CN111631312A - High-quality enzymolysis fermentation soybean meal - Google Patents
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Abstract
The invention relates to high-quality enzymolysis fermented soybean meal. The preparation method of the high-quality enzymolysis fermented soybean meal comprises the steps of crushing the peeled soybean meal, sieving the crushed soybean meal with a 80-mesh sieve, carrying out first-step enzymolysis by using alpha-galactosidase and glucoamylase, carrying out second-step enzymolysis by using papain, inactivating enzymes, inoculating composite strains of saccharomycetes, bacillus and lactobacillus, carrying out anaerobic fermentation after aerobic fermentation, and keeping active ingredients of a finished product to the maximum extent through spray drying. The prepared enzymolysis and fermentation soybean meal has the characteristics of special fermentation fragrance, protein content of about 55 percent, acid soluble protein content of more than 50 percent, basic removal of anti-nutritional factors such as antigen and the like, stable components, strong hydrophilicity, high water retention rate, strong palatability, obvious food calling for aquatic animals and the like. The feed is rich in functional small peptides, probiotics and metabolites thereof, is beneficial to the nutrition balance of intestinal tracts of cultured animals, solves the problem of nutritional diarrhea of the animals, is suitable for feeds of piglets, poultry, pets, aquatic products and the like, and is recommended to be used in a proportion of 2-5%.
Description
The application is a divisional application of an invention patent application on application date 2016.12.15, application number 201611163397.1 and invention name of 'high-quality enzymolysis fermented soybean meal and a preparation method and application thereof'.
Technical Field
The invention relates to high-quality enzymolysis fermented soybean meal and a preparation method and application thereof.
Background
The soybean meal is a byproduct obtained by extracting oil from soybeans, and is widely applied to the feed industry because the soybean meal contains rich protein (content: 43-48%), balanced amino acids, various mineral nutrient components and lysine (content: 2.5-2.8%) which is easy to be deficient in other vegetable feeds. However, in the oil extraction process, the obtained soybean meal is often subjected to high-temperature treatment, so that protein denaturation is serious, the solubility is poor, and digestion and absorption of animals are not facilitated. Meanwhile, the bean pulp also contains a plurality of anti-nutritional factors, such as phytic acid, oligosaccharide and glycinin, and the like which have thermal stability, so that the untreated bean pulp is easily caused to the problems of diarrhea, intestinal allergy and the like if being directly fed by animals, and is particularly not beneficial to young animals with incomplete intestinal functions. Therefore, the feeding value of the soybean meal is greatly influenced.
The enzymolysis soybean meal and the fermentation soybean meal are the mixture of soluble protein and small molecular polypeptide which is degraded by protease enzymolysis or microbial fermentation of soybean protein by using modern biotechnology. The protein treated by enzymolysis or fermentation is considered as an ideal plant protein of the feed for young animals due to the characteristics of easier absorption, low antigen and the like compared with the protein in the traditional soybeans. At present, related researches in China are numerous, and due to differences of design processes, differences of used enzymes or microbial strains, non-uniformity of parameters such as processing time and temperature and the like, products present diversified situations. The indexes of the finished product are greatly differentiated due to different treatment processes. The problems of unstable product quality, poor palatability, incomplete anti-nutritional factor treatment, substandard small peptide content and the like generally exist, fish meal cannot be effectively replaced, and the requirements of young animals on palatability, diarrhea resistance and high digestibility are really met. Products of the same type rely more on imports, such as: danish Hamlet and the like, but the use proportion in actual production is very small due to the problems of high price, unstable resource supply, long transportation period and the like of imported products. The quality of the enzymatic hydrolysis or fermentation soybean meal is improved, the functional soybean peptide with low antigen, easy absorption, good palatability and high activity retention rate is really realized, the batch production can be realized, and the method is a problem to be solved urgently by a plurality of scientific research institutions and manufacturers.
Disclosure of Invention
In order to solve the defects of the prior art, the invention aims to provide high-quality enzymolysis fermented soybean meal and a preparation method and application thereof.
The technical scheme of the invention is as follows:
a preparation method of high-quality enzymolysis fermented soybean meal comprises the following steps:
1) crushing the peeled soybean meal, sieving with a 80-mesh sieve, adding water which is 3-4 times of the weight of the soybean meal, and uniformly stirring;
2) adding alpha-galactosidase with the weight of 0.5-1 per mill of the weight of the soybean meal and saccharifying enzyme with the weight of 1-1.5 per mill of the weight of the soybean meal into the material obtained in the step 1) for enzymolysis; adding papain with the weight of 0.3-0.5 per mill of the soybean meal for enzymolysis after complete enzymolysis; heating to inactivate enzyme after complete enzymolysis;
3) cooling the material obtained in the step 2) to 30-35 ℃, inoculating mixed bacteria of saccharomycetes, bacillus and lactic acid bacteria, performing aerobic fermentation for 20-24h, performing anaerobic fermentation for 48-50h, and completing fermentation;
4) passing the material obtained in the step 3) through a colloid mill; spray drying to obtain the final product.
The content of crude protein in the soybean meal is more than or equal to 43.0 percent, and the soybean meal is preferably peeled, and the content of the crude protein is more than 45 percent.
The research of the invention finds that when liquid enzymolysis is carried out, if the water adding amount is too low (for example, less than 3 times of the weight of the soybean meal), the soybean meal is easy to agglomerate, the mixing uniformity of an enzyme preparation is influenced, and the enzymolysis effect is seriously influenced due to incomplete enzymolysis; if the water addition amount is too high (for example, more than 5 times of the weight of the soybean meal), the energy consumption is increased during the drying in the subsequent steps due to the too high water content, the production efficiency is affected, and the cost is increased. Based on a large amount of research, the optimal weight ratio of the soybean meal to the water is determined to be 1: 3-4.
