CN106148439A - A kind of method that power ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy prepares diglyceride - Google Patents

Abstract

The invention discloses a method for preparing diglyceride by catalyzing lard glycerol hydrolysis with ultrasonic pretreatment enzymatic method. In the bottle; 2. Add immobilized lipase Lipozyme RMIM to the mixture of lard and glycerin, place it in a constant temperature water bath for ultrasonic pretreatment, and then transfer it to a water bath constant temperature oscillator for reaction; 3. Remove fat from the above reaction mixture by filtration Enzymes, that is, the preparation of oils and fats containing diglycerides. The invention makes good use of lard, which is a cheap animal product processing by-product, improves the value of the by-product, is fast, efficient, has no solvent residue, and the product is easy to separate from the enzyme, providing a fast and efficient preparation of healthy A method for functionalizing diglycerides.

Description

A power ultrasonic pretreatment enzymatic method to catalyze glycerol hydrolysis of lard to prepare diacylglycerol method

technical field

The invention belongs to the technical field of lard deep processing, and relates to a method for preparing diglyceride, in particular to a method for preparing diglyceride by power ultrasonic pretreatment enzyme catalyzing lard glycerol hydrolysis in a solvent-free system.

Background technique

Lard, commonly known as lard, is a high-level saturated fatty acid glyceride. my country has very rich lard resources. With the change of people's dietary concepts and the improvement of living standards, the consumption of lard is decreasing due to the high cholesterol content of lard, and the phenomenon of "surplus" lard appears. It is of great significance to develop nutritious and healthy lard products.

Diacylglycerol (DAG) is a product of esterification of two hydroxyl groups in glycerol with fatty acids, and is a trace component present in various natural edible oils. After review by the U.S. Food and Drug Administration, DAG oil is recognized as a safe food ingredient. Experiments in animal and human models have shown that although diacylglycerols have similar digestibility and energy value to triglycerides, they can reduce postprandial blood lipid levels, inhibit weight gain and reduce visceral fat accumulation, so diacylglycerol is a Healthy functional oils. In recent years, diglycerides have been widely recognized as a functional oil for the prevention of obesity and some lifestyle-related diseases. And because the molecular structure of DAG contains a hydrophilic group -OH, it can show interfacial properties and surface activity, making it more suitable as an emulsifier and surfactant. At the same time, DAG also has many advantages such as safety, nutrition, high human compatibility, and good processing adaptability. Therefore, DAG can be used as a multifunctional additive, and has a wide range of applications in food, medicine, cosmetics, and chemical industries.

At present, the methods for synthesizing diglycerides mainly include chemical synthesis and biological enzyme catalysis. Chemical synthesis is a traditional method for preparing diglycerides. It is widely used in industrial production, but due to harsh reaction conditions, most of them require high temperature. Treatment, high cost, many by-products, poor sensory quality of the product, and easy pollution to the environment have limited the application of chemical synthesis methods. The bio-enzyme catalysis method has become the most researched method for preparing diglycerides due to the advantages of mild reaction conditions, low energy consumption, less environmental pollution, less side reactions, and better color and flavor of the resulting product, but the bio-enzyme method The reaction rate is slow, and it generally takes 10 h or more to complete the reaction.

The impact of ultrasound on enzyme-catalyzed reactions mainly affects mass transfer and molecular diffusion in the system through three effects: mechanical mass transfer, heating and cavitation. Appropriate ultrasonic assistance can not only reduce the mass transfer resistance, increase the contact probability between the enzyme and the substrate, but also change the conformation of the protein (enzyme) to make it more reasonable, thereby increasing the reaction rate and yield. Ultrasonic technology also has the advantages of high efficiency, easy acquisition, low price, and no pollution to the environment. Therefore, the application of ultrasonic technology in enzyme-catalyzed reactions has a good application prospect. In recent years, studies have shown that ultrasound-assisted enzymatic reactions can increase the rate and yield of the reaction, but there are few reports on the catalytic synthesis of diacylglycerol by ultrasonic pretreatment enzymatically.

Contents of the invention

The purpose of the present invention is to solve the problems existing in the prior art, and provide a method for preparing diglycerides by power ultrasonic pretreatment enzymatically catalyzing glycerol hydrolysis of lard. The method makes good use of lard, which is a cheap animal product processing by-product, increases the value of the by-product, is fast and efficient, has no solvent residue, and is easy to separate the product from the enzyme.

