CN111248383A - Preparation process of lactic acid fermentation rice juice fermentation liquor highly rich in aminobutyric acid - Google Patents

Preparation process of lactic acid fermentation rice juice fermentation liquor highly rich in aminobutyric acid Download PDF

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CN111248383A
CN111248383A CN202010209450.7A CN202010209450A CN111248383A CN 111248383 A CN111248383 A CN 111248383A CN 202010209450 A CN202010209450 A CN 202010209450A CN 111248383 A CN111248383 A CN 111248383A
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姜益军
杨锁华
王旭
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Synbiotech Biotechnology Yangzhou Co ltd
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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
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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
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52—Adding ingredients
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    • 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
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
    • A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
    • 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

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Abstract

The invention discloses a preparation process of lactic acid fermented rice juice fermentation liquor rich in aminobutyric acid, which comprises the steps of taking brown rice as a raw material, conducting induced germination to process the brown rice into germ rice, then conducting grinding and freezing spray grinding to obtain ultrafine powder with the particle size of less than or equal to 1000 meshes, then adding the ultrafine powder into water, conducting cooking, sequentially adding β -glucanase and neutral protease to conduct enzymolysis treatment, finally adding lactobacillus powder into the obtained enzymolysis liquid to conduct two-step fermentation, adding okra colloidal polysaccharide between the two-step fermentation, finally obtaining the fermentation liquor which is rich in nutrition, high in aminobutyric acid content, fine and smooth in taste and clear in appearance without precipitates.

Description

Preparation process of lactic acid fermentation rice juice fermentation liquor highly rich in aminobutyric acid
Technical Field
The invention relates to preparation of lactic acid fermented rice juice, in particular to a preparation process of lactic acid fermented rice juice fermentation liquor rich in aminobutyric acid. Belongs to the technical field of food processing.
Background
Rice is one of the most important grain crops in the world, common processing forms of the rice comprise rice starch, rice protein, rice wine and the like, but the processing forms can not fully utilize the nutritional ingredients of the rice, so that great waste is caused. Moreover, with the improvement of living standard and the acceleration of life rhythm, the demand of people for natural, green, healthy and convenient foods is more and more urgent. The lactic acid fermentation rice juice is a beverage product which is obtained by taking rice as a raw material through lactic acid fermentation, can effectively improve the nutrient composition, has rice fragrance of the rice, meets the health and refined diet requirements of modern people, and has good economic popularization value.
The brown rice has higher nutritive value than polished rice, and 60-70% of vitamins, minerals and a large amount of essential amino acids in the rice are accumulated in an outer layer tissue. The brown rice has good protein quality, mainly contains relatively complete components of protamine and amino acid, and is easy to digest and absorb by human body. However, the brown rice has a rough taste, and the direct preparation of the brown rice into a finished beverage has the problems of precipitation or suspended matters, so that the drinking taste is influenced.
Gamma-aminobutyric acid (GABA, aminobutyric acid), one of physiologically active ingredients present in the embryo and aleurone layers of brown rice, is a non-protein amino acid widely present in a free state in prokaryotes and eukaryotes, has various effects in the brain and spinal cord of mammals, and functions as an inhibitory neurotransmitter in the central nervous system. About 30% of central nervous synapse sites transmit GABA. Plays an important role in the cerebral cortex, hippocampus, thalamus, basal ganglia and cerebellum of a human body, and has a regulating effect on various functions of the body. When GABA is deficient in the human body, the emotions such as anxiety, uneasiness, tiredness, anxiety and the like are obviously generated.
However, the content of gamma-aminobutyric acid in rice is too low to achieve significant health and therapeutic effects. Therefore, the method can improve the content of the gamma-aminobutyric acid as much as possible in the rice juice fermentation process, and has great economic value. In addition, most of the existing rice juice products are thick paste type, and people are used to select refreshing type drinks when people are thirsty, which undoubtedly influences the sales volume of the rice juice.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid comprises the following specific steps in parts by weight:
(1) inducing germination: adding 1 part of brown rice into 10-12 parts of water, treating for 1-2 hours at the temperature of 20-25 ℃ and the magnetic field intensity of 0.1-0.2T, then treating for 2-4 hours at the temperature of-5-10 ℃, then treating for 5-6 hours at the temperature of 30-35 ℃ under the ozone atmosphere, and inducing to germinate to obtain germ rice;
(2) then sequentially grinding and freezing and spraying the milled rice with embryo buds to process the milled rice into superfine powder with the particle size of less than or equal to 1000 meshes;
(3) adding the superfine powder into 5-8 parts of water, cooking, sequentially adding β -glucanase and neutral protease for enzymolysis, inactivating enzymes, and filtering to obtain an enzymolysis solution;
(4) and finally, adding 0.001-0.002 part of lactobacillus powder into the enzymolysis liquid, performing first fermentation, adding 0.02-0.03 part of okra gum polysaccharide, performing second fermentation, and performing post-treatment to obtain the lactic acid fermented rice juice fermentation liquid rich in aminobutyric acid.
Preferably, in the step (2), the specific method of milling is as follows: the milling is realized by adopting an iron roller rice mill, and the process conditions are as follows: 4 rice mills are connected in series, and the ratio of the grinding pressure is 6: 4: 2: 1.
preferably, in the step (2), the specific method of the freeze jet milling is as follows: and (3) spraying the powder obtained after grinding at a high speed at a spraying speed of 70-75 m/s to collide with a freezing wall so as to obtain the ultrafine powder, wherein the temperature of the freezing wall is-15 to-18 ℃.