The invention uses alpha-galactosidase and glucoamylase to carry out enzymolysis on the soybean meal, and can effectively remove soybean oligosaccharides (stachyose, raffinose, sucrose and the like) in the soybean meal; the bean pulp is subjected to enzymolysis treatment by using papain, so that the soybean protein is decomposed into micromolecular protein and small peptide molecules, the content of small peptides in a finished product is effectively improved, and the soybean pulp is more beneficial to absorption and utilization of young animals.
The enzymolysis process of the invention is divided into two steps, alpha-galactosidase and glucoamylase are added in the first step, and papain is added for the second step of enzymolysis after the enzymolysis is completed. The three enzymes cannot be added simultaneously, otherwise, the papain can decompose the alpha-galactosidase and the saccharifying enzyme, and the removal effect of the oligosaccharide is influenced.
The complete standard of the first step of enzymolysis is as follows: the total content of stachyose and raffinose is less than 1%, and the detection can be carried out by the conventional method in the field.
The invention further optimizes the addition amount and enzymolysis conditions of alpha-galactosidase and glucoamylase in the first enzymolysis process:
preferably, the addition amount of the alpha-galactosidase is 0.5 per mill of the weight of the soybean meal;
preferably, the addition amount of the saccharifying enzyme is 1 per mill of the weight of the soybean meal;
the optimum pH6.0, the optimum enzymolysis temperature is 50-55 ℃; the enzymolysis time is preferably 2-3 h.
The complete standard of the second step of enzymolysis is as follows: the content of acid soluble protein (small peptide) is detected to be more than 35 percent, and the detection can be carried out by the conventional method in the field.
The invention further optimizes the addition amount and enzymolysis conditions of papain in the second step of enzymolysis:
preferably, the addition amount of the papain is 0.3 per mill of the weight of the soybean meal;
the optimum pH6.0, the optimum enzymolysis temperature is 50-55 ℃; the enzymolysis time is preferably 3-5 h.
The alpha-galactosidase, the saccharifying enzyme and the papain are general terms in the field, and all enzyme preparations can be commercially obtained. Wherein, the enzyme activity of the alpha-galactosidase is preferably 3000IU/g, the enzyme activity of the saccharifying enzyme is preferably 20 ten thousand IU/g, and the enzyme activity of the papain is preferably 80 ten thousand IU/g.
The heating enzyme deactivation is preferably carried out for 30 minutes by immediately heating to 85-90 ℃ after the enzymolysis is finished.
The invention uses the synergistic effect of the saccharomycetes, the bacillus and the lactic acid bacteria, improves the protein solubility of the soybean meal, and is beneficial to digestion and absorption; and the microorganisms further decompose the residual polysaccharide or oligosaccharide of the soybean meal in the fermentation process. According to the invention, active substances such as lactic acid, probiotics, glutamic acid, lactic acid, vitamins, UGF (unknown growth factors) and the like can be generated in the anaerobic fermentation stage, so that the flavor of the enzymatic fermentation soybean meal is improved, and the product has a certain aromatic smell and delicate flavor, has a certain feeding attraction and is good in palatability; is beneficial to intestinal health of the cultured animals.
The fermentation process of the invention is divided into two stages, the first stage is firstly carried out aerobic (or aerobic) fermentation, which is beneficial to the rapid proliferation of yeast and the utilization of inorganic nutrient elements by bacillus; the anaerobic fermentation is carried out in the second stage, which is beneficial to the yeast to generate metabolites, provides an anaerobic environment for the rapid proliferation of lactic acid bacteria, and generates organic acids such as lactic acid.
In order to meet the requirement of large increment of yeast in the aerobic fermentation stage, cane molasses with the weight of 1% of that of the raw material soybean meal is added as a carbon source, and the yeast in the anaerobic stage generates a large amount of metabolites, so that the intestinal health of cultured animals is facilitated.
The inoculation amount of the mixed bacteria is preferably 1-2 per mill of the weight of the dry matter of the soybean meal raw material.
Preferably, the effective viable count of yeast, bacillus and lactic acid bacteria in the mixed bacteria is 4 × 109CFU/g、5×109CFU/g、6×109CFU/g。
The yeast, bacillus and lactic acid bacteria can be purchased from the market.
The yeast is preferably Saccharomyces cerevisiae 2.1392; the bacillus is preferably bacillus foddensis Paenibacillus pabuli 1.3772; the lactic acid bacteria are preferably Pediococcus pentosaceus pendosaceus 1.2695; the above strains are purchased from China general microbiological culture Collection center.
The research of the invention finds that the colloid mill treatment is carried out before the spray drying, which can effectively avoid the blocking of the die hole caused by the agglomeration of the materials due to rich small peptide content in the enzymolysis and fermentation processes; effectively improving the product quality.
Preferably, the mixture enters a colloid mill, is sieved by a 100-mesh sieve, and is subjected to spray drying to obtain a finished product, wherein the finished product can directly pass through a 80-mesh sieve, has uniform fineness and can be directly packaged.
The invention uses spray drying technology, which is different from flash drying or boiling drying machine used in the same line. The drying mode is rapid in drying, can be directly dried into powder, omits the step of crushing after drying, and has uniform fineness and strong free-running property of the finished product; active substances of the enzymolysis and fermentation soybean meal can be retained to the maximum extent, the product quality is ensured, and the retention rate is extremely high for bioactive substances generated in the fermentation process, the dissolubility, the color, the fragrance, the taste and the like of the product.
