CN111690543A - Tremella aurantialba and application thereof in preparation of tremella aurantialba fermented soybean milk - Google Patents
Tremella aurantialba and application thereof in preparation of tremella aurantialba fermented soybean milk Download PDFInfo
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- CN111690543A CN111690543A CN202010618454.0A CN202010618454A CN111690543A CN 111690543 A CN111690543 A CN 111690543A CN 202010618454 A CN202010618454 A CN 202010618454A CN 111690543 A CN111690543 A CN 111690543A
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- 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
- C12R2001/645—Fungi ; Processes using fungi
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
- A23C11/106—Addition of, or treatment with, microorganisms
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Abstract
The invention relates to the technical field of food fermentation, and discloses tremella aurantialba and application thereof in preparing tremella aurantialba fermented soybean milk; the Tremella aurantialba is classified and named as Tremella aurantialba (Tremella aurantiaba), the strain name is NX-20, the Tremella aurantiaca is preserved in the China general microbiological culture Collection center, the preservation time is 2019, 10 and 15 days, and the preservation number is CGMCC NO. 18588. The invention breaks the limitation of single strain of fermented soybean milk in the market, creatively uses tremella aurantialba as a fermentation strain, and provides the preparation method of the health-care tremella aurantialba fermented soybean milk, which has the advantages of simple process, stable quality and easiness in large-scale production.
Description
Technical Field
The invention relates to the technical field of food fermentation, in particular to tremella aurantialba and application thereof in preparing tremella aurantialba fermented soybean milk.
Background
The fermented milk is an acidic milk product formed by fermenting milk and dairy products under the action of characteristic bacteria, and substances such as carbon dioxide, lactic acid, diacetyl and the like ensure that the traditional fermented milk has milk fragrance, sour taste and unique mouthfeel; the fermented milk has higher nutritional value and health care function compared with fresh milk due to the abundant substances such as protein, fatty acid, vitamins and the like, but in the aspect of strains, lactobacillus is mainly used for preparing the fermented milk, and a novel product based on new strains is not developed for a long time.
Tremella aurantialba (Tremella aurantiaca), belonging to Tremella, is a kind of plant phylum fungi, also called Tremella flavicans, Rumex japonicus (Tibetan language), Tremella aurantialba, Tremella flavicans, and Tremella cerebri. The golden fungus has better nourishing and nutritive value than colloidal fungi such as tremella, black fungus and the like, and is an ideal high-grade banquet delicacy and health care product. The main active component of the golden fungus is extracellular polysaccharide which has multiple functions of oxidation resistance, inflammation resistance, tumor resistance, hyperlipemia resistance and the like. The tremella aurantialba polysaccharide extract also has the function of enhancing the immunologic function of a human body, so that the tremella aurantialba polysaccharide extract is extremely high in nutritive value and is an ideal health-care product. Since Tremella aurantialba belongs to a kind of fungi, the Tremella aurantialba has the advantage of being capable of being fermented and cultured, but no research on the development of novel fermented milk beverage by using Tremella aurantialba as a fermentation strain has been carried out so far. Therefore, the fermented milk taking the tremella aurantialba as the fermentation strain has great market potential in novel dairy products.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to solve the technical problem of the prior art and provides tremella aurantialba and application thereof in preparing tremella aurantialba fermented soybean milk.
In order to solve the technical problems, the invention discloses a tremella aurantialba collected in Yuzhou Yupingshan of Kunming City in 2018 in 4 months. The strain is classified and named as Tremella aurantia (Tremella aurantiaba), has the strain name of NX-20, is preserved in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms, and has the preservation address: beijing, the preservation time is 2019, 10 months and 15 days, the preservation number is CGMCC NO.18588, and the ITS sequences (internal transcription spacers 1 and 2) are shown as the sequence table SEQ ID No. 1.
The application of the tremella aurantialba in preparing the fermented soybean milk is also within the protection scope of the invention.
