CN115058305B - Preparation method and application of probiotics rice distiller's yeast - Google Patents
Preparation method and application of probiotics rice distiller's yeast Download PDFInfo
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- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 114
- 235000009566 rice Nutrition 0.000 title claims abstract description 114
- 239000006041 probiotic Substances 0.000 title claims abstract description 105
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 105
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 82
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 240000007594 Oryza sativa Species 0.000 title 1
- 241000209094 Oryza Species 0.000 claims abstract description 113
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- 230000000529 probiotic effect Effects 0.000 claims abstract description 75
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- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 claims 1
- 150000004985 diamines Chemical class 0.000 claims 1
- 229910052739 hydrogen Inorganic materials 0.000 claims 1
- 239000001257 hydrogen Substances 0.000 claims 1
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 13
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- 244000199866 Lactobacillus casei Species 0.000 description 8
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- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 239000012086 standard solution Substances 0.000 description 4
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 3
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 240000002582 Oryza sativa Indica Group Species 0.000 description 2
- 241000235527 Rhizopus Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
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- 230000007760 free radical scavenging Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
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- 241000235342 Saccharomycetes Species 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 229910021536 Zeolite Inorganic materials 0.000 description 1
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
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- 230000008901 benefit Effects 0.000 description 1
- 238000013124 brewing process Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- CYDMQBQPVICBEU-UHFFFAOYSA-N chlorotetracycline Natural products C1=CC(Cl)=C2C(O)(C)C3CC4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-UHFFFAOYSA-N 0.000 description 1
- CYDMQBQPVICBEU-XRNKAMNCSA-N chlortetracycline Chemical compound C1=CC(Cl)=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-XRNKAMNCSA-N 0.000 description 1
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- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
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- 238000011010 flushing procedure Methods 0.000 description 1
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- 230000008821 health effect Effects 0.000 description 1
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- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
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- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
- 229940004208 lactobacillus bulgaricus Drugs 0.000 description 1
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- 230000031700 light absorption Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000005499 meniscus Effects 0.000 description 1
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- 230000001502 supplementing effect Effects 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
- C12G3/021—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn
- C12G3/022—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn of botanical genus Oryza, e.g. rice
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/245—Lactobacillus casei
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- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
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- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
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Abstract
The invention discloses a preparation method and application of probiotic rice distiller's yeast, wherein the preparation method comprises the following steps: preparing MRS liquid culture medium; inoculating 0.05% of probiotic bacteria powder into an MRS liquid culture medium for culturing to obtain probiotic bacteria liquid; selecting full glutinous rice, cleaning, adding enough water, and beginning soaking; draining the soaked glutinous rice, steaming in a container until the outside is hard and the inside is soft, and the inside is free of white cores and loose and not rotten, so as to obtain glutinous rice; cooling glutinous rice with cool boiled water, regulating water content, adding saccharifying enzyme and probiotic bacteria liquid into the glutinous rice, and stirring; fermenting glutinous rice in a constant temperature incubator, filtering rice mash to obtain probiotic rice wine; and mixing the probiotic rice wine with rice flour, and drying and crushing to obtain the probiotic rice wine yeast. The probiotic rice distiller's yeast provided by the invention can be suitable for industrial mass production of probiotic rice wine, has a plurality of probiotics, has a nutrition and health care effect on human bodies, and has extremely profound significance on innovation and development of the rice wine industry.
Description
Technical Field
The invention relates to the technical field of rice wine brewing, in particular to a preparation method and application of probiotic rice distiller's yeast.
Background
The rice wine is prepared by taking glutinous rice as a raw material, steaming and cooling the glutinous rice, and then adding distiller's yeast for fermentation. However, glutinous rice wine has the problems of light taste and poor stability, and is still produced in a small scale, and the variety process is single in traditional. Rice distiller's yeast is the main starter for rice wine brewing, and the interaction among microorganisms in the distiller's yeast can provide rich metabolites for rice wine brewing, so that the quality and taste of the wine are determined. At present, pure rhizopus and saccharomycetes are commonly adopted for starter propagation for rice wine production, and although fermentation quality is controlled and harmful microorganisms are removed, the starter propagation cannot have flavor and taste, and the prepared starter propagation has low nutritive value and is difficult to meet the demands of people on low-alcohol, nutritional and healthy fermented wine. Therefore, scientifically preparing the functional rice distiller's yeast can concentrate dominant strains, improve the quality of the distiller's yeast, homogenize the quality of the distiller's yeast and further promote the healthy development of the rice wine industry to become a trend.