The principle of the spray dryer is as follows: the spray dryer is heated to convert hot air into hot air, the hot air enters a hot air distributor arranged at the top of the drying chamber, then uniformly enters the drying chamber and rotates in a spiral shape, meanwhile, feed liquid is conveyed to a centrifugal atomizer arranged at the top of the drying chamber to be atomized into tiny atomized liquid drops, the feed liquid and the hot air are in parallel flow contact, moisture is rapidly evaporated, and the feed liquid is dried into a finished product in a short time. The finished product is discharged from the bottom of the drying tower and the cyclone separator, and the waste gas is pumped out by a fan and is emptied.
Preferably, the spray drying conditions are: the air inlet temperature is 240 ℃, and the air outlet temperature is 95 ℃.
The invention uses high-temperature enzyme deactivation and spray drying high-temperature treatment to play a good role in passivating and removing heat-sensitive factors such as protease inhibitor, soybean agglutinin, urease and the like.
Specifically, the preparation method of the high-quality enzymolysis fermented soybean meal comprises the following steps:
1) crushing the peeled soybean meal, sieving with a 80-mesh sieve, adding water which is 3-4 times of the weight of the soybean meal, and uniformly stirring;
2) adding alpha-galactosidase with the weight of 0.5 per thousand of that of the soybean meal and saccharifying enzyme with the weight of 1 per thousand of that of the soybean meal into the material obtained in the step 1), and carrying out enzymolysis for 2-3 h; then adding papain with the weight of 0.3 per mill of the weight of the soybean meal for enzymolysis for 2-3 h; immediately heating to 85-90 ℃ after complete enzymolysis, and inactivating enzyme for 30 minutes;
wherein the enzymolysis condition is pH6.0, and the enzymolysis temperature is 50-55 deg.C; the enzyme activities of the alpha-galactosidase, the glucoamylase and the papain are 3000IU/g, 20 ten thousand IU/g and 80 ten thousand IU/g respectively;
3) cooling the material obtained in the step 2) to 30-35 ℃, inoculating mixed bacteria of yeast, bacillus and lactobacillus with the inoculation amount being 1 per mill of the weight of the dry matter of the soybean meal raw material, performing aerobic fermentation for 20-22h, performing anaerobic fermentation for 48-50h, and completing fermentation;
the mixed strain is prepared by the conventional method in the field, and the effective viable count of the saccharomycetes, the bacillus and the lactic acid bacteria in the mixed strain is respectively 4 × 109CFU/g、5×109CFU/g、6×109CFU/g；
The yeast is Saccharomyces cerevisiae 2.1392; the bacillus is feed bacillus Paenibacillus pabuli 1.3772; the lactobacillus is Pediococcus pentosaceus 1.2695;
4) and (3) passing the material obtained in the step 3) through a colloid mill, sieving the material by a 100-mesh sieve, and performing spray drying to obtain a finished product.
The invention also comprises the enzymolysis fermented soybean meal prepared by the method.
The invention also provides application of the enzymolysis and fermentation soybean meal in preparation of animal feed.
The invention also comprises animal feed containing the enzymolysis and fermentation soybean meal.
The enzymolysis fermented soybean meal and the feed are suitable for the fields of livestock (such as piglets), poultry, pets, aquaculture and the like.
Preferably, the addition amount of the enzymolysis and fermentation soybean meal in the animal feed is 2-5% (weight ratio).
The enzymolysis and fermentation soybean meal product is light yellow to yellow powder, has the crude protein content of about 55 percent and the acid-soluble protein content of more than 50 percent, has special fermentation fragrance, and has the smell obviously superior to that of the common enzymolysis soybean meal.
The high-quality enzymolysis and fermentation soybean meal product is different from common enzymolysis soybean meal or fermentation soybean meal. Firstly, the antigen of the product is basically removed of beta-conglycinin and acid-glycinin, and the Base-glycinin is basically removed; and secondly, compared with the common enzymolysis method or the fermented soybean meal, the oligosaccharide is more completely removed. The content of stachyose and raffinose in the product is lower than 1%; thirdly, the crude protein content is about 55 percent and is increased by 15 to 20 percent compared with that before enzymolysis and fermentation, while the protein content of the common fermented soybean meal is only increased by 8 to 10 percent, and the protein rising ratio is far higher than that of the common enzymolysis or fermented soybean meal; fourthly, the macromolecular protein of the product is basically changed into a micromolecular peptide segment, and the molecular weight is all less than 10 Kda; the content of small peptides is high, the acid-soluble protein content is more than 50%, while the acid-soluble protein content of common fermented soybean meal is generally 9-15%, the acid-soluble protein content of common zymolytic protein is generally 20-30%, and the acid-soluble protein content of the product is far higher than that of common fermented soybean meal or zymolytic soybean meal.
Therefore, the high-quality enzymolysis and fermentation soybean meal product improves the digestibility and palatability of soybean protein, is beneficial to the nutrition balance of intestinal tracts of cultured animals, and solves the problem of nutritional diarrhea of the animals; the feed is rich in functional small peptides, probiotics and metabolites thereof (lactobacillus, butyric acid bacteria, saccharomyces boulardii, lactic acid and butyric acid), has good palatability and obvious food calling property, and can effectively improve the flavor of the feed so as to improve the health of cultured animals; meanwhile, the water-retaining agent has the characteristics of stable components, strong hydrophilicity, high water retention rate, no delamination in water and the like. And the common fermented soybean meal or enzymolysis soybean meal has poor hydrophilicity and obvious layering phenomenon in water.