Wherein the preparation method comprises the following steps: the tremella aurantialba fermented soybean milk is prepared by taking tremella aurantialba as a fermentation strain and soybean milk as a basic culture medium and performing microbial fermentation, and specifically, tremella aurantialba fermented soybean milk is prepared by inoculating tremella aurantialba seed liquid into the culture medium (soybean milk), adding a carbon source and fermenting.
The culture method of the tremella aurantialba seed liquid comprises the following steps: inoculating the slant-preserved Tremella aurantialba into a broth culture medium containing potato glucose, culturing at 20-28 deg.C under stirring and aeration for 3-5 days until OD of the culture solution600And obtaining the product when the value reaches 1.2-1.5.
Wherein, the tremella aurantialba seed liquid is inoculated into the soybean milk according to the inoculation amount of 2-10% v/v.
The preparation method of the soybean milk comprises the following steps: soaking cleaned semen glycines in water for 10-12 hr, filtering to remove water, pulping with pulping machine, and removing bean dregs with 200 mesh filter screen to obtain filtrate, i.e. soybean milk; wherein, the soaking is to add 3-5 g of soybeans into each mL of water.
Wherein the carbon source is any one or combination of sucrose, fructose, glucose, citric acid and xylitol; the addition amount of the carbon source was 40g/L of soybean milk.
Wherein the fermentation is carried out at 23-27 ℃, stirring, ventilating and fermenting for 5-7 days, and the fermentation is stopped when the viscosity value of the fermentation liquid reaches 3200-3500 mPa.s, thus obtaining the tremella aurantialba fermented soybean milk stock solution.
Wherein the stirring speed is 180-240 rpm, and the aeration intensity is controlled to be 1.5-2.0 vvm.
The tremella aurantialba fermented soybean milk stock solution prepared by the method can be further processed to obtain a finished product, and the method specifically comprises the following steps: adding water into the tremella aurantialba fermented soybean milk stock solution, performing high-pressure homogenization treatment to break tremella aurantialba cells, performing pasteurization, cooling to room temperature, and filling under aseptic conditions to obtain the tremella aurantialba fermented soybean milk finished product.
Wherein the dosage ratio of the tremella aurantialba fermented soybean milk added into water to the water is 1: 2 (v/v); the pressure of the high-pressure homogenization treatment is 20-30 MPa, and the treatment times are 1-3 times; the pasteurization temperature is 62-65 ℃, and the time is 30 min.
The function of the carbon source in the tremella aurantialba fermented soybean milk finished product, namely the nutritional and flavored fermented soybean milk, mainly provides the carbon source for the growth of tremella aurantialba cells, and can improve the flavor of the fermented soybean milk.
Has the advantages that: compared with the prior art, the invention has the following advantages:
1. the strains and raw materials related by the invention are natural food materials, wherein the tremella aurantialba strain is a rare edible fungus with homology of medicine and food. The product obtained by fermenting the tremella aurantialba strain has unique flavor and excellent taste and also has high health-care effect.
2. The raw materials added in the feed supplement of the invention are one or more of sucrose, fructose, glucose, citric acid and xylitol, and the feed supplement liquid can be used not only as a carbon source but also as a seasoning to increase the flavor of the tremella aurantialba fermented soybean milk.
3. The invention meets the requirements of national standard on milk-containing fermented beverage, so the tremella aurantialba fermented soybean milk is called as tremella aurantialba fermented soybean milk.
4. The invention breaks the limitation of single strain of fermented soybean milk in the market, creatively uses tremella aurantialba as a fermentation strain, and provides the preparation method of the health-care tremella aurantialba fermented soybean milk, which has the advantages of simple process, stable quality and easiness in large-scale production. Compared with traditional fermented milk, the tremella aurantialba fermented soybean milk provided by the invention provides a new choice for the development of novel strains in the fermented milk industry on one hand, and has a very high health care function due to the extremely high nutritive value of tremella aurantialba.
5. The method is simple and convenient because the main nutrient substance in the tremella aurantialba fermented soybean milk is tremella aurantialba polysaccharide which has various biological activities of oxidation resistance, tumor resistance, diabetes resistance and the like, and the tremella aurantialba polysaccharide is a pseudoplastic non-Newtonian fluid. The pseudoplastic non-Newtonian fluid has the characteristic of shear dilution, and the viscosity of the solution is increased along with the increase of the mass concentration, so that the concentration of the tremella aurantialba polysaccharide can be monitored according to the viscosity index, and the fermentation state of the tremella aurantialba fermented soybean milk can be monitored.
