CN116004346A - Brewing method of intestinal tract probiotic glutinous rice prebiotics - Google Patents

Abstract

The invention discloses a brewing method of intestinal probiotics glutinous rice prebiotics, which is prepared by taking black brown glutinous rice, white brown glutinous rice and red brown glutinous rice produced in the county of abaca, guangdong as main raw materials, wherein the glutinous rice keeps germs and chaff.

Description

Brewing method of intestinal tract probiotic glutinous rice prebiotics

Technical Field

The invention relates to a brewing method of health-care food, belonging to the technical field of biological food.

Background

The Hakka mother wine is also called Hakka yellow wine, has reddish brown transparent wine color, is fragrant and sweet, has the effects of tonifying qi and activating blood, activating collaterals and dredging channels, enriching blood and generating blood, warming intestines and stomach and the like, is deeply favored by families, and is greatly limited by applicable people due to higher alcohol content, so that the brewing process of the Hakka mother wine is improved, and the brewing process is widely applicable to people and has intestinal probiotics.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention provides a brewing method of intestinal tract probiotic waxy brewing prebiotics.

The technical scheme adopted for solving the technical problems is as follows:

a brewing method of intestinal tract probiotic glutinous rice prebiotics is prepared from black brown glutinous rice, white brown glutinous rice and red brown glutinous rice produced in the county of abaca, guangdong, wherein the black brown glutinous rice, white brown glutinous rice and red brown glutinous rice retain germs and chaff, and the brewing method comprises the following steps:

(1) Selecting materials: and (5) selecting black brown glutinous rice, white brown glutinous rice and red brown glutinous rice, mixing and cleaning to obtain brewing raw materials.

(2) Soaking: soaking the brewing materials in mineral water or purified water for more than 24 hr to allow the materials to fully absorb water.

(3) Steaming: steaming the raw materials in a steam pot at 100-110deg.C under 1.1-1.5Mpa for 2 hr.

(4) And (3) mixing bacteria: taking out the steamed raw materials, spreading and airing to 38 ℃, scattering the ML191 saccharomycetes, and uniformly stirring, wherein the dry weight ratio of the raw materials to the ML191 saccharomycetes is 100:1.

(5) Fermentation: putting the raw materials after mixing with bacteria into a ceramic jar, punching holes in the middle of the raw materials to the bottom of the jar, maintaining the temperature in the ceramic jar at 30-35 ℃, adjusting the pH value to 7.1-7.2, and fermenting for 24 hours until the raw materials are soft.

(6) Juice is taken: delivering the fermented raw materials into a squeezing device to squeeze juice.

(7) And (3) inactivating: heating the juice to above 100deg.C for 30 min for inactivating.

(8) And (5) storing: filtering the inactivated juice, and vacuum sealing and storing by adopting a ceramic or food grade stainless steel tank.

The preparation process of the ML191 saccharomycetes is as follows:

soaking sweet wine Qu Jiaquan in water or purified water for 2 hr to obtain suspension, adding the suspension into MS culture medium, culturing for 24 hr at 28deg.C with vibration frequency of 120 times/min, culturing to obtain ML191 yeast suspension, and naturally drying to obtain ML191 yeast.

The beneficial effects of the invention are as follows: the invention is brewed by taking black brown glutinous rice, white brown glutinous rice and red brown glutinous rice produced in the county of the abaca in Guangdong province as main raw materials, the glutinous rice keeps germs and chaff, the conversion rate of probiotic trisaccharide to hexasaccharide in brewed prebiotics reaches more than 20%, and the obtained prebiotics are rich in polydextrose, L-arabinose, other oligosaccharides and the like, and have certain intestinal tract probiotics.

Drawings

The invention will be further described with reference to the drawings and examples.

FIG. 1 is a mass spectrum of a prebiotic of the present invention after 5-fold dilution.

Figure 2 is a mass spectrometric view of the prebiotic of the present invention after purification.

Detailed Description

A brewing method of intestinal tract probiotic glutinous rice prebiotics is prepared by taking black glutinous rice, white glutinous rice and red glutinous rice produced in the county of abaca, guangdong as main raw materials, wherein the black glutinous rice, white glutinous rice and red glutinous rice retain germs and chaff, and the black glutinous rice, white glutinous rice and red glutinous rice of the property have the advantage of being rich in selenium, and the germs account for 2% -3.5% of the weight of rice grains and are nutrition essence of rice. The embryo bud contains more than 2% of protein and fat, multiple vitamins and microelements required by human body, wherein the amino acid composition is balanced, and the embryo bud is full-value protein. The fat in the germ accounting for 70-80% of the weight is unsaturated fatty acid, the natural vitamin E is up to 2-3 mg/g, and the germ also contains nutrient components such as plant sterol, oryzanol, crude protein, unsaturated acid of human body, folic acid, vitamins, ferment and the like.

