CN111972665A - Bamboo shoot extract containing compound amino acid and polysaccharide, preparation method thereof and bamboo salt prepared from bamboo shoot extract - Google Patents
Bamboo shoot extract containing compound amino acid and polysaccharide, preparation method thereof and bamboo salt prepared from bamboo shoot extract Download PDFInfo
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- CN111972665A CN111972665A CN201910422229.7A CN201910422229A CN111972665A CN 111972665 A CN111972665 A CN 111972665A CN 201910422229 A CN201910422229 A CN 201910422229A CN 111972665 A CN111972665 A CN 111972665A
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- A—HUMAN NECESSITIES
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- A23J1/006—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from vegetable materials
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- A—HUMAN NECESSITIES
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- C—CHEMISTRY; METALLURGY
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Abstract
The invention provides a bamboo shoot extract containing compound amino acid and polysaccharide, a preparation method thereof and bamboo salt prepared from the bamboo shoot extract, wherein the preparation method comprises the following steps: processing bamboo shoots, fermenting the processed bamboo shoots by trichoderma reesei, bacillus amyloliquefaciens and bacillus licheniformis to obtain bamboo shoot protein liquid, dividing the bamboo shoot protein liquid into three parts, fermenting one part of the bamboo shoot protein liquid by aspergillus niger to obtain a bamboo shoot fermentation starter material I, fermenting one part of the bamboo shoot fermentation starter material II by bacillus subtilis to obtain a bamboo shoot fermentation starter material II, mixing the remaining part of the bamboo shoot fermentation starter material I and acidic protease into the remaining part of the bamboo shoot fermentation starter material I and the remaining part of the bamboo shoot fermentation starter material II and the alkaline protease into the remaining part of the bamboo shoot fermentation starter material II and the remaining part of the bamboo shoot fermentation starter material. The method combines a biological fermentation method and an enzymolysis method to release amino acid and polysaccharide in the bamboo shoot protein to the maximum extent, has mild reaction, saves energy, protects the environment, has high utilization rate of the bamboo shoots, and can realize standardized production.
Description
Technical Field
The invention belongs to the field of food processing, and particularly relates to a bamboo shoot extracting solution containing compound amino acid and polysaccharide, a preparation method of the bamboo shoot extracting solution and bamboo salt prepared from the bamboo shoot extracting solution.
Background
The bamboo shoots have unique taste and flavor, are called as 'the mountain delicacies in cold soil', and are popular with people. Ancient bamboo shoots are not only a delicious food, but also a good health-care medicine, and modern Chinese medical science considers that bamboo shoots are sweet in taste, slightly cold and nontoxic, have the effects of clearing heat and reducing phlegm, tonifying qi and harmonizing stomach, treating diabetes, promoting diuresis, benefiting diaphragm and refreshing intestines and the like, and are used for treating inappetence, constipation and phlegm-saliva stagnation.
Modern nutrition research shows that bamboo shoots contain a large amount of protein, carotene, various vitamins, inorganic salts such as iron, phosphorus, magnesium and the like, 18 amino acids beneficial to health and various polysaccharide substances capable of preventing cancers. These components are the material basis of various pharmacological actions of bamboo shoots.
The traditional bamboo shoot processing mainly comprises canned bamboo shoots, special dried bamboo shoots, bamboo shoot juice beverages and the like, and has the advantages of low processing and utilization, low added value and single product. With the continuous development of extraction technology, the extraction of active ingredients from bamboo shoots is the development of bamboo shoot deep processing at present. The process for extracting active ingredients from bamboo shoots mainly comprises an acid-base method, a hot water extraction method, an enzymolysis method and the like. Many researchers adopt the methods to extract active ingredients from bamboo shoots and process the active ingredients into bamboo shoot extracts (liquid), and according to different purposes, the bamboo shoot extract is prepared into various products, bamboo shoot wine for health care, bamboo shoot facial masks for cosmetics and the like.
The Chinese patent CN102429087A adopts an acid-base method to hydrolyze bamboo shoot protein, although the process is simple and the cost is low, the extraction process needs to use a large amount of fresh water and acid-base salt, which belongs to an extraction method with high energy consumption, and the discharged acid-base salt can generate adverse effects on the surrounding ecological environment and is not in accordance with the concepts of green economy and sustainable development. Chinese patent CN103361233A adopts an extraction method and microwave treatment to extract mineral elements, amino acids and other active ingredients from bamboo shoots, bamboo shavings, tabasheer, bamboo flesh, bamboo stems, bamboo branches and bamboo leaves, and mixes the extracted bamboo shoot liquid with fermented wine to prepare bamboo shoot liquid wine. The Chinese patent CN105154486A adopts enzymolysis and ultrasonic treatment to the bamboo shoots, which obviously improves the content of free amino acids and oligopeptides in the raw materials, has mild reaction of the enzymolysis method, does not need to consume a large amount of fresh water, acid, alkali, salt and energy, can overcome the defects of the processes such as acid-base method, leaching method and the like, and is a modern mainstream extraction process.
The enzymolysis method has mild reaction conditions, does not cause great environmental pollution, realizes the positioned hydrolysis, and is easy to realize the control of hydrolysis, but the enzymolysis process is not a simple process and involves a plurality of steps, and the first step is to screen out proper enzyme. Because the enzyme not only affects the reaction rate and the product yield, but also directly affects the flavor, the physical and chemical properties and the physiological activity of the product; the second step is protein dissolution, and the protein is not easy to dissolve and can not be combined with enzyme for hydrolysis.
