CN109123647B - A kind of preparation method of black barley enzyme effectively enriching γ-aminobutyric acid and polyphenols - Google Patents

A kind of preparation method of black barley enzyme effectively enriching γ-aminobutyric acid and polyphenols Download PDF

Info

Publication number
CN109123647B
CN109123647B CN201811220405.0A CN201811220405A CN109123647B CN 109123647 B CN109123647 B CN 109123647B CN 201811220405 A CN201811220405 A CN 201811220405A CN 109123647 B CN109123647 B CN 109123647B
Authority
CN
China
Prior art keywords
black barley
enzymolysis
lactobacillus
aminobutyric acid
black
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201811220405.0A
Other languages
Chinese (zh)
Other versions
CN109123647A (en
Inventor
宋立华
蒋芮
管玘
丁信文
欧阳鹏凌
宋晨玮
吴佳怡
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Jiao Tong University
Original Assignee
Shanghai Jiao Tong University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Jiao Tong University filed Critical Shanghai Jiao Tong University
Priority to CN201811220405.0A priority Critical patent/CN109123647B/en
Publication of CN109123647A publication Critical patent/CN109123647A/en
Application granted granted Critical
Publication of CN109123647B publication Critical patent/CN109123647B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/50—Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
    • A23L7/10—Cereal-derived products
    • A23L7/104—Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
    • 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
    • A23V2400/11—Lactobacillus
    • A23V2400/169—Plantarum
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Nutrition Science (AREA)
  • Genetics & Genomics (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Mycology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Biomedical Technology (AREA)
  • Agronomy & Crop Science (AREA)
  • Botany (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

本发明公开了一种有效富集γ‑氨基丁酸及多酚的黑大麦酵素制备方法，包括以下步骤：1)黑大麦酶解液培养基的制备：包括黑大麦清洗，粉碎，酶解，灭菌，冷却，得到所述黑大麦酶解液培养基；2)产γ‑氨基丁酸的乳杆菌的菌种用培养基活化得到种子液；3)将步骤2)得到的种子液接种到步骤1)得到的黑大麦酶解液培养基中，发酵，得到黑大麦发酵液。本发明筛选到可用于黑大麦发酵、有效富集γ‑氨基丁酸及多酚的乳杆菌，建立了黑大麦酵素加工的培养基制备工艺。本发明可为进一步开发以黑大麦为主要原料的功能性谷物酵素食品提供新的选择，因此具有较好的应用前景。

Figure 201811220405

The present invention discloses a method for preparing black barley enzyme for effectively enriching γ-aminobutyric acid and polyphenols. Sterilize, cool, obtain described black barley enzymolysis liquid medium; 2) the bacterial classification of Lactobacillus producing γ-aminobutyric acid is activated with medium to obtain seed liquid; 3) the seed liquid obtained in step 2) is inoculated into Step 1) Fermentation in the obtained black barley enzymatic hydrolysate medium to obtain a black barley fermentation broth. The present invention selects lactobacilli that can be used for black barley fermentation and effectively enriches γ-aminobutyric acid and polyphenols, and establishes a culture medium preparation process for black barley enzyme processing. The present invention can provide a new option for further developing the functional grain enzyme food with black barley as the main raw material, so it has a good application prospect.