Detailed Description
The following examples illustrate the present invention by way of a tremella aurantialba fermented soy milk and a method for preparing the same.
In the following examples, the culture broth containing potato dextrose was (potato 200 g, dextrose 20 g, distilled water 1000 ml).
Example 1: preparation method of tremella aurantialba fermented soybean milk
(1) Cultivation of seed liquid
Inoculating Tremella Aurantialba preserved on slant into seeding tank containing potato glucose broth culture medium, ventilating at 26 deg.C and 200rpm under stirring, controlling ventilation strength at 2.0vvm, and culturing for 4 days to make OD of culture solution600The value reached 1.3.
(2) Preparation of soybean milk
The preparation method of the soybean milk comprises the following steps: soaking the cleaned soybeans in 3-5 times (g/mL) of water for 10-12h, filtering to remove water, pulping by using a pulping machine, and removing bean dregs through a 200-mesh filter screen to obtain filtrate, namely the soybean milk.
(3) Fermentation of soybean milk
Inoculating the tremella aurantialba seed liquid in the step (1) into a fermentation tank containing soybean milk according to the inoculation amount of 5% (v/v), simultaneously adding 40g of carbon source/L of soybean milk, stirring and ventilating at 26 ℃ and 200rpm, controlling the ventilation strength to be 2.0vvm, culturing for 6 days, taking the viscosity as a fermentation monitoring index, and ending the fermentation after the viscosity value reaches 3200 mPas.
The carbon source supplemented during fermentation is the combination of sucrose, fructose, glucose, citric acid and xylitol, and the dosage is respectively 5g/L +10g/L +10g/L +10g/L +5 g/L.
(4) Downstream processing and canning
Blending the fermented milk in the step (3) with purified water in a certain proportion, performing high-pressure homogenization treatment to break tremella aurantialba cells, performing pasteurization, cooling to room temperature, and filling under aseptic condition to obtain the tremella aurantialba fermented soybean milk finished product. The ratio of the fermented milk to the purified water is 1: 2 (v/v); the high-pressure homogenizing pressure is 20-30 MPa, the times are 3, and each time is 15 minutes. Sterilizing at 65 deg.C for 30min, and pasteurizing.
Example 2: component analysis of tremella aurantialba fermented soybean milk
2.1 samples
Control group: according to the soybean milk prepared in the example 1, substances such as corresponding carbon sources and the like are added into the soybean milk, and the post-treatment method and conditions are the same as those of the experimental group except that the tremella aurantialba strain is not added for fermentation;
experimental groups: fermented soy milk prepared according to example 1.
2.2 analysis method of nutrient content of tremella aurantialba fermented soybean milk
(1) And (5) measuring the moisture.
Reference is made to GB 5009.3-2016. Accurately weighing 10g of tremella aurantialba fermented soybean milk stock solution, evaporating to dryness in a boiling water bath, drying in a drying oven at 105 ℃, taking out, cooling, weighing by using an analytical balance, and calculating the water content.
(2) And (5) measuring ash content.
Reference is made to GB 5009.4-2016. Accurately weighing 10g of tremella aurantialba fermented soybean milk stock solution, evaporating to dryness in boiling water, and heating by an electric furnace until carbonization and no smoke exist. Subsequently, the mixture was placed in a muffle furnace, burned at 550 ℃ for 4 hours, cooled to below 200 ℃ and transferred to a desiccator for cooling, and after cooling to room temperature, it was precisely weighed using an analytical balance.
(3) And (4) determining total sugar.
Reference is made to GB/T15672-2009.
(4) And (4) determining reducing sugar.
Reference is made to GB 5009.7-2016.
(5) And (4) fat determination.
Reference is made to GB 5009.6-2016.
(6) And (4) dietary fiber determination.
Reference is made to GB 5009.88-2014.
