CN114617228B - Method for preparing flavored stinky tofu by using acid water fermented by bamboo shoots - Google Patents

Method for preparing flavored stinky tofu by using acid water fermented by bamboo shoots Download PDF

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CN114617228B
CN114617228B CN202210378566.2A CN202210378566A CN114617228B CN 114617228 B CN114617228 B CN 114617228B CN 202210378566 A CN202210378566 A CN 202210378566A CN 114617228 B CN114617228 B CN 114617228B
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bean curd
acid water
bamboo shoots
acid
fermented
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CN114617228A (en
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蒋立文
李跑
廖卢艳
刘洋
唐辉
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Hunan Agricultural University
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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
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/40—Pulse curds
    • A23L11/45—Soy bean curds, e.g. tofu
    • 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
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A23L19/20—Products from fruits or vegetables; Preparation or treatment thereof by pickling, e.g. sauerkraut or pickles
    • 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
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/10—Natural spices, flavouring agents or condiments; Extracts thereof
    • 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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • 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
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3562—Sugars; Derivatives thereof
    • 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
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3571—Microorganisms; Enzymes
    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation

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Abstract

The invention discloses a method for preparing flavored stinky tofu by utilizing acid water fermented by bamboo shoots, and relates to the field of food processing. The method comprises the following steps: anaerobic fermentation is carried out by taking bamboo shoots and water as raw materials to obtain acid water; and (3) performing curdling by utilizing the acid water in the making process of the bean curd blank, and then cooling the bean curd blank and performing compression molding to obtain the flavored stinky bean curd. According to the invention, the acid water fermented by the bamboo shoots is directly used for curdling, and the special flavor substances (p-methylphenol) in the acid water can enable all parts of the bean curd to have special taste and smell, so that the step that the bean curd blank is soaked in brine for fermentation in the traditional making of the strong-smelling preserved bean curd is reduced, and the production process is greatly simplified.

Description

Method for preparing flavored stinky tofu by using acid water fermented by bamboo shoots
Technical Field
The invention relates to the field of food processing, in particular to a method for preparing flavored stinky tofu by utilizing acid water fermented by bamboo shoots.
Background
The bean curd has a long history of production, and the production process generally comprises the steps of soybean soaking, impurity removal, pulping, pulp boiling, marinating, cooling, squeezing, forming and the like. The quality of the bean curd in different regions is different. Besides the direct influence of different raw material varieties on the quality of the bean curd, the difference of marinating technology has a larger influence on the final quality of the bean curd. Gypsum is adopted for curdling, and the tender moisture content of the bean curd is high; the bittern such as magnesium chloride is adopted for curdling, so that the bean curd has harder texture and slightly bitter taste; the glucolactone is adopted for curdling, and the bean curd is basically over 80 percent of moisture content and is very tender due to the high curdling temperature and natural cooling forming. The bean curd prepared by the plaster curdling method is suitable for frying, the bean curd prepared by the magnesium chloride and other bittern is suitable for cold dishes, hot pot, etc., while the lactone bean curd prepared by the glucolactone curdling is mainly used for preparing soup ingredients, etc.
Along with the development of technology, the curdling technology becomes an important core element for differentiation of bean curd products. The compound coagulant is widely used and industrialized in the bean curd product industry, then a sour slurry marinating (yellow slurry water after bean curd is squeezed is naturally acidified or is inoculated with lactobacillus for fermentation and acidification) product is produced, and raw materials are changed, such as black beans are used for producing black bean curd, so that product differentiation is formed. But the variety of the bean curd is relatively single in general, and the bean curd products are rich in nutrition and are easy to be polluted by microorganisms, and the whole process of the cold chain system is not in seamless joint although the cold chain system is supported, so that the fresh bean curd still cannot meet the diversified demands of the market.
The fermented bean curd is considered as a regional characteristic product of slightly fermented bean curd and having a specific flavor, the flavor substance of which is mainly p-methylphenol. However, the preparation process of the stinky tofu is that the tofu blank is firstly prepared, then the tofu blank is immersed in brine for light fermentation, the preparation process is complicated, the brine preparation time is long (at least 2 years), the quality is uncontrollable, and potential safety hazards exist.
Therefore, the method for preparing the flavor stinky tofu by utilizing the fermented acid water of the bamboo shoots is provided, so that the preparation process of the stinky tofu is simplified, and the method has important significance in the field of stinky tofu preparation.
