CN117987202A

CN117987202A - Method for extracting tea seed oil by enzyme method

Info

Publication number: CN117987202A
Application number: CN202311818228.7A
Authority: CN
Inventors: 刘仰玺; 龚震超; 龚欣悦; 龚振赵
Original assignee: Fujian Yike Tea Agricultural Technology Co ltd
Current assignee: Fujian Yike Tea Agricultural Technology Co ltd
Priority date: 2023-12-27
Filing date: 2023-12-27
Publication date: 2024-05-07

Abstract

The invention relates to the technical field of tea seed oil processing, in particular to a method for extracting tea seed oil by an enzymatic method. The method for extracting tea seed oil by the enzyme method comprises the following steps: s1: selecting tea seeds; s2: cleaning tea seeds; s3: grinding tea seeds; s4, enzymolysis of tea seeds S5: inactivating treatment; s6: centrifuging; s7: obtaining tea seed oil. By adding the complex enzyme preparation, the components such as protein, starch, cellulose and the like in the tea seeds are thoroughly decomposed, the fat hardness in the tea seeds is reduced, and the fat hydrolysis and the oleic acid formation are promoted, so that the extraction efficiency of the tea seed oil is improved, the quality of the tea seed oil is improved, the production cost is reduced, the economic benefit is improved, the production efficiency and the quality of the tea seed oil are improved, and the development of the tea seed oil industry is promoted.

Description

Method for extracting tea seed oil by enzyme method

Technical Field

The invention relates to the technical field of tea seed oil processing, in particular to a method for extracting tea seed oil by an enzymatic method.

Background

Tea seed oil is a natural oil with rich nutrition, and contains rich monounsaturated fatty acid-oleic acid and polyunsaturated fatty acid-linoleic acid and linolenic acid. These fatty acids play an important role in human health. Oleic acid can reduce cholesterol content in blood, and prevent cardiovascular diseases; linoleic acid and linolenic acid are helpful for lowering blood pressure and blood sugar, and have good health promotion effects on patients with diabetes and hypertension.

In addition, the tea seed oil also contains rich microelements such as vitamin E, vitamin A, zinc, calcium and the like, and the nutritional ingredients have good health care effects on the immune system, nervous system and skeletal system of a human body. Wherein, the vitamin E has antioxidant effect, and can delay aging and maintain skin elasticity and luster; vitamin a helps to maintain vision and the health of the immune system.

The prior patent application publication CN107653055B provides a method for extracting tea seed oil by using an ultrasonic-assisted deionized water enzymatic method. Firstly, extracting tea saponin in tea seeds by using different solvents assisted by ultrasound, further adopting cellulase and protease to carry out two-step deionized water enzymolysis on tea seed residues after ultrasonic treatment, and adding calcium ions (nCa2+: nsaponin=1:2) in the enzymolysis process, and carrying out enzymolysis reaction for 5 hours; according to the invention, tea saponin is removed by combining ultrasonic extraction with calcium salt precipitation, so that the generation of emulsion in the tea seed deionized water decomposition process is reduced or avoided, the traditional subsequent demulsification link is replaced, and the clear oil rate is improved. Therefore, the extraction method has the advantages of simple equipment, high efficiency, greenness, less damage of nutrient components and the like.

The patent discloses a method for improving the clear oil rate of tea seed oil. But the problems in terms of clean oil yield are not solved. Meanwhile, the method for removing tea saponin by combining ultrasonic extraction and calcium salt precipitation cannot ensure 100% tea saponin removal, and the extraction rate is relatively low. The equipment cost and the operation cost are higher, the process flow is complex, and the large-scale production is not facilitated.

Disclosure of Invention

(One) solving the technical problems

Aiming at the defects of the prior art, the invention provides a method for extracting tea seed oil by an enzyme method, which solves the problems of low yield of clear oil, complex process flow and high cost.