The purpose of the present invention is achieved through the following technical solutions:

A method for preparing diglyceride by catalyzing glycerololysis of lard with power ultrasonic pretreatment enzymatic method, using lard and glycerin as raw materials, adopting power ultrasonic pretreatment enzymatic method to catalyze glycerol hydrolysis of lard to prepare diglyceride, the specific steps are as follows:

1. Take a certain amount of lard and put it in a water bath to completely melt it, then mix it with glycerin in a conical flask at a molar ratio of 1:0.5-2;

2. Add 1-5% lard weight immobilized lipase Lipozyme RMIM to the mixture of lard and glycerol, place the Erlenmeyer flask in a constant temperature water bath, insert the ultrasonic probe at 4-6 mm below the reaction solution, and place it at 40 Ultrasonic treatment at an ultrasonic temperature of ~60 °C and an ultrasonic power of 100 ~ 300 W for 5 ~ 20 min, then transferred to a constant temperature oscillator in a water bath, and reacted under the conditions of 40 ~ 60 °C and a stirring speed of 100 ~ 200 r/min 2 ~6 hours;

Three, filter and remove lipase from above-mentioned reaction mixture, promptly make the fat containing diglyceride;

4. The composition of the obtained reaction mixture is qualitatively analyzed by thin layer chromatography, and the content of each component of the obtained reaction mixture is quantitatively calculated by normal phase high performance liquid chromatography, and the content of each component is calculated by the area normalization method.

During the propagation of ultrasonic waves in the medium, due to the interaction between ultrasonic waves and the medium, ultrasonic energy is released into the medium, thereby producing ultrasonic effects. According to different uses, the application of ultrasonic effects is mainly divided into detection ultrasonic technology (high frequency, low energy) and power ultrasonic technology (low frequency, high energy). The present invention compares power ultrasonic assisted lipase (the whole reaction process is carried out under ultrasonic assistance), power ultrasonic pretreatment lipase, power ultrasonic pretreatment substrate (lard and glycerin) three ultrasonic assisted ways to enzymatic glycerol hydrolysis The effect of preparing lard diglycerides, the study found that power ultrasonic pretreatment of lipase has the best effect, which may be because power ultrasonic can disperse the energy in the reaction system and promote the emulsification of the reaction system, but long-term power ultrasonic treatment will The structure of the enzyme is destroyed, thereby reducing the degree of glycerol hydrolysis.

The present invention provides a method for quickly and efficiently preparing healthy functional diglycerides. Compared with the prior art, it has the following advantages:

(1) The present invention uses lard and glycerin as raw materials, and uses the specific immobilized lipase Lipozyme RMIM as a catalyst, which not only makes good use of lard, a cheap by-product of animal product processing, but also improves the value of by-products , and there is no solvent residue in the reaction, and the product is easy to separate from the enzyme;

(2) The present invention combines ultrasonic waves and enzyme-catalyzed reactions to prepare diglycerides, comprehensively utilizes many advantages of ultrasonic technology and enzyme-catalyzed reactions, not only has mild reaction conditions, low energy consumption, less environmental pollution, high specificity, It is green and environmentally friendly, and the reaction rate and yield are improved, and the reaction time is greatly shortened. Only a short time is needed, and diglyceride with a higher yield can be produced.

Description of drawings

Fig. 1 is a process flow diagram for the preparation of diglycerides by power ultrasonic pretreatment enzymatically catalyzing glycerololysis of lard.

detailed description

The technical solution of the present invention will be further described below in conjunction with the examples, but it is not limited thereto. Any modification or equivalent replacement of the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention should be covered by the present invention within the scope of protection.

Example 1:

1. Take a certain amount of lard and put it in a water bath to completely melt it, then mix it with glycerin in a conical flask according to 1:1 (molar ratio);

2. Add 4% lard weight of immobilized lipase Lipozyme RMIM to the mixture of lard and glycerin, place the Erlenmeyer flask in a constant temperature water bath, insert the ultrasonic probe about 5 mm below the reaction solution, and ultrasonically Under the conditions of temperature and ultrasonic power of 200 W, ultrasonic treatment was performed at a certain stirring speed for 5 min, transferred to a constant temperature oscillator in a water bath, and reacted at 50 °C and a stirring speed of 150 r/min for 4 h;

Three, filter and remove lipase from above-mentioned reaction mixture, promptly make the fat containing diglyceride;

Four, utilize thin layer chromatography to carry out qualitative analysis to the composition of gained reaction mixture, utilize normal phase high performance liquid chromatography to carry out quantitative calculation to the content of each component of gained reaction mixture, detect through high performance liquid chromatography, the DAG content in the product is 31.18 %.

Example 2:

1. Take a certain amount of lard and put it in a water bath to completely melt it, then mix it with glycerin in a conical flask according to 1:1 (molar ratio);

2. Add 3% lard weight of immobilized lipase Lipozyme RMIM to the mixture of lard and glycerin, place the Erlenmeyer flask in a constant temperature water bath, insert the ultrasonic probe about 5 mm below the reaction solution, and ultrasonically Under the conditions of temperature and ultrasonic power of 200 W, ultrasonic treatment was performed at a certain stirring speed for 5 min, transferred to a constant temperature oscillator in a water bath, and reacted at 50 °C and a stirring speed of 150 r/min for 4 h;

Three, filter and remove lipase from above-mentioned reaction mixture, promptly make the fat containing diglyceride;

Four, utilize thin layer chromatography to carry out qualitative analysis to the composition of gained reaction mixture, utilize normal phase high performance liquid chromatography to carry out quantitative calculation to the content of each component of gained reaction mixture, detect through high performance liquid chromatography, the DAG content in the product is 29.50 %.