Figure 201811220405

Description

Preparation method of black barley enzyme capable of effectively enriching gamma-aminobutyric acid and polyphenol
Technical Field
The invention relates to the technical field of grain and functional food processing, in particular to a preparation method of black barley enzyme capable of effectively enriching gamma-aminobutyric acid and polyphenol.
Background
Whole grains are grains that are dehulled and then composed of a cortex, germ, and endosperm. The cortex contains a large amount of dietary fiber and mineral substances, and the embryo contains a large amount of nutrient substances, so that the whole grain has richer nutrient components. Research has shown that the whole grain food can effectively reduce the risk of cardiovascular and cerebrovascular diseases, diabetes, malignant tumor and other related chronic diseases. However, because the whole grain cortex contains a large amount of cellulose, the whole grain cortex is directly eaten as staple food and is not beneficial to normal digestion and absorption of a human body; in addition, the whole grain food has poor water absorption and expansibility, long cooking time and poor taste, so the whole grain food is not easy to be accepted by people. The whole grain ferment obtained by fermentation can not only fully utilize the original nutrient components, improve some original bad flavors of the fermentation substrate, but also can generate some new functional components after fermentation, thereby further increasing the nutrient value of the whole grain.
The whole grain ferment is a nutrient mixture obtained by adding a small amount of honey into whole grains as a substrate and fermenting by using probiotics such as active baker's yeast, lactobacillus and the like. At present, the research on the cereal enzymes is few in products on the market at home, and the products are different in quality, so that the industrial production is not formed. Although the enzyme products such as the high-brown rice enzyme have been produced in advance in this respect in japan, the "black barley enzyme" product using black barley as a fermentation substrate and the invention thereof have not been reported yet.
The existing research results show that the grain ferment fermentation process conditions are different, and the types and the contents of the gamma-aminobutyric acid and other functional active substances are greatly different, so that how to obtain the whole grain ferment food with higher active ingredients such as gamma-aminobutyric acid (GABA), polyphenol and the like through the optimization of a culture medium, the screening of strains and raw materials and the optimization of the fermentation process is a problem worthy of deep research.
Therefore, those skilled in the art have made an effort to develop a method for preparing black barley enzyme that is effectively enriched in γ -aminobutyric acid and polyphenols.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention provides a method for efficiently enriching active ingredients such as γ -aminobutyric acid and polyphenols from black barley.
The invention discloses a preparation method of black barley enzyme for effectively enriching gamma-aminobutyric acid and polyphenol, which comprises the following steps:
1) preparing a black barley enzymolysis liquid culture medium: comprises the steps of cleaning, crushing, enzymolysis, sterilization and cooling of the black barley to obtain the black barley enzymolysis liquid culture medium;
2) activating the strain of lactobacillus producing gamma-aminobutyric acid by using a culture medium to obtain a seed solution; the Lactobacillus producing gamma-aminobutyric acid is Lactobacillus hilgardii (Lactobacillus hilgardii) DSMZ 20051, Lactobacillus newcastle (Lactobacillus diolivorans) DSMZ 14421 and Lactobacillus kisonensis (Lactobacillus kisonensis) JCM15041 which are separated in 2002;
3) inoculating the seed liquid obtained in the step 2) into the black barley enzymolysis liquid culture medium obtained in the step 1), and fermenting to obtain black barley fermentation liquid.
Among them, Lactobacillus Kisonensis (Lactobacillus Kisonensis) is named by the name of Kisonensis, the finder thereof.
Further, the black barley is cleaned, crushed, sieved by a 40-mesh sieve and then subjected to enzymolysis.