In addition, the main steps of the technological process of the invention, namely enzymolysis and fermentation, can be carried out in an enzymolysis tank, each production link is strictly controlled by a temperature control system, the problems that the traditional solid fermentation is greatly influenced by the environment, the indexes of the stacking solid fermentation are not uniform, cross contamination is easy to occur and the like are avoided, the quality uniformity of finished products can be effectively ensured, the indexes of the finished products are stable, and continuous batch production can be realized.
Therefore, the soybean meal subjected to enzymolysis and fermentation is low in antigen and small in molecular weight, beneficial to effective direct absorption of animal intestinal tracts, stable in quality and controllable in processing flow, can be used as functional protein to replace imported fish meal, and effectively reduces the use proportion of the imported fish meal in the formula. The production cost of the method is about 2000 yuan/T, the raw material cost is about 3500 yuan/T, the total is about 5500 yuan/T, the price of the normal fish meal with equal protein is about 8000 yuan/T, and the market price of the Hamlet of the imported enzymolysis soybean meal is about 9500 yuan/T, so that the 2-5% of the product is used for replacing the fish meal in the feed formula, the feed cost can be effectively saved by about 50-100 yuan/T, the breeding benefit is obviously improved, and higher economic value is created.
Drawings
FIG. 1 is a diagram showing the results of detection of SDS-PAGE antigen protein in experiment 3.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or instruments used are conventional products available from regular distributors, not indicated by the manufacturer.
The peeled soybean meal contains crude protein more than or equal to 45%, α -galactosidase from Guangdong Yiduoli company and having enzyme activity of 3000IU/g, saccharifying enzyme from Hunan Lierkang company and having enzyme activity of 20 ten thousand IU/g, papain from Jiangmen Tiancheng company and having enzyme activity of 80 ten thousand IU/g, Saccharomyces cerevisiae 2.1392, Bacillus subtilis Paenibacillus Paenibacillus pauli 1.3772, Lactobacillus pentosaceus Pentococcus pentosaceus 1.2695, and the strains are from China general microorganism strain preservation management center, and the effective viable count of the yeast, the bacillus and the lactobacillus in the mixed bacteria are respectively 4 × 109CFU/g、5×109CFU/g、6×109CFU/g。
Example 1
A preparation method of high-quality enzymolysis fermented soybean meal comprises the following steps:
1) crushing 1000kg of peeled soybean meal, sieving with a 80-mesh sieve, adding water with the weight 3-4 times of the weight of the soybean meal, and uniformly stirring;
2) adding alpha-galactosidase with the weight of 0.5 per thousand of that of the soybean meal and saccharifying enzyme with the weight of 1 per thousand of that of the soybean meal into the material obtained in the step 1), and carrying out enzymolysis for 2-3 h; then adding papain with the weight of 0.3 per mill of the weight of the soybean meal for enzymolysis for 2-3 h; immediately heating to 85-90 ℃ after complete enzymolysis, and inactivating enzyme for 30 minutes;
wherein the enzymolysis condition is pH6.0, and the enzymolysis temperature is 50-55 deg.C; the enzyme activities of the alpha-galactosidase, the glucoamylase and the papain are 3000IU/g, 20 ten thousand IU/g and 80 ten thousand IU/g respectively;
3) cooling the material obtained in the step 2) to 30-35 ℃, inoculating mixed bacteria of yeast, bacillus and lactobacillus with the inoculation amount of 1 per mill of the weight of the dry matter of the soybean meal raw material, performing aerobic fermentation for 20-22h, performing anaerobic fermentation for 48-50h, and completing fermentation, wherein the effective viable count of the yeast, the bacillus and the lactobacillus in the mixed bacteria is 4 × 10 respectively9CFU/g、5×109CFU/g、6×109CFU/g；
4) And (3) feeding the material obtained in the step 3) into a colloid mill, sieving the material by a 100-mesh sieve, and performing spray drying to obtain a finished product.
Example 2
A preparation method of high-quality enzymolysis fermented soybean meal comprises the following steps:
1) crushing 1000kg of peeled soybean meal, sieving with a 80-mesh sieve, adding water with the weight 3-4 times of the weight of the soybean meal, and uniformly stirring;
2) adding alpha-galactosidase with the weight of 0.3 per thousand of that of the soybean meal and saccharifying enzyme with the weight of 1 per thousand of that of the soybean meal into the material obtained in the step 1), and carrying out enzymolysis for 1.5-2 h; then adding papain with the weight of 0.2 per mill of the weight of the soybean meal for enzymolysis for 1.5-2 h; immediately heating to 85-90 ℃ after complete enzymolysis, and inactivating enzyme for 30 minutes;
wherein the enzymolysis condition is pH6.0, and the enzymolysis temperature is 50-55 deg.C; the enzyme activities of the alpha-galactosidase, the glucoamylase and the papain are 3000IU/g, 20 ten thousand IU/g and 80 ten thousand IU/g respectively;
3) cooling the material obtained in the step 2) to 30-35 ℃, inoculating mixed bacteria of yeast, bacillus and lactic acid bacteria (same as the embodiment 1), wherein the inoculation amount is 2 per mill of the weight of the dry matter of the soybean meal raw material, performing aerobic fermentation for 18-20h, and performing anaerobic fermentation for 48-50h to complete the fermentation;
4) and (3) feeding the material obtained in the step 3) into a colloid mill, sieving the material by a 100-mesh sieve, and performing spray drying to obtain a finished product.