(7) Determination of total acids in food.
Reference is made to GB/T12456-.
(8) Determination of amino acids in food products.
Reference is made to GB 5009.88-2014. An amino acid full-automatic analyzer (model: S-433D, manufactured by Sykam, Germany) was used.
(9) Measurement of uronic acid
Accurately weighing 25mg of sample, fully dissolving the sample by using distilled water, and fixing the volume in a 50mL volumetric flask to prepare a crude polysaccharide sample solution of 0.5 mg/mL. Absorbing 100 mu L of each group of samples into a test tube, then adding 500 mu L of sodium tetraborate monosulfuric acid solution, cooling in ice water, mixing uniformly on a vortex oscillator after cooling, heating in boiling water bath for 5min, adding 100g of meta-diphenyl solution after cooling in ice water, mixing uniformly, replacing a sample to be tested with 1.0mL of distilled water as a blank control, measuring the light absorption value at 520nm, carrying out experiments for three times in parallel, and taking an average value. And substituting the measured light absorption value (average value) of each group of samples into the standard curve to calculate the uronic acid content in the samples.
2.3 results of measurement
(1) The main nutrient components of the tremella aurantialba fermented soybean milk are shown in table 1.
TABLE 1 Tremella Aurantialba fermented Soybean milk essential nutrient
The results are shown in Table 1, wherein the control group was soy milk before fermentation and the experimental group was soy milk after fermentation. The tremella aurantialba fermented soybean milk is rich in main nutritional ingredients and contains sugars, crude protein and crude fat which have nutritional value to human bodies. In addition, it also contains polysaccharide with antioxidant and antitumor effects.
(2) The detection results of the golden fungus fermented soybean milk free amino acids are shown in table 2.
TABLE 2 amino acid types of fermented milks of Tremella Aurantialba
The amino acid components of the tremella aurantialba functional fermented soybean milk are shown in table 2, 17 free amino acids are detected, wherein 7 essential amino acids are included, the essential amino acids comprise limited amino acids such as lysine, threonine and the like which have key positions in human bodies, the total content of the free amino acids is 3.284 mu g/mL, and the total content of the essential amino acids is 19% of the total content of the amino acids. After the tremella aurantialba strain is adopted for fermentation, the content of the essential amino acid is increased by 47.4%, so that the tremella aurantialba strain adopted by the invention can effectively increase the amount of the essential amino acid. The main amino acid in the tremella aurantialba fermented soybean milk beverage is histidine, and the secondary amino acid is aspartic acid, glutamic acid, arginine, alanine and the like, and the content of the methionine is the minimum.
Example 3: evaluation of flavor of tremella aurantialba fermented soybean milk
3.1 flavor evaluation method
(1) Electronic tongue evaluation
Firstly, 0.01 mol.L is used-1And 0.01 mol. L of an aqueous sodium chloride solution-1The sensor of the electronic tongue (model: ZX-43, manufactured by INSENT) was diagnosed with 0.01 mol.L-1The hydrochloric acid is used for pre-balancing the sensor, the control group and experimental group products in the embodiment 2 are detected after the sensor passes the balancing, the acquisition time of each sample by the electronic tongue is 120 seconds, 1 data is acquired every second, each sample needs to be acquired 9 times, then the sample needs to be cleaned for 30 seconds, and finally the average data of the last 3 times is taken as the result of the test.
(2) Sensory evaluation
The sensory score refers to the evaluation plan of the implementation of the sensory quality evaluation of the national industry standard of GBT 21732-. Ten technical experts formed a fermented milk evaluation group for sensory evaluation of the fermented milk beverages, and the total score was 100, and the evaluation results were averaged.
TABLE 3 organoleptic scoring sheet for milk-containing beverage
3.2 flavor evaluation results
(1) The electronic tongue response values of the tremella aurantialba fermented soybean milk are shown in table 4.