Preferably, in the step (3), the cooking process conditions are as follows: stirring for 30-40 minutes at 65-70 ℃.
Preferably, the specific method in the step (3) comprises the steps of adding the ultrafine powder into water, cooking, cooling to 45-50 ℃, adjusting the pH to 4.0-5.5, adding β -glucanase, performing enzymolysis for 30-40 minutes, adjusting the pH to 7-8, adding neutral protease, performing enzymolysis for 80-100 minutes, inactivating enzymes, and filtering to obtain an enzymolysis solution, wherein the adding amount of β -glucanase and the adding amount of neutral protease are respectively 10-12U/g and 22-25U/g based on the weight of the brown rice.
Preferably, in the step (4), the okra gum polysaccharide is prepared by the following method: chopping fresh okra, adding the chopped okra into 5-8 times of absolute ethyl alcohol, leaching for 2-3 times, centrifuging, and drying the obtained solid at 30-40 ℃ for 12-24 hours to obtain the okra extract.
Preferably, in the step (4), the process conditions of the first fermentation step are as follows: anaerobic fermentation is carried out for 20-25 hours at the temperature of 40-45 ℃; the process conditions of the second step of fermentation are as follows: anaerobic fermentation is carried out for 10-12 hours at 40-45 ℃.
Preferably, in the step (4), the lactobacillus powder is prepared from bifidobacterium, lactobacillus acidophilus and streptococcus thermophilus according to a mass ratio of 1: 0.4-0.6: 0.1-0.2 fermenting separately to obtain each strain fermentation liquor, mixing and fermenting, and post-treating.
Further preferably, MRS solid medium is used for both single fermentation and mixed fermentation, and the conditions of the single fermentation or the mixed fermentation are as follows: culturing at 36-38 deg.C for 2-3 days.
Further preferably, the amount of inoculated strain is 4 to 5% by volume at the time of fermentation.
Further preferably, the post-treatment method comprises the following specific steps: firstly, flocculating the obtained fermentation liquor by using disodium hydrogen phosphate and calcium chloride, wherein the addition amount of the disodium hydrogen phosphate and the addition amount of the calcium chloride are respectively 1-2% and 0.5-0.8% of the mass of the fermentation liquor; and then performing filter pressing by adopting a plate frame, drying at 45-50 ℃ until the water content is 20-30%, performing cyclone separation, crushing by using a crusher, and sieving by using a 30-mesh sieve to obtain the lactobacillus powder.
Preferably, in the step (4), the post-treatment is performed by the following specific method: centrifuging at 10000-12000 rpm for 5-8 minutes, taking supernatant, and pasteurizing to obtain the lactic acid fermentation rice juice fermentation liquor.
The lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid is obtained by the preparation process.
The application of the lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid in preparing rice juice drinks.
A rice juice beverage is prepared by adding 8-10 times of water into the fermentation liquor.
The invention has the beneficial effects that:
the invention uses brown rice as raw material, firstly induces germination and processes into germ rice, then the germ rice is milled, frozen, sprayed and ground to obtain ultrafine powder with the grain diameter less than or equal to 1000 meshes, then the ultrafine powder is added into water and cooked, β -glucanase and neutral protease are sequentially added for enzymolysis treatment, finally lactobacillus powder is added into the obtained enzymolysis liquid for two-step fermentation, okra colloidal polysaccharide is added between the two-step fermentation, finally the obtained fermentation liquid has rich nutrition, high aminobutyric acid content, fine and smooth mouthfeel and clear appearance without precipitation.
The induced germination is divided into three steps, firstly, magnetic field induction is carried out at the temperature of 20-25 ℃, then low-temperature treatment is carried out at the temperature of-5-10 ℃, finally ozone induction is carried out at the temperature of 30-35 ℃, glutamate decarboxylase is fully released in the germination process, β -glucanase and neutral protease are added in the subsequent steps for enzymolysis treatment, protein contained in brown rice can be decomposed into a large number of micromolecule amino acids, a large number of glutamic acids are generated through enzymolysis through matching of the two, α -carboxyl of the glutamic acids is removed under the action of the glutamate decarboxylase to obtain gamma-aminobutyric acid, and the content of the gamma-aminobutyric acid in the product is greatly improved.
The milled germinated rice is processed into superfine powder with the grain size of less than or equal to 1000 meshes through grinding and freezing jet milling, the grain size is small, the dispersion in water is facilitated, the stability is improved, the generation of precipitation is avoided, β -glucanase can also enable high molecular viscous glucan to be decomposed into low viscosity isomaltose and isomaltotriose, the viscosity is facilitated to be reduced, the filtering speed is improved, and the clarity of the obtained product is better.
Detailed Description
The present invention will be further illustrated by the following examples, which are intended to be merely illustrative and not limitative.
The bifidobacterium related to the invention, ATCC29521, is purchased from Shanghai Fuxiang biotechnology Limited; lactobacillus acidophilus, ATCC4356, purchased from north noro biotechnology limited, shanghai; streptococcus thermophilus, ATCC19258, was purchased from North Noro Biotech, Inc. of Shanghai.
