[0030]Example 1 Preparation of magnetically responsive lipase A with 1,3 position specificity

[0031] Specifically refers to the use of 10nm nano-Fe 3 o 4 Particle-modified Thermomyces lanuginosus lipase (Lipase from Thermomyces lanuginosus , TLL) preparation: Take 1mL nano-Fe with carboxyl functional groups on the surface 3 o 4 solution (10nm, iron content 155mg / L), add 10μL of 10mmol / L carbodiimide (1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide, EDC) solution and 25mmol / L N- Hydroxysulfosuccinimide ( N -hydroxysulfosuccinimide, Sulfo-NHS) solution, shaking reaction at room temperature for 10min, and centrifuging to collect the nano-Fe after carboxyl activation 3 o 4 Particles; the activated nano-Fe 3 o 4 The particles were added to 200 μL of TLL lipase solution with a concentration of 5 mg / mL, shaken at room temperature for 2 hours, magnetically separated, washed repeatedly with distilled water, removed uncoupled free lipase, and collected nano-Fe 3 o 4 G...

Embodiment 2

[0032] Example 2 Preparation of magnetically responsive lipase A with 1,3 position specificity

[0033] Specifically refers to the use of 10nm nano-Fe 3 o 4 Particle-modified Pseudomonas fluorescens lipase (Lipase from Pseudomonas fluorescens , PLL) preparation: Take 1 mL of nano-Fe with carboxyl functional groups on the surface 3 o 4 solution (10nm, iron content 155mg / L), add 10μL EDC solution with a concentration of 10mmol / L and Sulfo-NHS solution with a concentration of 25mmol / L, shake and react at room temperature for 10min, and centrifuge to collect nano-Fe after carboxyl activation 3 o 4 Particles; the activated nano-Fe 3 o 4 The particles were added to 200 μL of PFL lipase solution with a concentration of 1 mg / mL, shaken at room temperature for 2 hours, magnetically separated, washed repeatedly with distilled water, removed uncoupled free lipase, and collected nano-Fe 3 o 4 Granule-lipase complex.

Embodiment 3

[0034] Example 3 Preparation of magnetically responsive lipase A with 1,3 position specificity

[0035] Specifically refers to the use of 15nm nano-Fe 3 o 4 Particle-modified Thermomyces lanuginosus lipase (Lipase from Thermomyces lanuginosus , TLL) preparation: Take 1mL nano-Fe with carboxyl functional groups on the surface 3 o 4 Solution (15nm, iron content 100mg / L), add 10μL of EDC solution with a concentration of 10mmol / L and 20μL of Sulfo-NHS solution with a concentration of 25mmol / L, shake and react at room temperature for 30min, centrifuge to collect carboxyl-activated Nano Fe 3 o 4 Particles; the activated nano-Fe 3 o 4 Particles were added to 1 mL of 5 mg / mL Thermomyces lanuginosus In the lipase solution, shake and react at room temperature for 2 hours, magnetically separate, wash repeatedly with distilled water, remove uncoupled free lipase, and collect nano-Fe 3 o 4 Granule-lipase complex.

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Abstract

The invention relates to a preparation method of magnetic response composite lipase, and preparation of biodiesel by using the magnetic response composite lipase, and belongs to the technical field of preparation of biodiesel by using immobilized enzyme. According to the preparation method, magnetic nano Fe3O4 is adopted to modify lipase A with 1,3-position specificity, an immobilization technique is adopted to prepare the lipase A with the 1,3-position specificity and lipase B without position specificity into a composite enzyme double-enzyme system, as a method of composite lipase concerted catalysis is adopted, the substrate specificity of single lipase is overcome, the transesterification efficiency in preparing biodiesel by using an enzyme method is improved, recycling and repeated use of enzyme are achieved, and the enzyme activity loss in the reaction process is reduced.

Description

technical field [0001] The invention relates to the technical field of biodiesel preparation by enzymatic method, in particular to the preparation of an immobilized composite double enzyme system with magnetic responsiveness by using two lipases with different catalytic properties and its application in biodiesel synthesis. Background technique [0002] Compared with traditional petrochemical diesel, biodiesel has attracted extensive attention because of its renewable, biodegradable, and environmentally friendly properties. Biodiesel is mainly a long-chain fatty acid ester (fatty acid methyl or ethyl ester) prepared from animal and vegetable oils through transesterification. At present, the preparation methods of biodiesel mainly include chemical method and biological enzymatic method. Chemical methods, mainly alkali-catalyzed methods, have relatively mature production processes and are commonly used methods for preparing biodiesel; however, chemical methods have disadvanta...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/10C12N11/04C12P7/64
CPCY02E50/10
Inventor 李静李文英于伟泳
Owner TAIYUAN UNIV OF TECH
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