TABLE 4 electronic tongue response values of tremella aurantialba fermented soymilk
It is apparent from table 4 that the difference between the experimental group and the control group is large, wherein the control group is soybean milk before fermentation, and the experimental group is soybean milk after fermentation. The sour taste of the fermented rice is greatly increased compared with the fermented rice after fermentation; the bitter taste is reduced slightly; the rising amplitude of the sweet taste and the astringent taste is small; the umami taste and the fullness are slightly reduced. It is obvious that the tremella aurantialba fermented milk not only reduces the bitterness but also greatly increases the sourness while maintaining the umami and sweet taste of natural soybeans, and the reason for this is that the fermented milk produces uronic acid after fermentation, the content of uronic acid before fermentation is 0g/L, and the content of uronic acid after fermentation reaches 1.51g/L, whereas the content of uronic acid produced by lactic acid bacteria fermentation of the prior art is about 0.1-0.5 g/L. This adds flavour to the fermented milk. Therefore, the tremella aurantialba fermented milk has wide industrial prospect in the field of soybean milk.
(2) The sensory scores of the tremella aurantialba fermented soymilk are shown in table 5.
TABLE 5 golden fungus function fermented soybean milk sensory evaluation table
As can be seen from Table 5, the tremella aurantialba fermented soybean milk has a sensory evaluation score of 88 at the highest and 65 at the lowest, and is divided into 77.8 average scores, and the tremella aurantialba fermented soybean milk is slightly sweet in overall taste, harmonious in flavor, proper in smell, uniform in texture, and good in development prospect.
The invention provides a tremella aurantialba and an application method thereof in preparing fermented soybean milk, and a method and a way for realizing the technical scheme are many, the above description is only a preferred embodiment of the invention, and it should be noted that for a person skilled in the art, a plurality of improvements and decorations can be made without departing from the principle of the invention, and the improvements and decorations should also be regarded as the protection scope of the invention. All the components not specified in the present embodiment can be realized by the prior art.
Sequence listing
<110> Nanjing university of industry
<120> Tremella aurantialba and application thereof in preparation of Tremella aurantialba fermented soybean milk
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a 481
Claims (10)
1. The Tremella aurantialba is classified and named as Tremella aurantialba (Tremella aurantiaba), has a strain name of NX-20, is preserved in the China general microbiological culture Collection center, has the preservation time of 2019, 10 and 15 days, and has the preservation number of CGMCC NO. 18588.
2. Use of the Tremella aurantialba as claimed in claim 1 for preparing fermented soybean milk.
3. The use according to claim 2, wherein the preparation method comprises: inoculating the tremella aurantialba seed liquid into the soybean milk, adding a carbon source, and fermenting to obtain the tremella aurantialba fermented soybean milk.
4. The use of claim 3, wherein the culture method of the Tremella aurantialba seed liquid comprises: inoculating Tremella aurantialba into broth culture medium containing potato glucose, culturing at 20-28 deg.C under stirring and aeration for 3-5 days, and adjusting OD of culture solution600And obtaining the product when the value reaches 1.2-1.5.
5. The use according to claim 3, wherein the Tremella aurantialba seed fluid is inoculated into the soybean milk at an inoculum size of 2-10% v/v.
6. The use according to claim 5, wherein the soy milk is prepared by a process comprising: soaking cleaned semen glycines in water for 10-12 hr, filtering to remove water, pulping with pulping machine, and removing bean dregs with 200 mesh filter screen to obtain filtrate.
7. The use according to claim 3, wherein the carbon source is any one or more of sucrose, fructose, glucose, citric acid and xylitol; the addition amount of the carbon source was 40g/L of soybean milk.
8. The application of claim 3, wherein the fermentation is performed by stirring, ventilating and fermenting at 23-27 ℃ for 5-7 days, and the fermentation is stopped when the viscosity of the fermentation liquid reaches 3200-3500 mPa.s.
9. The use according to claim 4 or 8, wherein the stirring speed is 180-240 rpm, and the aeration intensity is controlled to be 1.5-2.0 vvm.
10. The use of claim 3, wherein the tremella aurantialba fermented soybean milk is prepared by adding water into tremella aurantialba fermented soybean milk, homogenizing under high pressure, sterilizing by pasteurization, cooling to room temperature, and packaging under aseptic condition.
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