阅读说明：本技术 一种富含γ-氨基丁酸的芦笋助眠凝胶棒及其制备方法 (Asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid and preparation method thereof ) 是由 李大刚 李艳梅 于 2021-02-01 设计创作，主要内容包括：本发明提供一种富含γ-氨基丁酸的芦笋助眠凝胶棒及其制备方法,制备方法如下：鲜芦笋加水研磨后,添加谷氨酸、乳酸钙、VB-6和蔗糖获得芦笋匀浆液；将枯草芽孢杆菌的菌种保藏液加入至灭菌的培养基中培养,离心后收集菌饼；将菌饼加入到芦笋匀浆液中,恒温培养,过滤得到发酵清液；向发酵清液中添加魔芋粉、卡拉胶粉和果胶粉,煮沸并降温后罐装入长条形制胶容器中,冷却得到富含γ-氨基丁酸的芦笋助眠凝胶棒。本发明利用芦笋酶和枯草芽孢杆菌的共同作用,提升了发酵液中γ-氨基丁酸物质的含量,发酵产品具有较好的生物安全性和助眠效果,酸甜可口,带有芦笋独特清香,将发酵产品制备成凝胶棒具有使用的简便性和高效性。(The invention provides an asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid and a preparation method thereof, wherein the preparation method comprises the following steps: grinding fresh Germinatus Phragmitis with water, adding glutamic acid, calcium lactate, and VB 6 Mixing with sucrose to obtain homogenized asparagus slurry; adding a strain preservation solution of bacillus subtilis into a sterilized culture medium for culturing, and collecting a bacterial cake after centrifugation; adding the mushroom cake into the asparagus homogenate, culturing at constant temperature, and filtering to obtain a fermentation clear liquid; adding konjac flour, carrageenan powder and pectin powder into the fermented clear liquid, boiling, cooling, filling into a strip-shaped gel making container, and cooling to obtain the asparagus sleep-aiding gel stick rich in gamma-aminobutyric acid. The invention utilizes the combined action of the aspartase and the bacillus subtilis to improve the content of the gamma-aminobutyric acid substance in the fermentation liquid, the fermentation product has better biological safety and sleep-aiding effect, is sour, sweet and delicious, has unique and faint scent of the asparagus, and the preparation of the fermentation product into the gel stick has the simplicity and the high efficiency in use.)

1. A preparation method of an asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid is characterized by comprising the following steps:

step 1, adding water into fresh asparagus, and grinding and homogenizing the fresh asparagus by a colloid mill, wherein the asparagus (kg) is prepared by mixing water (L) 1:2-1: 10;

step 2, glutamic acid, calcium lactate and VB are added into the serous fluid obtained in the step 1₆Mixing with sucrose, stirring and mixing to obtain asparagus homogenate;

step 3, preparation of bacteria for fermentation: adding a bacillus subtilis strain preservation solution with the preservation number of CCTCCM2020746 into a sterilized trypsin soybean culture medium, wherein the volume ratio of the bacillus subtilis strain preservation solution to the sterilized trypsin soybean culture medium is 1:20-1:100, and performing shake culture to obtain an overnight culture bacterial solution;

step 4, centrifuging the overnight culture liquid, removing supernatant, and collecting to obtain a fungus cake;

step 5, adding the fungus cake into the asparagus homogenate, wherein the ratio of the fungus cake to the asparagus homogenate is that every 100mL of fungus liquid is collected, the fungus cake is added into 1-10L of asparagus homogenate, and the fungus cake and the asparagus homogenate are uniformly mixed to obtain initial fermentation liquid;

step 6, statically culturing the initial fermentation liquor at constant temperature to obtain asparagus fermentation liquor;

and 7, filtering the asparagus fermentation liquor, removing residues to obtain a fermentation clear liquid, adding konjac flour, carrageenan powder and pectin powder into the fermentation clear liquid, stirring and dissolving, uniformly mixing, boiling for 1-5min, cooling to 50-60 ℃, canning into a strip-shaped glue making container, and cooling to room temperature to obtain the asparagus sleep-aiding gel stick rich in gamma-aminobutyric acid.

2. The method for preparing a sleep-aiding gel bar of asparagus rich in gamma-aminobutyric acid as claimed in claim 1, wherein in step 1, water (L) is added to asparagus (kg) in an amount of 1:4 to 1: 8.