Disclosure of Invention
The invention aims to provide a method for extracting amino acid and polysaccharide in bamboo shoots, which combines a biological enzyme method and a biological fermentation technology to release the amino acid and the polysaccharide in bamboo shoot protein to the maximum extent to prepare a bamboo shoot extracting solution. The bamboo shoot extract can be used for preparing bamboo salt, and the method has the advantages of mild reaction, energy conservation, environmental protection and standardized production.
The invention relates to a method for extracting amino acids and polysaccharides in bamboo shoots, which comprises the following steps:
(1) washing bamboo shoot, draining, cutting into pieces (with particle size of 5-15mm, for example), pulverizing (with particle size of 20-60 mesh, preferably about 30-50 mesh), decocting for one or more times, and mixing decoctions;
(2) filtering the decoction in the step (1) to obtain filter residue I and filter residue I, standing the filter residue I, performing suction filtration through a vacuum extraction tank to obtain filtrate II and filter residue II, mixing the filter residue I and the filter residue II, and uniformly stirring with water (for example, distilled water with the volume of 1-4 times of the total filter residue) to obtain a bamboo shoot residue suspension;
(3) adding the bamboo shoot residue suspension obtained in the step (2) into a fermentation tank, adding a mixed bacterium comprising trichoderma reesei, bacillus amyloliquefaciens and bacillus licheniformis for fermentation (the adding amount of the mixed bacterium is 0.05-1.0 vol% of the bamboo shoot residue suspension, and further 0.2-0.5 vol%), sterilizing the fermentation liquid after the fermentation is finished, cooling, mixing with the suction filtration liquid II obtained in the step (2) to obtain a bamboo shoot protein liquid, and dividing the bamboo shoot protein liquid into three parts (the weight ratio of the first part to the second part to the third part can be 1: 0.2-5: 4-12, further 1: 0.5-2: 6-10, and further 1: 1: 8).
(4) Adding the first part of the bamboo shoot protein solution I obtained in the step (3) into a fermentation tank A, adding a carbon source, a nitrogen source, inorganic salt and Aspergillus niger liquid into the fermentation tank A, fermenting (preferably constant-temperature oscillation fermentation for 60-100 hours, preferably 70-80 hours), centrifuging the fermentation liquor, removing the precipitate, and taking the supernatant to obtain a bamboo shoot fermentation starter material I;
(5) adding the second part of the bamboo shoot protein solution II obtained in the step (3) into a fermentation tank B, adding a carbon source, a nitrogen source, inorganic salt and bacillus subtilis solution into the fermentation tank B, fermenting (preferably, fermenting at constant temperature under oscillation for 24-60 hours, such as 30-45 hours), centrifuging the fermentation liquor, removing the precipitate, and taking the supernatant to obtain a bamboo shoot fermentation starter material II;
(6) adding the third part of the bamboo shoot protein liquid III in the step (3) into an enzymolysis tank, adding a bamboo shoot fermentation starter material I and acid protease, adjusting the pH to 4-5, preferably 4.2-4.8, performing enzymolysis (preferably keeping the temperature at 45-60 ℃ for 2.5-3h), then adjusting the pH of the liquid to 8.0-10.5, preferably about 8.3-9, adding a bamboo shoot fermentation starter material II and alkali protease, and performing further enzymolysis (for example keeping the temperature at 40-60 ℃ for 2-4h) to obtain an enzymolysis liquid;
(7) heating the enzymolysis liquid obtained in the step (6) to inactivate enzyme, cooling (for example to 40-60 ℃), centrifuging to obtain filter residue III and filtrate III, adjusting pH of the filtrate III to 4-6, preferably about 5-6, decolorizing (for example adding activated carbon and stirring), and filtering to obtain filtrate IV and obtain bamboo shoot extract crude product.
Further, in the step (1), the pulverized product of bamboo shoot is mixed with water, for example, 1 (2-10), preferably 1: (3-5) decocting in water for several times, which means that the supernatant is taken after one-time decoction, and the remaining precipitated residue solids are further decocted in water in an amount of 2-10 times, preferably 3-5 times, the amount of the water added to the residue.
Preferably, the method of the present invention further comprises:
(8) and (3) passing the filtrate IV obtained in the step (7) through a micro-filter and a Dalton ultrafiltration membrane to obtain an ultrafiltrate with the molecular weight of less than 3000 Dalton, and passing the ultrafiltrate through a nanofiltration membrane concentrator to obtain a nanofiltration concentrated solution, namely the bamboo shoot extract.
Preferably, in the step (1), the bamboo shoot crushed material is transferred to a sandwich reaction tank for decoction, and the reaction temperature at the upper part of the sandwich reaction tank is controlled to be 90-105 ℃, and the reaction temperature at the lower part is controlled to be 110-125 ℃.
Further, in the mixed bacteria in the step (3), the volume ratio of trichoderma reesei to bacillus amyloliquefaciens to bacillus licheniformis is 1:0.2-1.5:0.5-4, preferably 1: 0.5-0.8: 1-2, more preferably 1: 0.6-0.7: 1.2-1.4, the colony concentration of Trichoderma reesei is 3X 1010-5×1010cfu/ml, preferably at a colony concentration of 3X 1010-4×1010cfu/ml, colony concentration of Bacillus amyloliquefaciens 2X 10 10-4×1010cfu/ml, preferably at a colony concentration of 2X 1010-3×1010cfu/ml, colony concentration of Bacillus licheniformis 3X 1010-5×1010cfu/ml, preferably at a colony concentration of 3X 1010-4×1010cfu/ml。
Further, the fermentation conditions of the fermentation tank in the step (3) are that the ratio of the oxygen amount to the volume of the feed liquid is 0.3-0.6, preferably 0.3-0.4, the pH value of the feed liquid is controlled to be 4-6, preferably 4.5-5.5, more preferably 4.5-4.8, the fermentation temperature is controlled to be 40-60 ℃, preferably 45-55 ℃, and the fermentation time is 24-90h, preferably 36-72h, more preferably 48-64 h.