Disclosure of Invention
The invention aims to provide a method for preparing flavor stinky tofu by utilizing acid water fermented by bamboo shoots, which solves the problems in the prior art and simplifies the preparation process of the stinky tofu.
In order to achieve the above object, the present invention provides the following solutions:
according to one of the technical schemes, the method for preparing the flavored stinky tofu by utilizing the acid water fermented by the bamboo shoots comprises the following steps of:
anaerobic fermentation is carried out by taking bamboo shoots and water as raw materials to obtain acid water;
and (3) performing curdling by utilizing the acid water in the making process of the bean curd blank, and then cooling the bean curd blank and performing compression molding to obtain the flavored stinky bean curd.
No auxiliary fermentation substances such as strains, salt and the like are needed to be added in the whole anaerobic fermentation process. The bamboo shoot fermentation process is a large principal of lactobacillus, and the relative abundance of the lactobacillus in the fermentation process is more than 90%. The lactobacillus has rich metabolites and good health care effect and nutrition effect on human body. Meanwhile, organic acid generated in the bamboo shoot fermentation process is mainly organic acid such as lactic acid and acetic acid, and acid water formed by fermentation contains rich lactic acid bacteria, polysaccharide, flavone and dietary fiber, so that the fermentation process is safe and beneficial to human bodies.
The special volatile odor substances formed by the fermentation of the bamboo shoots are mainly phenol substances. Compared with the strong pungent taste of the flavor substances phenol substances and indole substances formed by common brine fermentation, the characteristic smell of the bamboo shoots needs to be formed by evaporation after a certain temperature, and the smell is more healthful and softer than the common brine.
Metazoan refers to biologically active compounds (including probiotic metabolites, cellular components or mixtures thereof) produced by probiotics during fermentation that are beneficial to health, such as short chain fatty acids (short chain fatty acids, SCFA), tryptamine, peptides, teichoic acids, peptidoglycans, polysaccharides, organic acids and lipids, and the like. Compared with probiotics, the metazoan has the advantages of clear chemical structure, high safety dosage, long shelf life and the like; the metaplasia has the biological activities of anti-inflammatory, antibacterial, immunoregulation, antioxidation, anti-obesity, antihypertensive, hypolipidemic, liver protection, wound healing promotion and the like, and has wide prospect in the aspect of developing novel health functional foods. The amino acid differential metabolites selected from acid bamboo shoots fermented by 30d, 60d and 90d in the bamboo shoot fermentation process comprise 5-hydroxy-L-tryptophan, N-alpha-acetyl-L-ornithine, N-ethylglycine, N-acetyl-L-phenylalanine, N-acetyl-L-aspartic acid, N-acetylornithine, beta-tyrosine, epsilon-acetyl-L-lysine, methionine, lysine, tyrosine, tryptophan, threonine and aspartic acid, wherein lysine, tryptophan and threonine are three essential amino acids for human body. During the fermentation process of the bamboo shoots, the relatively active flavor substances are mostly esters, phenols and acids, the alcohol content is in a trend of increasing firstly and then decreasing secondly, and the content is up to 13.94% at the time of 30 d; the content of esters always tends to decrease, and the content is up to 27.24 mug/g at 0 d; the phenol content tends to increase and decrease, and the content is up to 64.89 mug/g when 90d is fermented; the p-cresol is the flavor substance with the highest content in the whole fermentation process, the relative content of the p-cresol reaches 55.4+/-0.01 mug/g when 90d is fermented, the p-cresol has pungent smell, most of the components are excellent metazoan, acid water of the fermented bamboo shoots (organic acid, lactobacillus, flavonoids and phenols in acid water are metazoan components with clear structures and definite functions) is utilized for curdling, wherein the content of the total acid is in an ascending trend in the process of fermenting 0d to 90d, the total acid is stable after 90d, and the content is up to 11.05 g/kg; the content of flavone tends to increase and decrease with the increase of fermentation time, and the highest content reaches 3.56mg/g at the time of 90d fermentation. Lactic acid and acetic acid become organic acids which greatly contribute to sour taste, and the relative content of lactic acid is up to 3.71+/-0.03 mg/g when the lactic acid is fermented by 90 d; the acetic acid content is secondary, the relative content reaches 3.07+/-0.01 mg/g when 90d is fermented, so that the nutritional functional components and metaplasia of the bean curd can be increased, and different physiological functions and actions are brought to the bean curd. Meanwhile, the bamboo shoot fermentation product, namely the metaplasia, is fully utilized to produce the bean curd, so that the special flavor of the bean curd can be formed, the traditional production process of the strong-smelling preserved bean curd is greatly simplified, and meanwhile, the innovative product meets the living demands of people.
