CN111728205B - Method for increasing content of free amino acids in enzyme, enzyme prepared by using method and application of enzyme - Google Patents
Method for increasing content of free amino acids in enzyme, enzyme prepared by using method and application of enzyme Download PDFInfo
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- CN111728205B CN111728205B CN202010664817.4A CN202010664817A CN111728205B CN 111728205 B CN111728205 B CN 111728205B CN 202010664817 A CN202010664817 A CN 202010664817A CN 111728205 B CN111728205 B CN 111728205B
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/125—Casei
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/531—Lactis
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Preparation Of Fruits And Vegetables (AREA)
Abstract
The invention provides a method for improving the content of free amino acids in ferment, the ferment prepared by the method and application of the ferment. The preparation method is simple, the saccharomycetes, the lactobacillus casei and the lactobacillus bifidus are added in the fermentation process, the sugar is used as a carbon source, the fruit is used as a fermentation substrate, the three strains are mutually matched, the anaerobic environment can be accelerated to form, the growth of anaerobic lactobacillus is promoted, the generation and metabolism of important amino acid in the fermentation process are promoted, the yield and the release amount of free amino acid in the fermentation raw material are increased, the nutritional ingredients in the fruit are fully decomposed, and the content of the free amino acid in the ferment is improved.
Description
Technical Field
The invention belongs to the technical field of microbial fermentation, and relates to a fermentation method of fruit ferment, in particular to a method for improving the content of free amino acid in the ferment, the ferment prepared by the method and application of the ferment.
Background
The ferment is generally a high molecular substance with a biocatalytic function, and is generally prepared by fermenting various plant raw materials such as fruits, vegetables and the like under the action of beneficial microorganisms. The ferment is rich in nutrition, and is rich in vitamins, amino acids, enzymes, minerals and microelements. After entering human body, the enzyme can be quickly absorbed and utilized by human body, and participates in various physiological and biochemical reactions of human body life activities, and is an indispensable substance for human body. The ferment is also an important medium of human metabolism, and can regulate human body function, thereby achieving the effects of strengthening physique and improving human immunity. Researches show that the enzyme has good antioxidant performance, can remove redundant free radicals in human bodies, relieves the damage of the free radicals to the bodies, and has good health care function.
The grape ferment is a common ferment, and the compounds such as polyphenol contained in the grape ferment have the function of reducing blood fat, and the grape ferment also has the functions of antioxidation, can prevent aging and keep the normal metabolism of the body, so the grape ferment can play a better role in the aspects of beauty treatment, face nursing, food health care and the like,
CN108077826A discloses a process for producing grape ferment, which utilizes the existing grape wine production equipment, takes fresh grapes and wine grapes as raw materials, according to the characteristic of high sugar content of the grapes, by optimizing the fermentation process, adding a fermentation regulator consisting of inulin, lactose and natamycin, inoculating lactobacillus plantarum HX1, and developing a new process for producing the grape ferment. However, in the process of preparing ferments, the microbial composition of the surface of vegetables and fruits as the fermentation raw material is very diverse due to the differences of the fermentation raw material, the fermentation conditions and the beneficial microbes, and thus, the components and functions of the ferments obtained are also greatly different.
Beneficial microorganisms play a very important role in the fermentation process, and are the key to determining the components and functions of the ferment product. At present, although the grape ferment prepared by commercial ferment bacteria has better antioxidant activity, the key nutrient contents such as free amino acid contained in the grape ferment are lower, and the free amino acid refers to a single amino acid molecule existing in a free state and can be directly absorbed and utilized.
Therefore, screening beneficial microorganisms and combinations thereof with good fermentation performance has important significance in improving the content of free amino acids in the ferment, improving the nutritional value of the ferment and improving the application prospect.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a method for improving the content of free amino acids in ferment, the ferment prepared by using the method and application of the ferment.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the present invention provides a method for increasing the content of free amino acids in ferments, comprising the steps of:
mixing the crushed fruits, sugar and water in a fermentation container, adding a microbial agent, and performing anaerobic fermentation to obtain the enzyme; the microbial agent comprises yeast, lactobacillus casei and lactobacillus bifidus.