Example 1:
a preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid comprises the following specific steps in parts by weight:
(1) inducing germination: adding 1 part of brown rice into 10 parts of water, treating for 2 hours at the temperature of 25 ℃ and the magnetic field intensity of 0.1T, then treating for 4 hours at the temperature of-5 ℃, then treating for 6 hours at the temperature of 30 ℃ under the ozone atmosphere, and inducing to germinate to obtain germ rice;
(2) then sequentially grinding and freezing and spraying the milled rice with embryo buds to process the milled rice into superfine powder with the particle size of less than or equal to 1000 meshes;
(3) adding the superfine powder into 5 parts of water, cooking, sequentially adding β -glucanase and neutral protease for enzymolysis, inactivating enzymes, and filtering to obtain an enzymolysis solution;
(4) and finally, adding 0.002 part of lactobacillus powder into the enzymolysis liquid, performing first-step fermentation, adding 0.02 part of okra gum polysaccharide, performing second-step fermentation, and performing post-treatment to obtain the lactic acid fermented rice juice fermentation liquid rich in aminobutyric acid.
Wherein, in the step (2), the specific method of milling is as follows: the milling is realized by adopting an iron roller rice mill, and the process conditions are as follows: 4 rice mills are connected in series, and the ratio of the grinding pressure is 6: 4: 2: 1.
in the step (2), the specific method of freezing, spraying and grinding is as follows: the powder obtained after milling was sprayed at a high speed at a spraying speed of 75m/s to collide against a freezing wall having a temperature of-15 c, thereby obtaining an ultrafine powder.
In the step (3), the cooking process conditions are as follows: stirring for 30 minutes at 70 ℃.
The specific method of the step (3) is that the ultrafine powder is added into water, cooked, cooled to 50 ℃, the pH value is adjusted to 4.0, β -glucanase is added, enzymolysis is carried out for 40 minutes, the pH value is adjusted to 7, neutral protease is added, enzymolysis is carried out for 100 minutes, enzyme is deactivated, and the enzymolysis liquid is obtained after filtration, wherein the adding amount of β -glucanase and the adding amount of neutral protease are respectively 10U/g and 25U/g based on the weight of the brown rice.
In the step (4), the okra gum polysaccharide is prepared by the following method: cutting fresh okra, adding into 5 times of anhydrous ethanol, leaching for 3 times, centrifuging, and drying the obtained solid at 30 deg.C for 24 hr.
In the step (4), the process conditions of the first fermentation step are as follows: anaerobic fermentation at 40 deg.C for 25 hr; the process conditions of the second step of fermentation are as follows: anaerobic fermentation at 40 deg.C for 12 hr.
In the step (4), the lactobacillus powder is prepared by mixing the strains of bifidobacterium, lactobacillus acidophilus and streptococcus thermophilus according to the mass ratio of 1: 0.4: 0.2 fermenting separately to obtain each strain fermentation liquor, then mixing and fermenting, and post-processing.
MRS solid culture medium is used for both single fermentation and mixed fermentation, and the conditions of the single fermentation or the mixed fermentation are as follows: cultured at 36 ℃ for 3 days. The inoculum size of the strain was 4% by volume during fermentation. The specific method of post-treatment is as follows: firstly, flocculating the obtained fermentation liquor by utilizing disodium hydrogen phosphate and calcium chloride, wherein the addition amount of the disodium hydrogen phosphate and the addition amount of the calcium chloride are respectively 2% and 0.5% of the mass of the fermentation liquor; and then, performing filter pressing by adopting a plate frame, drying at 50 ℃ until the water content is 20%, performing cyclone separation, crushing by using a crusher, and sieving by using a 30-mesh sieve to obtain the lactobacillus powder.
In the step (4), the post-treatment method comprises the following specific steps: centrifuging at 12000 r/min for 5 min, collecting supernatant, and pasteurizing to obtain the lactic acid fermented rice juice fermentation liquid.
Example 2:
a preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid comprises the following specific steps in parts by weight:
(1) inducing germination: adding 1 part of brown rice into 12 parts of water, treating for 1 hour at the temperature of 20 ℃ and the magnetic field intensity of 0.2T, then treating for 2 hours at the temperature of-10 ℃, then treating for 5 hours at the temperature of 35 ℃ under the ozone atmosphere, and inducing to germinate to obtain germ rice;
(2) then sequentially grinding and freezing and spraying the milled rice with embryo buds to process the milled rice into superfine powder with the particle size of less than or equal to 1000 meshes;
(3) adding the superfine powder into 8 parts of water, cooking, sequentially adding β -glucanase and neutral protease for enzymolysis, inactivating enzymes, and filtering to obtain an enzymolysis solution;
(4) and finally, adding 0.001 part of lactobacillus powder into the enzymolysis liquid, performing first-step fermentation, adding 0.03 part of okra gum polysaccharide, performing second-step fermentation, and performing post-treatment to obtain the lactic acid fermented rice juice fermentation liquid rich in aminobutyric acid.
Wherein, in the step (2), the specific method of milling is as follows: the milling is realized by adopting an iron roller rice mill, and the process conditions are as follows: 4 rice mills are connected in series, and the ratio of the grinding pressure is 6: 4: 2: 1.
in the step (2), the specific method of freezing, spraying and grinding is as follows: the powder obtained after milling was sprayed at a high speed at a spraying speed of 70m/s to collide against a freezing wall having a temperature of-18 c, thereby obtaining an ultrafine powder.
In the step (3), the cooking process conditions are as follows: stirring at 65 deg.C for 40 min.