Further, in the steps (4) and (5), the inoculation amount of the aspergillus niger bacterial liquid accounts for 5-15V%, preferably 8-12V%, of the bamboo shoot protein liquid I, and the inoculation amount of the bacillus subtilis bacterial liquid accounts for 2-10V%, preferably 4-8V%, of the bamboo shoot protein liquid II.
The preparation method of the Aspergillus niger liquid in the step (4) comprises the following steps: preparing an Aspergillus niger culture medium by using 1-5%, preferably about 2% of soluble starch, 0.5-2%, preferably about 1% of glucose, 0.05-0.3%, preferably about 0.1% of potassium dihydrogen phosphate solution, 0.05-0.2%, preferably about 0.1% of magnesium sulfate solution, 0.1-0.3%, preferably about 0.2% of sodium nitrate and water according to the mass fraction of the Aspergillus niger bacterial liquid, cooling to room temperature after sterilization, inoculating Aspergillus niger into the culture medium, and performing constant-temperature shaking culture at the constant temperature of 120-200r/min, preferably about 160r/min for 60-80h, preferably about 72h at the temperature of 25-30, preferably about 28 ℃ for 60-200 r/min, preferably about 160r/min to obtain the Aspergillus niger bacterial liquid.
The preparation method of the bacillus subtilis liquid in the step (5) comprises the following steps: uniformly mixing 1-5%, preferably about 2% of glucose, 0.05-0.3%, preferably about 0.1% of peptone and 0.3-0.7%, preferably about 0.5% of sodium chloride by mass of the bacillus subtilis liquid, adding water to prepare an aspergillus niger culture medium, adjusting the pH to 7.0-7.5, preferably about 7.2, cooling to room temperature after sterilization, inoculating the bacillus subtilis into the culture medium, and carrying out constant-temperature shaking culture at 25-35 ℃, preferably about 30 ℃ at the rotating speed of 120-200r/min, preferably about 160r/min for 16-36h, preferably about 24h to obtain the bacillus subtilis liquid.
Further, in steps (4) and (5), the carbon source of the fermentation tank A or B is glucose, and accounts for 1-3 wt%, preferably about 2 wt% of the respective bamboo shoot protein liquid, the nitrogen source is beef extract, and accounts for 0.1-0.5 wt%, preferably about 0.3 wt% of the respective bamboo shoot protein liquid, and the inorganic salt is sodium chloride, and accounts for 0.3-0.7 wt%, preferably about 0.5 wt% of the respective bamboo shoot protein liquid.
Further, in the steps (4) and (5), the fermentation conditions are oscillated at constant temperature, the fermentation temperature of the aspergillus niger is 27-30 ℃, the rotation speed is 150-160r/min, the fermentation temperature of the bacillus subtilis is 33-36 ℃, and the rotation speed is 150-160 r/min.
Further, in the step (6), the adding amount of the bamboo shoot fermentation starter I, II is 0.5-3 wt%, preferably 1-2 wt% of the bamboo shoot protein liquid III, and the adding amount of the acid protease or the alkali protease is 0.5-3 wt%, preferably 1-2 wt% of the bamboo shoot protein liquid III.
Further, in the step (6), the acid protease is a commercially available protease, such as pepsin, and the alkaline protease is commercially available pancreatin or other alkaline protease.
Further, in the step (7), filtering the activated carbon by adopting a plate-and-frame filter, wherein the proportion of the activated carbon in the enzymolysis solution is that 0.5-2g of activated carbon is added into each 100-150ml of enzymolysis solution, and the decoloring time is 20-40 min.
Further, in the step (8), the size of the microfilter is 0.4-1.5 μm, preferably 0.5-1.0 μm, the size of the ultrafiltration membrane is 1000-3000 dalton, preferably 1800-2800 dalton, and the size of the nanofiltration membrane concentrator is 150-250 dalton, preferably 200-220 dalton.
The invention further relates to the bamboo shoot extract prepared by the method, which is detected according to the detection methods of GB 5009.124 determination of amino acids in national food standard for food safety and NY/T1676 determination of crude polysaccharide content in edible fungi, wherein the amino acid content is more than 2.5g/100ml, and the polysaccharide content is more than 0.6g/100 ml.
The invention also provides bamboo salt which comprises edible salt and the bamboo shoot extracting solution, wherein the bamboo shoot extracting solution accounts for 0.1-2.5 wt% of the weight of the raw salt (edible salt), and preferably 0.5-1.0 wt%.
The invention also provides a method for preparing bamboo salt by using the obtained bamboo shoot extract, which comprises the following steps:
(1) pouring the refined salt raw material into a hopper of a bucket elevator, lifting the raw salt into a raw salt bin by the elevator, adding the raw salt into a mixer after the raw salt is metered by a metering device at a discharge port at the lower end of the raw salt bin, and starting the mixer;
(2) manually putting the bamboo shoot extracting solution into a liquid storage tank, and adding the bamboo shoot extracting solution into a mixer through a bamboo shoot solution metering pump;
(3) stopping stirring when the materials are uniformly mixed by the mixer to obtain mother salt;
(4) opening a discharge port at the lower end of the raw material salt bin, quantitatively feeding refined raw material salt into a mixing machine through a metering device, and stopping stirring when the mixing machine continuously stirs the materials until the materials are fully and uniformly mixed to obtain wet bamboo salt;
(5) and conveying the wet bamboo salt to a drying bed for drying, conveying the dried bamboo salt to a bamboo salt finished product bin, and packaging to obtain the finished product bamboo salt.