Further, the bamboo shoots are cut into filaments or small pieces of 1-2mm×1-2mm×5-8mm or the whole bamboo shoots are soaked for fermentation.
Further, the acidity of the acid water is 0.8-1.2%, and the number of lactobacillus is not less than 10 5 On the order of CFU/mL.
Further, the anaerobic fermentation specifically comprises: fermenting at 25-35deg.C for 30-90d.
Further, the bamboo shoots account for 60-80% of the total mass of the raw materials.
Further, the water accounts for 20-40% of the total weight of the raw materials.
Further, the addition amount of the acid water in the soybean milk spot-adding process is 10-20% of the volume of the soybean milk.
According to the second technical scheme, the flavored stinky tofu prepared by the method is used.
The invention discloses the following technical effects:
(1) The bamboo shoots are good dietary fiber food materials and contain rich mineral substances, dietary fibers and flavonoid substances. The acid water (i.e. the coagulant for bean curd curdling) prepared by anaerobic fermentation of the bamboo shoots serving as the raw material has the advantages of short fermentation time, direct use for curdling, recycling, no solid, and certain smell (p-methylphenol) and nutrient components (organic acid, lactobacillus, flavonoids and phenols).
(2) In the method, only acid water fermented by bamboo shoots is used as a coagulant, so that the method has the advantages of high coagulation speed and fine mouthfeel, and the product meets the relevant standard. Compared with the traditional gypsum spot slurry, the flocculation rate is increased by 30-60 seconds after the gypsum spot slurry is added according to the proportion, the water content is increased by about 5% under the same pressure, the bean curd yield is increased by 5-10% compared with the traditional method, the surface of the fried stinky bean curd is crisp, the interior is tender, the special taste of the product is more obvious, and the requirements of consumers with strong partial preference are met.
(3) The flavored fermented bean curd prepared by the acid water spot slurry fermented by the bamboo shoots comprises main components (moisture, protein, inorganic salt and the like) contained in conventional bean curd, and also contains various products fermented by the acid bamboo shoots, such as polysaccharide, flavone, lactobacillus and metabolites thereof. Meanwhile, the special flavor substance p-methylphenol brought by acid water fermented by bamboo shoots is released along with the rising of temperature in the frying or frying process of the prepared flavor strong-smelling preserved bean curd, so that the product has special fermented 'smelly' flavor.
(4) According to the invention, the acid water fermented by the bamboo shoots is directly used for curdling, and the special flavor substances (p-methylphenol) in the acid water can enable all parts of the bean curd to have special taste and smell, so that the step that the bean curd blank is soaked in brine for fermentation in the traditional making of the strong-smelling preserved bean curd is reduced, and the production process is greatly simplified.
(5) The flavored fermented bean curd prepared by the invention has lower acidity, contains lactobacillus plantarum and bamboo shoot polysaccharide which are beneficial to the quality guarantee of the product under the same preservation condition, the quality guarantee period of the product is prolonged by more than 1 time, the taste of the product is changed after the product is preserved for 2-3 days at the temperature of 0-5 ℃ in general, the product can be preserved for 5-7 days at the same temperature, the taste and the color are stable, the microorganism quantity is not obviously increased, and the process advantage is reflected.
(6) The solid matters after the bamboo shoots are fermented can be processed into the special pickle bag for seasonings or snails powder, so that comprehensive utilization is realized.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows bacterial diversity distribution of sour bamboo shoots under different fermentation time conditions.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
The "percentage" in the invention refers to mass percent unless otherwise specified.
The soybean used in the embodiment of the invention is non-transgenic soybean with protein content of more than 40%.
The water quality used for soaking the soybeans in the embodiment of the invention is soft water or pure water.