Probiotics are active microorganisms beneficial to a host, can improve the micro-ecological balance of the host, and have the effects of promoting nutrient absorption and keeping intestinal health. Chinese patent No. CN 114058467A discloses a method for preparing probiotic rice distiller's yeast, the method for preparing reinforced rice distiller's yeast comprises the following steps: sterilizing indica rice flour at 100-120 deg.c, inoculating rhizopus, adding sterile water in the amount of 85% of indica rice flour, adding lactobacillus casei, streptococcus thermophilus, lactobacillus bulgaricus, lactobacillus reuteri and lactobacillus acidophilus, and co-culturing to obtain the probiotic distiller's yeast. The method adds a large amount of ingredients, greatly increases the input cost of raw materials, and the rice wine prepared by using the rice wine yeast has the defects of complex steps, low content of probiotics and the like, and is not beneficial to popularization and application.
In view of this, it is still required to search how to research the distiller's yeast produced by the finished product of the probiotic rice wine at present, develop a high-activity probiotic rice distiller's yeast with stable quality, the simple technological process when using it in rice wine brewing, make people eat rice wine, can obtain dual nutrition health effects of rice wine and probiotic especially important, so offer the reference for innovation and sustainable development of rice wine trade.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a preparation method of probiotic rice distiller's yeast and application thereof, and the probiotic rice distiller's yeast is further prepared by adding high-activity probiotics into cooked and cooled glutinous rice, and the probiotic rice distiller's yeast is used for brewing rice wine without adding other materials, so that the nutritional value of the rice wine can be improved, and the probiotic rice wine with high activity can be obtained.
In order to achieve the above purpose, the present invention provides the following technical solutions: a preparation method of probiotic rice distiller's yeast, comprising the following steps:
s1, preparing an MRS liquid culture medium;
s2, inoculating 0.05% of probiotic bacteria powder into an MRS liquid culture medium for culturing to obtain probiotic bacteria liquid;
s3, selecting the glutinous rice with full grains, cleaning, adding enough water, and beginning to soak;
s4, draining the soaked glutinous rice, steaming in a container until the glutinous rice is hard outside and soft inside, and has no white core inside and is loose but not rotten, so as to obtain glutinous rice;
s5, cooling the glutinous rice with cool boiled water, regulating the water content of the glutinous rice, respectively adding saccharifying enzyme and probiotics liquid into the glutinous rice, and uniformly stirring;
s6, placing the glutinous rice subjected to the step S5 into a constant temperature incubator for fermentation, and filtering rice mash to obtain probiotic rice wine;
and S7, mixing the probiotic rice wine and rice flour, and drying and crushing to obtain the probiotic rice wine yeast.
Preferably, the preparation of the MRS liquid culture medium comprises the following steps: weighing 10g of peptone, 10g of beef extract, 5g of yeast extract and K 2 HPO 4 2g，KH 2 PO 4 2g, diamine hydrogen citrate 2g, sodium acetate 5g, mgSO 4 ·7H 2 O 0.58g，MnSO 4 0.25g of Tween 80, 1mL of water 1000mL, pH of 6.2-6.4 and sterilizing at 121 ℃ for 15min to obtain the MRS liquid culture medium.
Preferably, the probiotic bacteria powder in the step S2 is lactobacillus rhamnosus BV, lactobacillus salivarius AP or lactobacillus casei CS; the culture was anaerobic at 35℃until the cell growth logarithmic phase.
Preferably, the washing time in the step S3 is 2-3 times, and the soaking time is 8-12 hours.
Preferably, in the step S5, the temperature of the glutinous rice is reduced to 35 ℃, the moisture is regulated to 55% -65%, and 0.2% -0.3% of saccharifying enzyme and 1% -1.5% of probiotics bacterial liquid are respectively added into the glutinous rice.
Preferably, the fermentation temperature in the constant temperature incubator of the step S6 is 28-32 ℃ and the fermentation time is 36-48 h.
Preferably, in the step S7, the mass ratio of the probiotic rice wine to the rice flour is 1:3, and the drying specifically comprises: drying at 30deg.C until the water content is below 10%.
In addition, in order to achieve the above purpose, the present invention further provides the following technical solutions: an application of probiotic rice distiller's yeast in probiotic rice wine.