In the invention, the microbial agent containing saccharomycetes, lactobacillus casei and lactobacillus bifidus is added in the anaerobic fermentation process, and the obtained ferment has higher content of free amino acid. When the yeast, the lactobacillus casei and the lactobacillus bifidus are fermented, sugar is used as a carbon source, fruits are used as fermentation substrates, different strains are matched with each other in a long-time fermentation process, the yeast can accelerate the formation of an anaerobic environment and promote the growth of anaerobic lactic acid bacteria, and the lactobacillus casei and the lactobacillus bifidus fully decompose nutritional ingredients in the fruits through the metabolism of some important amino acids and improve the content of free amino acids in the ferment.
During fermentation, if the three probiotics are not added, one of the probiotics is randomly reduced or replaced, for example, the lactobacillus casei is not added or the lactobacillus casei is replaced by other types of probiotics, the content of amino acid in the obtained ferment is obviously reduced, which indicates that the yeast, the lactobacillus casei and the lactobacillus bifidus have certain synergistic effect besides the respective functions, the reaction efficiency in the fermentation process is increased through cell factors or cell metabolites generated among cells, the production and release of free amino acid are increased, and the content of the free amino acid in the obtained ferment is increased.
In a preferred embodiment of the present invention, the amount of the microbial inoculum is 0.8 to 1.2% of the total volume of the fruit, sugar and water, and may be, for example, 0.8%, 0.85%, 0.9%, 0.95%, 0.98%, 1%, 1.05%, 1.08%, 1.1%, 1.15%, 1.2%, or the like.
Preferably, the microorganisms exist in a liquid microbial inoculum, for example, yeasts exist in a yeast microbial inoculum, the yeast microbial inoculum can be purchased or prepared from a conventional channel, and the content of the strains can be calculated according to the density of the strains in the microbial inoculum; the usage amount of the lactobacillus casei and the lactobacillus bifidus is 1 (0.8-3), and can be 1.
According to the invention, the content of amino acid in the obtained ferment can be influenced by the usage amount of each strain, when the usage amount ratio of the saccharomycetes, the lactobacillus casei and the lactobacillus bifidus is 1 (0.8-3) to (0.5-5), the content of free amino acid in the obtained ferment is higher, and if the usage amount of any one of the microbial inoculum is lower, the coordination effect among the strains can not be realized, so that the content of the amino acid can not be increased. For example, if the content of yeast is only 30% of lactobacillus casei or 15% of lactobacillus bifidus, the yeast may not maintain normal growth and reproduction, nor increase the amino acid content in the fermentation product through interaction between cells or interaction between cell products during fermentation; similarly, similar situations can be generated under the condition that the use amount of other strains is too much or too little, and the effect of better improving the content of amino acid can be generated only when the three strains are reasonably proportioned and can play the self-role to complete fermentation.
In a preferred embodiment of the present invention, the fruit is berry fruit or stone fruit.
Preferably, the fruit comprises any one of apple, peach, pear, citrus or grape, preferably grape.
Due to the fact that the types of fruits are multiple, and the nutritional ingredients and the surface microbial environment of the fruits are different, the enzyme preparation method provided by the invention has the advantages that the content of free amino acids in the obtained product is high when the grape enzyme is prepared, and the effect is good.
Preferably, the sugar comprises any one of rock sugar, white sugar or honey or a combination of at least two of them.
As a preferred embodiment of the present invention, the weight ratio of the fruit to the sugar is (2-5): 1, for example, 2:1, 2.5.
Preferably, the mass ratio of the fruit to the water is (0.3 to 0.8) 1, and may be, for example, 0.3.
In a preferred embodiment of the present invention, the time for the anaerobic fermentation is 20 to 40 days, and may be, for example, 20 days, 22 days, 25 days, 28 days, 30 days, 32 days, 35 days, 38 days, or 40 days.
Preferably, the temperature of the anaerobic fermentation is 20 to 30 ℃, for example, 20 ℃, 21 ℃, 22 ℃, 23 ℃, 24 ℃, 25 ℃, 26 ℃, 27 ℃, 28 ℃, 29 ℃ or 30 ℃ and the like.
As a preferable technical scheme of the invention, in the process of anaerobic fermentation, the fermentation container is kept closed or one-way exhaust is carried out. The fermentation container can be a generally closed plastic barrel or a container with a top cover provided with a one-way exhaust valve.
Preferably, the anaerobic fermentation period also comprises the operation of shaking exhaust.