Further, the content of the bamboo shoot extract in the wet bamboo salt is 0.1 to 2.5 wt%, preferably 0.5 to 1.0 wt%, based on 1000kg of the raw salt.
The invention has the beneficial effects that:
(1) before the enzymolysis of the bamboo shoot protein, mixed bacteria consisting of trichoderma reesei, bacillus amyloliquefaciens, bacillus subtilis and bacillus licheniformis are adopted for fermentation, and cellulose, hemicellulose and the like in the bamboo shoot pulp can be degraded into micromolecules such as polysaccharide and the like, so that the constraint of the cellulose and the hemicellulose on the protein is avoided, the protein is separated, the full contact between the protein and protease is increased, the enzymolysis effect of the protein is improved, and the yield is ensured.
(2) The bamboo shoot protein is used as a fermentation substrate to carry out fermentation induction to produce the enzyme, and the enzyme is combined with commercial enzyme to hydrolyze the bamboo shoot protein, so that the enzyme which is consistent with the characteristics of the raw material and has higher specificity can be obtained by using a biological fermentation method, and the activity of the commercial enzyme with high enzyme is combined to realize the control of enzymolysis, improve the utilization rate of the raw material and reduce the production cost.
(3) Bamboo shoot protein is used as a main raw material for microbial fermentation, different moulds are introduced by adopting a fermentation process, a wider enzyme system, particularly acid protease and alkaline protease are generated, and the enzyme system and the acid protease and the alkaline protease are respectively cut from different parts of the protein and are cooperated to make up for deficiencies. Firstly, aspergillus niger is inoculated to ferment, on one hand, the aspergillus niger can fully utilize the nutrient content of bamboo shoot protein, promote the growth of the aspergillus niger, metabolize to generate rich enzyme systems, such as acid protease, pectinase, cellulase and xylanase, especially the acid protease with stronger activity, and can cut peptide chains from two ends. Secondly, bacillus subtilis is inoculated and fermented to produce alkaline protein which is mainly endonuclease, the activity of the endonuclease is very strong, and the alkaline protein can be hydrolyzed from the inside of the protein to cut off a peptide chain, so that micromolecular amino acid is generated.
The method adopts a combined technology of the biological enzyme method and the biological fermentation to deeply develop and utilize the bamboo shoot protein resource, and has the advantages of higher safety, mild conditions and easy control of the hydrolysis process. The fermentation and enzymolysis can decompose macromolecular protein into small molecular amino acid, thereby improving the absorption, nutrition and functional properties of the original protein. Therefore, the method has great economic and social significance by using the bamboo shoots or the bamboo shoot processing leftovers as raw materials and combining biological fermentation and enzymolysis technology to extract amino acid and polysaccharide in the bamboo shoots.
Drawings
FIG. 1 is a process flow chart of extracting amino acids and polysaccharides from bamboo shoots.
FIG. 2 is a flow chart of a process for preparing bamboo salt containing bamboo shoot extract.
Detailed Description
In order that the invention may be readily understood and appreciated, reference will now be made to the following examples and accompanying drawings. Herein,% represents mass unless otherwise specified.
Example 1
As shown in figure 1, the method for extracting amino acids and polysaccharides from bamboo shoots comprises the following steps:
(1) pretreatment: removing bamboo shoot shells and soil from 5kg of bamboo shoots, putting the bamboo shoots in a drum-type fruit washing machine, cleaning, draining, pressing the bamboo shoots into fragments with the thickness not more than 8mm by using a crusher, crushing the fragments by using a crusher, and sieving the fragments by using a 40-mesh sieve; and transferring the sieved bamboo shoot granules into a sandwich reaction tank, heating to the upper part of the reaction tank to 100 ℃, heating to the lower part of the reaction tank to 120 ℃, decocting for 2 times and 2 hours each time under the stirring condition (the stirring speed is 80r/min), and combining 30L of decoction liquid (15 kg of water is added for each time, and after the decoction, the decocted precipitation dregs are decocted again by adding water).
(2) Preparing a bamboo shoot protein solution: filtering the decoction with 10 mesh plate frame and 100 mesh coarse gauze respectively to obtain filtrate I and residue I. Standing the filtrate I for 10h, transferring into a vacuum extraction tank, and performing vacuum filtration at the vacuum degree of 400Pa and the temperature of 45 ℃ to obtain a filtrate II and a filter residue II; mixing the filter residue I and the filter residue II, and uniformly stirring with distilled water with the volume of 1 time to prepare a bamboo shoot residue suspension; adding the bamboo shoot residue suspension into a fermentation tank, and adding a mixture of the bamboo shoot residue suspension and the bamboo shoot residue suspension in a volume ratio of 1: 0.6: 1.2 Trichoderma reesei (Shandong Yiyuan kang Yuan Biotech Co., Ltd., viable count 3X 10)10cfu/ml), Bacillus amyloliquefaciens (Shandong Yiyuan kang Yuan Biotech Co., Ltd., viable count 2X 10)10cfu/ml), Bacillus licheniformis (Shandong LvLong Biotech Co., Ltd., viable count 3X 10)10cfu/ml) The total volume of the mixed bacteria accounts for 0.3% of the bamboo shoot residue suspension, then 0.3% of oxygen is introduced, the pH is controlled to be 4.5 by citric acid or potassium hydroxide, and the mixed bacteria are fermented for 72 hours at the temperature of 45 ℃. After fermentation, sterilizing at 120 ℃ for 10min, cooling to room temperature, and uniformly mixing with the filtrate II to obtain the bamboo shoot protein solution.