The specific method of the step (3) is that the ultrafine powder is added into water and cooked, the temperature is reduced to 45 ℃, the pH value is adjusted to 5.5, β -glucanase is added, enzymolysis is carried out for 30 minutes, the pH value is adjusted to 8, neutral protease is added, enzymolysis is carried out for 80 minutes, enzyme is deactivated, and the enzymolysis liquid is obtained after filtration, wherein the adding amount of β -glucanase and the adding amount of neutral protease are 12U/g and 22U/g respectively based on the weight of the brown rice.
In the step (4), the okra gum polysaccharide is prepared by the following method: cutting fresh okra, adding into 8 times of anhydrous ethanol, leaching for 2 times, centrifuging, and drying the obtained solid at 40 deg.C for 12 hr.
In the step (4), the process conditions of the first fermentation step are as follows: anaerobic fermentation at 45 deg.c for 20 hr; the process conditions of the second step of fermentation are as follows: anaerobic fermentation is carried out for 10 hours at the temperature of 45 ℃.
In the step (4), the lactobacillus powder is prepared by mixing the strains of bifidobacterium, lactobacillus acidophilus and streptococcus thermophilus according to the mass ratio of 1: 0.6: 0.1 fermenting separately to obtain each strain fermentation liquid, then mixing and fermenting, and post-processing.
MRS solid culture medium is used for both single fermentation and mixed fermentation, and the conditions of the single fermentation or the mixed fermentation are as follows: cultured at 38 ℃ for 2 days. The inoculum size of the strain was 5% by volume during fermentation. The specific method of post-treatment is as follows: firstly, flocculating the obtained fermentation liquor by utilizing disodium hydrogen phosphate and calcium chloride, wherein the addition amount of the disodium hydrogen phosphate and the addition amount of the calcium chloride are respectively 1 percent and 0.8 percent of the mass of the fermentation liquor; and then, performing filter pressing by adopting a plate frame, drying at 45 ℃ until the water content is 30%, performing cyclone separation, crushing by using a crusher, and sieving by using a 30-mesh sieve to obtain the lactobacillus powder.
In the step (4), the post-treatment method comprises the following specific steps: centrifuging at 10000 r/min for 8 min, collecting supernatant, and pasteurizing to obtain the lactic acid fermented rice juice fermentation liquid.
Example 3:
a preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid comprises the following specific steps in parts by weight:
(1) inducing germination: adding 1 part of brown rice into 11 parts of water, treating for 1.5 hours at the temperature of 23 ℃ and the magnetic field intensity of 0.1T, then treating for 3 hours at the temperature of-8 ℃, then treating for 5.5 hours at the temperature of 33 ℃ under the ozone atmosphere, and inducing to germinate to obtain germ rice;
(2) then sequentially grinding and freezing and spraying the milled rice with embryo buds to process the milled rice into superfine powder with the particle size of less than or equal to 1000 meshes;
(3) adding the superfine powder into 6 parts of water, cooking, sequentially adding β -glucanase and neutral protease for enzymolysis, inactivating enzymes, and filtering to obtain an enzymolysis solution;
(4) and finally, adding 0.0015 part of lactobacillus powder into the enzymolysis liquid, performing first-step fermentation, adding 0.025 part of okra gum polysaccharide, performing second-step fermentation, and performing post-treatment to obtain the lactic acid fermented rice juice fermentation liquid rich in aminobutyric acid.
Wherein, in the step (2), the specific method of milling is as follows: the milling is realized by adopting an iron roller rice mill, and the process conditions are as follows: 4 rice mills are connected in series, and the ratio of the grinding pressure is 6: 4: 2: 1.
in the step (2), the specific method of freezing, spraying and grinding is as follows: the powder obtained after milling was sprayed at a high speed at a spraying speed of 72m/s to collide against a freezing wall having a temperature of-17 c to obtain an ultrafine powder.
In the step (3), the cooking process conditions are as follows: stirring at 68 ℃ for 35 minutes.
The specific method of the step (3) is that the ultrafine powder is added into water, cooked, cooled to 48 ℃, the pH value is adjusted to 5, β -glucanase is added, enzymolysis is carried out for 35 minutes, the pH value is adjusted to 7.5, neutral protease is added, enzymolysis is carried out for 90 minutes, enzyme is deactivated, and the enzymolysis liquid is obtained after filtration, wherein the adding amount of β -glucanase and the adding amount of neutral protease are respectively 11U/g and 23U/g based on the weight of the brown rice.
In the step (4), the okra gum polysaccharide is prepared by the following method: cutting fresh okra, adding into 6 times of anhydrous ethanol, leaching for 3 times, centrifuging, and drying the obtained solid at 35 deg.C for 18 hr.
In the step (4), the process conditions of the first fermentation step are as follows: anaerobic fermentation at 42 deg.c for 23 hr; the process conditions of the second step of fermentation are as follows: anaerobic fermentation at 42 deg.C for 11 hr.
In the step (4), the lactobacillus powder is prepared by mixing the strains of bifidobacterium, lactobacillus acidophilus and streptococcus thermophilus according to the mass ratio of 1: 0.5: 0.15 fermenting separately to obtain each strain fermentation liquid, mixing and fermenting, and post-treating.