The brewing method comprises the following steps:

(1) Selecting materials: and (5) selecting black brown glutinous rice, white brown glutinous rice and red brown glutinous rice, mixing and cleaning to obtain a brewing raw material. The dosage of the black brown glutinous rice, the white brown glutinous rice and the red brown glutinous rice is not strictly required, and is generally 1:1:1, black brown glutinous rice, white brown glutinous rice and red brown glutinous rice are preferably produced in the same year, and the activity of the raw materials can be ensured.

(2) Soaking: soaking the brewing material in mineral water or purified water for 24-28 hr to make the material fully absorb water (in this invention, all are normal temperature and normal pressure, the water surface is higher than the material, so that the material fully absorbs water and swells.

(3) Steaming: the raw materials are put into a steam pot for steaming, the pressure in the steam pot is 1.1-1.5Mpa, the steam temperature is 100-110 ℃, and the steaming time is 2 hours, so that the raw materials are well-cooked and no scorching and caking conditions exist.

(4) And (3) mixing bacteria: taking out the steamed raw materials, spreading and airing to 38 ℃, scattering the ML191 saccharomycetes while stirring, and uniformly stirring, wherein the dry weight ratio of the raw materials to the ML191 saccharomycetes is 100:1.

(5) Fermentation: placing the mixed raw materials into a ceramic jar, punching holes in the middle of the raw materials to the bottom of the jar, maintaining the temperature in the ceramic jar at 30-35 ℃, adjusting the pH to 7.1-7.2 by adopting a food-grade pH regulator, fermenting for 24 hours until the raw materials are soft, converting the raw materials into various monosaccharides and oligosaccharides with probiotics, and generating prebiotics substances such as xylo-oligosaccharide, L-arabinose and other oligosaccharides.

(6) Juice is taken: the fermented raw materials are sent into a squeezing device to squeeze juice, no special requirements are imposed on a juicer for squeezing the juice, and the juice is squeezed by a general food juicer, so that solid impurities with larger particles are filtered.

(7) And (3) inactivating: heating the juice to above 100deg.C for 30 min for inactivating.

(8) And (5) storing: filtering the inactivated juice with 200 mesh sieve, and vacuum sealing with ceramic or food grade stainless steel tank. The vacuum degree is not strictly required, and the negative pressure in the tank is ensured.

The preparation process of the ML191 saccharomycetes is as follows:

soaking sweet wine Qu Jiaquan in water or purified water for 2 hours to obtain a suspension, adding the suspension into an MS culture medium for culturing for 24 hours at 28 ℃ with the vibration frequency of 120 times/min, obtaining ML191 saccharomycete suspension after culturing, and naturally drying to obtain the ML191 saccharomycete. The sweet distiller's yeast is commercially available at present, and the sweet distiller's yeast adopted by the invention is sweet distiller's yeast produced by Jiangsu Xinshenao biotechnology limited company, and has the product standard number: Q/XSASW0001S. After the re-culture, the sweet distiller's yeast can convert maltose, panose and glucose generated by fermentation into probiotic trisaccharides to hexasaccharides in the fermentation process, and the conversion rate reaches more than 20%.

The experimental conditions for the fermentation of ML191 yeasts are as follows:

from the matrix materials of different sources, 8 strains of positive strains which can generate glucan and metabolize citric acid are separated, 8 strains of strains which can convert glucose into glucan and 6 strains of positive strains of citric acid metabolism are separated. Based on the glucan production, the production speed is determined by preliminary measurement results of polysaccharide and ML191 saccharomycetes activity in rice, 6 strains with higher glucan activity relative to the ML191 saccharomycetes are determined, glucose is used as a carbon source, rice starch and yeast extract are used as nitrogen sources, the pH control and vibration oxygenation in the fermentation process can greatly improve the proliferation speed and cell biomass of the monilia, the yield of the ML191 saccharomycetes can be obviously improved, and the activity stability of the ML191 saccharomycetes can be obviously improved. Thus, it was determined that the MS medium was a moniliforme oligosaccharide optimized fermentation medium whose enzyme-producing optimized fermentation conditions were 28℃and 120 times/min shaking culture.