(3) Preparation of Aspergillus niger liquid and Bacillus subtilis liquid
Weighing 2% of soluble starch, 1% of glucose and 0.1% of KH (potassium hydrogen chloride)2PO4、0.1％MgSO4、0.2％NaNO3And in addition, the total volume of the culture medium is 1.2L with water, the pH is required to be adjusted to 7.2, the culture medium is sterilized at 120 ℃, then cooled to room temperature, black koji (Arctine Yiyuancuan biological technology Co., Ltd., inoculum size is 0.03%) is inoculated into the culture medium, and the culture medium is subjected to constant-temperature shaking culture at the rotation speed of 160r/min at the temperature of 28 ℃ for 72h to obtain an Aspergillus niger liquid.
Weighing 2% of glucose, 0.5% of NaCl, 0.1% of peptone and water in parts by mass, uniformly mixing, adjusting the pH value of the culture medium to 7.2, sterilizing at 120 ℃, cooling to room temperature, inoculating bacillus subtilis (Yiyuyuan Kongyuan Biotech Co., Ltd., inoculation amount of 0.02%) into the culture medium, and performing constant-temperature oscillation culture at 30 ℃ at the rotation speed of 160r/min for 24 hours to obtain the bacillus subtilis liquid.
(4) Preparation of bamboo shoot yeast material
The bamboo shoot protein solution is divided into three parts (the ratio of the bamboo shoot protein solution I, the bamboo shoot protein solution II and the bamboo shoot protein solution III is 1: 1: 8). And (3) placing the bamboo shoot protein solution I3L in a fermentation tank A, taking the bamboo shoot protein solution I as a mass reference, adding 2% of glucose, 0.3% of beef extract and 0.5% of NaCl, sterilizing at 120 ℃, cooling to room temperature, adding 10% of Aspergillus niger liquid obtained in the step (3), rotating at 160r/min, oscillating at 27 ℃ for fermentation for 72h, and centrifuging at 5000r/min for 10min to obtain a supernatant serving as the prepared bamboo shoot fermentation starter material I.
And (3) placing the bamboo shoot protein liquid II 3L in a fermentation tank B, taking the bamboo shoot protein liquid II as a mass reference, adding 2% of glucose, 0.3% of beef extract and 0.5% of NaCl, sterilizing at 120 ℃, cooling to room temperature, adding 5% of the bacillus subtilis liquid obtained in the step (3), performing oscillation fermentation at the rotation speed of 160r/min and the temperature of 36 ℃ for 38h, and centrifuging at the rotation speed of 5000r/min for 10min to obtain a supernatant, thus obtaining the bamboo shoot fermentation starter material II.
(5) Enzymolysis of bamboo shoot protein liquid
And (3) placing the remaining 24L of bamboo shoot protein liquid in an enzymolysis tank. Taking the bamboo shoot protein solution III as a mass, adding 1% of bamboo shoot fermentation starter I and 1% of pepsin, adjusting the pH to 4.2, and carrying out enzymolysis at 45 ℃ for 2.5h to obtain a bamboo shoot enzymolysis solution I; and regulating the pH value of the bamboo shoot enzymolysis liquid I to 8.3 by using potassium hydroxide, adding 1% of bamboo shoot fermentation starter II and 1% of trypsin, and carrying out enzymolysis for 3 hours at the temperature of 45 ℃ to obtain a bamboo shoot enzymolysis liquid II.
(6) Bamboo shoot enzymolysis liquid goes out enzyme, decoloration and removes flavor
Heating the bamboo shoot enzymolysis liquid II obtained in the step (5) to 120 ℃, preserving the temperature for 10min to inactivate enzyme, then cooling to 50 ℃, centrifugally separating at the rotating speed of 4000r/min, and separating residues from liquid to obtain filtrate III and filter residues III. The filter residue III can be used as feed. Adjusting pH of filtrate III to 5, adding active carbon (1g active carbon/100 ml filtrate), decolorizing at rotation speed of 100r/min for 30min, and filtering to obtain filtrate IV.
(7) Preparation of bamboo shoot extract
Circulating the filtrate IV obtained in the step (6) through a 1.0 μm microfilter and a 3000 dalton ultrafiltration membrane under 0.5MPa by using a circulating pump for 30min, and controlling the flow rate to be 60L/min to obtain an ultrafiltrate with the molecular weight of less than 3000 dalton; and (2) allowing the ultrafiltrate to pass through a 250 dalton nanofiltration membrane concentrator at 0.1MPa by using a circulating pump, observing that the liquid before the membrane is turbid and stops concentrating when precipitates are separated out to obtain nanofiltration concentrated solution, namely extracting solution rich in amino acid and polysaccharide, wherein the amino acid content is more than 2.5g/100ml and the polysaccharide content is more than 0.6g/100ml according to the detection methods of GB 5009.124 'determination of amino acid in national food safety standard' and NY/T1676 'determination of crude polysaccharide content in edible fungi'.
Example 2
As shown in figure 1, the method for extracting amino acids and polysaccharides from bamboo shoots comprises the following steps:
(1) pretreatment: removing bamboo shoot shells and soil from 10kg of bamboo shoots, placing the bamboo shoots in a drum type fruit washing machine, cleaning, draining, pressing to fragments with the thickness of not more than 6mm by using a crusher, then crushing by using a crusher, sieving by using a 20-mesh sieve, transferring bamboo shoot particles into a sandwich reaction tank, heating to the upper part of 100 ℃, decocting for 3 times and 2.5 hours each time under the stirring condition (the stirring speed is 60r/min), and combining 90L of decoction liquid (30 kg of water is added for each time, and after the decoction, the decocted sediment dregs are decocted by adding water again for decoction).