The beneficial effects of the invention are as follows:
1) According to the preparation method of the probiotic rice distiller's yeast, the preparation of the probiotic rice distiller's yeast is completed by cooking glutinous rice, mixing with yeast, culturing bacteria, saccharifying, fermenting, adding rice flour into the probiotic rice wine. The number of the live probiotics is easily influenced by fermentation process conditions such as ethanol concentration, acidity and the like, and therefore, the probiotics can reach the optimal peak value through the limited control of the optimal temperature and time of the probiotics during the bacterial activation. In order to make the probiotics more suitable for survival and growth in the rice wine brewing process, the method adopts the production process of the probiotic rice wine to prepare the probiotic rice wine yeast, so that the probiotics can be suitable for the rice wine fermentation environment in advance, and the survival rate of the probiotics is further improved. Therefore, the probiotics rice wine yeast of the invention contains much higher viable count than other probiotics rice wine yeast.
2) The probiotic rice distiller's yeast is the integration of probiotics and rice distiller's yeast, and can be directly used for brewing the probiotic rice wine without adding rice distiller's yeast additionally, the preparation process is simple, the cost is greatly saved, and industrialization is easy to realize. The rice wine prepared by the probiotics rice wine starter has mellow taste, better quality, green and safe performance, and can enhance the immune function and disease resistance of human organisms after long-term eating.
Detailed Description
In the following, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
A preparation method of probiotic rice distiller's yeast comprises the following specific steps:
(1) Activating strains: (1) preparing MRS liquid culture medium; (2) inoculating probiotic bacteria powder into MRS culture medium for culturing to obtain mature probiotic bacteria liquid;
(2) The rice wine is prepared by (1) soaking rice: selecting full glutinous rice, cleaning, adding enough water, and soaking for 8-12 h; (2) and (3) cooking: draining glutinous rice, steaming in a steamer until the glutinous rice is hard outside and soft inside, and has no white core inside, and is loose but not rotten; (3) and (3) mixing yeast: cooling the glutinous rice to 35 ℃ with cool boiled water, regulating the water content of the glutinous rice to 55% -65%, adding saccharifying enzyme and probiotics bacterial liquid according to 0.2% -0.3% and 1% -1.5% of raw materials respectively, and uniformly stirring. (4) Fermentation: placing the cooked glutinous rice into a constant temperature incubator, fermenting at 28-32 deg.C for 36-48 h.
(3) The rice wine starter is prepared by (1) mixing fermented rice wine and rice flour at a mass ratio of 1:3, and oven drying at 30deg.C until water content is below 10%; (2) and drying and crushing to obtain the probiotic rice distiller's yeast.
Further, preparation of MRS liquid culture medium: weighing eggBai 10g, beef extract 10g, yeast extract 5g, K 2 HPO 4 2g，KH 2 PO 4 2g, diamine hydrogen citrate 2g, sodium acetate 5g, mgSO 4 ·7H 2 O 0.58g，MnSO 4 0.25g of Tween 80, 1mL of water 1000mL, pH of 6.2-6.4 and sterilizing at 121 ℃ for 15min to obtain the MRS liquid culture medium.
The liquid culture medium comprises various carbon sources, nitrogen sources, vitamins and trace elements for growth of probiotics.
Further, the probiotics used for fermentation, including but not limited to lactobacillus rhamnosus BV, lactobacillus salivarius AP or lactobacillus casei CS, should be guaranteed to have an exact health efficacy and thus exert a beneficial effect.
Further, the method comprises the steps of activating and culturing the probiotics for fermentation before fermenting and culturing the probiotics powder for fermentation in rice wine, inoculating the probiotics for fermentation on an MRS culture medium for culturing to a logarithmic phase, and inoculating the probiotics for fermentation in glutinous rice for fermenting and culturing.
Further, the glutinous rice is full and full grains, and the glutinous rice is cleaned for 2 to 3 times;
further, the glutinous rice is soaked for 8-12 h, water is poured first when the glutinous rice is soaked, then the glutinous rice is soaked in water, the water surface is higher than the rice flour, and the soaked glutinous rice requires the particles to be kept complete.
Further, the drained glutinous rice is placed in a steamer drawer and steamed for about 30min by intense fire, so that the cooked rice is hard outside and soft inside without white core inside and loose without rotting.