3. According to claim 1The preparation method of the asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid is characterized in that 0.01 wt% -0.5 wt% of glutamic acid, 0.02 wt% -0.2 wt% of calcium lactate and 0.05 wt% -0.5 wt% of wtVB are added into the serous fluid in the step 2₆And 2% wt to 8% wt of sucrose.

4. The method for preparing a sleep-aiding gel bar of asparagus rich in gamma-aminobutyric acid as claimed in claim 1, wherein the shake culture in step 3 is performed at 37 ℃ and a rotation speed of 190-220rpm for 12-24h to obtain an overnight culture solution.

5. The method for preparing a sleep-aiding gel bar with asparagus rich in gamma-aminobutyric acid as claimed in claim 1, wherein the rotation speed of the centrifugation in the step 4 is 2000-4000 rpm, and the centrifugation time is 10-20 min.

6. The method for preparing a dormancy aid gel bar of asparagus with rich gamma-aminobutyric acid as claimed in claim 1, wherein the ratio of the mushroom cake in step 5 to asparagus homogenate is 4-8L asparagus homogenate per 100mL mushroom solution collected mushroom cake.

7. The method for preparing a sleep-aiding gel bar of asparagus rich in gamma-aminobutyric acid as claimed in claim 1, wherein the static culture in step 6 is a static culture at 30-40 ℃ for 12-24 h.

8. The method for preparing a sleep-aiding gel bar with asparagus rich in gamma-aminobutyric acid as claimed in claim 1, wherein step 7 is to add 1-4g of konjac flour, 3-10g of carrageenan powder and 1-4g of pectin powder to each liter of fermentation clear liquid.

9. An asparagus sleep-aiding gel stick rich in gamma-aminobutyric acid, characterized in that the asparagus sleep-aiding gel stick is prepared by the method of any one of claims 1 to 8.

Technical Field

The invention relates to a food biotechnology, in particular to an asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid and a preparation method thereof.

Background

Asparagus officinalis is seedling of Asparagus officinalis of Asparagus, and is an edible vegetable with low sugar, low fat and high protein. The asparagus contains chemical components such as flavonoid, oligosaccharide, sulfuric acid, triterpenic acid, steroid saponin, amino acid derivatives and the like, and has the activities of resisting mutation, tumors, fungi, blood fat, fatigue and the like. In addition, the asparagus is rich in free amino acid, which accounts for about 70-90 wt% of the total protein content, and is one of the highest varieties of known fruits and vegetables. Large-area asparagus is planted in Shandong, Jiangsu, Shanxi, Henan, Fujian and other provinces in China, the yield is 1000 kilograms per mu at the peak period of the yield, 500 kilograms per mu is obtained at low time, and the total annual yield is about three hundred million and fifty thousand kilograms.

Gamma-aminobutyric acid (GABA) is a naturally occurring nonprotein amino acid which is an important inhibitory neurotransmitter substance in the central nervous system of mammals, and GABA has anxiolytic, nervousness relieving, mood regulating, memory improving and sleep improving effects. The human body can naturally synthesize GABA, regulate emotion and inhibit nervous system excitation, but with the increase of age, the metabolic function of the human body is reduced, the content of GABA is gradually reduced, and physiological phenomena such as anxiety, stress, insomnia and the like can be generated. The proper amount of GABA is supplemented, so that the emotions of anxiety, tension and the like can be relieved, the sleep is regulated, and the deep sleep is promoted. Multinational studies have shown that adults can supplement GABA at 20-50mg per day to meet physical needs, and the daily intake of GABA should not be higher than 550 mg. The asparagus juice contains natural GABA, but the content is low (<0.1mg/mL), and sufficient GABA is difficult to supplement only by eating the asparagus.

Disclosure of Invention

The invention aims to provide a preparation method of an asparagus sleep-aiding gel rod rich in gamma-aminobutyric acid, which utilizes the combined action of asparaginase and bacillus subtilis to improve the content of gamma-aminobutyric acid in fermentation liquor, so that the fermentation product has better biological safety and sleep-aiding effect. The preparation method is simple, efficient, safe and rapid, the fermented product prepared by the method is sour, sweet and delicious, has unique faint scent of asparagus, and the preparation of the fermented product into a gel stick has simplicity and convenience in use and high efficiency.