MRS solid culture medium is used for both single fermentation and mixed fermentation, and the conditions of the single fermentation or the mixed fermentation are as follows: cultured at 37 ℃ for 2 days. The inoculum size of the strain was 4.5% by volume during fermentation. The specific method of post-treatment is as follows: firstly, flocculating the obtained fermentation liquor by utilizing disodium hydrogen phosphate and calcium chloride, wherein the addition amount of the disodium hydrogen phosphate and the addition amount of the calcium chloride are respectively 1.5 percent and 0.6 percent of the mass of the fermentation liquor; and then performing filter pressing by adopting a plate frame, drying at 48 ℃ until the water content is 25%, performing cyclone separation, crushing by using a crusher, and sieving by using a 30-mesh sieve to obtain the lactobacillus powder.
In the step (4), the post-treatment method comprises the following specific steps: centrifuging at 11000 r/min for 7 min, taking supernatant, and pasteurizing to obtain the lactic acid fermented rice juice fermentation liquid.
Comparative example 1
A preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid comprises the following specific steps in parts by weight:
(1) inducing germination: adding 1 part of brown rice into 10 parts of water, treating for 12 hours at the temperature of 30 ℃, and inducing to germinate to obtain germ rice;
(2) then sequentially grinding and freezing and spraying the milled rice with embryo buds to process the milled rice into superfine powder with the particle size of less than or equal to 1000 meshes;
(3) adding the superfine powder into 5 parts of water, cooking, sequentially adding β -glucanase and neutral protease for enzymolysis, inactivating enzymes, and filtering to obtain an enzymolysis solution;
(4) and finally, adding 0.002 part of lactobacillus powder into the enzymolysis liquid, performing first-step fermentation, adding 0.02 part of okra gum polysaccharide, performing second-step fermentation, and performing post-treatment to obtain the lactic acid fermented rice juice fermentation liquid rich in aminobutyric acid.
Wherein, in the step (2), the specific method of milling is as follows: the milling is realized by adopting an iron roller rice mill, and the process conditions are as follows: 4 rice mills are connected in series, and the ratio of the grinding pressure is 6: 4: 2: 1.
in the step (2), the specific method of freezing, spraying and grinding is as follows: the powder obtained after milling was sprayed at a high speed at a spraying speed of 75m/s to collide against a freezing wall having a temperature of-15 c, thereby obtaining an ultrafine powder.
In the step (3), the cooking process conditions are as follows: stirring for 30 minutes at 70 ℃.
The specific method of the step (3) is that the ultrafine powder is added into water, cooked, cooled to 50 ℃, the pH value is adjusted to 4.0, β -glucanase is added, enzymolysis is carried out for 40 minutes, the pH value is adjusted to 7, neutral protease is added, enzymolysis is carried out for 100 minutes, enzyme is deactivated, and the enzymolysis liquid is obtained after filtration, wherein the adding amount of β -glucanase and the adding amount of neutral protease are respectively 10U/g and 25U/g based on the weight of the brown rice.
In the step (4), the okra gum polysaccharide is prepared by the following method: cutting fresh okra, adding into 5 times of anhydrous ethanol, leaching for 3 times, centrifuging, and drying the obtained solid at 30 deg.C for 24 hr.
In the step (4), the process conditions of the first fermentation step are as follows: anaerobic fermentation at 40 deg.C for 25 hr; the process conditions of the second step of fermentation are as follows: anaerobic fermentation at 40 deg.C for 12 hr.
In the step (4), the lactobacillus powder is prepared by mixing the strains of bifidobacterium, lactobacillus acidophilus and streptococcus thermophilus according to the mass ratio of 1: 0.4: 0.2 fermenting separately to obtain each strain fermentation liquor, then mixing and fermenting, and post-processing.
MRS solid culture medium is used for both single fermentation and mixed fermentation, and the conditions of the single fermentation or the mixed fermentation are as follows: cultured at 36 ℃ for 3 days. The inoculum size of the strain was 4% by volume during fermentation. The specific method of post-treatment is as follows: firstly, flocculating the obtained fermentation liquor by utilizing disodium hydrogen phosphate and calcium chloride, wherein the addition amount of the disodium hydrogen phosphate and the addition amount of the calcium chloride are respectively 2% and 0.5% of the mass of the fermentation liquor; and then, performing filter pressing by adopting a plate frame, drying at 50 ℃ until the water content is 20%, performing cyclone separation, crushing by using a crusher, and sieving by using a 30-mesh sieve to obtain the lactobacillus powder.
In the step (4), the post-treatment method comprises the following specific steps: centrifuging at 12000 r/min for 5 min, collecting supernatant, and pasteurizing to obtain the lactic acid fermented rice juice fermentation liquid.
Comparative example 2
A preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid comprises the following specific steps in parts by weight:
(1) inducing germination: adding 1 part of brown rice into 10 parts of water, treating for 2 hours at the temperature of 25 ℃ and the magnetic field intensity of 0.1T, then treating for 4 hours at the temperature of-5 ℃, then treating for 6 hours at the temperature of 30 ℃ under the ozone atmosphere, and inducing to germinate to obtain germ rice;
(2) then sequentially grinding the milled rice with embryo and processing the milled rice into powder;
(3) adding the powder into 5 parts of water, cooking, sequentially adding β -glucanase and neutral protease for enzymolysis, inactivating enzymes, and filtering to obtain an enzymolysis solution;
(4) and finally, adding 0.002 part of lactobacillus powder into the enzymolysis liquid, performing first-step fermentation, adding 0.02 part of okra gum polysaccharide, performing second-step fermentation, and performing post-treatment to obtain the lactic acid fermented rice juice fermentation liquid rich in aminobutyric acid.