EXAMPLE 1 preparation of coagulant for curdling of tofu
Removing outer layer of fresh bamboo shoot, cutting into blocks, placing in a barrel, adding water to cover the bamboo shoot blocks (no salt, water quality)1.5kg and bamboo shoot mass of 3.5 kg), anaerobic fermentation is carried out at room temperature; the fermentation temperature is 30 ℃ (25-35 ℃ can achieve the technical effect similar to that of 30 ℃), the fermentation time is 60d (30 d-90d can be achieved), the acid pulp water obtained by filtering after the fermentation is finished is the coagulator for bean curd curdling (the acidity of the coagulator is 0.8-1.2% (total acid is titrated), and the number of lactic acid bacteria is 10) 5 On the order of CFU/mL, i.e., meets the use requirements as a coagulant for curdling of tofu).
The major changes in the relative amounts of specific flavor substances of bamboo shoots at different fermentation times (labeled GX-day, e.g., GX-0 at day 0, and GX-10 at day 10, and so on) are shown in table 1.
TABLE 1 variation of the relative content of characteristic flavor substances during bamboo shoot fermentation (%)
As can be seen from Table 1, the relative content of p-methylphenol in the volatile components reached 70-90%.
The microbial flora change of the bamboo shoots at different fermentation time is measured, specifically:
(1) Extraction of total DNA from a sample
Sampling at intervals of 5, 20, 40, 60 and 90d (for example, GX-0 at fermentation time of 0 day, GX-5 at fermentation time of 5 days, and the like), uniformly stirring the fermentation liquor and the bamboo shoot sample (1:1) to obtain about 50mL each, and performing suction filtration by using a water system microporous filter membrane with a pore diameter of 0.45 μm. Cutting the filter membrane after the suction filtration is completed, putting the cut filter membrane into a micro centrifuge tube, and extracting total DNA according to the specification of the DNA kit.
(2) High-throughput sequencing method for acid bamboo shoot 16SrDNA
Bacterial 16sDNA V3-V4 region amplification was performed using the total DNA of the sample as a template. The primers are 338F (5'-ACTCCTACGGGAGGCAGCAG-3') and 806R (5 '-GGACTACHVGGGTWTCTAAT-3')
25. Mu.L of PCR amplification system containing 25ng of DNA template, 2.5. Mu.L of each of the upstream and downstream primers, and 12.5. Mu.L of PCR premix. The PCR amplification conditions are that the PCR amplification is performed for 3 min at 98 ℃; denaturation at 98℃for 10 s, annealing at 54℃for 30 s, extension at 72℃for 45 s, and cycling for 35 times; finally, the extension is carried out for 10 min at 72 ℃. And (3) taking the PCR product, detecting by using 1% agarose gel electrophoresis, and sending a qualified sample to Hangzhou Liangchuan biotechnology Co-Ltd for high-throughput sequencing.
The microbial flora structure distribution (relative proportion,%) of the bamboo shoots at different fermentation times is shown in table 2.
TABLE 2 relative proportions of Lactobacillus in the main fermentation stage (%)
As can be seen from Table 2, the number of the microorganisms of the lactobacillus is absolute, the relative content of the lactobacillus is more than 60% within 30-90d, and the acid bamboo shoot water fermented for 60-90d can meet the production requirement in consideration of the actual production requirement, and the relative content of the lactobacillus reaches 90%.
The significant differential metabolites of bamboo shoot fermentations 30d and 90d are shown in table 3.
TABLE 3 variation of major metabolites of bamboo shoot fermentations 30d and 90d
30d and 90d, 21 metabolites were up-regulated; 13 metabolites were down-regulated. The up-regulating substances include styrene oxide, 5-hydroxy-L-tryptophan, indole-3-pyruvic acid, ricinoleic acid, D-erythro-N-stearoyl sphingosine, phytosphingosine, formononetin, 1-acyl-sn-glycero-3-phosphorylcholine, sphingomyelin 3-hydroxy benzyl alcohol, methionine, xanthine, orotic acid, L-3-phenyllactic acid, lactic acid, N-acetyl-L-aspartic acid, N-acetyl-L-phenylalanine, flavone, hexadecanediacid; down-regulated substances are pyrazinamide, N-acetylornithine, ε -acetyl-L-lysine, jasmonic acid, benzoin, oleamide, 1-methyladenosine, 7-methyladenosine, 1-acyl-sn-glycero-3-phosphoethanolamine, glutamic acid, 4-hydroxybenzoic acid, 3-hydroxycinnamic acid.
As can be seen from Table 3, during 30-90 days of fermentation, a large amount of physiologically active substances are generated in the fermentation system, the substance content is in the rising stage, the system contains a large amount of active substances such as flavone, amino acid and the like, and the acid bamboo shoot water with a certain acidity and 60d fermentation period can meet the requirement of a coagulant suitable for bean curd curdling in consideration of production needs.