Preferably, the fruit is washed with sterile water, ground and sterilized before mixing the fruit, sugar and water.
As a preferred technical scheme of the invention, the method comprises the following steps:
(1) Washing grapes with sterile water, grinding and sterilizing, and mixing the grapes and the sugar with water in a fermentation container, wherein the weight ratio of the grapes to the sugar is (2-5) to 1, and the mass ratio of the grapes to the water is (0.3-0.8) to 1;
(2) And (2) adding a microbial agent into the fermentation container, wherein the inoculation amount of the microbial agent is 0.8-1.2% of the total volume of the glucose, the sugar and the water, the usage amount of the yeast, the lactobacillus casei and the lactobacillus bifidus is 1 (0.8-3) and 0.5-5), and performing anaerobic fermentation at 20-30 ℃ for 20-40 days to obtain the enzyme.
In a second aspect, the present invention provides a ferment prepared by the method of the first aspect.
In the grape ferment obtained by the preparation method provided by the invention, the content of aspartic acid (Asp) is 27.8-64.2 mu g/mL, the content of glycine (Gly) is 21.3-25.1 mu g/mL, the content of alanine (Ala) is 37.9-88.0 mu g/mL, the content of valine (Val) is 22.3-25.6 mu g/mL, the content of leucine (Leu) is 13.5-27.8 mu g/mL, the content of lysine (Lys) is 33.7-51.3 mu g/mL, and the content of total free amino acid is 375.4-435.6 mu g/mL.
In a third aspect, the present invention further provides a use of the method of the first aspect or the ferment of the second aspect in preparing a health product.
The recitation of numerical ranges herein includes not only the above-recited numerical values, but also any numerical values between non-recited numerical ranges, and is not intended to be exhaustive or to limit the invention to the precise numerical values encompassed within the range for brevity and clarity.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention provides a method for improving the content of free amino acid in ferment, which is simple in preparation method, and the yield and release amount of the free amino acid in a fermentation raw material are increased by adding saccharomycetes, lactobacillus casei and lactobacillus bifidus in a certain proportion in the fermentation process and mutually matching different strains of the saccharomycetes, the lactobacillus casei and the lactobacillus bifidus in the long-time fermentation process, so that the content of the free amino acid in a ferment product can be obviously improved compared with the content of the free amino acid in a commercially available mainstream fruit ferment bacterium;
(2) When the grape ferment is obtained by fermenting grapes serving as a raw material, the content of free amino acid in the product is higher, the content of total amino acid in the ferment is 375.4-435.6 mug/mL, wherein the content of aspartic acid is 27.8-64.2 mug/mL, the content of glycine is 21.3-25.1 mug/mL, the content of alanine is 37.9-88.0 mug/mL, the content of valine is 22.3-25.6 mug/mL, the content of leucine is 13.5-27.8 mug/mL, and the content of lysine is 33.7-51.3 mug/mL.
Detailed Description
The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.
In the following examples, the room temperature was 25 ℃; the fruits are all available from conventional sources, wherein the grapes are from the theme park of the Mallotus philippinensis in Jiading district, shanghai city.
Example 1
This example provides a method for increasing the content of free amino acids in grape ferment.
The method comprises the following specific steps:
(1) Cleaning grapes with clear water, chopping, adding 750g of grapes, 250g of rock sugar and 1500g of water into a glass fermentation bottle, and uniformly mixing;
(2) Adding 1% by volume of microbial agent (wherein yeast, lactobacillus casei and lactobacillus bifidus are mixed in a ratio of 1;
(3) And (3) carrying out anaerobic fermentation at room temperature, slightly shaking to open the cover and exhaust air at irregular time in the fermentation process, and fermenting for 30 days to obtain the grape ferment.
Example 2
This example provides a method for increasing the content of free amino acids in grape ferment.
The difference from example 1 is that the yeast, lactobacillus casei and lactobacillus bifidus are mixed in a ratio of 1.
Example 3
This example provides a method for increasing the content of free amino acids in grape ferment.
The difference from example 1 is that the yeast, lactobacillus casei and lactobacillus bifidus are mixed in a ratio of 1.
Comparative example 1
The comparative example provides a preparation method of grape ferment.
The only difference from example 1 is that no microbial agent is added in the process; the remaining conditions were the same as in example 1.
Comparative example 2
The comparative example provides a preparation method of grape ferment.