Wherein, in the step (2), the specific method of milling is as follows: the milling is realized by adopting an iron roller rice mill, and the process conditions are as follows: 4 rice mills are connected in series, and the ratio of the grinding pressure is 6: 4: 2: 1.
in the step (3), the cooking process conditions are as follows: stirring for 30 minutes at 70 ℃.
The specific method of the step (3) is that the powder is added into water and cooked, the temperature is reduced to 50 ℃, the pH value is adjusted to 4.0, β -glucanase is added, enzymolysis treatment is carried out for 40 minutes, the pH value is adjusted to 7, neutral protease is added, enzymolysis treatment is carried out for 100 minutes, enzyme is deactivated, and filtration is carried out, so as to obtain enzymolysis liquid, wherein the adding amount of β -glucanase and the adding amount of neutral protease are respectively 10U/g and 25U/g based on the weight of the brown rice.
In the step (4), the okra gum polysaccharide is prepared by the following method: cutting fresh okra, adding into 5 times of anhydrous ethanol, leaching for 3 times, centrifuging, and drying the obtained solid at 30 deg.C for 24 hr.
In the step (4), the process conditions of the first fermentation step are as follows: anaerobic fermentation at 40 deg.C for 25 hr; the process conditions of the second step of fermentation are as follows: anaerobic fermentation at 40 deg.C for 12 hr.
In the step (4), the lactobacillus powder is prepared by mixing the strains of bifidobacterium, lactobacillus acidophilus and streptococcus thermophilus according to the mass ratio of 1: 0.4: 0.2 fermenting separately to obtain each strain fermentation liquor, then mixing and fermenting, and post-processing.
MRS solid culture medium is used for both single fermentation and mixed fermentation, and the conditions of the single fermentation or the mixed fermentation are as follows: cultured at 36 ℃ for 3 days. The inoculum size of the strain was 4% by volume during fermentation. The specific method of post-treatment is as follows: firstly, flocculating the obtained fermentation liquor by utilizing disodium hydrogen phosphate and calcium chloride, wherein the addition amount of the disodium hydrogen phosphate and the addition amount of the calcium chloride are respectively 2% and 0.5% of the mass of the fermentation liquor; and then, performing filter pressing by adopting a plate frame, drying at 50 ℃ until the water content is 20%, performing cyclone separation, crushing by using a crusher, and sieving by using a 30-mesh sieve to obtain the lactobacillus powder.
In the step (4), the post-treatment method comprises the following specific steps: centrifuging at 12000 r/min for 5 min, collecting supernatant, and pasteurizing to obtain the lactic acid fermented rice juice fermentation liquid.
Comparative example 3
A preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid comprises the following specific steps in parts by weight:
(1) inducing germination: adding 1 part of brown rice into 10 parts of water, treating for 2 hours at the temperature of 25 ℃ and the magnetic field intensity of 0.1T, then treating for 4 hours at the temperature of-5 ℃, then treating for 6 hours at the temperature of 30 ℃ under the ozone atmosphere, and inducing to germinate to obtain germ rice;
(2) then sequentially grinding and freezing and spraying the milled rice with embryo buds to process the milled rice into superfine powder with the particle size of less than or equal to 1000 meshes;
(3) adding the superfine powder into 5 parts of water, cooking, sequentially adding β -glucanase for enzymolysis, inactivating enzyme, and filtering to obtain an enzymolysis solution;
(4) and finally, adding 0.002 part of lactobacillus powder into the enzymolysis liquid, performing first-step fermentation, adding 0.02 part of okra gum polysaccharide, performing second-step fermentation, and performing post-treatment to obtain the lactic acid fermented rice juice fermentation liquid rich in aminobutyric acid.
Wherein, in the step (2), the specific method of milling is as follows: the milling is realized by adopting an iron roller rice mill, and the process conditions are as follows: 4 rice mills are connected in series, and the ratio of the grinding pressure is 6: 4: 2: 1.
in the step (2), the specific method of freezing, spraying and grinding is as follows: the powder obtained after milling was sprayed at a high speed at a spraying speed of 75m/s to collide against a freezing wall having a temperature of-15 c, thereby obtaining an ultrafine powder.
In the step (3), the cooking process conditions are as follows: stirring for 30 minutes at 70 ℃.
The specific method of the step (3) is to add the ultrafine powder into water, cook, cool to 50 ℃, adjust the pH to 4.0, add β -glucanase, carry out enzymolysis for 140 minutes, inactivate enzyme and filter to obtain an enzymolysis liquid, wherein, the adding amount of β -glucanase and neutral protease is 10U/g and 25U/g respectively based on the weight of the brown rice.
In the step (4), the okra gum polysaccharide is prepared by the following method: cutting fresh okra, adding into 5 times of anhydrous ethanol, leaching for 3 times, centrifuging, and drying the obtained solid at 30 deg.C for 24 hr.
In the step (4), the process conditions of the first fermentation step are as follows: anaerobic fermentation at 40 deg.C for 25 hr; the process conditions of the second step of fermentation are as follows: anaerobic fermentation at 40 deg.C for 12 hr.
In the step (4), the lactobacillus powder is prepared by mixing the strains of bifidobacterium, lactobacillus acidophilus and streptococcus thermophilus according to the mass ratio of 1: 0.4: 0.2 fermenting separately to obtain each strain fermentation liquor, then mixing and fermenting, and post-processing.