The specific reaction band of the strain ML191 yeast is obtained after SDS-PAGE separation, and the result shows that the strain ML191 yeast can produce molecular weight glucanase with the molecular weight of 183kDa and 142kDa respectively, the molecular weight of the strain ML191 yeast exists in the supernatant and on the cell surface at the same time, and the molecular weight of the strain ML191 yeast can exist on the cell surface only: strain PC12 can produce three ML191 yeasts of molecular weight 148 kDa,138 kDa and 115kDa, respectively, the former two belonging to extracellular enzymes, the latter being associated with cells. Similar to the strain ML191 yeast, the strain L4 can produce two kinds of ML191 yeast which exist on the supernatant and the cell surface at the same time, the molecular weight is 145 kDa and 136kDa respectively, and the dextran sucrase with the molecular weight of 115kDa exists only in the supernatant. Enzyme activity tests show that the glucansucrase activities in the whole cell fermentation broths and supernatants of the strains ML191 saccharomycetes, DM1-2, PC13 and L4 are 5.204U and 710U/mL, 1.638U and 1.352U/mL,3.244 and 1.118U/mL, 3.250U and 1.006U/mL respectively; and the proportion of extracellular enzymes is 90.5%,82.5%,34.5% and 31%, respectively.

The dextran sucrase in the supernatant is separated and concentrated by adopting an ammonium sulfate precipitation-centrifugal separation-dialysis-polyethylene glycol concentration-freeze-drying method, and the activity of the dextran ML191 yeast in the strain ML191 yeast, the DM1-2, the pc13 and the L4 crude enzyme powder is respectively 10.7U/mg, 2.7U/mg, 2.8U/mg and 2.5U/mg according to the dry weight of cells. The enzyme activities of the 4 strains are 24.4U/mg, 6.5U/mg, 4.2U/mg and 4.0U/mg respectively according to the measurement of the protein content. Recovery rates of enzyme activities of the respective strains were 32%,32%,44% and 43%, respectively. After 2 weeks of storage at-18 ℃, the crude enzyme powder activities of the 4 strains were reduced by 31.6%,48.2%,41.2% and 37.2%, respectively. The dextran ML191 yeast in the supernatant was purified by low temperature ultrafiltration (100 kDa cut-off) to obtain concentrated enzyme solutions, and the strain ML191 yeast was obtained from 500mL of fermentation broth, with concentrated enzyme forces of DM1-2, PC13 and L4 of 75.8U/mL, 18.7U/mL, 15.8U/mL and 13.9U/mL, respectively, and with dextran sucrase activity recovery rates of 76%,74%,69% and 73%, respectively. After 2 weeks of storage at-18 ℃, the concentrated enzyme activities decreased by 6.6%,11.5%,9.1% and 6.4%, respectively. Monitoring of the substrate and product of the fermentation of monilia to oligosaccharides at a glucose to panose ratio of 2:1 showed that sucrose and maltose were rapidly consumed in the logarithmic growth phase with significant amounts of oligosaccharides being produced, thus determining the optimal medium fermentation parameters for the ML191 yeast: taking amylopectin glutinous rice as a main raw material, thoroughly steaming and spreading the glutinous rice to be cool to 30 degrees, then mixing the glutinous rice with yeast, adding 1% of ML191 yeast, and fermenting for 24 hours to obtain the high-yield probiotic low-glucose.

The initial pH value, inoculation amount and culture temperature of the produced glucose oligomer are researched through a single factor experiment, the pH7.1-pH7.2 is determined through response surface analysis, the inoculation amount is 1%, the fermentation temperature is 25.0-35.0 ℃ and is the proper culture condition for the optimal production of the glucose oligomer by the ML191 yeast, and the theoretical yield of the total prebiotics of the glucose oligomer reaches 32g/L.

Through the analysis, the fermentation conditions selected by the invention are as follows: fermenting at 30-35deg.C and pH of 7.1-7.2 for 24 hr. The prebiotics of the invention have extremely low alcohol content (less than 5 percent, less than 5.5 percent is the category of alcoholic beverage), and are applicable to wider crowds.