Further, the enzymolysis conditions are as follows: the mass ratio of the black barley to the water, namely the feed-liquid ratio is 1:5-10, preferably 1: 5; the enzyme used for enzymolysis is alpha-high temperature amylase, and further, the addition amount of the enzyme is 10U/g; the enzymolysis temperature is 65-75 ℃, and the preferable temperature is 70 ℃; the enzymolysis time is 20-60min, preferably 40 min.
Further, the enzymolysis termination conditions are as follows: after the enzymolysis is finished, the temperature is quickly raised to 100 ℃, and the mixture is boiled for 15 min.
Further, the enzymolysis liquid is sterilized after the enzymolysis is finished.
Preferably, the sterilization conditions of the enzymolysis liquid are as follows: cooling at 115 deg.C for 15min to obtain black barley enzymolysis liquid culture medium.
Further, the strain of lactobacillus producing gamma-aminobutyric acid is activated by MRS culture medium to obtain seed liquid.
Further, the activated lactobacillus seed liquid for producing the gamma-aminobutyric acid is inoculated to a black barley enzymolysis liquid culture medium, wherein the inoculation amount is 3-9% by volume percent, and is preferably 3-5%.
Further, the fermentation time is 24-72h, preferably 48 h.
Preferably, Lactobacillus kisonensis (Lactobacillus kisonensis) JCM15041 is inoculated into the black barley enzymolysis liquid medium.
Preferably, the inoculation amount is 3 percent by volume, the fermentation temperature is 30 ℃, and the fermentation time is 24 hours.
In some embodiments, step 1) is specifically: cleaning and crushing black barley, sieving the black barley with a 40-mesh sieve, adding pure water, heating and stirring the materials according to the mass ratio of 1:5 when pasting, adding alpha-high-temperature amylase according to the addition amount of 10U/g when the enzymolysis temperature reaches 70 ℃, carrying out enzymolysis for 40 minutes, then quickly heating to 100 ℃, boiling for 15 minutes, sterilizing at 115 ℃ for 15 minutes, and cooling to obtain a black barley enzymolysis liquid culture medium; the step 3) is specifically as follows: inoculating Lactobacillus kisonensis (Lactobacillus kisonensis) JCM15041 into the black barley zymolysis solution culture medium obtained in the step 1), wherein the inoculation amount is 3%, and fermenting for 24 hours at 30 ℃.
Furthermore, the screened lactobacillus is not only suitable for the black barley, but also can be widely applied to the fermentation process of brown rice, quinoa, beans and other foods.
Further, the preparation method of the black barley enzyme can obviously increase the content of other functional components such as essential fatty acid gamma-linoleic acid and the like, and adds procyanidine B2And the like.
The invention firstly screens lactobacillus which can be used for fermenting the black barley and effectively enrich gamma-aminobutyric acid and polyphenol, and establishes a culture medium preparation process and an optimal fermentation process condition for processing the black barley enzyme. Under the condition of the fermentation process, the active ingredients such as gamma-linoleic acid, malic acid and epicatechin are obviously increased, and catechin, coumaric acid and procyanidine B are added2And the like. The invention can provide a new choice for further developing the functional cereal ferment food taking the black barley as the main raw material, and has better application prospect.
The conception, the specific steps, and the technical effects produced by the present invention will be further described in conjunction with the accompanying drawings to fully understand the objects, the features, and the effects of the present invention.
Drawings
FIG. 1 is a process flow of ferment fermentation of black barley according to a preferred embodiment of the present invention;
FIG. 2 shows the result of qualitative detection of gamma-aminobutyric acid by paper chromatography according to a preferred embodiment of the present invention;
FIG. 3 shows the result of quantitative determination of gamma-aminobutyric acid by the amino acid analyzer according to the preferred embodiment of the present invention;
FIG. 4 shows active ingredients in the fermented product of black barley according to a preferred embodiment of the present invention in significantly varying amounts;