(II) technical scheme

In order to achieve the above purpose, the application provides a method for extracting tea seed oil by an enzymatic method, which adopts the following technical scheme:

A method for extracting tea seed oil by enzyme method comprises fermenting with complex enzyme preparation. The preparation of the complex enzyme preparation requires the following materials: 100 parts by mass of phosphate buffer solution, 0.2-1 part by mass of polyethylene glycol, 1-3 parts by mass of protease, 6-9 parts by mass of amylase, 2-5 parts by mass of cellulase, 1-2 parts by mass of phospholipase and 2-4 parts by mass of lipase; and is prepared by the following method:

a1: 100ml of phosphate buffer pH7.2 was added to the beaker.

A2: polyethylene glycol is added into the phosphate buffer solution, and the mixture is stirred and mixed uniformly.

A3: adding protease, amylase, cellulase, phospholipase and lipase into the prepared solution while stirring in sequence, and gently stirring uniformly.

A4: the compound enzyme preparation is obtained, sealed and stored in a refrigeration way in a dark place.

Illustratively, the process is performed in a sterile operating station.

Preferably, the mass ratio of the protease, the amylase, the cellulase, the phospholipase and the lipase is 1-3:6-9:2-5:1-2:2-4.

Preferably, the mass ratio of the protease to the amylase to the cellulase to the phospholipase to the lipase is 2:7:4:2:3.

The specific explanation is that during the process of extracting tea seed oil, protease can decompose protein in cell walls and cell membranes, so that grease is easier to release from the cells; the amylase breaks down starch and saccharide substances in cells, and reduces the viscosity of the cells, thereby being beneficial to the flow and extraction of grease; the cellulase breaks the cell wall, so that the grease is easier to release from the cells; the phospholipase can decompose phospholipid in cell membrane to increase permeability of cell membrane, thereby facilitating release and extraction of oil; the lipase can help to decompose the fat in the cells, so that the fat is easier to release from the cells, and meanwhile, the lipase can also play a role in degreasing, remove impurities in the tea seed oil and improve the quality of the fat.

Preferably, the phosphate buffer is a buffer.

The phosphate buffer solution has strong buffering capacity, can effectively maintain the stable pH value of a reaction system, and can prevent the loss of enzyme activity caused by pH change. Meanwhile, the method can adapt to the optimal pH values of different enzymes, is favorable for the synergistic effect of various enzymes in the compound enzyme preparation, can ensure the stability of the enzymes, and can not inhibit the activity of the enzymes. In addition, the complex has better compatibility with various enzymes, forms stable complex with the enzymes, and is beneficial to improving the stability and activity of the complex enzyme preparation.

Preferably, the phosphate buffer has a pH of 7.2.

The protease, amylase, cellulase, phospholipase and lipase have good activity in a weak alkaline environment, and promote enzymolysis in the reaction process.

Preferably, the polyethylene glycol acts as a stabilizer.

The polyethylene glycol can form a layer of protective deionized water film on the surface of the enzyme to wrap the enzyme molecules, reduce the influence of the outside on the enzyme and maintain the conformation and activity of the enzyme; simultaneously, the surface tension around the enzyme is reduced, the collision and loss are reduced, and the natural state of the enzyme is maintained.

Preferably, the mass fraction of the polyethylene glycol is 2% -3% of the total amount of the enzyme.

In particular, it is noted that low concentrations of polyethylene glycol can increase the stability of the enzyme while maintaining the enzyme activity.

The method specifically needs to be explained, the enzyme is prevented from being damaged due to the fact that stirring is gently and evenly conducted, and the enzyme is prevented from being damaged due to the fact that stirring is conducted forcefully, so that the effect of the enzyme is lost, the test result is affected, and the expected test purpose cannot be achieved.

A method for extracting tea seed oil by enzyme method comprises the following raw materials: 100 parts by mass of tea seeds, 50-100 parts by mass of 20% n-propanol solution, 200-300 parts by mass of deionized water and 50-100 parts by mass of complex enzyme preparation.

The enzymatic extraction method of tea seed oil comprises the following steps:

s1: fresh, mildew-free, odorless and full tea seeds are selected.