MRS solid culture medium is used for both single fermentation and mixed fermentation, and the conditions of the single fermentation or the mixed fermentation are as follows: cultured at 36 ℃ for 3 days. The inoculum size of the strain was 4% by volume during fermentation. The specific method of post-treatment is as follows: firstly, flocculating the obtained fermentation liquor by utilizing disodium hydrogen phosphate and calcium chloride, wherein the addition amount of the disodium hydrogen phosphate and the addition amount of the calcium chloride are respectively 2% and 0.5% of the mass of the fermentation liquor; and then, performing filter pressing by adopting a plate frame, drying at 50 ℃ until the water content is 20%, performing cyclone separation, crushing by using a crusher, and sieving by using a 30-mesh sieve to obtain the lactobacillus powder.
In the step (4), the post-treatment method comprises the following specific steps: centrifuging at 12000 r/min for 5 min, collecting supernatant, and pasteurizing to obtain the lactic acid fermented rice juice fermentation liquid.
Comparative example 4
A preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid comprises the following specific steps in parts by weight:
(1) inducing germination: adding 1 part of brown rice into 10 parts of water, treating for 2 hours at the temperature of 25 ℃ and the magnetic field intensity of 0.1T, then treating for 4 hours at the temperature of-5 ℃, then treating for 6 hours at the temperature of 30 ℃ under the ozone atmosphere, and inducing to germinate to obtain germ rice;
(2) then sequentially grinding and freezing and spraying the milled rice with embryo buds to process the milled rice into superfine powder with the particle size of less than or equal to 1000 meshes;
(3) adding the superfine powder into 5 parts of water, cooking, sequentially adding β -glucanase and neutral protease for enzymolysis, inactivating enzymes, and filtering to obtain an enzymolysis solution;
(4) and finally, adding 0.002 part of lactobacillus powder into the enzymolysis liquid, and performing first-step fermentation, second-step fermentation and post-treatment to obtain the lactic acid fermentation rice juice fermentation liquid rich in aminobutyric acid.
Wherein, in the step (2), the specific method of milling is as follows: the milling is realized by adopting an iron roller rice mill, and the process conditions are as follows: 4 rice mills are connected in series, and the ratio of the grinding pressure is 6: 4: 2: 1.
in the step (2), the specific method of freezing, spraying and grinding is as follows: the powder obtained after milling was sprayed at a high speed at a spraying speed of 75m/s to collide against a freezing wall having a temperature of-15 c, thereby obtaining an ultrafine powder.
In the step (3), the cooking process conditions are as follows: stirring for 30 minutes at 70 ℃.
The specific method of the step (3) is that the ultrafine powder is added into water, cooked, cooled to 50 ℃, the pH value is adjusted to 4.0, β -glucanase is added, enzymolysis is carried out for 40 minutes, the pH value is adjusted to 7, neutral protease is added, enzymolysis is carried out for 100 minutes, enzyme is deactivated, and the enzymolysis liquid is obtained after filtration, wherein the adding amount of β -glucanase and the adding amount of neutral protease are respectively 10U/g and 25U/g based on the weight of the brown rice.
In the step (4), the process conditions of the first fermentation step are as follows: anaerobic fermentation at 40 deg.C for 25 hr; the process conditions of the second step of fermentation are as follows: anaerobic fermentation at 40 deg.C for 12 hr.
In the step (4), the lactobacillus powder is prepared by mixing the strains of bifidobacterium, lactobacillus acidophilus and streptococcus thermophilus according to the mass ratio of 1: 0.4: 0.2 fermenting separately to obtain each strain fermentation liquor, then mixing and fermenting, and post-processing.
MRS solid culture medium is used for both single fermentation and mixed fermentation, and the conditions of the single fermentation or the mixed fermentation are as follows: cultured at 36 ℃ for 3 days. The inoculum size of the strain was 4% by volume during fermentation. The specific method of post-treatment is as follows: firstly, flocculating the obtained fermentation liquor by utilizing disodium hydrogen phosphate and calcium chloride, wherein the addition amount of the disodium hydrogen phosphate and the addition amount of the calcium chloride are respectively 2% and 0.5% of the mass of the fermentation liquor; and then, performing filter pressing by adopting a plate frame, drying at 50 ℃ until the water content is 20%, performing cyclone separation, crushing by using a crusher, and sieving by using a 30-mesh sieve to obtain the lactobacillus powder.
In the step (4), the post-treatment method comprises the following specific steps: centrifuging at 12000 r/min for 5 min, collecting supernatant, and pasteurizing to obtain the lactic acid fermented rice juice fermentation liquid.
Test examples
The performance of the fermentation liquids obtained in examples 1 to 3 and comparative examples 1 to 4 was examined, and the results are shown in Table 1.
And (2) carrying out gamma-aminobutyric acid detection by referring to a liquid chromatography method in NY/T2890-2016, wherein a sample extracting solution directly uses fermentation liquor obtained in examples 1-3 and comparative examples 1-4, the sample dosage is 1mL, and the calculation formula is as follows: ω ═ ρ × V2/V1Omega is the content of gamma-aminobutyric acid (mg/L), rho is the measured mass concentration (mg/L), V1Volume of fermentation broth (L), V2To a constant volume (L).
The content of flavone (rutin equivalent) is detected by spectrophotometry.