Further, cooling glutinous rice to 35 ℃ with cool boiled water, regulating the water content to 55% -65%, adding saccharifying enzyme (50000U/g) and probiotics bacterial liquid according to 0.2% -0.3% and 1% -1.5% of raw materials, and stirring uniformly to obtain the mixture.
Further, the fermentation temperature in the constant temperature incubator is 28-32 ℃ and the fermentation time is 36-48 h.
Further, the probiotic rice wine and rice flour are mixed according to the mass ratio of 1:3, dried at 30 ℃ until the water content is lower than 10%, and crushed to obtain the probiotic rice wine yeast.
The following are the following methods for measuring the number of living bacteria, physical and chemical indexes, DPPH radical scavenging rate, hydroxyl radical scavenging rate and total reducing force:
(1) Number of viable bacteria
Sterilizing the culture medium by using a detection culture medium of the viable count of the MRS culture medium, cooling to a certain temperature, and adding 25 mg/mL-30 mg/mL of the chlorotetracycline before pouring the culture medium into a flat plate; diluting the fermentation broth to a gradient of 10 -4 ～10 -6 The viable count is measured, and the obtained product is placed on an anaerobic workbench for culturing for 48 hours at 37 ℃ and then counted.
(2) Physical and chemical index
(1) The total acid adopts a pH potentiometric titration method, and the formula is as follows:
wherein, V-the volume of NaOH standard solution is consumed in the measurement of the sample, mL;
V 0 in blank test, the volume of NaOH standard solution is consumed, and the volume is mL;
c-concentration of NaOH standard solution, mol/L;
0.090-1 mL of 1mol/L NaOH standard solution is consumed, corresponding to grams of lactic acid, g/mmol;
V 1 the volume of the sample is aspirated, mL.
(2) The total sugar adopts the cheap-Ainong method, and the formula is as follows:
wherein, each 5mL of F-Fi Lin Jiaye and B solution is equal to the mass of glucose, g;
V 1 -the volume of sample dilution consumed during titration, mL;
V 2 the volume of the sample is aspirated, mL.
(3) The alcohol degree adopts a distillation method, and the operation is as follows:
accurately measuring 100mL of rice wine in a 500mL distillation flask by using a measuring cylinder, flushing the volumetric flask with 50mL of water for three times, merging the washing liquid into the 500mL distillation flask, adding a plurality of zeolite, connecting a snake-shaped condenser pipe, starting cooling water, slowly heating for distillation, collecting distillate, supplementing water to the scale when the distillate is close to the scale, uniformly mixing, putting into a clean and dry alcohol meter, inserting a thermometer, reading the scale indication value at the tangent position of a meniscus, and recording the temperature. Based on the measured alcohol measurement value and temperature, the alcohol measurement temperature concentration conversion table is checked and converted into alcohol degree at 20 ℃.
(3) DPPH radical scavenging rate
Adding 2mL of DPPH solution (with final concentration of 60 μmol/L) into 2mL of rice wine, mixing, standing at room temperature for light-shielding reaction for 30min, centrifuging at 7800r/min for 10min, collecting supernatant, and measuring absorbance A of sample at wavelength of 517nm S Each sample 3 groups were parallel. The formula is as follows: :
in which A s 2mL DPPH reagent is added with 2mL sample liquid;
A b 2mL of absolute ethanol is added with 2mL of sample liquid;
A c 2mL of DPPH reagent was added to 2mL of absolute ethanol.