Example 3
A preparation method of high-quality enzymolysis fermented soybean meal comprises the following steps:
1) crushing 1000kg of peeled soybean meal, sieving with a 80-mesh sieve, adding water with the weight 3-4 times of the weight of the soybean meal, and uniformly stirring;
2) adding alpha-galactosidase with the weight of 0.5 per thousand of that of the soybean meal and saccharifying enzyme with the weight of 1 per thousand of that of the soybean meal into the material obtained in the step 1), and carrying out enzymolysis for 1.5-2 h; then adding papain with the weight of 1.0 per mill of the weight of the soybean meal for enzymolysis for 1.5-2 h; immediately heating to 85-90 ℃ after complete enzymolysis, and inactivating enzyme for 30 minutes;
wherein the enzymolysis condition is pH6.0, and the enzymolysis temperature is 50-55 deg.C; the enzyme activities of the alpha-galactosidase, the glucoamylase and the papain are 3000IU/g, 20 ten thousand IU/g and 80 ten thousand IU/g respectively;
3) cooling the material obtained in the step 2) to 30-35 ℃, inoculating mixed bacteria of yeast, bacillus and lactic acid bacteria (same as the embodiment 1), wherein the inoculation amount is 2 per mill of the weight of the dry matter of the soybean meal raw material, performing aerobic fermentation for 18-20h, and performing anaerobic fermentation for 48-50h to complete the fermentation;
4) and (3) feeding the material obtained in the step 3) into a colloid mill, sieving the material by a 100-mesh sieve, and performing spray drying to obtain a finished product.
Experimental example 1
The finished products of the enzymolysis and fermentation soybean meal of the examples 1 to 3 are subjected to conventional detection, and the detection method and indexes are shown in table 1.
The detection method comprises the following steps:
the crude protein detection method comprises the following steps: GB/T6432-
The water content detection method comprises the following steps: GB/T6435-
The ash content detection method comprises the following steps: GB/T6438-2007
The acid soluble protein detection method comprises the following steps: GB/T22492-
Protein solubility detection method: DB 13/T812-
The detection method of amino nitrogen comprises the following steps: GB/T5009.39-2003
The pH value detection method comprises the following steps: GB/T8884-2007.
TABLE 1
Table 1 shows that, in example 2, the papain addition ratio is reduced, the fermentation time is shortened, the cost can be saved, and the production efficiency can be improved, but the finished product indexes of acid soluble protein, amino nitrogen and crude protein are obviously lower than those in examples 1 and 3; example 3 the ratio of papain to strain added is further increased on the basis of example 1, the detection index of the finished product is similar to that of example 1, and the cost is obviously increased, therefore, example 1 is the preferred scheme of the invention.
Comparative example 1
A preparation method of common fermented soybean meal comprises the following steps:
1) 1000kg of peeled soybean meal is taken;
2) the mixed strain (same as the embodiment 1) is placed in warm water of about 5kg and 35 ℃ and stirred for 5 minutes to activate the strain; the inoculation amount is 1 per mill of the dry matter weight of the soybean meal raw material;
3) taking 300g of papain (80 ten thousand IU/g) (the adding proportion is as follows: 0.3 per mill), placing in 2kg of 50 ℃ warm water, and carrying out enzyme activation;
4) placing the activated bacteria liquid and the activated enzyme liquid into 500kg of warm water at 35 ℃, uniformly stirring, and uniformly mixing with 1000kg of soybean meal;
5) bagging, placing in a special breathing bag for fermented soybean meal, wherein each bag is 40 kg;
6) standing at about 30 deg.C for 72 hr for fermentation;
7) drying in a drum dryer;
8) and (6) crushing and packaging.
The working principle of the rotary drum dryer is as follows: the heat source of the dryer comes from a combustion device, and the dryer adopts a concurrent heating mode. Therefore, the materials to be dried enter the cylinder body from the feeding box and the feeding chute, and are pushed backwards by the spiral shoveling plate. Because the dryer is placed in an inclined mode, materials flow to the rear end under the action of gravity and rotation on the one hand, and on the other hand, the materials are repeatedly picked up by the picking plate and are brought to the upper end to be continuously scattered, so that the materials form an even curtain in the cylinder, the materials fully exchange heat with hot air in the cylinder, and the contained moisture is gradually dried due to repeated scattering of the materials, so that the drying purpose is achieved.
Comparative example 2
A preparation method of common fermented soybean meal comprises the following steps:
1) 1000kg of peeled soybean meal is taken;
2) the mixed strain (same as the embodiment 1) is placed in warm water of about 5kg and 35 ℃ and stirred for 5 minutes to activate the strain; the inoculation amount is 2 per mill of the dry matter weight of the soybean meal raw material;
3) taking 300g of papain (80 ten thousand IU/g) (the adding proportion is as follows: 0.3 per mill), placing in 2kg of 50 ℃ warm water, and carrying out enzyme activation;
4) placing the activated bacteria liquid and the activated enzyme liquid into 500kg of warm water at 35 ℃, uniformly stirring, and uniformly mixing with 1000kg of soybean meal;
5) bagging, placing in a special breathing bag for fermented soybean meal, wherein each bag is 40 kg;
6) standing at about 30 deg.C for 72 hr for fermentation;
7) drying in a drum dryer;
8) and (6) crushing and packaging.