Example 3:

1. Take a certain amount of lard and put it in a water bath to completely melt it, then mix it with glycerin in a conical flask according to 1:1 (molar ratio);

2. Add 3% lard weight of immobilized lipase Lipozyme RMIM to the mixture of lard and glycerin, place the Erlenmeyer flask in a constant temperature water bath, insert the ultrasonic probe about 5 mm below the reaction solution, and ultrasonically Temperature and ultrasonic power of 250 W, ultrasonic treatment at a certain stirring speed for 5 minutes, transferred to a constant temperature oscillator in a water bath, and reacting for 4 hours at 50 °C and a stirring speed of 150 r/min;

Three, filter and remove lipase from above-mentioned reaction mixture, promptly make the fat containing diglyceride;

4. Utilize thin-layer chromatography to qualitatively analyze the composition of the resulting reaction mixture, and utilize normal phase high-performance liquid chromatography to quantitatively calculate the contents of each component of the resulting reaction mixture. After detection by high-performance liquid chromatography, the DAG content in the product is 37.87 %.

Example 4:

1. Take a certain amount of lard and put it in a water bath to completely melt it, then mix it with glycerin in a conical flask according to 1:1 (molar ratio);

2. Add 3% lard weight of immobilized lipase Lipozyme RMIM to the mixture of lard and glycerol, place the Erlenmeyer flask in a constant temperature water bath, insert the ultrasonic probe about 5 mm below the reaction solution, and ultrasonically Temperature and ultrasonic power of 250 W, ultrasonic treatment at a certain stirring speed for 5 minutes, transferred to a constant temperature oscillator in a water bath, and reacting for 4 hours at 50 °C and a stirring speed of 150 r/min;

Three, filter and remove lipase from above-mentioned reaction mixture, promptly make the fat containing diglyceride;

Four, utilize thin layer chromatography to carry out qualitative analysis to the composition of gained reaction mixture, utilize normal phase high performance liquid chromatography to carry out quantitative calculation to the content of each component of gained reaction mixture, detect through high performance liquid chromatography, the DAG content in the product is 46.43 %.

Claims (10)

1. the method that a ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy prepares diglyceride, it is characterised in that described method walks Rapid as follows:

One, take after a certain amount of Adeps Sus domestica is placed in and is completely melt in water-bath, with glycerol according to 1:0.5～2 mixed in molar ratio in In conical flask；

Two, in the mixture of Adeps Sus domestica and glycerol, Adeps Sus domestica weight 1～the immobilized-lipase Lipozyme RMIM of 5% is added, will Conical flask is placed in water bath with thermostatic control, under ultrasonic probe insertion reaction liquid, at the ultrasonic temperature of 40～60 DEG C, 100～300 W Under ultrasonic power, supersound process 5～20 min, is then transferred in water-bath constant temperature oscillator, 40～60 DEG C, mixing speed 2～6 h are reacted under conditions of 100～200 r/min；

Three, from the reactions above mixture is filtered to remove lipase, i.e. prepares the oils and fats containing diglyceride.

The method that ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy the most according to claim 1 prepares diglyceride, it is special Levy and be that described Adeps Sus domestica is 1:1 with the mol ratio of glycerol.

The method that ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy the most according to claim 1 prepares diglyceride, it is special Levy and be that the addition of described immobilized-lipase Lipozyme RMIM is Adeps Sus domestica weight 4%.

The method that ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy the most according to claim 1 prepares diglyceride, it is special Levy and be that the addition of described immobilized-lipase Lipozyme RMIM is Adeps Sus domestica weight 3%.

The method that ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy the most according to claim 1 prepares diglyceride, it is special Levy and be under described ultrasonic probe insertion reaction liquid at 4～6 mm.

The method that ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy the most according to claim 1 prepares diglyceride, it is special Levy and be that described ultrasonic temperature is 45 DEG C.

The method that ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy the most according to claim 1 prepares diglyceride, it is special Levy and be that described ultrasonic temperature is 55 DEG C.

The method that ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy the most according to claim 1 prepares diglyceride, it is special Levy and be that described ultrasonic power is 250 W.

The method that ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy the most according to claim 1 prepares diglyceride, it is special Levy and be that described ultrasonic power is 200 W.

The method that ultrasonic pretreatment enzyme law catalysis Adeps Sus domestica glycerol rhizolomy the most according to claim 1 prepares diglyceride, it is special Levy and be that described ultrasonic time is 5min.