(4) Hydroxyl radical scavenging rate
1mL of 1, 10-phenanthroline (2.5 mmol/L), 1mL of PBS (0.01 mol/L, pH 7.4) and 1mL of distilled water are taken, fully and uniformly mixed, and 1mL of FeSO is added 4 (2.5 mmol/L), and 1mL of H was added 2 O 2 (20 mmol/L), centrifuged at 600 r/min for 10min in a 37℃water bath for 1.5h, and the supernatant was assayed at 536nm for absorbance A 1 The method comprises the steps of carrying out a first treatment on the surface of the 1mL of distilled water was used in place of 1mL of H 2 O 2 Is A 0 The method comprises the steps of carrying out a first treatment on the surface of the 1mL of distilled water is replaced by 1mL of sample to be measured to be A 2 . The formula is as follows:
in which A 2 Containing samples and H 2 O 2 ；
A 1 -no sample, H-containing 2 O 2 ；
A 0 -no sample and H 2 O 2 。
(5) Total reducing force measurement
Will V C The concentrations were formulated to be 5. Mu.g/mL, 10. Mu.g/mL, 30. Mu.g/mL, 50. Mu.g/mL, 70. Mu.g/mL, respectively. Respectively sucking 1mL of each gradient into a test tube, adding 1mL of 1% potassium ferricyanide solution and 1mL of 0.1mol/L PBS (pH 6.6), uniformly mixing, reacting in a water bath at 50 ℃ for 20min, rapidly cooling in an ice-water bath, adding 1mL of 10% trichloroacetic acid solution, uniformly mixing, transferring into a 5mL centrifuge tube, centrifuging at 3000r/min for 5min, taking 1mL of supernatant, adding 1mL of water and 1mL of 0.1% ferric trichloride solution, uniformly mixing, measuring the light absorption value under the wavelength condition of 700nm, drawing a standard curve, and calculating the reducing force according to a formula obtained by the standard curve.
The sensory scoring criteria for the wine are shown in Table 1
Table 1 organoleptic scoring criteria for rice wine
Example 2
A preparation method of probiotic rice distiller's yeast comprises the following steps:
inoculating 0.05% lactobacillus rhamnosus BV on MRS culture medium, anaerobic culturing at 35deg.C for 8-10 hr until the logarithmic phase (OD) of thallus growth 600 =1.0), the number of viable bacteria is about (1 to 3) ×10 8 CFU/mL。
Selecting full glutinous rice, washing 2-3 times with clear water, soaking for 8-12 hours, draining, placing the glutinous rice in a steamer tray, steaming for about 30 minutes with strong fire, cooling the glutinous rice to 35 ℃ with cool boiled water, adjusting the moisture of the glutinous rice to 55-65%, adding saccharifying enzyme (50000U/g) and probiotics bacterial liquid respectively according to 0.2-0.3% and 1-1.5% of raw materials, stirring uniformly, making into a non-loose nest shape by using the viscosity of the rice grains, and sealing and fermenting for 36-48 hours at 28-32 ℃.
Taking 10kg of prepared rice wine per 10kg of glutinous rice, adding 30kg of rice flour, uniformly mixing, drying at 30 ℃ until the water content is lower than 10%, and crushing to obtain the probiotic rice wine yeast. The distiller's yeast obtained under the condition is milky white to pale yellow powder, has special distiller's yeast smell, and has no foreign matter visible to normal vision. The quality index of the detected probiotic rice distiller's yeast is shown in table 2.
Table 2 example 2 method rice wine yeast quality index
An application of probiotic rice distiller's yeast in probiotic rice wine: rice wine fermentation test of the probiotic rice wine yeast prepared in example 2.
The rice wine without lactobacillus rhamnosus BV rice wine yeast is used as a control group, and the test group is rice wine prepared by only adding lactobacillus rhamnosus BV rice wine yeast, and the result shows that the addition of lactobacillus rhamnosus BV rice wine yeast can improve the total acid and the alcohol degree of the rice wine and reduce the total sugar content in the rice wine, wherein the total acid is 6.13+/-0.08 g/L, and the total acid is improved by about 1.93g/L; the alcohol content was 5.9% vol. In addition, the DPPH free radical clearance of the lactobacillus rhamnosus BV rice wine is 70.12+/-0.05%, the hydroxyl free radical clearance is 73.16 +/-0.03 g/L, the total reducing force is 20.34+/-0.32 mug/mL, and the total number of viable bacteria is (1.94+/-0.08) multiplied by 10 8 The rice wine prepared by using the rice wine yeast has good palatability and has a sensory evaluation score of 86+/-2 minutes as shown in tables 3 and 4.
TABLE 3 physicochemical index and sensory score results of Rice wine according to example 2 method
TABLE 4 antioxidant Activity results of Rice wine according to example 2 method
Example 3
A preparation method of probiotic rice distiller's yeast comprises the following steps:
inoculating 0.05% Lactobacillus salivarius AP onto MRS culture medium, anaerobic culturing at 35deg.C for 15-17 hr, and culturing until the logarithmic phase (OD) 600 =1.0), the number of viable bacteria is about (5 to 7) ×10 7 CFU/mL。
Selecting full glutinous rice, washing 2-3 times with clear water, soaking for 8-12 h, draining, spreading the rice in a steamer tray, steaming for about 30min with strong fire, cooling the glutinous rice to 35 ℃ with cool boiled water, regulating the water content of the glutinous rice to 55-65%, adding saccharifying enzyme (50000U/g) and probiotics bacterial liquid according to 0.2-0.3% and 1-1.5% of raw materials respectively, stirring uniformly, making into a non-loosening nest shape by using the viscosity of the rice grains, and sealing and fermenting for 36-48 h at 28-32 ℃.