FIG. 5 shows the types and proportions of the newly added active ingredients in the fermented product of black barley according to a preferred embodiment of the present invention.
Detailed Description
The technical contents of the preferred embodiments of the present invention will be more clearly and easily understood by referring to the drawings attached to the specification. The present invention may be embodied in many different forms of embodiments and the scope of the invention is not limited to the embodiments set forth herein.
Example 1
The present example is the preparation of a black barley enzymolysis liquid culture medium:
as shown in table 1, the influence of the gelatinized material-liquid ratio, the enzymolysis temperature and the enzymolysis time on the content of reducing sugar in the black barley zymolyte is examined by adopting a three-factor three-level orthogonal test, and the preparation process conditions for optimally establishing the black barley zymolyte culture medium are as follows: crushing black barley, sieving the crushed black barley with a 40-mesh sieve, and sequentially mixing the raw materials according to a material-liquid ratio of 1:5(m/m) adding pure water, heating, raising the temperature and stirring; when the temperature reaches 70 deg.C, adding alpha-high temperature amylase (10U/g, purchased from Shanghai Aladdin Biotechnology, Ltd.) for 40 min; after the enzymolysis time is over, quickly heating to 100 ℃, and boiling for 15 min; finally, sterilizing at 115 ℃ for 15min, and cooling for later use. The influence of 70 ℃ and 75 ℃ on the enzymolysis result is not very different, and the enzymolysis temperature can be 70 ℃ from the cost-saving perspective.
TABLE 1 factor orthogonal test L for affecting reducing sugar content of black barley zymolyte9(34) Results
Figure BDA0001834616810000031
Figure BDA0001834616810000041
Example 2
This example is a screening of a lactobacillus producing gamma-aminobutyric acid:
the black barley enzymolysis liquid is used as a culture medium to establish a black barley fermentation process flow, as shown in figure 1. Activating the strain with MRS culture medium to form seed liquid, and inoculating to cooled black barley enzymolysis liquid culture medium for fermentation to obtain black barley fermentation liquid. And using gamma-aminobutyric acid as an index, using the black barley enzymolysis liquid as a culture medium, and respectively inoculating 21 lactobacillus strains in the black barley culture medium, wherein 11 lactobacillus plantarum strains (the numbers are respectively 11, 35, 25, 294, 2519, 34, 3736, 16, 234, 30 and 01) are separated and stored from food microbiology laboratory cornel of Shanghai traffic university (wherein the number 34 is lactobacillus casei, and the rest are lactobacillus plantarum); the origin and provenance of the other 10 strains are shown in Table 2.
TABLE 2 sources and sources of part of the fermentation tubes used in the present invention
Figure BDA0001834616810000042
Figure BDA0001834616810000051
Note: number 1 in this table is not the same strain as number 01 isolated and stored from cornel, a food microbiology laboratory of shanghai university of transportation.
The experimental results show that the bacteria No. 7, 9 and 10 have certain capacity of enriching the gamma-aminobutyric acid, as shown in the figure 2 and the figure 3. The "+" in fig. 3 indicates significant difference compared to the blank (unfermented black barley zymolyte broth), P < 0.05. On the basis, through orthogonal experiments, the fermentation process conditions of the black barley capable of effectively enriching the gamma-aminobutyric acid are optimized and established. 1%, 3% and 5% of activated bacteria No. 7, 9 and 10 are respectively inoculated in a black barley zymolyte liquid culture medium, the samples are sampled and tested for gamma-aminobutyric acid after the culture is carried out for 24 hours under the conditions of initial pH 6.5 and 30 ℃, and the results of orthogonal experiments are shown in Table 3. The orthogonal experiment result shows that: the optimal fermentation condition is strain No. 10, the fermentation time is 24 hours, and the inoculation amount is 5%. Under the condition, the enrichment amount of the gamma-aminobutyric acid is 43.56mg/100g, and is improved by 1.34 times compared with an unfermented control group (18.59mg/100 g).