The acid water in the invention adopts MRS culture medium combined with transparent ring characteristics for producing calcium carbonate, and meanwhile, the similarity of the lactobacillus plantarum identified after physiological and biochemical tests is very high, which indicates that the lactobacillus plantarum in the fermentation system takes absolute advantage.
Example 2 preparation of flavored fermented Bean curd
(1) Cleaning soybeans: removing impurities such as soil, stones, grass scraps and metal scraps mixed in the soybean raw materials, and selecting the soybeans which are free of mildew points, bright in color, full in seeds and stored for more than 3 months for cleaning.
(2) Soaking: soaking soybean in water at 20deg.C for 6 hr (6-10 hr can achieve similar technical effect as 6 hr; soaking time is 8-15 hr when soaking temperature is about 5deg.C; soaking time is 4-6 hr when soaking temperature is about 25deg.C), wherein soaking water consumption is 1 weight ratio of soybean to water: 2.3.
(3) Pulping: pulping with a high-speed pulping machine. The soybean milk is repeatedly ground for three to four times so as to fully extract the proteins in the soybean, the bean dregs can be directly and continuously ground for three times so as to fully extract the proteins in the soybean, and the mesh number of the filtered soybean milk is more than 200 meshes.
(4) Boiling pulp: pouring the squeezed raw soybean milk into a pot to be boiled with strong fire (ensuring that soybean milk is heated uniformly), and skimming foam on the surface while boiling without covering a pot cover. The soybean milk is boiled to a temperature above 100 ℃, and the concentration of the boiled soybean milk is controlled to be 7.5-8.5 Baume degrees.
(5) And (3) curdling: the coagulant prepared in example 1 was added to make a slurry, the coagulant accounted for 15% of the mass of the soybean milk (10-20% could achieve a technical effect similar to 15%), and stirring was carried out while adding the coagulant until coagulated particles appeared.
(6) Cooling: after the coagulated particles appear in the bittern, the bittern slowly coagulates into blocks along with the cooled bean curd, and separation begins to appear, and the water is slightly yellowish.
(7) And (3) press molding: the beancurd is separated by using a wrapping cloth, the beancurd jelly of each layer is uniformly splashed into a die (the water in the beancurd jelly is mostly wrapped in a protein network and is not easy to be discharged automatically, so that the formed beancurd jelly is properly crushed to remove a part of water contained in the formed beancurd jelly), and the beancurd jelly is directly pressurized by using a heavy object (or is finished by using a special machine, and a dried beancurd machine can finish the splashing and pressing simultaneously). The flavored fermented bean curd is obtained by pressing, controlling the water content to 70-85% (the bean curd is tender and cannot be too dry, and the later frying foamability and taste of the fermented bean curd are affected).
(8) Recovery of yellow serofluid: the squeezed yellow serofluid can be directly transferred into a closed fermentation tank for cooling, and then the sour bamboo shoot water is properly added for continuous fermentation so as to be used for next bean curd curdling.
The acidity of the flavored fermented bean curd prepared by the embodiment is 0.85%, the pH value is between 4.5 and 5.5, and the flavored fermented bean curd can be preserved for 5 to 7 days at the temperature of 0 to 5 ℃ and has a shelf life which is 1 time longer than that of the flavored fermented bean curd which is preserved for 2 to 3 days at the temperature of 0 to 5 ℃ commonly used in the prior art. The coagulating agent for curdling is prepared by fermenting bamboo shoots, contains rich substances such as lactobacillus, polysaccharide, organic acid, flavone and the like, is favorable for separating out bean curd proteins, increases bean curd nutrient components (lactobacillus, flavone and the like), and can prolong the shelf life of bean curd.
The organic acid, lactic acid bacteria, flavonoids and phenols in the flavored fermented bean curd prepared in this example were detected in the following manner.
The detection method of the organic acid comprises the following steps:
accurately weighing fermented sour bamboo shoot blocks 10 g, adding 10 g distilled water, and uniformly mixing; mashing with juicer for 3 min, homogenizing every 10 s, and pulverizing completely. Accurately weighing 1.000 and g, homogenizing into a 10 mL centrifuge tube, adding 9 g distilled water, and ultrasonically extracting for 30 min at 25deg.C. Centrifuge at 1000 r/min,15℃for 15 min. Taking supernatant into a 25 mL volumetric flask, and fixing the volume by distilled water; the solution was passed through a 0.2 μm hydrophilic microporous membrane to a 2 mL ampoule with a syringe, which was numbered and then run on a machine to determine the organic acid concentration, 3 replicates were run for each sample.