In order to achieve the purpose, the invention adopts the technical scheme that: a preparation method of asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid comprises the following steps:

step 1, adding water into fresh asparagus, grinding the fresh asparagus into homogenate by a colloid mill, wherein the asparagus (kg) and the water (L) are 1:2-1:10 (mass-volume ratio);

step 2, glutamic acid, calcium lactate and VB are added into the serous fluid obtained in the step 1₆And the cane sugar are stirred and mixed evenly to obtain asparagus homogenate;

step 3, preparation of bacteria for fermentation: adding the strain preservation solution of the bacillus subtilis with the preservation number of CCTCCM2020746 into a sterilized trypsin soybean culture medium (TSB), wherein the volume ratio of the strain preservation solution of the bacillus subtilis to the sterilized trypsin soybean culture medium is 1:20-1:100(ml: ml), and performing shake culture to obtain an overnight culture solution;

step 4, centrifuging the overnight culture liquid, removing supernatant, and collecting to obtain a fungus cake;

step 5, adding the fungus cake into the asparagus homogenate, wherein the using amount ratio of the fungus cake to the asparagus homogenate is that the fungus cake obtained by collecting each 100mL of bacterial liquid is added into 1-10L of asparagus homogenate and is uniformly mixed to obtain initial fermentation liquid;

step 6, statically culturing the initial fermentation liquor at the constant temperature of 30-40 ℃ for 12-24h to obtain asparagus fermentation liquor;

and 7, filtering the asparagus fermentation liquor, removing residues to obtain a fermentation clear liquid, adding konjac flour, carrageenan powder and pectin powder into each liter of fermentation clear liquid, stirring and dissolving, uniformly mixing, boiling for 1-5min, cooling to 50-60 ℃, canning into a strip-shaped gel making container, and cooling to room temperature to obtain the asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid.

Further, in step 1, asparagus (kg) and water (L) are 1:4-1: 8.

Further, 0.01 wt% to 0.5 wt% glutamic acid, 0.02 wt% to 0.2 wt% calcium lactate, 0.05 wt% to 0.5 wt% wtVB are added to the slurry in step 2₆And 2% wt to 8% wt sucrose; preferably, 0.05 wt% to 0.2 wt% glutamic acid, 0.05 wt% to 0.2 wt% calcium lactate, 0.1 wt% to 0.2 wt% VB are added to the slurry in step 2₆And 4% wt to 6% wt sucrose.

Further, the shaking culture in the step 3 is performed for 12-24h at 37 ℃ and 190-220rpm, so as to obtain overnight culture solution. The preferred earthquake swing culture time is 14-18 h.

Further, the rotation speed of the centrifugation in the step 4 is 2000-4000 revolutions per minute, and the centrifugation time is 10-20 min.

Further, the using amount ratio of the fungus cake and the asparagus homogenate in the step 5 is that every 100mL of fungus liquid collected fungus cake is added into 4-8L of asparagus homogenate.

Further, step 6 static culture is static culture at 30-40 deg.C for 12-24 h. Preferably, step 6 the initial fermentation broth is incubated statically at 35-37 ℃ for 14-16 h.

Further, step 7, adding 1-4g of konjac flour, 3-10g of carrageenan powder and 1-4g of pectin powder into each liter of fermentation clear liquid; preferably, step 7, adding 2-3g of konjac flour, 5-8g of carrageenan powder and 2-3g of pectin powder into each liter of fermentation clear liquid, stirring for dissolving, mixing uniformly, boiling for 2-3min, cooling to 54-56 ℃, and canning.

The invention also discloses an asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid, which is prepared by adopting the method.

The invention relates to a preparation method of an asparagus sleep-aiding gel stick rich in gamma-aminobutyric acid, which is characterized in that a microorganism fermentation technology is adopted, fresh asparagus juice is used as a fermentation substrate, bacillus subtilis is used as a fermentation strain, and the asparagus gel stick rich in gamma-aminobutyric acid and having a certain sleep-aiding function is produced. Specifically, the present invention has the following advantages compared to the prior art:

1) the gamma-aminobutyric acid GABA obtained by the invention is biosynthesized rather than chemically synthesized, namely, is synthesized by glutamate decarboxylase (one part is from asparagus juice and the other part is from bacillus subtilis) under a proper substrate (glutamic acid) and environmental conditions, so that the gamma-aminobutyric acid GABA does not contain other chemical synthesis byproducts, and has better biological safety and availability.