(2) Preparing a bamboo shoot protein solution: filtering the decoction with 10 mesh plate frame and 150 mesh coarse gauze to obtain filtrate I and residue I, standing for 11 hr, transferring to vacuum extraction tank (vacuum degree: 420Pa), vacuum filtering at 50 deg.C to obtain filtrate II and residue II; mixing the filter residue I and the filter residue II, and uniformly stirring with distilled water with the volume of 2 times of that of the mixture to prepare a bamboo shoot residue suspension; adding the bamboo shoot residue suspension into a fermentation tank, and adding a mixture of the bamboo shoot residue suspension and the bamboo shoot residue suspension in a volume ratio of 1: 0.6: 1.4 Trichoderma reesei (Shandong Yiyuan kang Yuan Biotech Co., Ltd., viable count 4X 10)10cfu/ml), Bacillus amyloliquefaciens (Shandong Yiyuan kang Yuan Biotech Co., Ltd., viable count 3X 10)10cfu/ml), Bacillus licheniformis (Shandong LvLong Biotech Co., Ltd., viable count 3X 10)10cfu/ml), the total volume of the mixed bacteria accounts for 0.4 percent of the bamboo shoot residue suspension, then 0.4 percent of oxygen is introduced, the pH value is controlled to be 4.8 by citric acid or potassium hydroxide, and the mixed bacteria are fermented for 64 hours at the temperature of 50 ℃. After fermentation, sterilizing at 120 ℃ for 12min, cooling to room temperature, and uniformly mixing with the filtrate II to obtain the bamboo shoot protein solution.
(3) Preparation of Aspergillus niger liquid and Bacillus subtilis liquid
Weighing 2% of soluble starch, 1% of glucose and 0.1% of KH (potassium hydrogen chloride)2PO4、0.1％MgSO4、0.2％NaNO3And in addition, 1.8L of water and total culture medium are added, the pH of the culture medium is adjusted to 7.2, the culture medium is sterilized at 120 ℃, then cooled to room temperature, Aspergillus niger (Arctia-Yiyuan-Kangyuan-Biotech Co., Ltd., inoculum size of 0.05%) is inoculated into the culture medium, and the Aspergillus niger liquid is obtained after constant-temperature shaking culture at 28 ℃ and the rotation speed of 160r/min for 72 h.
Weighing 2% of glucose, 0.5% of NaCl, 0.1% of peptone and water in parts by mass, uniformly mixing, adjusting the pH value of the culture medium to 7.2, sterilizing at 120 ℃, cooling to room temperature, inoculating bacillus subtilis (Yiyuyuan Kongyuan biological technology Co., Ltd., inoculation amount of 0.03%) into the culture medium, and performing constant-temperature oscillation culture at 30 ℃ at the rotation speed of 160r/min for 24 hours to obtain the bacillus subtilis liquid.
(4) Preparation of bamboo shoot yeast material
The bamboo shoot protein liquid is divided into three parts (the proportion of each part is 1: 1: 8). Placing bamboo shoot protein solution I9L in a fermentation tank A, taking the bamboo shoot protein solution I as a mass reference, adding 2% of glucose, 0.3% of beef extract and 0.5% of NaCl, sterilizing at 120 ℃, cooling to room temperature, adding 10% of Aspergillus niger liquid obtained in the step (3), performing oscillation fermentation at the rotation speed of 160r/min and the temperature of 28 ℃ for 72h to obtain bamboo shoot fermentation starter I, and centrifuging at the rotation speed of 6000r/min for 12min to obtain supernatant liquid to obtain the bamboo shoot fermentation starter I.
And (3) placing the bamboo shoot protein liquid II 9L in a fermentation tank B, taking the bamboo shoot protein liquid II as a mass reference, adding 2% of glucose, 0.3% of beef extract and 0.5% of NaCl, sterilizing at 120 ℃, cooling to room temperature, adding 5% of the bacillus subtilis liquid obtained in the step (3), performing oscillation fermentation at the rotating speed of 160r/min and the temperature of 35 ℃ for 38h, and centrifuging at the rotating speed of 6000r/min for 12min to obtain a supernatant, thus obtaining the bamboo shoot fermentation starter material II.
(5) Enzymolysis of bamboo shoot protein liquid
And (3) placing the remaining bamboo shoot protein liquid III 72L in an enzymolysis tank. Adding 2% of bamboo shoot yeast I and 2% of pepsin by mass of the bamboo shoot protein liquid III, adjusting the pH to 4.5, and carrying out enzymolysis at 48 ℃ for 3h to obtain a bamboo shoot enzymolysis liquid I; and regulating the pH value of the bamboo shoot enzymolysis liquid I to 8.6 by using potassium hydroxide, adding 2% of bamboo shoot starter II and 2% of trypsin, and carrying out enzymolysis at 48 ℃ for 3.5 hours to obtain a bamboo shoot enzymolysis liquid II.
(6) Bamboo shoot enzymolysis liquid goes out enzyme, decoloration and removes flavor
Heating the bamboo shoot enzymolysis liquid II obtained in the step (5) to 125 ℃, preserving the temperature for 11min for enzyme deactivation, then cooling to 55 ℃, centrifugally separating at the rotating speed of 5000r/min, and separating residues from liquid to obtain filtrate III and filter residues III. The filter residue III can be used as feed. Adjusting pH of filtrate III to 5.4, adding activated carbon (1g activated carbon/120 ml filtrate), decolorizing at rotation speed of 110r/min for 35min, and filtering to obtain filtrate IV.