Chromatographic conditions: chromatographic column: welch Ultimate AQ-C18 column (4.6 mm X250 mm, 5 μm). Mobile phase: with 0.1% phosphoric acid solution: methanol (97:3, V: V) ratio was eluted with a flow equality for 10 min, then methanol was allowed to reach 100% and equilibrate for 5 min with a short time gradient, and the mobile phase was adjusted to 0.1% phosphoric acid solution: methanol=93: 3 (V: V), equilibrated for 5 min. Column temperature: 40 ℃. Detection wavelength: 210 nm. And the sample feeding amount is 20 mu L.
Drawing a standard curve: (1) standard stock solution: the standard substances of acetic acid 40 mg, lactic acid 50 mg, oxalic acid 40 mg, citric acid 40 mg, malic acid 40 mg, tartaric acid 20 mg and succinic acid 125 mg are respectively weighed to be accurate to 0.0001 g, and distilled water is used for constant volume to 10 mL volumetric flask, and the mixture is uniformly mixed and stored at 4 ℃. (2) standard use solution: standard stock solutions 0.2 mL, 0.4 mL, 0.8 mL, 2.0 mL, 4.0 mL were drawn into 10 mL volumetric flasks, respectively, and fixed to volume with 0.1% phosphoric acid.
The content of the main organic acid is found to be 3.71mg/g.
The detection method of the flavonoid substances comprises the following steps:
the determination is carried out by referring to the method in GB 34/T2743-2016 "spectrophotometry for determining the content of total flavonoids in Sophora flower buds and products thereof".
Sample treatment: the sour bamboo shoot samples under different fermentation time are placed in a 60 ℃ oven for drying and are crushed into powder. Accurately weighing the uniformly crushed sour bamboo shoot sample 5 g to 100 mL beaker, adding 30 mL of 60% ethanol solution, ultrasonically extracting for 30 min, and fixing the volume to a 50mL volumetric flask by using the 60% ethanol solution.
The content of flavonoid substances in the detected sample is 3.56mg/g at maximum.
The method for detecting the phenol substances comprises the following steps:
(1) The sample treatment method comprises the following steps: before the solid-phase microextraction head is used, the solid-phase microextraction head must be aged at a sample inlet, and 85 mu mCAR/PDMS extraction head is activated at 240 ℃ and kept for 40 min until no impurity peak appears on a chromatographic detection chart. Taking a sample of 2 mL by a pipetting gun, placing the sample into a headspace bottle, sealing the headspace bottle by a sealing gasket, placing the headspace bottle on a magnetic stirrer, preheating the headspace bottle at 60 ℃, keeping the temperature for 20 min, setting the stirring speed of magnetic particles at 200 r/min, inserting an activated extraction head into the sample bottle, pushing out a fiber head, keeping the distance between an adsorption probe and a liquid surface at about 1 cm, retracting the fiber head after 40 min of adsorption, pulling out the extraction head after the adsorption is finished, inserting the adsorbed extraction head into a gas chromatography sample inlet, pushing out the fiber head, and analyzing the fiber head at 240 ℃ for 5 min.
(2) GC conditions: the chromatographic column is a CD-WAX elastic quartz capillary column (30 m ×0.25 mm ×0.25 μm); the carrier gas is high-purity (99.999%) helium, and the flow rate of the helium is controlled to be 1.0 mL/min; the temperature of the sample inlet is set to 240 ℃; and selecting not to split and sample. Setting a temperature raising program: the column temperature is 50 ℃, the temperature is kept for 2 min, the temperature is increased to 200 ℃ at 5 ℃/min, the temperature is kept for 14 min, and the temperature is increased to 240 ℃ at 15 ℃/min; mass spectrometry conditions: the ion source is an electron ionization (electron ionization, el) source, and the temperature of the ion source is 200 ℃; electron energy 70 eV; the emission current is 150 muA; multiplier voltage 1037V; the interface temperature is 220 ℃; the mass scanning range is 45-500 m/z.