TABLE 3 orthogonal test results for fermentation enrichment of gamma-aminobutyric acid using black barley liquid medium
Figure BDA0001834616810000052
Figure BDA0001834616810000061
Example 3
The embodiment is the establishment and optimization of technological parameters for simultaneously enriching polyphenol and gamma-aminobutyric acid in black barley fermentation:
the optimized fermentation process orthogonal experiment is established by taking polyphenol as a research index, and the experimental result is shown in table 4. The result shows that the technological parameters for optimally enriching the polyphenol in the black barley fermentation product are as follows: bacterium No. 10, inoculum size 9%, feed liquid ratio 1: fermenting at 30 deg.C for 24 h.
TABLE 4 orthogonal experimental results of polyphenol enrichment by fermentation of black barley liquid medium
Figure BDA0001834616810000062
Figure BDA0001834616810000071
Considering comprehensively that both the gamma-aminobutyric acid and the experimental result are better than that of the bacterium No. 10, the enrichment conditions of the polyphenol and the gamma-aminobutyric acid of the bacterium No. 10 under the conditions of the inoculation amounts of 3%, 6% and 9% are further verified, and the results are shown in tables 5 and 6, wherein the enrichment effect of the 3% inoculation amount is higher than that of the 6% and 9%, and the enrichment effect of the 3% inoculation amount is as follows: the polyphenol content is 88mg/100g, which is improved by 1.05 times compared with the unfermented group (43mg/100 g); the gamma-aminobutyric acid also reaches the maximum 65.14mg/100g, which is improved by 2.42 times compared with the unfermented group (19.06mg/100 g). In consideration of the actual production cost, 10 bacteria was used, and the inoculum size was 3%.
TABLE 5 results of verifying polyphenol content in fermented product of black barley under optimum conditions (fungus No. 10)
Figure BDA0001834616810000072
P <0.0001vs unfermented control group
TABLE 6 content of gamma-aminobutyric acid (mg/100g) under optimal fermentation process conditions for polyphenol production
Figure BDA0001834616810000073
In conclusion, the process conditions for effectively enriching the active ingredients such as the gamma-aminobutyric acid, the polyphenol and the like by taking the black barley enzymolysis liquid as the culture medium and the lactobacillus as the fermentation strain are as follows: crushing black barley, sieving the black barley with a 40-mesh sieve, sequentially adding pure water according to a designed proportion, heating and stirring the mixture according to a ratio of 1:5(m/m) of material liquid during gelatinization, adding alpha-high-temperature amylase (with the addition amount of 10U/g) when the mixture reaches an enzymolysis temperature of 70 ℃, carrying out enzymolysis for 40min, then quickly heating the mixture to 100 ℃, boiling the mixture for 15min, sterilizing the mixture at 115 ℃ for 15min, cooling the mixture, inoculating No. 10 bacteria, wherein the inoculation amount is 3%, and the fermentation time is 24h, under the condition, the content of gamma-aminobutyric acid is 65.14mg/100g, which is 2.42 times higher than that of an unfermented control group (19.06mg/100g), the polyphenol content is 88mg/100g, and is 1.05 times higher than that of a control group (43mg/100 g).
In addition to the measurement of polyphenol content, UPLC-MS analysis of black barley fermented product showed that the content of other active ingredients changed after fermentation, and fig. 4 shows the portion of active ingredients with a large content change after fermentation. It can be seen that the functional active components such as essential fatty acid gamma-linoleic acid, malic acid, hesperetin and epicatechin are remarkably increased after fermentation.
In addition to the above active ingredients, UPLC-MS analysis also found that new active substances not contained in the grain before fermentation could be produced after the black barley was fermented, as shown in fig. 5. Mainly comprises catechin, salvianolic acid, coumaric acid, and procyanidin B2And the like.
The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations could be devised by those skilled in the art in light of the present teachings without departing from the inventive concepts. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.