Taking 10kg of prepared rice wine per 10kg of glutinous rice, adding 30kg of rice flour, uniformly mixing, drying at 30 ℃ until the water content is lower than 10%, and crushing to obtain the probiotic rice wine yeast. The distiller's yeast obtained under the condition is milky white to pale yellow powder, has special distiller's yeast smell, and has no foreign matter visible to normal vision. The quality index of the detected probiotic rice distiller's yeast is shown in Table 5.
TABLE 5 example 3 method Rice distiller's yeast quality index
Example 3 Rice wine fermentation test of probiotic rice distiller's yeast
The rice wine without lactobacillus salivarius AP rice wine yeast is used as a control group, the test group is rice wine prepared by adding only lactobacillus salivarius AP, and the result shows that the adding of lactobacillus salivarius AP rice wine yeast can improve the total sugar and the alcohol degree of the rice wine, wherein the alcohol degree is 3.4+/-0.13% Vol, the total sugar content is 240+/-20 g/L, and the total acid content is 5.5+/-0.15 g/L. In addition, the DPPH radical clearance of the lactobacillus salivarius AP rice wine is 68.26+/-0.05%, the hydroxyl radical clearance is 71.51+/-0.01%, and the total reducing power is 19.45+ -0.32 μg/mL, total viable count of (7.94+ -0.08). Times.10 7 CFU/mL, the rice wine prepared by using the rice wine yeast has mellow taste, and the sensory evaluation score is 90+ -4 minutes, as shown in tables 6 and 7.
TABLE 6 physicochemical index and sensory score results of Rice wine according to example 3 method
TABLE 7 antioxidant Activity results of Rice wine according to example 3 method
Example 4
A preparation method of probiotic rice distiller's yeast comprises the following steps:
inoculating 0.05% Lactobacillus casei CS on MRS culture medium, anaerobic culturing at 35deg.C for 6-8 hr, and culturing until the logarithmic phase (OD) 600 =1.0), the number of viable bacteria is about (1 to 3) ×10 7 CFU/mL。
Selecting 10kg of full glutinous rice, washing 2-3 times with clear water, soaking for 8-12 hours, draining, spreading the rice in a steamer tray, steaming for about 30 minutes with strong fire, cooling the glutinous rice to 35 ℃ with cool boiled water, adjusting the moisture of the glutinous rice to 55-65%, adding saccharifying enzyme (50000U/g) and probiotics bacteria liquid according to 0.2-0.3% and 1-1.5% of raw materials respectively, stirring uniformly, making into a non-loose nest shape by using the viscosity of the rice grains, and sealing and fermenting for 36-48 hours at 28-32 ℃.
Taking 10kg of prepared rice wine per 10kg of glutinous rice, adding 30kg of rice flour, uniformly mixing, drying at 30 ℃ until the water content is lower than 10%, and crushing to obtain the probiotic rice wine yeast. The distiller's yeast obtained under the condition is milky white to pale yellow powder, has special distiller's yeast smell, and has no foreign matter visible to normal vision. The quality index of the detected probiotic rice distiller's yeast is shown in Table 8.
Table 8 example 4 method Rice distiller's yeast quality index
Example 4 Rice wine fermentation test of probiotic rice distiller's yeast
The rice wine without lactobacillus casei CS rice wine yeast is used as a control group, and the test group is only rice wine prepared by adding lactobacillus casei CS rice wine yeast, and the result shows that the adding lactobacillus casei CS rice wine yeast can reduce the alcoholic strength and the total sugar content of the rice wine, and reduce the total acid content of the rice wine, wherein the alcoholic strength is 2.5+/-0.17%Vol, the total sugar content is 232+/-21 g/L, and the total acid content is 2.8+/-0.28 g/L. In addition, the DPPH free radical scavenging rate of the lactobacillus casei CS rice wine is 68.26+/-0.05%, the hydroxyl free radical scavenging rate is 71.51+/-0.01%, the total reducing power is 19.45+/-0.32 mug/mL, and the total number of viable bacteria is (7.94+/-0.08). Times.10 7 CFU/mL, the rice wine prepared by using the rice wine yeast has mellow taste, and the sensory evaluation score is 87+ -4 minutes, as shown in tables 9 and 10.