2) The zymocyte used in the invention is bacillus subtilis with the preservation number of CCTCCM2020746, and is a safe strain for food fermentation. The strain can effectively promote the generation of GABA in the fermentation process, and the self metabolism generates organic acid, thereby maintaining the stability of GABA in the solution.

3) The asparagus fermentation liquor obtained by the invention also contains more than 10 free protein amino acids under the condition of containing gamma-aminobutyric acid (GABA), and has a better nutritional effect of quickly supplementing amino acids.

4) The invention utilizes the glutamic acid decarboxylase system contained in the asparagus and has the capability of catalyzing and converting the glutamic acid into GABA. The bacillus subtilis also has a glutamic acid decarboxylase system, can catalyze the conversion of glutamic acid to GABA, and simultaneously, an acidic environment is spontaneously formed in the fermentation process of the bacillus subtilis, so that the dissociation balance of GABA monomers is facilitated, and the formation of GABA dimers or oligomers is reduced.

5) The asparagus fermentation liquor obtained by the invention keeps the flavor of the asparagus, the free amino acid in the asparagus enriches the amino acid content in the fermentation liquor, and the organic acid generated by fermentation improves the taste of the asparagus juice, so that the fermentation liquor is sour, sweet and delicious. The invention also considers the consumer demand, and the fermentation liquor is made into the gel stick, which has the advantages of convenient transportation, carrying and use, and better market potential and competitiveness.

In conclusion, 1 strain of bacillus subtilis (the strain preservation number is CCTCCM2020746) capable of efficiently promoting GABA production is screened from more than 20 food fermentation strains. Under the premise of supplementing a proper amount of substrates and providing proper fermentation conditions, the GABA content in the asparagus juice can be increased by 2-5 times to reach 0.2-0.5mg/mL by fermenting the asparagus juice through the strain. The final asparagus fermented product is rich in GABA and more than 10 free amino acids, has certain functions of antianxiety, sleep aiding and amino acid nutrition, is suitable in sour and sweet taste and fragrant in taste, is convenient to eat and carry after being prepared into a gel stick, and is a high-quality sleep aiding functional asparagus fermented product.

Detailed Description

The invention is further illustrated by the following examples:

example 1

The embodiment discloses a preparation method of an asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid, which specifically comprises the following steps:

picking fresh asparagus in the step (1), cutting into sections, adding purified water according to the proportion of the asparagus to water of 1:5 (mass kg: volume L), and grinding and homogenizing by a colloid mill to obtain asparagus homogenate;

step (2) adding 0.05% wt of glutamic acid, 0.1% wt of calcium lactate and 0.1% wt of vb to the homogenate of asparagus obtained in step (1)₆And 4 wt% of cane sugar, stirring and mixing uniformly;

step (3) preparing bacteria for fermentation: taking 4mL of strain preservation solution of bacillus subtilis with the preservation number of CCTCCM2020746, adding the strain preservation solution into sterilized 200mL of trypsin soybean culture medium (TSB), and performing shake-flask culture for 16h at 37 ℃ and the rotation speed of 190rpm to obtain overnight culture solution.

Step (4) centrifuging the overnight culture liquid obtained in the step (3) at the rotating speed of 2000 rpm for 20min, discarding the supernatant, and collecting the obtained bacterial cake;

step (5) adding all the fungus cakes obtained in the step (4) into 4L of the fresh asparagus homogenate prepared in the step (1), and uniformly mixing to obtain initial fermentation liquor;

step (6), statically culturing the initial fermentation liquor obtained in the step (5) at 37 ℃ for 16h to obtain asparagus fermentation liquor;

and (7) filtering the fermentation liquor obtained in the step (6), removing residues to obtain a fermentation clear liquid, adding 3g of konjac flour, 8g of carrageenan powder and 3g of pectin powder into each liter of fermentation clear liquid, stirring and dissolving, uniformly mixing, boiling for 3min, cooling to 55 ℃, canning into a strip-shaped gel making container, packaging, and cooling to room temperature to obtain the asparagus sleeping aid gel stick.

In the asparagus gel stick finally obtained in the embodiment, the GABA concentration is 0.301mg/g, the free amino acid concentration is 3.526mg/g, and the total amino acid concentration is 6.471 mg/g.