Comparative example 3
A preparation method of common fermented soybean meal comprises the following steps:
1) 1000kg of peeled soybean meal is taken;
2) the mixed strain (same as the embodiment 1) is placed in warm water of about 5kg and 35 ℃ and stirred for 5 minutes to activate the strain; the inoculation amount is 1 per mill of the dry matter weight of the soybean meal raw material;
3) 500g of papain (80 ten thousand IU/g) is taken (the adding proportion is as follows: 0.5 per mill), placing in 2kg of 50 ℃ warm water, and carrying out enzyme activation;
4) placing the activated bacteria liquid and the activated enzyme liquid into 500kg of warm water at 35 ℃, uniformly stirring, and uniformly mixing with 1000kg of soybean meal;
5) bagging, placing in a special breathing bag for fermented soybean meal, wherein each bag is 40 kg;
6) standing at about 30 deg.C for 72 hr for fermentation;
7) drying in a drum dryer;
8) and (6) crushing and packaging.
Experimental example 2
The finished products of the enzymolysis and fermentation soybean meal of the comparative examples 1 to 3 are subjected to conventional detection, the detection method is the same as that of the experiment 1, and the detection indexes are shown in a table 2.
TABLE 2
Comparative examples 1-3 are conventional solid fermented soybean meal with basically similar indexes, comparative example 2 increases the strain addition ratio and reduces the papain addition ratio, and the acid soluble protein content is obviously reduced; the proportion of the papain is increased in the comparative example 3, and the detection result of the finished product is not much different from that in the comparative example 1.
Experimental example 3
The high-quality enzymolysis fermented soybean meal prepared in example 1, the common fermented soybean meal and the peeled soybean meal prepared in comparative example 1 were respectively detected as follows:
1) antigen detection
The results of protein detection by SDS-PAGE are shown in FIG. 1, in which: m1, Marker; m2, low molecular weight Marker; 0, peeling soybean meal raw material; 1, common fermented soybean meal (prepared in comparative example 1); 2, high quality enzymatically fermented soybean meal (prepared in example 1).
As can be seen in fig. 1, the enzymatic fermented soybean meal prepared in example 1 completely removes beta-conglycinin and acid-glycinin, and the removal of Base-glycinin is obviously better than that of the common fermented soybean meal, so that the removal is complete; the enzymolysis fermentation soybean meal has the advantages that small molecular proteins are small, the proteins are aggregated together and cannot be separated, so that the small molecular proteins cannot be observed as bands are same as markers, but can be determined to be below 10000da, while the common fermentation soybean meal protein is not completely degraded, the molecular weight is partially below 10000da, and the majority is above 10000 da. The results show that: the enzymolysis and fermentation soybean meal is obviously superior to the common fermentation soybean meal in the antigen treatment of the soybean meal.
2) Oligosaccharide sugar content detection, and oligosaccharide detection results of the peeled soybean meal, the common fermented soybean meal (comparative example 1) and the high-quality enzymatic fermented soybean meal (example 1) are shown in table 3.
TABLE 3
As can be seen from Table 3, raffinose is less than 0.15% and stachyose is less than 0.7% in the high-quality enzymolysis fermented soybean meal; the degradation rate of the high-quality enzymolysis and fermentation soybean meal on raffinose in oligosaccharide in the soybean meal is 99.03%, and the degradation rate on stachyose is 94.38%; while the degradation rate of the common fermented soybean meal to raffinose is 66.3 percent, and the degradation rate to stachyose is 65.17 percent; the results show that: the degradation rate of the high-quality enzymolysis and fermentation soybean meal for resisting the nutritional factor oligosaccharide is obviously superior to that of common fermentation soybean meal.
Experimental example 4
The high-quality enzymatic fermented soybean meal prepared in example 1, the common fermented soybean meal and the peeled soybean meal in comparative example 1 were respectively subjected to conventional index detection, and the results are shown in table 4 (the detection method is the same as in experiment 1).
TABLE 4
Table 4 shows that the acid soluble protein (small peptide) content of the common fermented soybean meal is superior to that of the soybean meal raw material, while the high-quality enzymolysis fermented soybean meal is significantly higher than that of the common fermented soybean meal and the soybean meal, and the protein ratio is also significantly increased, and the results show that: the proportion of anti-nutritional factors of the high-quality enzymolysis and fermentation soybean meal is reduced, the proportion of protein is increased, and protein macromolecules are degraded into small molecular peptide fragments or free amino acids, so that the acid soluble protein is obviously increased.
Experimental example 5
The content of beneficial bacteria in the high-quality enzymatic fermented soybean meal prepared in example 1 and the content of beneficial bacteria in the common fermented soybean meal in comparative example 1 were respectively detected, and the results are shown in table 5.
TABLE 5
Table 5 shows that, in example 1, the retention rate of beneficial bacteria is significantly higher than that of the common fermented soybean meal in comparative example 1 by spray drying of the high-quality enzymatic fermented soybean meal, the traditional fermented soybean meal adopts a drum drying mode to obviously destroy the beneficial bacteria in the fermented soybean meal, only a small amount of bacillus is retained, and yeast and lactic acid bacteria hardly exist.