Table 9 physicochemical index and sensory score results of rice wine according to example 4 method
Table 10 results of antioxidant activity of rice wine according to example 4 method
The rice wine prepared by taking the glutinous rice as the raw material and only adding the probiotic rice wine yeast has better taste, and the rice wine prepared by the process has a certain antioxidant capacity and can increase the nutritive value of the probiotic rice wine. The probiotic rice distiller's yeast can be suitable for industrialized mass production of probiotic rice wine, has multiple probiotics, and has nutrition and health promotion effects on human body. The invention provides a new idea for generating economic benefit by using rice wine yeast, and has extremely profound significance for innovation and development of rice wine industry.
Although the present invention has been described with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described, or equivalents may be substituted for elements thereof, and any modifications, equivalents, improvements and changes may be made without departing from the spirit and principles of the present invention.
Claims (7)
1. A preparation method of probiotic rice distiller's yeast, which is characterized by comprising the following steps:
s1, preparing an MRS liquid culture medium;
s2, inoculating 0.05% lactobacillus rhamnosus BV into an MRS liquid culture medium for culturing to obtain probiotic bacteria liquid, wherein the number of viable bacteria is 1-3 multiplied by 10 8 CFU/mL；
S3, selecting the glutinous rice with full grains, cleaning, adding enough water, and beginning to soak;
s4, draining the soaked glutinous rice, steaming in a container until the glutinous rice is hard outside and soft inside, and has no white core inside and is loose but not rotten, so as to obtain glutinous rice;
s5, cooling the glutinous rice to 35 ℃ by using cool boiled water, regulating the water content of the glutinous rice to 55% -65%, respectively adding 0.2% -0.3% of saccharifying enzyme and 1% -1.5% of probiotic bacteria liquid into the glutinous rice, and uniformly stirring;
s6, fermenting the glutinous rice obtained in the step S5 in a constant temperature incubator, filtering the rice mash to obtain the probiotic rice wine, wherein the DPPH free radical clearance is 70.12+/-0.05%, the hydroxyl free radical clearance is 73.16 +/-0.03 g/L, the total reducing force is 20.34+/-0.32 mug/mL, and the total number of viable bacteria is 1.94+/-0.08 multiplied by 10 8 CFU/mL；
And S7, mixing the probiotic rice wine and rice flour, and drying and crushing to obtain the probiotic rice wine yeast.
2. The probiotic rice wine yeast according to claim 1The preparation method of (2) is characterized in that: preparation of the MRS liquid culture medium: weighing peptone 10g, beef extract 10g, yeast extract 5g, K 2 HPO 4 2 g，KH 2 PO 4 2g, hydrogen diamine citrate 2g, sodium acetate 5g, mgSO 4 ·7H 2 O 0.58 g，MnSO 4 0.25g, tween 80 1mL, water 1000mL,pH 6.2~6.4 and sterilizing at 121 ℃ for 15min to obtain the MRS liquid culture medium.
3. The method for preparing the probiotic rice wine yeast according to claim 1, wherein the method comprises the following steps: the culture in the step S2 is anaerobic culture at 35 ℃ until the cell growth logarithmic phase.
4. The method for preparing the probiotic rice wine yeast according to claim 1, wherein the method comprises the following steps: the cleaning times in the step S3 are 2-3 times, and the soaking time is 8-12 hours.
5. The method for preparing the probiotic rice wine yeast according to claim 1, wherein the method comprises the following steps: and the fermentation temperature in the constant-temperature incubator in the step S6 is 28-32 ℃, and the fermentation time is 36-48 h.
6. The method for preparing the probiotic rice wine yeast according to claim 1, wherein the method comprises the following steps: in the step S7, the mass ratio of the probiotic rice wine to the rice flour is 1:3, and the drying is specifically as follows: drying at 30deg.C until the water content is below 10%.
7. Use of a probiotic rice wine yeast prepared by the method for preparing a probiotic rice wine yeast according to any one of claims 1-6 in a probiotic rice wine.
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