Claims (5)

1.一种有效富集γ-氨基丁酸及多酚的黑大麦酵素制备方法，其特征在于，包括以下步骤：1. a black barley enzyme preparation method for effectively enriching γ-aminobutyric acid and polyphenol, is characterized in that, comprises the following steps: 1)黑大麦酶解液培养基的制备：包括黑大麦清洗，粉碎，酶解，灭菌，冷却，得到所述黑大麦酶解液培养基；1) preparation of black barley enzymolysis liquid medium: comprising black barley washing, pulverization, enzymolysis, sterilization, cooling, to obtain described black barley enzymolysis liquid medium; 2)产γ-氨基丁酸的乳杆菌的菌种用培养基活化得到种子液；所述产γ-氨基丁酸的乳杆菌为希氏乳杆菌(Lactobacillus hilgardii)DSMZ 20051、2002年分离得到的乳杆菌新种丘氏乳杆菌(Lactobacillus diolivorans)DSMZ 14421或kisonensis乳杆菌(Lactobacillus kisonensis)JCM15041；2) the bacterial classification of the lactobacillus producing γ-aminobutyric acid is activated with a medium to obtain seed liquid; the lactobacillus producing γ-aminobutyric acid is obtained by Lactobacillus hilgardii (Lactobacillus hilgardii) DSMZ 20051, 2002 Lactobacillus new species Lactobacillus diolivorans DSMZ 14421 or Lactobacillus kisonensis JCM15041; 3)将所述步骤2)得到的种子液接种到所述步骤1)得到的黑大麦酶解液培养基中，发酵，得到黑大麦发酵液；3) inoculating the seed liquid obtained in the step 2) into the black barley enzymolysis liquid medium obtained in the step 1), and fermenting to obtain a black barley fermentation liquid; 其中，所述步骤1)中的酶解条件为黑大麦与水的质量比，即料液比为1:5-10；所述步骤1)中的酶解所用的酶为α-高温淀粉酶；所述步骤1)中的酶解温度为65-75℃；酶解时间为20-60min；所述步骤3)中的接种量为体积百分比3％-9％；所述步骤3)中的发酵时长是24-72h。Wherein, the enzymolysis condition in the step 1) is the mass ratio of black barley and water, that is, the ratio of solid to liquid is 1:5-10; the enzyme used in the enzymolysis in the step 1) is α-high temperature amylase ; the enzymolysis temperature in the step 1) is 65-75°C; the enzymolysis time is 20-60min; the inoculation amount in the step 3) is 3%-9% by volume; the The fermentation time is 24-72h. 2.如权利要求1所述的黑大麦酵素制备方法，其特征在于，所述步骤1)中的粉碎后过40目筛后再进行酶解。2. The black barley enzyme preparation method as claimed in claim 1, is characterized in that, the enzymolysis is carried out after passing through a 40-mesh sieve after pulverizing in the step 1). 3.如权利要求1所述的黑大麦酵素制备方法，其特征在于，所述步骤2)中的培养基为MRS培养基。3. black barley enzyme preparation method as claimed in claim 1, is characterized in that, the substratum in described step 2) is MRS substratum. 4.如权利要求1所述的黑大麦酵素制备方法，其特征在于，所述步骤1)具体为：将黑大麦清洗、粉碎后过40目筛后，加入纯水，糊化时料液质量比1:5，加热升温并搅拌，到达酶解温度70℃时，按10U/g的添加量加入α-高温淀粉酶，酶解时间40min，后迅速升温至100℃，煮沸15min，115℃、15min杀菌、冷却，得到黑大麦酶解液培养基。4. black barley ferment preparation method as claimed in claim 1, is characterized in that, described step 1) is specially: after black barley is cleaned, pulverized and crossed after 40 mesh sieves, pure water is added, and the quality of feed liquid during gelatinization The ratio of 1:5, heating and stirring, when reaching the enzymatic hydrolysis temperature of 70 ℃, add α-high temperature amylase according to the addition amount of 10U/g, the enzymatic hydrolysis time is 40min, then quickly heat up to 100 ℃, boil for 15min, 115 ℃, Sterilize and cool for 15 minutes to obtain a black barley enzymatic hydrolyzate medium. 5.如权利要求1所述的黑大麦酵素制备方法，其特征在于，所述步骤3)具体为：接种kisonensis乳杆菌(Lactobacillus kisonensis)JCM15041到所述步骤1)得到的黑大麦酶解液培养基中，接种量为3％，30℃下，发酵24h。5. black barley enzyme preparation method as claimed in claim 1, is characterized in that, described step 3) is specially: inoculate kisonensis lactobacillus (Lactobacillus kisonensis) JCM15041 to the black barley enzymolysis liquid culture that described step 1) obtains In the base, the inoculum amount was 3%, and the fermentation was carried out at 30°C for 24h.
CN201811220405.0A 2018-10-19 2018-10-19 A kind of preparation method of black barley enzyme effectively enriching γ-aminobutyric acid and polyphenols Active CN109123647B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201811220405.0A CN109123647B (en) 2018-10-19 2018-10-19 A kind of preparation method of black barley enzyme effectively enriching γ-aminobutyric acid and polyphenols