In conclusion, the finished product of the embodiment 1 of the invention has thorough treatment on the harmful antigens of the soybean meal; the stachyose and raffinose with the intestinal loss comparison are removed more thoroughly; protein macromolecules are well modified, so that the content of small peptides and the content of free amino acids are obviously increased; in addition, the retention rate of beneficial bacteria is greatly improved by adopting a spray drying mode, so the preferred proposal of the invention is embodiment 1.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A preparation method of high-quality enzymolysis fermented soybean meal is characterized by comprising the following steps:
1) crushing the peeled soybean meal, sieving with a 80-mesh sieve, adding water which is 3-4 times of the weight of the soybean meal, and uniformly stirring;
2) adding alpha-galactosidase with the weight of 0.5-1 per mill of the weight of the soybean meal and saccharifying enzyme with the weight of 1-1.5 per mill of the weight of the soybean meal into the material obtained in the step 1) for enzymolysis; adding papain with the weight of 0.3-0.5 per mill of the soybean meal for enzymolysis after complete enzymolysis; heating to inactivate enzyme after complete enzymolysis; the enzyme activity of the alpha-galactosidase is 3000IU/g, the enzyme activity of the glucoamylase is 20 ten thousand IU/g, and the enzyme activity of the papain is 80 ten thousand IU/g;
wherein, in the enzymolysis process of the alpha-galactosidase and the saccharifying enzyme, the optimum pH6.0 is realized, and the optimum enzymolysis temperature is 50-55 ℃; the preferable enzymolysis time is 2-3 h; in the enzymolysis process of the papain, the optimum pH value is 6.0, and the optimum enzymolysis temperature is 50-55 ℃; the preferable enzymolysis time is 3-5 h;
3) cooling the material obtained in the step 2) to 30-35 ℃, inoculating mixed bacteria of saccharomycetes, bacillus and lactic acid bacteria, performing aerobic fermentation for 20-24h, performing anaerobic fermentation for 48-50h, and completing fermentation;
the yeast is Saccharomyces cerevisiae 2.1392; the bacillus is feed bacillus Paenibacillus pabuli 1.3772; the lactobacillus is Pediococcus pentosaceus 1.2695;
the inoculation amount of the mixed strain is 1-2 per mill of the weight of the dry matter of the soybean meal raw material;
4) and (3) passing the material obtained in the step 3) through a colloid mill, and performing spray drying to obtain a finished product.
2. The preparation method of the high-quality enzymatic hydrolysis fermented soybean meal as claimed in claim 1, wherein the content of crude protein in the soybean meal is not less than 43.0%; preferably, the soybean meal is peeled soybean meal, and the crude protein is more than 45%.
3. The method for preparing high-quality enzymatic fermented soybean meal according to claim 1 or 2, wherein the effective viable count of yeast, bacillus and lactic acid bacteria in the mixed bacteria is 4 × 109CFU/g、5×109CFU/g、6×109CFU/g。
4. The method for preparing high-quality enzymatic fermented soybean meal according to any one of claims 1 to 3, wherein the amount of the alpha-galactosidase added is 0.5% by weight of the soybean meal; the adding amount of the saccharifying enzyme is 1 per mill of the weight of the soybean meal; the addition amount of the papain is 0.3 per mill of the weight of the soybean meal.
5. The method for preparing high-quality enzymatically fermented soybean meal according to any one of claims 1 to 4, wherein the spray drying conditions are: the air inlet temperature is 240 ℃, and the air outlet temperature is 95 ℃; and/or the presence of a gas in the gas,
and the heating enzyme deactivation is to immediately heat to 85-90 ℃ after the enzymolysis is finished, and carry out enzyme deactivation for 30 minutes.
6. The method for preparing high-quality enzymatic fermented soybean meal according to claim 1, characterized by comprising the steps of:
1) crushing the peeled soybean meal, sieving with a 80-mesh sieve, adding water which is 3-4 times of the weight of the soybean meal, and uniformly stirring;
2) adding alpha-galactosidase with the weight of 0.5 per thousand of that of the soybean meal and saccharifying enzyme with the weight of 1 per thousand of that of the soybean meal into the material obtained in the step 1), and carrying out enzymolysis for 2-3 h; then adding papain with the weight of 0.3 per mill of the weight of the soybean meal for enzymolysis for 2-3 h; immediately heating to 85-90 ℃ after complete enzymolysis, and inactivating enzyme for 30 minutes;
3) cooling the material obtained in the step 2) to 30-35 ℃, inoculating mixed strains of yeast, bacillus and lactobacillus, wherein the inoculation amount is 1 per mill of the weight of the dry matter of the soybean meal raw material, performing aerobic fermentation for 20-22h, performing anaerobic fermentation for 48-50h, and completing fermentation;
the effective viable count of yeast, Bacillus and lactobacillus in the mixed bacteria is 4 × 109CFU/g、5×109CFU/g、6×109CFU/g；
4) And (3) passing the material obtained in the step 3) through a colloid mill, sieving the material by a 100-mesh sieve, and entering a spray dryer to obtain a finished product.
7. The method for preparing high-quality enzymatic fermented soybean meal according to claim 1, characterized by comprising the steps of:
1) crushing 1000kg of peeled soybean meal, sieving with a 80-mesh sieve, adding water with the weight 3-4 times of the weight of the soybean meal, and uniformly stirring; the content of crude protein in the peeled soybean meal is more than or equal to 45 percent;
2) adding alpha-galactosidase with the weight of 0.3 per thousand of that of the soybean meal and saccharifying enzyme with the weight of 1 per thousand of that of the soybean meal into the material obtained in the step 1), and carrying out enzymolysis for 1.5-2 h; then adding papain with the weight of 0.2 per mill of the weight of the soybean meal for enzymolysis for 1.5-2 h; immediately heating to 85-90 ℃ after complete enzymolysis, and inactivating enzyme for 30 minutes;
wherein the enzymolysis condition is pH6.0, and the enzymolysis temperature is 50-55 deg.C;
3) cooling the material obtained in the step 2) to 30-35 ℃, inoculating mixed bacteria of yeast, bacillus and lactobacillus with the inoculation amount of 2 per mill of the weight of the dry matter of the soybean meal raw material, performing aerobic fermentation for 18-20h, performing anaerobic fermentation for 48-50h, and completing fermentation;
4) and (3) feeding the material obtained in the step 3) into a colloid mill, sieving the material by a 100-mesh sieve, and performing spray drying to obtain a finished product.