(3) Qualitative and relative content analysis: the total ion flow chromatogram is subjected to qualitative identification of components by searching a standard spectrum library of American national institute of standards and technology (National Institute of Standards and Technology, NIST for short) 2014s, and relative qualitative analysis is carried out by adopting an area normalization method to obtain the relative content of volatile flavor substances of each component.
The measurement finds that: the relative content of the p-methylphenol substance reaches 40-60ug/g.
Measurement of lactic acid bacteria: the method is carried out by adopting a combination of qualitative and quantitative modes.
The method comprises the steps of performing high-throughput sequencing on total DNA of acid bamboo shoot extracted microorganisms under different fermentation time conditions by using an Illumina Miseq technology, researching bacterial diversity of acid bamboo shoots fermented by 30d, 60d and 90d, and researching the acid bamboo shoot fermentation time of 90d, wherein the results are shown in figure 1Its relative abundance is highest, with up to 99.57%. Furthermore, by the use of the common probiotics produced during fermentation: lactobacillus genusLactobacillus) Lactococcus genusLactococcus) And Weissella genusWeissella) Statistical analysis of the sum of the relative abundance of the three materials shows that the relative abundance is highest when the sour bamboo shoots are fermented 90d. In conclusion, from the standpoint of lactobacillus distribution, the number of lactobacillus is the largest when the sour bamboo shoots are fermented by 90d. The lactobacillus quantity reaches 10 in combination with the food microbiology inspection of national food safety standard of GB 4789.35-2016 6 -10 8 CFU/mL。
Total acid determination: the method in GB/T123456-2008 "determination of total acids in food" is used for determination, and the total acids of sour bamboo shoots fermented for about 60-90 days are 0.9-1.2%.
In the bean curd prepared by curdling the coagulant rich in organic acid, lactobacillus, flavonoids and phenols prepared in example 1, the total amount of organic acid, lactobacillus, flavonoids and phenols respectively reaches 1.47mg/g (30% of lactic acid), 10% 3 -10 5 CFU/g, 0.78mg/g and 37.8 percent (volatile component content is 28 ug/g), and the pH value of the sour bamboo shoot water fermented for about 60 days is 1.0 percent, thereby meeting the requirements of a bean curd coagulant and improving the nutritional value of bean curd.
The main flavor substance of the strong-smelling preserved bean curd is p-methylphenol, the content of the p-methylphenol in the strong-smelling preserved bean curd prepared in the embodiment is detected by GC-MS detection, the detection result is 10-30ug/g, after frying treatment, the characteristics of smelling and smelling the odor and eating the fragrance are obvious, which indicates that the flavored stinky tofu prepared by the invention is the stinky tofu when the tofu blank is prepared, the step of immersing the tofu blank in brine for fermentation is omitted, and the process flow is saved.
The applicant has proved through innumerable experiments that substances similar to the flavor of the stinky tofu can be produced only by the bamboo shoots in the fermentation process, and substances similar to the flavor of the stinky tofu cannot be produced by fermenting other fruits and vegetables under the same conditions, so that the invention adopts the acid water fermented by the bamboo shoots as brine, so that the tofu has the flavor of the stinky tofu in the pulp-dropping process, and further, the step of soaking the prepared tofu blank in the brine again for fermentation is avoided.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.

Claims (4)

1. A method for preparing flavored stinky tofu by using acid water fermented by bamboo shoots is characterized by comprising the following steps:
anaerobic fermentation is carried out by taking bamboo shoots and water as raw materials to obtain acid water;
the acid water is utilized to carry out curdling in the making process of the bean curd blank, and then the bean curd blank formed by cooling and pressing is the flavored stinky bean curd;
auxiliary fermentation substances are not added in the anaerobic fermentation process;
the auxiliary fermentation material comprises strains and salts; the total acid content of the acid water is 0.8-1.2%, and the lactobacillus amount is not less than 10 5 CFU/mL order of magnitude;
the anaerobic fermentation specifically comprises the following steps: fermenting at 25-35deg.C for 30-90d.
2. The method for producing flavored fermented bean curd using acid water derived from bamboo shoots as claimed in claim 1, wherein the bamboo shoots account for 60-80% of the total mass of the raw materials.
3. The method for preparing flavored fermented bean curd by using acid water fermented from bamboo shoots according to claim 1, wherein the acid water is added in an amount of 10-20% of the volume of the soybean milk in the process of curdling.
4. A flavored fermented bean curd prepared by the method according to any one of claims 1 to 3.
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