S2: and (3) carrying out ultrasonic cleaning on the selected tea seeds, and placing the tea seeds in an oven to lay flat and set the drying surface at 35 ℃.

S3: the tea seeds were crushed using a crusher and then ground for 20min.

S4: and (3) placing the tea seeds subjected to pulp grinding into a fermentation tank, adding n-butanol solution, deionized water and a compound enzyme preparation, uniformly mixing, and starting fermentation.

S5: and (5) inactivating the mixed solution at 90 ℃ for 10min after fermentation.

S6: the inactivated mixture was placed in a centrifuge at 12000rpm at 4℃for 20min.

S7: collecting supernatant, and steaming at 50deg.C for 30min to obtain tea seed oil.

Preferably, the fermentation is preceded by pulp milling.

The specific explanation is that after the pulp is ground, the surface area of the tea seeds contacted with the compound enzyme preparation is increased, so that the mixed fermentation is facilitated, and the reaction is more sufficient.

Preferably, the complex enzyme preparation is prepared from phosphate buffer, polyethylene glycol, protease, amylase, cellulase, phospholipase and lipase.

The complex enzyme preparation can degrade protein and phospholipid substances in tea seeds, decompose starch and cellulose in the tea seeds, and reduce the combination of protein and grease, so that the quality and stability of the grease are improved, and the extraction of the grease is promoted.

(III) beneficial effects

The invention provides a method for extracting tea seed oil by an enzymatic method, which has the following beneficial effects:

(1) According to the method for extracting the tea seed oil by the enzyme method, through the cooperation of protease, amylase, cellulase, phospholipase and lipase, the cell wall and the cell membrane are damaged under the synergistic effect of a plurality of enzymes, so that the grease is easier to release from the cells, the extraction of the grease is facilitated, the extraction efficiency is improved, the quality of the tea seed oil is improved, and more tea seed oil products with high quality are obtained.

(2) According to the method for extracting the tea seed oil by the enzyme method, the compound enzyme preparation is added to remove impurities in the tea seed oil, so that the deionized water decomposition and oleic acid formation of the oil are promoted, the acid value and the peroxide value are reduced, the oil yield and the oil purity of the tea seed oil are improved, the quality and the stability of the oil are further improved, the nutritive value of the tea seed oil is improved, and the effect of improving the quality of the tea seed oil is achieved.

(3) According to the method for extracting the tea seed oil by the enzyme method, based on the prior art, the enzyme is used for preparing and mixing, enzymolysis is carried out layer by layer according to plant attributes of the tea seeds, and the structure of the tea seeds is gradually decomposed, so that the effect of extracting the tea seed oil is better achieved, the efficiency of extracting the tea seed oil is improved, the high-quality tea seed oil is further extracted, and the further development of the product industry is promoted.

Detailed Description

All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Preparation examples 1 to 3

A complex enzyme preparation, the preparation raw materials and the corresponding components are shown in the following table 1, and is prepared by the following preparation method:

a1: 100ml of phosphate buffer pH7.2 was added to the beaker.

Illustratively, the process is performed in a sterile operating station.

Table 1: the components and the components of the substances in preparation examples 1 to 3

Example 1

In particular to a method for extracting tea seed oil by an enzyme method, which comprises the following raw materials: 100 parts by weight of tea seeds, 50 parts by weight of 20% n-propanol solution, 200 parts by weight of deionized water and 50 parts by weight of a complex enzyme preparation. The method for extracting tea seed oil by the enzyme method comprises the following steps:

s1: fresh, mildew-free, odorless and full tea seeds are selected.

S3: the tea seeds were crushed using a crusher and then ground for 20min.

S4: and (3) placing the tea seeds subjected to pulp grinding into a fermentation tank, adding n-butanol solution, deionized water and a compound enzyme preparation, uniformly mixing, and setting the fermentation time to be 2 hours and 50 ℃.