TABLE 1 examination of nutrient composition and appearance of fermentation broths
Content of gamma-aminobutyric acid (mg/L) Flavone content (mg/L) Appearance state
Example 1 12.35 123.12 Clear without precipitation
Example 2 12.33 123.32 Clear without precipitation
Example 3 12.59 123.66 Clear without precipitation
Comparative example 1 6.35 71.78 Clear without precipitation
Comparative example 2 10.45 100.03 With uniformly suspended small particles
Comparative example 3 7.87 80.03 With uniformly suspended small particles
Comparative example 4 12.34 123.12 Clear without precipitation
The fermentation liquids obtained in examples 1 to 3 and comparative examples 1 to 4 were sealed and left standing at 25 ℃ for 12 months, and changes in appearance state were observed, and the results are shown in Table 2.
TABLE 2 change in appearance after 12 months of standing
Appearance state
Example 1 Clear without precipitation
Example 2 Clear without precipitation
Example 3 Clear without precipitation
Comparative example 1 Clear without precipitation
Comparative example 2 Turbid with sediment at the bottom
Comparative example 3 Turbid with sediment at the bottom
Comparative example 4 Turbid with sediment at the bottom
As can be seen from tables 1 and 2, the fermentation liquids obtained in examples 1 to 3 have high contents of gamma-aminobutyric acid and flavone, clear appearance without precipitation, no change in appearance after 12 months, and good stability. Comparative example 1 treatment such as magnetic field, freezing, ozone and the like is omitted in the germination induction step, comparative example 2 omits the freezing spray grinding step, and comparative example 3 only adopts neutral protease in the enzymolysis treatment, so that the quality of the product is deteriorated; the product stability of comparative examples 2 and 3 also became significantly worse. In the comparative example 4, the okra gum polysaccharide is omitted, and the stability of the product is obviously poor.
Although the present invention has been described with reference to the specific embodiments, it is not intended to limit the scope of the present invention, and various modifications and variations can be made by those skilled in the art without inventive changes based on the technical solution of the present invention.

Claims (10)

1. A preparation process of lactic acid fermentation rice juice fermentation liquor rich in aminobutyric acid is characterized by comprising the following specific steps in parts by weight:
(1) inducing germination: adding 1 part of brown rice into 10-12 parts of water, treating for 1-2 hours at the temperature of 20-25 ℃ and the magnetic field intensity of 0.1-0.2T, then treating for 2-4 hours at the temperature of-5-10 ℃, then treating for 5-6 hours at the temperature of 30-35 ℃ under the ozone atmosphere, and inducing to germinate to obtain germ rice;
(2) then sequentially grinding and freezing and spraying the milled rice with embryo buds to process the milled rice into superfine powder with the particle size of less than or equal to 1000 meshes;
(3) adding the superfine powder into 5-8 parts of water, cooking, sequentially adding β -glucanase and neutral protease for enzymolysis, inactivating enzymes, and filtering to obtain an enzymolysis solution;
(4) and finally, adding 0.001-0.002 part of lactobacillus powder into the enzymolysis liquid, performing first fermentation, adding 0.02-0.03 part of okra gum polysaccharide, performing second fermentation, and performing post-treatment to obtain the lactic acid fermented rice juice fermentation liquid rich in aminobutyric acid.
2. The preparation process according to claim 1, wherein in the step (2), the specific method of milling is as follows: the milling is realized by adopting an iron roller rice mill, and the process conditions are as follows: 4 rice mills are connected in series, and the ratio of the grinding pressure is 6: 4: 2: 1.
3. the process according to claim 1, wherein in the step (2), the specific method of the freeze jet milling is: and (3) spraying the powder obtained after grinding at a high speed at a spraying speed of 70-75 m/s to collide with a freezing wall so as to obtain the ultrafine powder, wherein the temperature of the freezing wall is-15 to-18 ℃.
4. The process according to claim 1, wherein in the step (3), the cooking conditions are as follows: stirring for 30-40 minutes at 65-70 ℃.
5. The preparation process of claim 1, wherein the step (3) comprises the specific steps of adding the ultrafine powder into water, cooking, cooling to 45-50 ℃, adjusting the pH value to 4.0-5.5, adding β -glucanase, performing enzymolysis for 30-40 minutes, adjusting the pH value to 7-8, adding neutral protease, performing enzymolysis for 80-100 minutes, inactivating the enzyme, and filtering to obtain an enzymolysis solution, wherein the adding amount of β -glucanase and the adding amount of neutral protease are respectively 10-12U/g and 22-25U/g based on the weight of the brown rice.
6. The preparation process according to claim 1, wherein in the step (4), the process conditions of the first fermentation step are as follows: anaerobic fermentation is carried out for 20-25 hours at the temperature of 40-45 ℃; the process conditions of the second step of fermentation are as follows: anaerobic fermentation is carried out for 10-12 hours at 40-45 ℃.
7. The process according to claim 1, wherein in the step (4), the lactobacillus powder is prepared from bifidobacterium, lactobacillus acidophilus and streptococcus thermophilus according to a mass ratio of 1: 0.4-0.6: 0.1-0.2 fermenting separately to obtain each strain fermentation liquor, mixing and fermenting, and post-treating.
8. A lactic acid fermented rice juice fermentation liquid rich in aminobutyric acid obtained by the preparation process of any one of claims 1 to 7.
9. The use of a lactic acid fermented rice juice broth with high aminobutyric acid content according to claim 8 for preparing rice juice beverage.
10. A rice juice beverage prepared by adding 8-10 times of water to the fermentation liquid of claim 8.
CN202010209450.7A 2020-03-23 2020-03-23 Preparation process of lactic acid fermentation rice juice fermentation liquor highly rich in aminobutyric acid Pending CN111248383A (en)

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