(7) Preparation of bamboo shoot extract
Circulating the filtrate IV obtained in the step (6) through a 0.8 mu m microfilter and a 2800 Dalton ultrafiltration membrane under 0.55MPa for 30min by using a circulating pump, and controlling the flow rate to be 60L/min to obtain an ultrafiltrate with the molecular weight of less than 2800 Dalton; and (2) allowing the ultrafiltrate to pass through a 200 dalton nanofiltration membrane concentrator at 0.12MPa by using a circulating pump, observing that the liquid before the membrane is turbid and stops concentrating when precipitates are separated out to obtain nanofiltration concentrated solution, namely extracting solution rich in amino acid and polysaccharide, wherein the amino acid content is more than 2.5g/100ml and the polysaccharide content is more than 0.6g/100ml according to the detection methods of GB 5009.124 'determination of amino acid in national food safety standard' and NY/T1676 'determination of crude polysaccharide content in edible fungi'.
Example 3
As shown in FIG. 2, a method for preparing bamboo salt containing the extract solution of bamboo shoot of example 1 comprises the following steps:
(1) 1000kg of refined salt raw material (hereinafter referred to as raw salt) is poured into a hopper of a bucket elevator with the power of 10kw, and the raw salt is lifted to a raw salt bin by the elevator; manually putting the bamboo shoot extract into a 50L liquid storage tank;
(2) after the raw material salt is metered for 2 times by a metering device (100 kg/time) at a discharge port at the lower end of a raw material salt bin, 200kg of refined raw material salt is put into a mixer, and the mixer is started (the power is 35kw, and the stirring speed is 20 r/min);
(3) The bamboo shoot extract was measured by a measuring pump (20L/h) for 0.5h, and 10L of the bamboo shoot extract of example 1 was added to the mixer;
(4) mixing for 30min by a mixer, and stopping stirring to obtain mother salt;
(5) opening a discharge port at the lower end of the raw material salt bin, metering for 8 times by a metering device (100 kg/time), adding the remaining 800kg of refined raw material salt into a mixer, continuously stirring and mixing for 20min by the mixer, and stopping stirring to obtain wet bamboo salt;
(6) opening the discharge port of the mixer, and conveying the wet bamboo salt to a heating area of 6m by a screw conveyer with the power of 5kw2Is dried by the drying bedThe temperature of hot air is controlled at 130 ℃, the temperature of the tail gas of the drying bed is controlled at 80 ℃, the pressure in the drying bed is controlled at-10 Pa, and the thickness of a material layer in the drying bed is controlled at 250 mm;
(7) conveying the dried bamboo salt to a bamboo salt finished product bin through a conveying lifter, and then automatically packaging by using a packaging machine to obtain the finished product bamboo salt (the net packaging content is 0.25 kg).
Claims (14)
1. A preparation method of a bamboo shoot extracting solution comprises the following steps:
(1) cutting bamboo shoot into pieces, pulverizing, decocting for one or more times, mixing decoctions,
preferably, the ratio of the bamboo shoot crushed material to water 1: (2-10), preferably 1: (3-5) decocting in water for several times, which means that after decocting once, the supernatant is taken, and the remaining precipitated residue solids are further decocted in water, preferably in an amount of 2-10 times, more preferably 3-5 times, the amount of the precipitated residue;
(2) Filtering the decoction in the step (1) to obtain filter residue I and filtrate I, standing the filtrate I, performing suction filtration to obtain filtrate II and filter residue II, mixing the filter residue I and the filter residue II, and uniformly stirring with water to obtain a bamboo shoot residue suspension;
(3) adding the bamboo shoot residue suspension obtained in the step (2) into a fermentation tank, adding mixed bacteria comprising trichoderma reesei, bacillus amyloliquefaciens and bacillus licheniformis for fermentation, sterilizing fermentation liquor after fermentation is completed, cooling to room temperature, mixing with the suction filtration liquid II obtained in the step (2) to obtain bamboo shoot protein liquid, dividing the bamboo shoot protein liquid into three parts,
preferably, the weight ratio of the first part, the second part and the third part in the bamboo shoot protein liquid can be 1: 0.2-5: 4-12, further 1: 0.5-2: 6-10, further for example 1: 1: 8;
(4) adding the first part of the bamboo shoot protein solution I obtained in the step (3) into a fermentation tank A, adding a carbon source, a nitrogen source, inorganic salt and Aspergillus niger bacterial liquid into the fermentation tank A, oscillating and fermenting at constant temperature, centrifuging fermentation liquor, removing precipitate, and taking supernatant to obtain a bamboo shoot fermentation starter material I;
(5) adding the second part of the bamboo shoot protein liquid II obtained in the step (3) into a fermentation tank B, adding a carbon source, a nitrogen source, inorganic salt and bacillus subtilis liquid into the fermentation tank B, oscillating and fermenting at constant temperature, centrifuging fermentation liquor, removing precipitate, and taking supernatant to obtain a bamboo shoot fermentation starter material II;
(6) Adding the third part of the bamboo shoot protein liquid III in the step (3) into an enzymolysis tank, adding a bamboo shoot fermentation starter material I and acid protease, adjusting the pH to 4-5, preferably 4.2-4.8, carrying out enzymolysis, then adjusting the pH of the feed liquid to 8.0-10.5, preferably about 8.3-9, adding a bamboo shoot fermentation starter material II and alkali protease, and further carrying out enzymolysis to obtain an enzymolysis liquid;
(7) heating the enzymolysis liquid obtained in the step (6) to inactivate enzyme, cooling, performing centrifugal separation to obtain filter residue III and filtrate III, adjusting pH of the filtrate III to 4-6, preferably about 5-6, decolorizing, and filtering to obtain filtrate IV, to obtain bamboo shoot extract crude product.
2. The method of manufacturing of claim 1, wherein the method further comprises:
(8) and (3) passing the filtrate IV obtained in the step (7) through a micro-filter and a Dalton ultrafiltration membrane to obtain an ultrafiltrate with the molecular weight of less than 3000 Dalton, and passing the ultrafiltrate through a nanofiltration membrane concentrator to obtain a nanofiltration concentrated solution, namely the bamboo shoot extract.