Example 2

In particular to a method for extracting tea seed oil by an enzyme method, which comprises the following raw materials: 100 parts by weight of tea seeds, 50 parts by weight of 20% n-propanol solution, 300 parts by weight of deionized water and 50 parts by weight of a complex enzyme preparation.

Preparation procedure reference example 1, differing from example 1 in the ratio of raw materials to feed solution being 1:4.

Example 3

In particular to a method for extracting tea seed oil by an enzyme method, which comprises the following raw materials: 100 parts by weight of tea seeds, 100 parts by weight of 20% n-propanol solution, 300 parts by weight of deionized water and 100 parts by weight of a compound enzyme preparation.

Preparation procedure reference example 1, differing from example 1 in the ratio of raw materials to feed solution being 1:5.

Example 4

In particular to a method for extracting tea seed oil by an enzyme method, which comprises the following raw materials: 100 parts by weight of tea seeds, 50 parts by weight of 20% n-propanol solution, 200 parts by weight of deionized water and 50 parts by weight of a complex enzyme preparation.

Preparation procedure reference example 1, differing from example 1 in that the fermentation parameters were set at 2.5h,50 ℃.

Example 5

Preparation procedure with reference to example 1, the difference from example 1 is that the ratio of raw materials to feed liquid is 1:4, and the fermentation parameters are set at 50℃for 2.5 h.

Example 6

Preparation procedure with reference to example 1, the difference from example 1 is that the ratio of raw materials to feed liquid is 1:5, and the fermentation parameters are set at 50℃for 2.5 h.

Example 7

Preparation procedure reference example 1, differing from example 1 in that the fermentation parameters were set at 3h,50 ℃.

Example 8

The preparation procedure is described in example 1, with the difference from example 1 that the ratio of raw materials to feed solution is 1:4 and the fermentation parameters are 3h at 50 ℃.

Example 9

Preparation procedure with reference to example 1, the difference from example 1 is that the ratio of raw materials to feed solution is 1:5, and the fermentation parameters are set at 3h,50 ℃.

Example 10

Preparation procedure reference example 1, differing from example 1 in that the fermentation parameters were set at 2h,55 ℃.

Example 11

Preparation procedure with reference to example 1, the difference from example 1 is that the ratio of raw materials to feed solution is 1:4, and the fermentation parameters are set at 2h,55 ℃.

Example 12

Preparation procedure with reference to example 1, the difference from example 1 is that the ratio of raw materials to feed solution is 1:5, and the fermentation parameters are set at 2h,55 ℃.

Example 13

Preparation procedure reference example 1, differing from example 1 in that the fermentation parameters were set at 2.5h,55 ℃.

Example 14

Preparation procedure referring to example 1, the difference from example 1 is that the ratio of raw materials to feed solution is 1:4, and the fermentation time is set to 2.5h at 55 ℃.

Example 15

Preparation procedure referring to example 1, the difference from example 1 is that the ratio of raw materials to feed solution is 1:5, and the fermentation time is set to 2.5h at 55 ℃.

Example 16

Preparation procedure reference example 1, differing from example 1 in that the fermentation parameters were set at 3h,55 ℃.

Example 17

Preparation procedure with reference to example 1, the difference from example 1 is that the ratio of raw material to feed solution is 1:4, and the fermentation time is set at 3h,55 ℃.

Example 18

Preparation procedure with reference to example 1, the difference from example 1 is that the ratio of raw material to feed solution is 1:5, and the fermentation time is set at 3h,55 ℃.

Comparative example 1

Comparative example 1, identical to example 1, differs in that: the complex enzyme preparation is replaced by protease.

Comparative example 2

Comparative example 1, identical to example 1, differs in that: the complex enzyme preparation is replaced by lipase.

Comparative example 3

Comparative example 1, identical to example 1, differs in that: after the enzyme reaction, no inactivation treatment was performed.