The only difference from example 1 is that the microbial agent only contains lactobacillus casei and lactobacillus bifidus in a volume ratio of 1:1, and the inoculation amount is kept to be 1%; the remaining conditions were the same as in example 1.
Comparative example 3
The comparative example provides a preparation method of grape ferment.
The difference from the embodiment 1 is only that the microbial agent only contains yeast and lactobacillus casei with the volume ratio of 1:1, and the inoculation amount is kept to be 1%; the remaining conditions were the same as in example 1.
Comparative example 4
The comparative example provides a preparation method of grape ferment.
The only difference from example 1 is that the microbial agent only contains lactobacillus casei, and the inoculation amount is kept at 1%; the remaining conditions were the same as in example 1.
Comparative example 5
The comparative example provides a preparation method of grape ferment.
Only differs from example 1 in that the microbial agent contains only yeast, the inoculum size remaining 1%; the remaining conditions were the same as in example 1.
Comparative example 6
The comparative example provides a preparation method of grape ferment.
The only difference from example 1 is that the microbial agent was prepared by replacing Lactobacillus bifidus with Aspergillus, and the inoculum size was kept at 1%; the remaining conditions were the same as in example 1.
Comparative example 7
The comparative example provides a preparation method of grape ferment.
The only difference from example 1 is that the microbial agent added in the method is commercial ferment bacteria A, which only contains lactobacillus plantarum, and the inoculation amount is kept at 1%; the remaining conditions were the same as in example 1.
Comparative example 8
The comparative example provides a preparation method of grape ferment.
The only difference from example 1 is that the microbial agent added in the method is a commercial ferment bacterium B, which only contains lactobacillus casei, and the inoculation amount is kept to be 1%; the remaining conditions were the same as in example 1.
Free amino acid content test
The content of free amino acids in the ferments prepared according to the methods provided in examples 1 to 3 and comparative examples 1 to 8 was examined. Wherein, the content of free amino acid is detected by a full-automatic amino acid analyzer (Hitachi);
the above examples and comparative examples were all fermented under the same conditions with the same batch of grapes, and the test results were averaged after three measurements and the results obtained are shown in table 1 below:
TABLE 1 content of free amino acids in the grape ferments prepared in different ways (μ g/mL)
As can be seen from the above table, when the grapes, the rock candy and the water provided in example 1 are mixed in a mass ratio of 3;
and as can be seen from comparison of examples 1-3, the fermentation effect is better when the usage amount of the yeast, the lactobacillus casei and the lactobacillus bifidus is 1 (0.8-1.2) and 1-1.5; that is, the ratio of the yeast, lactobacillus casei and lactobacillus bifidus is important for increasing the content of free amino acids.
As can be seen from the comparison between example 1 and comparative examples 2 to 6, the yeast, lactobacillus casei and lactobacillus bifidus are all indispensable, and the synergistic cooperation effect exists among the three strains, and if any one of the three strains is reduced or replaced, the effect of improving the content of free amino acid cannot be achieved;
as can be seen from the comparison of example 1 with comparative examples 1, 7 and 8, the contents of various free amino acids and total free amino acids in grape ferment can be significantly increased by adding the beneficial microorganisms described in the present invention, as compared to the natural fermentation group and the commercially available ferment bacteria A and ferment bacteria B.
Meanwhile, the preparation method provided by the invention is also suitable for preparing other fruits, and can also improve the content of free amino acids in the obtained ferment, but the improvement effect is not obvious compared with the preparation of the grape ferment, and the method is not repeated for reasons of brevity and conciseness.
In conclusion, the method for improving the content of the free amino acid in the ferment provided by the invention has simpler steps, and the yield and release amount of the free amino acid in the fermentation raw material can be obviously improved and the content of the free amino acid in the ferment product can be improved by the synergistic cooperation of the saccharomycetes, the lactobacillus casei and the lactobacillus bifidus.
The applicant declares that the above description is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be understood by those skilled in the art that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are within the scope and disclosure of the present invention.
Claims (17)
1. A method for increasing the content of free amino acids in ferment, which is characterized by comprising the following steps:
mixing the crushed fruits, sugar and water in a fermentation container, adding a microbial agent, and performing anaerobic fermentation to obtain the enzyme;
the microbial agent comprises yeast, lactobacillus casei and lactobacillus bifidus;
the usage amount of the yeast, the lactobacillus casei and the lactobacillus bifidus is 1 (1.2 to 3) to 0.5 to 1.5;
the inoculation amount of the microbial agent is 0.8 to 1.2 percent of the total volume of the fruits, the sugar and the water.