Example 2

The embodiment discloses a preparation method of an asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid, which specifically comprises the following steps:

picking fresh asparagus in the step (1), cutting into sections, adding purified water according to the proportion of the asparagus to water of 1:2 (mass kg: volume L), and grinding and homogenizing by a colloid mill to obtain asparagus homogenate;

step (2) adding 0.01 wt% of glutamic acid, 0.02 wt% of calcium lactate and 0.05 wt% of wtVB into the asparagus homogenate obtained in step (1)₆And 2 wt% of cane sugar, stirring and mixing uniformly;

step (3) preparing bacteria for fermentation: taking 1mL of strain preservation solution of bacillus subtilis with the preservation number of CCTCCM2020746, adding the strain preservation solution into sterilized 100mL of trypsin soybean culture medium (TSB), and performing shake-flask culture for 12h at 37 ℃ and the rotation speed of 200rpm to obtain overnight culture solution.

Step (4) centrifuging the overnight culture bacterial liquid obtained in the step (3) at the rotating speed of 3000 r/min for 15min, removing the supernatant, and collecting to obtain a bacterial cake;

step (5) adding all the fungus cakes obtained in the step (4) into 1L of the fresh asparagus homogenate prepared in the step (1), and uniformly mixing to obtain initial fermentation liquor;

step (6), statically culturing the initial fermentation liquor obtained in the step (5) at 30 ℃ for 12 hours to obtain asparagus fermentation liquor;

and (7) filtering the fermentation liquor obtained in the step (6), removing residues to obtain a fermentation clear liquid, adding 1g of konjac flour, 3g of carrageenan powder and 1g of pectin powder into each liter of fermentation clear liquid, stirring and dissolving, uniformly mixing, boiling for 1min, cooling to 50 ℃, canning into a strip-shaped gel making container, packaging, and cooling to room temperature to obtain the asparagus sleeping aid gel stick.

In the asparagus gel stick finally obtained in the embodiment, the GABA concentration is 0.492mg/g, the free amino acid concentration is 5.583mg/g, and the total amino acid concentration is 9.218 mg/g.

Example 3

The embodiment discloses a preparation method of an asparagus sleep-aiding gel bar rich in gamma-aminobutyric acid, which specifically comprises the following steps:

picking fresh asparagus in the step (1), cutting into sections, adding purified water according to the proportion of the asparagus to water of 1:10 (mass kg: volume L), and grinding and homogenizing by a colloid mill to obtain asparagus homogenate;

step (2) adding 0.5% wt of glutamic acid, 0.2% wt of calcium lactate and 0.5% wt of wTVB to the asparagus homogenate obtained in step (1)₆And 8 wt% of cane sugar, and stirring and mixing uniformly;

step (3) preparing bacteria for fermentation: taking 5mL of strain preservation solution of bacillus subtilis with the preservation number of CCTCCM2020746, adding the strain preservation solution into sterilized 100mL of trypsin soybean culture medium (TSB), and performing shake-flask culture at 37 ℃ and the rotation speed of 220rpm for 24h to obtain overnight culture solution.

Step (4) centrifuging the overnight culture liquid obtained in the step (3) at the rotating speed of 4000 revolutions per minute for 10 minutes, removing the supernatant, and collecting to obtain a fungus cake;

step (5) adding all the fungus cakes obtained in the step (4) into 10L of the fresh asparagus homogenate prepared in the step (1), and uniformly mixing to obtain initial fermentation liquor;

step (6), statically culturing the initial fermentation liquor obtained in the step (5) at 40 ℃ for 24 hours to obtain asparagus fermentation liquor;

and (7) filtering the fermentation liquor obtained in the step (6), removing residues to obtain a fermentation clear liquid, adding 4g of konjac flour, 10g of carrageenan powder and 4g of pectin powder into each liter of fermentation clear liquid, stirring and dissolving, uniformly mixing, boiling for 5min, cooling to 60 ℃, canning into a strip-shaped gel making container, packaging, and cooling to room temperature to obtain the asparagus sleeping aid gel stick.

In the asparagus gel stick finally obtained in the embodiment, the GABA concentration is 0.124mg/g, the free amino acid concentration is 1.317mg/g, and the total amino acid concentration is 2.768 mg/g.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.

Sequence listing

<110> Panjin Ding Xin Bai Cao Yuan Co Ltd

Panjin Dingxin kitchen Co.,Ltd.

<120> asparagus sleep-aiding gel stick rich in gamma-aminobutyric acid and preparation method thereof

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