The various parameters of the enzymatic extraction of tea seed oil of examples 1-18 and comparative examples 1-3 were recorded separately and gave the following results, see in particular tables 2, 3:

table 2: examples 1-18 and comparative examples 1-3 each test parameters were compared to each other

Table 3: examples 1-18 and comparative examples 1-3 each test parameters were compared to each other

As can be seen from comparison of the test data of the examples and the comparative examples in tables 2 and 3, the tea seed oil prepared by the technical scheme of the application has the characteristics of excellent quality and high nutritive value. The method for extracting tea seed oil improves extraction efficiency and quality of tea seed oil.

As can be seen from comparison of the test data of examples and comparative examples 1 and 2 in tables 2 and 3, the enzymes in the complex enzyme preparation act synergistically to destroy cell walls and cell membranes, so that the grease is more easily released from the cells. Meanwhile, impurities in the tea seed oil can be removed, and the acid value and the peroxide value are reduced, so that the quality and stability of the grease are improved.

Compared with the traditional extraction method, the complex enzyme preparation can remarkably improve the extraction efficiency. The protease can decompose proteins in cell walls and cell membranes, the amylase can decompose starch and saccharide substances in cells, the cellulase can destroy cell walls, the phospholipase can decompose phospholipids, phospholipid impurities in the grease are removed, and the acid value of the grease is reduced. The lipase can decompose fatty glyceride, remove free fatty acid and other impurities, and improve the stability and oxidation stability of the grease. The enzymes in the compound enzyme preparation act synergistically to decompose macromolecular substances, prevent the macromolecular substances from depositing in grease, and further improve the quality of the grease.

Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. A method for extracting tea seed oil by an enzymatic method is characterized in that: the method comprises the following steps:

s1: selecting fresh, mildew-free, odorless and full tea seeds;

s2: ultrasonically cleaning selected tea seeds, and paving the tea seeds on the surface of a baking oven;

S3: crushing tea seeds by using a crusher, and then pulping;

s4: placing the tea seeds after pulping into a fermentation tank, adding an organic solvent, deionized water and a compound enzyme preparation, uniformly mixing, and starting fermentation;

S5: inactivating the fermented mixed solution;

S6: placing the inactivated mixed solution in a centrifuge, and centrifuging and layering;

s7: collecting supernatant to obtain tea seed oil.

2. The method for extracting tea seed oil by an enzymatic method according to claim 1, wherein the method comprises the following steps: the organic solvent is 20% n-propanol solution.

3. The method for extracting tea seed oil by an enzymatic method according to claim 1, wherein the method comprises the following steps: the complex enzyme preparation is prepared from phosphate buffer solution, polyethylene glycol, protease, amylase, cellulase, phospholipase and lipase.

4. The method for extracting tea seed oil by an enzymatic method according to claim 1, wherein the method comprises the following steps: and adding the organic solvent and deionized water, and then adding the complex enzyme preparation.

5. The method for extracting tea seed oil by an enzymatic method according to claim 1, wherein the method comprises the following steps: the mass ratio of the protease to the amylase to the cellulase to the phospholipase to the lipase is 1-3:6-9:2-5:1-2:2-4.

6. A method for extracting tea seed oil by an enzymatic method according to claim 3, wherein: the complex enzyme preparation consists of 100 parts by mass of phosphate buffer solution, 0.2-1 part by mass of polyethylene glycol, 1-3 parts by mass of protease, 6-9 parts by mass of amylase, 2-5 parts by mass of cellulase, 1-2 parts by mass of phospholipase and 2-4 parts by mass of lipase; the preparation method comprises the following steps:

a1: 100ml of phosphate buffer with pH7.2 was added to the beaker;

a2: adding polyethylene glycol into phosphate buffer solution, and stirring and mixing uniformly;

A3: adding protease, amylase, cellulase, phospholipase and lipase into the prepared solution while stirring in sequence, and gently stirring uniformly;

The operations were described as being performed in a sterile console.

7. The method for extracting tea seed oil by an enzymatic method according to claim 1, wherein the method comprises the following steps: the fermentation time is 2-3 h and 50-55 ℃; the centrifugation parameters were 12000rpm,20min,4 ℃.