8. The method for preparing high-quality enzymatic fermented soybean meal according to claim 1, characterized by comprising the steps of:
1) crushing 1000kg of peeled soybean meal, sieving with a 80-mesh sieve, adding water with the weight 3-4 times of the weight of the soybean meal, and uniformly stirring; the content of crude protein in the peeled soybean meal is more than or equal to 45 percent;
2) adding alpha-galactosidase with the weight of 0.5 per thousand of that of the soybean meal and saccharifying enzyme with the weight of 1 per thousand of that of the soybean meal into the material obtained in the step 1), and carrying out enzymolysis for 1.5-2 h; then adding papain with the weight of 1.0 per mill of the weight of the soybean meal for enzymolysis for 1.5-2 h; immediately heating to 85-90 ℃ after complete enzymolysis, and inactivating enzyme for 30 minutes;
wherein the enzymolysis condition is pH6.0, and the enzymolysis temperature is 50-55 deg.C;
3) cooling the material obtained in the step 2) to 30-35 ℃, inoculating mixed bacteria of yeast, bacillus and lactobacillus with the inoculation amount of 2 per mill of the weight of the dry matter of the soybean meal raw material, performing aerobic fermentation for 18-20h, performing anaerobic fermentation for 48-50h, and completing fermentation;
4) and (3) feeding the material obtained in the step 3) into a colloid mill, sieving the material by a 100-mesh sieve, and performing spray drying to obtain a finished product.
9. High quality enzymatically fermented soybean meal produced by the method of any one of claims 1 to 8.
10. The use of the high-quality enzymatically fermented soybean meal of claim 9 in the preparation of animal feed; preferably, the addition amount of the high-quality enzymolysis and fermentation soybean meal in the animal feed is 2-5% (weight ratio).
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| CN106615618A (en) * | 2017-01-16 | 2017-05-10 | 武汉轻工大学 | Method of producing fermented soybean meal by using multi-strain mixed solid secondary fermentation technology and application thereof |
| CN107950753B (en) * | 2017-12-28 | 2024-02-06 | 广东省农业科学院动物科学研究所 | Method for producing biological feed by aerobic and anaerobic coupled fermentation |
| CN108464393A (en) * | 2018-04-17 | 2018-08-31 | 佛山市雷米高动物营养保健科技有限公司 | A kind of pet composition and preparation method thereof containing fermented bean dregs |
| CN110663820A (en) * | 2018-07-02 | 2020-01-10 | 辽宁波尔莱特农牧实业有限公司 | Method for preparing fermented bran through bacterium-enzyme synergistic fermentation |
| CN109673860A (en) * | 2019-03-04 | 2019-04-26 | 江西正邦农业科学研究院 | A kind of high digestion, high immune, high food calling protein concentrate raw material production method |
| CN109965084A (en) * | 2019-05-14 | 2019-07-05 | 华中农业大学 | A kind of method of solid fermentation dregs of beans production high protein feed |
| CN110367375A (en) * | 2019-06-11 | 2019-10-25 | 浙江博仕佳生物科技有限公司 | A kind of Radix Astragali, honeysuckle and dregs of beans combined fermentation product and beasts, birds and aquatic products breeding feed |
| CN110200131A (en) * | 2019-07-01 | 2019-09-06 | 博益德（北京）生物科技有限公司 | A kind of single enzymatic hydrolysis mixed fermentation prepares the method and application of functional fiber feed |
| JP6661121B1 (en) * | 2019-07-26 | 2020-03-11 | ハイアマウント株式会社 | Method for producing fermentation liquor containing short-chain fatty acids |
| CN110583854A (en) * | 2019-10-10 | 2019-12-20 | 北京挑战农业科技有限公司 | High-acid-soluble protein and high-lactic acid fermented soybean meal and preparation method thereof |
| CN111466485B (en) * | 2020-05-22 | 2022-08-02 | 济南百斯杰生物工程有限公司 | Application of alpha-galactosidase in enzymatic hydrolysis of soybean meal |
| CN112335885A (en) * | 2020-11-05 | 2021-02-09 | 苏州微克生活科技有限公司 | Method for preparing probiotic dietary supplement through multi-bacterium combined fermentation |
| CN113528420B (en) * | 2021-07-28 | 2022-12-02 | 四川润格生物科技有限公司 | Yeast hydrolysate and preparation method and application thereof |
| CN114767757B (en) * | 2022-04-14 | 2023-07-25 | 江西省科学院生物资源研究所 | Accurate fermentation method of compound Chinese herbal medicine byproducts and application thereof |
| CN114847395A (en) * | 2022-04-21 | 2022-08-05 | 成都盟盛生物科技有限公司 | Refined peptide as raw material of primary intestinal motility protein and processing technology thereof |
| CN115137023A (en) * | 2022-06-16 | 2022-10-04 | 广东省农业科学院动物卫生研究所 | Feed containing deep-fermented soybean meal and preparation method and application thereof |
| CN115886135A (en) * | 2022-12-16 | 2023-04-04 | 安徽天邦饲料科技有限公司 | Production method of low-antigen enzymolysis fermented soybean meal feed |
| CN116035191A (en) * | 2023-01-09 | 2023-05-02 | 华南理工大学 | Method for preparing soybean flavor material by combining enzymolysis and fermentation technology and application |
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