3. The production method according to claim 1 or 2, wherein the composition of the mixed bacteria in step (3) is Trichoderma reesei, Bacillus amyloliquefaciens, Bacillus licheniformis,
the volume ratio of trichoderma reesei to bacillus amyloliquefaciens to bacillus licheniformis is 1: 0.5-0.8: 1-2, preferably 1: 0.6: 1.2-1.4, the colony concentration of Trichoderma reesei is 3X 10 10-5×1010cfu/ml, preferably 3X 1010-4×1010cfu/ml, colony concentration of Bacillus amyloliquefaciens 2X 1010-4×1010cfu/ml, preferably 2X 1010-3×1010cfu/ml, colony concentration of Bacillus licheniformis 3X 1010-5×1010cfu/ml, preferably 3X 1010-4×1010cfu/ml; and/or
The fermentation conditions of the fermentation tank in the step (3) are that the ratio of the oxygen amount introduced per minute to the volume of the feed liquid is 0.3-0.6, preferably 0.3-0.4, the pH value of the feed liquid is controlled to be 4.5-5.5, preferably 4.5-4.8, the fermentation temperature is controlled to be 45-55 ℃, preferably 45-50 ℃, and the fermentation time is 36-72 hours, preferably 48-64 hours.
4. The preparation method according to any one of claims 1 to 3, wherein in the steps (4) and (5), the inoculation amount of the Aspergillus niger bacterial liquid accounts for 5-15V%, preferably 8-12V%, and the inoculation amount of the Bacillus subtilis bacterial liquid accounts for 2-10V%, preferably 4-8V%, of the bamboo shoot protein liquid II.
5. The production method according to any one of claims 1 to 4, wherein in steps (4) and (5), the carbon source of the fermentor A or B is glucose, which accounts for 1 to 2 wt% of the respective bamboo shoot protein solution, the nitrogen source is beef extract, which accounts for 0.2 to 0.3 wt% of the respective bamboo shoot protein solution, and the inorganic salt is sodium chloride, which accounts for 0.3 to 0.5 wt% of the respective bamboo shoot protein solution.
6. The process according to any one of claims 1 to 5, wherein the fermentation conditions in steps (4) and (5) are oscillated at constant temperature, the temperature for the fermentation of Aspergillus niger is 27 to 30 ℃, the rotation speed is 150 to 160r/min, the temperature for the fermentation of Bacillus subtilis is 33 to 36 ℃, and the rotation speed is 150 to 160 r/min.
7. The method according to any one of claims 1 to 6, wherein in the step (6), the bamboo shoot fermentation starter I or II is added in an amount of 1 to 3 wt%, preferably 1 to 2 wt%, of the bamboo shoot protein solution III, and the acidic protease or the alkaline protease is added in an amount of 1 to 3 wt%, preferably 1 to 2 wt%, of the bamboo shoot protein solution III.
8. The production method according to any one of claims 1 to 7, wherein, in step (6), the acidic protease is pepsin and the alkaline protease is pancreatin.
9. The preparation method according to any one of claims 1 to 8, wherein in the step (7), the filtering activated carbon is filtered by a plate-and-frame filter, the ratio of the activated carbon in the enzymolysis solution is 1 to 2g of activated carbon added in each 100 to 150ml of enzymolysis solution, and the decoloring time is 30 to 40 min.
10. The method of any one of claims 1-9, wherein in step (8), the microfilter is between 0.4 and 1.0 μm, preferably between 0.5 and 0.8 μm, the ultrafiltration membrane is between 1000 and 3000 daltons, preferably between 1800 and 2800 daltons, and the nanofiltration membrane concentrator is between 150 and 250 daltons, preferably between 200 and 220 daltons.
11. The extract solution of bamboo shoot obtained by the method according to any one of claims 1 to 10, which is measured according to the methods specified in GB 5009.124 "determination of amino acids in food safety national standards" and NY/T1676 "determination of crude polysaccharide content in edible fungi", has an amino acid content of more than 2.5g/100ml and a polysaccharide content of more than 0.6g/100 ml.
12. A bamboo salt comprising an edible salt and the bamboo shoot extract solution as claimed in claim 11, wherein the bamboo shoot extract solution is contained in an amount of 0.1 to 2.5 wt%, preferably 0.5 to 1.0 wt%, based on the weight of the raw salt (edible salt).
13. A method for preparing bamboo salt using the bamboo shoot extract liquid of claim 11, comprising the steps of:
(1) pouring the refined salt raw material into a hopper of a bucket elevator, lifting the raw salt into a raw salt bin by the elevator, adding the raw salt into a mixer after the raw salt is metered by a metering device at a discharge port at the lower end of the raw salt bin, and starting the mixer;
(2) manually putting the bamboo shoot extracting solution into a liquid storage tank, and adding the bamboo shoot extracting solution into a mixer through a bamboo shoot solution metering pump;
(3) stopping stirring when the materials are uniformly mixed by the mixer to obtain mother salt;
(4) opening a discharge port at the lower end of the raw material salt bin, quantitatively feeding refined raw material salt into a mixing machine through a metering device, and stopping stirring when the mixing machine continuously stirs the materials until the materials are fully and uniformly mixed to obtain wet bamboo salt;
(5) and conveying the wet bamboo salt to a drying bed for drying, conveying the dried bamboo salt to a bamboo salt finished product bin, and packaging to obtain the finished product bamboo salt.
14. The method according to claim 13, wherein the wet bamboo salt in the step (4) comprises 0.1-2.5 wt%, preferably 1.5-2.0 wt% of bamboo shoot extract solution based on 1000kg of raw salt.
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