The mass spectrum detection result (negative ion mode [ M-H ] of the waxy brewing prebiotics obtained by the invention after being diluted by 5 times] ^- ) As shown in FIG. 1, the detection results of the purification mass spectrum are shown in FIG. 2, and in FIG. 2, monosaccharides 179.0557, disaccharides 341.1080, trisaccharides 503.1639, tetrasaccharides 665.2149 and pentasaccharides 827.2719, the characteristic molecular weight of the oligosaccharides with the polymerization degree of 1-5 can be found in the mass spectrum, which indicates the content of the oligosaccharides (prebiotics) in the sample, and the content of the oligosaccharides with the probiotics is up to 32g/l through re-verification.

The applicant Guangdong Mei Lingquan ecological food limited company entrusts Guangdong province food safety key laboratory to research the glutinous rice prebiotics obtained by the invention, and the Guangdong province food safety key laboratory provides an experimental report in 2022, wherein the original text of the experimental report conclusion is extracted as follows:

in this experimental investigation, it was found that 88% of subjects had little stress during the "Mei Lingquan" waxy prebiotic intervention but could solve by themselves, 80% of subjects had meals on schedule and had healthy diets, 48% of subjects moved weekly, these health habits might be important reasons for their ability to maintain intestinal health and weight, however, poor night stay habits (78%) also had a negative risk. Meanwhile, body cold conditions and other body characterizations related to dysmenorrhea are investigated, and the interference of the wine can effectively improve the cold conditions of menstrual period hands and feet of 51% (18/35) of subjects, the cold conditions of menstrual period abdomen of 64% (18/28) of subjects and the acne growth conditions of menstrual period faces of 48% (11/23) of subjects, so that the problems related to menstrual cold conditions, poor body characterization and the like can be effectively improved by regularly drinking the 'Mei Lingquan' glutinous rice prebiotics.

In addition, the research shows that 80% (4/5) of the diarrhea conditions of the subjects are improved after the 'Mei Lingquan' glutinous rice prebiotics are intervened, 100% (6/6) of the abdominal distension conditions of the subjects are improved, and 100% (6/6) of the constipation conditions of the subjects are improved, so that the 'Mei Lingquan' glutinous rice prebiotics have a certain intestinal tract probiotics effect.

The above embodiments do not limit the scope of the invention, and various changes and modifications may be made therein without departing from the spirit and scope of the invention as claimed.

Claims (1)

1. A brewing method of intestinal tract probiotic glutinous rice prebiotics is characterized in that the intestinal tract probiotic glutinous rice prebiotics are brewed by taking black glutinous rice, white glutinous rice and red glutinous rice produced by the county of abaca in Guangdong as main raw materials, and the black glutinous rice, the white glutinous rice and the red glutinous rice retain germs and chaff, and the brewing method is as follows:

(1) Selecting materials: mixing black brown glutinous rice, white brown glutinous rice and red brown glutinous rice, and cleaning to obtain brewing raw materials;

(2) Soaking: soaking the brewing materials in mineral water or purified water for more than 24 hours to allow the materials to fully absorb water;

(3) Steaming: steaming the raw materials in a steam pot at 100-110deg.C under 1.1-1.5Mpa for 2 hr;

(4) And (3) mixing bacteria: taking out the steamed raw materials, spreading and airing to 38 ℃, scattering the ML191 saccharomycetes, and uniformly stirring, wherein the dry weight ratio of the raw materials to the ML191 saccharomycetes is 100:1, a step of;

(5) Fermentation: putting the raw materials after mixing with bacteria into a ceramic jar, punching holes in the middle of the raw materials to the bottom of the jar, maintaining the temperature in the ceramic jar at 30-35 ℃, adjusting the pH value to 7.1-7.2, and fermenting for 24 hours until the raw materials are soft;

(6) Juice is taken: delivering the fermented raw materials into squeezing equipment to squeeze juice;

(7) And (3) inactivating: heating the juice to above 100deg.C for 30 min for inactivating;

(8) And (5) storing: filtering the inactivated juice, and vacuum-pumping and sealing the juice for storage by adopting a ceramic or food-grade stainless steel tank;

the preparation process of the ML191 saccharomycetes is as follows:

soaking sweet wine Qu Jiaquan in water or purified water for 2 hr to obtain suspension, adding the suspension into MS culture medium, culturing for 24 hr at 28deg.C with vibration frequency of 120 times/min, culturing to obtain ML191 yeast suspension, and naturally drying to obtain ML191 yeast.

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