2. The method of claim 1, wherein the fruit is a berry-like fruit or a stone-like fruit.
3. The method of claim 1, wherein the fruit comprises any one of apple, peach, pear, citrus, or grape.
4. The method according to claim 3, wherein the fruit is grapes.
5. The method of claim 1, wherein the sugar comprises any one or a combination of at least two of crystal sugar, white sugar or honey.
6. The method of claim 1, wherein the weight ratio of fruit to sugar is (2~5): 1.
7. The method of claim 6, wherein the weight ratio of fruit to sugar is 3:1.
8. The method according to claim 1, wherein the mass ratio of the fruit to the water is (0.3 to 0.8): 1.
9. The method according to claim 8, wherein the fruit and water mass ratio is 0.5.
10. The method of claim 1, wherein the anaerobic fermentation time is 20 to 40 days.
11. The method according to claim 1, wherein the temperature of the anaerobic fermentation is 20 to 30 ℃.
12. The method of claim 1, wherein the fermentation vessel is kept closed or vented in one direction during the anaerobic fermentation.
13. The method of claim 1, wherein the anaerobic fermentation is further performed with a shake venting operation.
14. The method of claim 1, further comprising the steps of washing the fruit with sterile water, grinding and sterilizing prior to mixing the fruit, sugar and water.
15. Method according to claim 1, characterized in that it comprises the following steps:
(1) Washing grapes with sterile water, grinding and sterilizing, and mixing the grapes and sugar with water in a fermentation container, wherein the weight ratio of the grapes to the sugar is (2~5): 1, and the mass ratio of the grapes to the water is (0.3 to 0.8): 1;
(2) And adding a microbial inoculum into the fermentation container, wherein the inoculation amount of the microbial inoculum is 0.8 to 1.2 percent of the total volume of the glucose, the sugar and the water, the usage amount of the yeast, the lactobacillus casei and the lactobacillus bifidus is 1 (1.2 to 3) (0.5 to 1.5), and the enzyme is obtained after anaerobic fermentation at the temperature of 20 to 30 ℃ for 20 to 40 days.
16. An enzyme prepared by the method of any one of claims 1-15.
17. Use of the method according to any one of claims 1 to 15 or the ferment according to claim 16 for the preparation of a health product.
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| CN108887674A (en) * | 2018-07-10 | 2018-11-27 | 集美大学 | A kind of preparation method of lycium ruthenicum composite enzyme liquid |
| CN108991498A (en) * | 2018-06-26 | 2018-12-14 | 菁英生命科学（深圳）有限公司 | Ferment and preparation method thereof |
| CN109953321A (en) * | 2017-12-25 | 2019-07-02 | 海南合生源生物科技有限公司 | The preparation method of grape ferment |
| CN111057637A (en) * | 2019-12-26 | 2020-04-24 | 江南大学 | Multi-strain fermented apple vinegar rich in free amino acids and production process thereof |
| CN111362736A (en) * | 2020-04-16 | 2020-07-03 | 上海市农业科学院 | Preparation method of grape waste fruit enzyme for fertilizer, prepared grape waste fruit enzyme and application of grape waste fruit enzyme |
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| CN109953321A (en) * | 2017-12-25 | 2019-07-02 | 海南合生源生物科技有限公司 | The preparation method of grape ferment |
| CN108991498A (en) * | 2018-06-26 | 2018-12-14 | 菁英生命科学（深圳）有限公司 | Ferment and preparation method thereof |
| CN108887674A (en) * | 2018-07-10 | 2018-11-27 | 集美大学 | A kind of preparation method of lycium ruthenicum composite enzyme liquid |
| CN111057637A (en) * | 2019-12-26 | 2020-04-24 | 江南大学 | Multi-strain fermented apple vinegar rich in free amino acids and production process thereof |
| CN111362736A (en) * | 2020-04-16 | 2020-07-03 | 上海市农业科学院 | Preparation method of grape waste fruit enzyme for fertilizer, prepared grape waste fruit enzyme and application of grape waste fruit enzyme |
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