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201811220405.0A CN109123647B (en) 2018-10-19 2018-10-19 A kind of preparation method of black barley enzyme effectively enriching γ-aminobutyric acid and polyphenols

Publications (2)

Publication Number Publication Date
CN109123647A CN109123647A (en) 2019-01-04
CN109123647B true CN109123647B (en) 2021-10-01

Family

ID=64808840

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201811220405.0A Active CN109123647B (en) 2018-10-19 2018-10-19 A kind of preparation method of black barley enzyme effectively enriching γ-aminobutyric acid and polyphenols

Country Status (1)

Country Link
CN (1) CN109123647B (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109805374A (en) * 2019-01-23 2019-05-28 武汉轻工大学 A kind of collagen-rich peach gum ferment and the preparation method and application thereof
CN110522034A (en) * 2019-08-21 2019-12-03 上海交通大学 A functional food containing Lactobacillus niger fermented product and its application
CN112931865A (en) * 2021-03-29 2021-06-11 山东百德生物科技有限公司 Edible amino acid enzyme rich in gamma-aminobutyric acid and preparation method thereof
CN114568687A (en) * 2022-03-10 2022-06-03 上海交通大学 Preparation method of quinoa compound whole grain enzyme
CN117770317B (en) * 2023-06-15 2026-04-28 江南大学 Preparation method of oat-based fermented milk enriched with polyphenols based on combined enzymatic hydrolysis-fermentation technology

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105942084A (en) * 2016-04-28 2016-09-21 天津科技大学 Method for producing probiotic functional food through buckwheat fermentation

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ITRM20070398A1 (en) * 2007-07-17 2009-01-18 Giuliani Spa PROCEDURE FOR THE PREPARATION OF BUTIRRIC GAMMA-AMINO ACID (GABA) THROUGH LACTIC BACTERIA (LAB) ON AGRO-FOOD SURPLUS.

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105942084A (en) * 2016-04-28 2016-09-21 天津科技大学 Method for producing probiotic functional food through buckwheat fermentation

Also Published As

Publication number Publication date
CN109123647A (en) 2019-01-04

Similar Documents

Publication Publication Date Title
US12144363B2 (en) Method for preparing Cordyceps militaris ferment by two-stage fermentation and complex enzymatic hydrolysis
Nile The nutritional, biochemical and health effects of makgeolli–a traditional Korean fermented cereal beverage
CN104694371B (en) Citrus fruit vinegar prepared by composite strain mixed fermentation and preparation method thereof
CN101519631A (en) Production and preparation method of red jujube vinegar
CN107495043B (en) A kind of sweet corn whole milk lactic acid bacteria drink and preparation method thereof
JP6288837B2 (en) Lactic acid bacteria fermented product of rice bran cereal powder or rice bran saccharified product and method for producing the same
TW201244641A (en) Very low salt soy sauce and method for producing same
CN114606152B (en) A strain of Bacillus Velez, microbial agent and its application
CN115644332B (en) Rosa roxburghii and coix seed composite beverage and preparation method thereof
CN110747145A (en) Lactobacillus for high yield of gamma-aminobutyric acid and isolated culture method and application thereof
CN109123647A (en) A kind of black barley ferment preparation method of effective enriching gamma-aminobutyric and polyphenol
CN109486645B (en) Method for brewing and blending fragrant mature vinegar by applying immobilized targeted multi-microbe strain
CN103789140B (en) A kind of brewing method of red date naked oats composite wine
CN108308481A (en) A kind of preparation process of probiotics fermention citrus beverages
CN112980646A (en) Kefir source composite probiotic fermented pear juice and oat viable bacteria vinegar drink and preparation method thereof
CN104068371B (en) A kind of broad bean chili sauce accelerating flavouring fermenting agent and preparation method thereof
CN113528380B (en) Multi-strain composite brown rice solid-state fermentation microbial inoculum, and preparation method and application thereof
CN112544831A (en) Preparation method of highland barley enzyme beverage rich in natural gamma-aminobutyric acid
CN117770317B (en) Preparation method of oat-based fermented milk enriched with polyphenols based on combined enzymatic hydrolysis-fermentation technology
AU2005231013B2 (en) Method of Manufacturing Liquid Koji
JP2005312438A (en) FOOD MATERIAL WITH HIGH gamma-AMINOBUTYRIC ACID CONTENT AND METHOD FOR PRODUCING THE SAME
CN107904185B (en) Lactobacillus and application thereof
CN110583932A (en) Preparation method of fermented corn germ meal beverage
KR102289292B1 (en) Method for manufacturing of draft rice wine with an increased shelf life through the separation processing of lees and draft rice wine using the same
Jemima et al. Microbes in production of Non-alcoholic Beverages

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant