CN114276955A - Microbial agent for producing protein feed by solid state fermentation of potato residues - Google Patents
Microbial agent for producing protein feed by solid state fermentation of potato residues Download PDFInfo
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Abstract
The invention provides a microbial agent for producing protein feed by solid state fermentation of potato residues, which can be applied to the production of protein feed by the solid state fermentation of the potato residues. The invention discloses a Bacillus subtilis BCSW-21 with the preservation number of CGMCC 23931 which is preserved in the China general microbiological culture collection center. The strain can be compounded with Saccharomyces cerevisiae BCGQ2107 and alteromonas sp G1-3 to form a microbial agent, which not only can improve the content of protein produced by solid fermentation of potato residues, but also can effectively inhibit common pathogenic bacteria. The potato residue treated by the microbial inoculum has strong bouquet and certain feeding nutritive value, can recycle potato residue waste, and reduces environmental pollution.
Description
Technical Field
The invention relates to the technical field of feed design, in particular to a microbial agent for fermenting protein feed.
Background
The potato has a long cultivation history in China and is an important grain and economic crop. In recent years, the potato planting production in China is stably developed, and the coverage area is continuously expanded. Meanwhile, the stem leaf garbage attached to the potato production and the potato residue waste in the processing are also greatly increased. The resources of the wastes are difficult to process and recycle, and the wastes pollute the environment after being placed for a long time, thereby causing adverse results. The potato residue is one of agricultural wastes obtained in the starch production process, and is rich in nutrients such as starch, cellulose, hemicellulose, pectin, protein, free amino acid, salt and the like. If the potato residues are utilized to prepare the protein feed, the additional benefit of a potato starch processing factory can be improved, and the discharge of wastes can be reduced.
At present, the utilization form of the potato residue is mainly divided into: fermentation, physical and chemical methods and mixing methods. The fermentation method comprises the steps of adding a microbial agent into a culture medium by using potato residue as a substrate to ferment, and fermenting the mixed culture into a novel biomass product and an organic product; the physical and chemical method is to utilize an enzyme preparation to treat the potato residue and extract recyclable substances from the potato residue; the mixing method combines fermentation method and physical and chemical method.
The feeding treatment of the potato residue is more biased to a fermentation method, and the potato residue is fermented to generate required substances such as microbial protein, enzyme and the like by adding a microbial agent so as to form high-quality feed. The production of livestock protein feed by solid state fermentation is one of the important development directions of potato residue feeding. The solid fermented feed is animal feed prepared by fermenting raw materials with low added value, and is characterized by low cost, more raw materials, easy processing and the like. After potato residue is inoculated with thalli for fermentation, nutritional factors such as a carbon source and a nitrogen source in the potato residue are converted into the thalli, and protein is accumulated, so that the protein content in the potato residue is improved, and an active protein feed can be produced through fermentation, so that the potato residue is better in nutritional quality and more favorable for digestion and absorption of animals.
The yeast mainly comprises saccharomyces cerevisiae, candida utilis, beer yeast and the like, can enable the fermented feed to generate wine fragrance, improve the palatability of the feed, improve the flavor of the feed, increase the protein yield and increase the utilization rate of the feed because the yeast is mycoprotein. The bacillus mainly comprises bacillus subtilis, bacillus licheniformis, bacillus coagulans, bacillus cereus and the like, can degrade anti-nutritional factors and toxic substances, secretes cellulase and protease to degrade cellulose and macromolecular protein, and regulates the intestinal health of animals. The method for producing the protein feed by solid-state fermentation of the potato residues by adopting the microbial inoculum can improve the crude protein content of the feed and the palatability of livestock, and compared with the direct discharge of the potato residues, the method not only improves the economic benefit of enterprises, but also reduces the environmental pollution.
Disclosure of Invention
The invention provides a microbial agent capable of producing protein feed by applying solid state fermentation of potato residues, and the microbial agent can be used for improving mycoprotein by a solid state fermentation technology, so that the crude protein content of the potato residue feed is improved. The microbial agent fermented feed can generate wine fragrance, improve the palatability of the feed and improve the flavor of the feed. Has the double effects of improving economic benefit and reducing waste discharge.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a biological material BCSW-21 which is classified and named as Bacillus subtilis, the classified and named latin is Bacillus subtilis and is preserved in China general microbiological culture collection center, the preservation address is No. 3 Siro No. 1 of Beijing Korean-Yangtze district, the preservation date is 2021 year, 11 month and 18 day, the preservation number is: CGMCC 23931. The method has an obvious effect when applied to the solid state fermentation of the potato residues to produce the protein.
The invention also provides a microbial agent for producing protein feed by solid state fermentation of potato residues, which contains the bacillus subtilis with the preservation number of CGMCC 23931, and also comprises saccharomyces cerevisiae with the preservation number of CGMCC 23129 and alteromonas with the preservation number of CGMCC 14457.
Wherein, the biological material BCGQ2107 is classified and named as Saccharomyces cerevisiae, the classified and named Latin is named as Saccharomyces cerevisiae, the biological material is preserved in China general microbiological culture collection center, the preservation address is No. 3 of Xilu No. 1 of Beijing Shangyang Beichen, the preservation date is 2021 year, 8 month and 6 days, and the preservation number is: CGMCC 23129;
the biological material G1-3 is classified and named as alteromonas, the classified and named Latin is named as Altreomonas sp, and is preserved in the China general microbiological culture Collection center, the preservation address is No. 3 of Xilu No. 1 of Beijing Kogyo-Yang district, the preservation date is 2017, month 25, and the preservation number is: CGMCC 14457.
Preferably, the microbial agent for producing the protein feed by solid state fermentation of the potato residues comprises 1-3 parts by volume of saccharomyces cerevisiae, 1-3 parts by volume of bacillus subtilis and 1-2 parts by volume of alteromonas.
The microbial agent can inhibit Escherichia coli, Staphylococcus aureus and Streptomyces scabies.
Based on the microbial agent, the invention also provides a preparation and use method for preparing protein feed by solid state fermentation of potato residues, which comprises the following steps: respectively culturing, optimizing and expanding saccharomyces cerevisiae, bacillus subtilis and alteromonas, and mixing to prepare a microbial agent; mixing the microbial agent with a fermentation substrate, and carrying out sealed fermentation to obtain a fermented feed; drying and crushing to obtain the fermented feed.
In the microbial agent, a culture medium of saccharomyces cerevisiae comprises 0.3 to 0.5 part of molasses, 0.3 to 0.5 part of potassium sulfate, yeast extract and ammonium sulfate according to parts by weight, and yeastThe mass ratio of the paste to the ammonium sulfate is 4: 1. The culture condition of the yeast microbial inoculum is that the yeast microbial inoculum is shake-cultured for 30 hours at the temperature of 28-30 ℃ and the rotating speed of 180-200 r/min until OD is reached6001.5 to 1.7.
The culture medium of the bacterial microbial agent (bacillus subtilis and alteromonas) comprises molasses 0.1-0.3 parts by mass, potassium sulfate 0.3-0.5 parts by mass, yeast extract and ammonium chloride, and the mass ratio of the yeast extract to the ammonium chloride is 3: 1. The culture condition of the bacterial microbial inoculum is that the bacterial microbial inoculum is shake-cultured for 28 hours at the temperature of 35-37 ℃ and the rotating speed of 180-200 r/min until OD is reached6000.8 to 1.0.
And mixing the microbial agent with a fermentation substrate according to the addition amount of 4-5% by volume, and then performing solid state fermentation to prepare the protein feed.
The fermentation substrate comprises, by mass, 55-85% of potato residues, 0-10% of straws, 0-15% of bean pulp, 8-20% of bran, 5-10% of caragana microphylla powder, 2% of ammonium sulfate and 0.2% of potassium sulfate.
Further, the preferable fermentation substrate comprises, by mass, 60-80% of potato residues, 0-10% of straws, 0-15% of bean pulp, 10-15% of bran, 5-10% of corn caragana microphylla powder, 2% of ammonium sulfate and 0.2% of potassium sulfate.
Further, the temperature of sealed fermentation is 25-40 ℃, and more preferably 25-37 ℃; the time for the sealed fermentation is preferably 2-10 d, and more preferably 3-5 d.
Further, the pH of the fermentation substrate before solid state fermentation is 5.0-7.8, preferably 5.5-6.5; the water content is 45-55 wt.%, and the preferred water content is 50-52.5 wt.%.
After fermentation, drying for 24h at 80 ℃, crushing and sieving with a 40-mesh sieve.
And finally, measuring various indexes of the potato residue protein feed.
The invention has the beneficial effects that:
1. the raw material for producing the protein feed is potato residue, and with the increasing development of the potato starch industry, the potato residue is taken as a byproduct, and the yield is also very considerable. The potato residue is used as a raw material of the feed, has extremely high nutritive value, is rich in nutritive substances such as starch, cellulose, hemicellulose, pectin, protein, free amino acid, salt and the like, and has very high reuse value.
The Bacillus subtilis with the preservation number of CGMCC 23931 is screened out, has good effect on producing protein by solid state fermentation of potato residues, and the microbial agent compounded with Saccharomyces cerevisiae with the preservation number of CGMCC 23129 and Altreomonas sp with the preservation number of CGMCC 14457 obviously improves the protein content after solid state fermentation and contributes to reduction of heavy metals in the fermentation product. The amino acid produced by solid state fermentation has high content, and the composition is more beneficial to preventing diseases and improving the nutritive value of poultry.
2. The method for producing the protein feed is a solid-state fermentation method, the solid-state fermentation has the advantages of low cost, less investment, wide raw materials, convenient operation, less pollution and the like, and the post-treatment of the fermentation is simple. The microorganism is easy to grow in the solid fermentation culture medium, and the enzyme activity is relatively high.
3. The microbial agent for producing the protein feed comprises saccharomyces cerevisiae, bacillus subtilis and alteromonas, and the strains are nontoxic, harmless and easy to obtain. The prepared microbial inoculum can also inhibit the growth of mixed bacteria, can effectively inhibit common pathogenic bacteria staphylococcus aureus and escherichia coli, and has a certain inhibiting effect on the streptomyces scabicus susceptible to the potatoes.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments will be briefly described below.
FIG. 1 is a graph showing the results of examining the antagonistic activity against Staphylococcus aureus in the preferred embodiment 2 of the present invention.
FIG. 2 is a graph showing the results of measurement of the antagonistic activity against Escherichia coli in the preferred embodiment 2 of the present invention.
FIG. 3 is the result of the measurement of the antagonistic activity against Streptomyces scabies in the preferred embodiment 2 of the present invention.
FIG. 4 is a flow chart of potato pulp protein feed preparation in accordance with a preferred embodiment 3 of the present invention.
FIG. 5 is a graph comparing the protein feed products obtained in preferred example 4 and example 5 of the present invention.
Detailed Description
The following detailed description of the present invention is provided in conjunction with examples to better illustrate the aspects of the present invention and its advantages. It should be noted that the following description of the specific embodiments and examples is intended to be illustrative only and not limiting.
Potato residue, bran, straw, soybean meal and corn caragana microphylla powder are all commercially available. The strains used in the invention are all provided by the Biotechnology Limited company of Baichuan of North Tianjin. The protein content of the protein feed is determined by a Kjeldahl method according to GB T6432-1994 feed crude protein determination, the water content of the feed is determined by GBT6435-2014 feed water determination, and the crude fiber of the feed is determined by GBT5009.10-2003 crude cellulose determination.
Example 1
Example 1 is a method for preparing the microbial agent and optimizing the expansion culture.
The strain preparation culture medium comprises the following components:
the slant culture medium of the bacillus subtilis and the alteromonas is an LB solid culture medium, and the formula of the culture medium comprises 0.5% of yeast extract powder, 1% of peptone, 1% of NaCl, 2.5% of agar, and 7.2-7.4 of pH (mass percentage).
The liquid culture medium of the bacillus subtilis and the alteromonas is an LB culture medium, and the formula of the culture medium comprises 0.5% of yeast extract powder, 1% of peptone, 1% of NaCl, and 7.2-7.4 (in percentage by mass).
The optimized culture medium for the enlarged culture of the bacillus subtilis and the alteromonas comprises the following components in percentage by mass: molasses 0.3 parts, potassium sulfate 0.3 parts, and yeast extract and ammonium chloride in a 3:1 ratio.
The slant culture medium of the saccharomyces cerevisiae is a YEPD solid culture medium, and the formula of the culture medium comprises 1% of yeast extract powder, 2% of glucose, 2% of peptone, 2.5% of agar and pH 6.0 (in percentage by mass).
The liquid culture medium of the saccharomyces cerevisiae is a liquid YEPD culture medium, and the formula of the culture medium comprises 1% of yeast extract powder, 2% of glucose, 2% of peptone and 6.0% of pH (mass percentage).
The optimized culture medium for the enlarged culture of the saccharomyces cerevisiae comprises the following components in percentage by weight: 0.3 part of molasses, 0.3 part of potassium sulfate, and the yeast extract and ammonium sulfate are added according to a composite ratio of 4 to 1 (mass percent).
The culture method of the microbial inoculum comprises the following steps:
respectively carrying out slant strain culture, liquid first-stage strain culture and liquid second-stage strain culture on saccharomyces cerevisiae, bacillus subtilis and alteromonas.
Slant culture of strain: taking out the strain from an ultralow temperature refrigerator at minus 80 ℃ for later use, inoculating the strain into slant culture by adopting a streaking inoculation method, and then placing the slant culture in an incubator at 28-37 ℃ for dark culture for 24-48 h to obtain a single bacterial colony, wherein the test tube is preferably stored in the refrigerator at 4 ℃ if the single bacterial colony is not directly used.
Liquid first-level strain culture: and subpackaging the liquid culture medium into 100mL triangular flasks, wherein each flask is filled with 40-50 mL of the liquid culture medium, dipping a single colony obtained by slant culture of the strain, inoculating the single colony into the triangular flasks by using an inoculating loop, and culturing on a shaking table at the culture temperature of 28-37 ℃ at the rotation speed of 180-200 rpm for 24-48 h to obtain the first-level strain.
Liquid secondary strain culture: selecting a culture medium corresponding to the strain, preparing according to an optimized culture medium formula, subpackaging into 1000mL of large triangular flasks, wherein each flask is filled with 300-400 mL of the culture medium, inoculating the cultured primary strain, wherein the inoculum size is 1.0-1.5%, placing the strain on a shaking table for culturing at the culture temperature of 28-37 ℃, the rotation speed of 180-200 rpm and the culture time of 24-48 h to obtain the secondary liquid strain.
The obtained 3 kinds of secondary strains are mixed into the microbial agent. The microbial agent is obtained by mixing the obtained secondary bacterial liquid according to the volume ratio of 2 parts of saccharomyces cerevisiae, 2 parts of bacillus subtilis and 1 part of alteromonas. More excellentOptionally, mixing the obtained secondary bacterial liquid according to the volume ratio of 1 part of saccharomyces cerevisiae, 1 part of bacillus subtilis and 1 part of alteromonas to obtain a microbial agent, mixing the microbial agent to obtain a microbial agent, and performing shake culture at the temperature of 32-39 ℃ and the rotating speed of 180-200 r/min until the OD is reached600The obtained microbial agent can be used for solid state fermentation of protein feed in the next step to reach 1.347.
Example 2
The antagonistic action of the microbial inoculum of the invention on pathogenic bacteria is verified, and the experimental method comprises the following steps:
the culture medium of the escherichia coli and the staphylococcus aureus is an LB culture medium. The culture medium of the streptomyces scabies is an SP culture medium, and the formula comprises 0.4% of yeast extract powder, 1% of malt extract, 0.4% of glucose and 7.0-7.2% of pH (mass percentage).
Pouring sterilized LB and YEPD culture mediums (agar addition amount is 2%) into plates, pouring 15mL to 20mL of each plate, solidifying to obtain a lower layer culture medium, placing a sterilized oxford cup on the surface of the lower layer culture medium, taking LB or SP culture medium with agar addition amount of 1.5%, adding 3% of pathogenic indicator bacteria (one of escherichia coli, staphylococcus aureus and streptomyces scabies), uniformly mixing and pouring onto the lower layer culture medium, solidifying to obtain an upper layer culture medium. The Oxford cup was removed to expose the well, 0.1mL of the bacterial suspension of the test microorganism (more preferred microbial preparation obtained by mixing in example 1) was injected into the well, and the mixture was incubated in a biochemical incubator at 37 ℃ or 30 ℃ for 18 hours, and the plate number indicating the presence or absence of the transparent ring or the microbial dilution was observed and recorded. The experimental result shows that the compound has inhibition effect on three pathogenic bacteria. The bacteriostatic effect is shown in figure 1, figure 2 and figure 3.
Example 3
Embodiment 3 provides an application method of the microbial fermentation inoculum in the technical scheme in preparation of potato residue fermented feed. And (3) mixing the microbial agent and a fermentation substrate in the embodiment 1, and sealing and fermenting to obtain the fermented feed.
The potato residue is provided by inner Mongolian Monsson agricultural science and technology Co., Ltd, and bran, soybean meal and corn caragana microphylla powder are all sold in the market. The addition amount of the microbial agent is 4%, the main raw material of the fermentation substrate is potato residue, and the fermentation substrate comprises the following components in percentage by mass: 80% of potato residue, 2% of straw, 4% of bean pulp, 8% of bran, 6% of caragana microphylla powder, 2% of ammonium sulfate and 0.2% of potassium sulfate. The temperature of the sealed fermentation is 37 ℃, the time of the sealed fermentation is 3d, the pH of the substrate is adjusted to 6.5 before the solid state fermentation of the mixed materials, and the water content is 52.5 wt.%. The standards of the fermentation substrate obtained by mixing are as follows: it can be held by hand to form a mass, and can disperse when falling to the ground, and is slightly moist and light yellow in color. The solid-state fermentation method comprises the steps of mixing the mixed material with a microbial agent, sealing and fermenting, and sealing and fermenting in a fermentation tank or a fermentation bag. The end standard of solid state fermentation is that the fermented feed is brownish yellow and gives off wine aroma. The flow chart of the potato residue protein feed preparation is shown in figure 4.
Example 4
A blank group preparation method of the potato residue feed by solid state fermentation according to the method.
The potato residue is provided by inner Mongolian Monsson agricultural science and technology Co., Ltd, and bran, soybean meal and corn caragana microphylla powder are all sold in the market. The fermentation substrate consists of 80% of potato residues, 2% of straws, 4% of bean pulp, 8% of bran and 6% of caragana microphylla powder, and is uniformly mixed after 2% of ammonium sulfate and 0.2% of potassium sulfate are added. And drying the stirred fermentation substrate for 24 hours at 80 ℃, crushing and sieving the fermentation substrate with a 40-mesh sieve to be tested.
Example 5
Example 5 is a laboratory set preparation method for solid state fermentation of potato pulp feed as described.
The potato residue is provided by inner Mongolian Monsson agricultural science and technology Co., Ltd, and bran, soybean meal and corn caragana microphylla powder are all sold in the market. The fermentation substrate consists of 80% of potato residues, 2% of straws, 4% of soybean meal, 8% of bran and 6% of caragana microphylla powder, 2% of ammonium sulfate and 0.2% of potassium sulfate are added and then uniformly mixed, and the mixed microbial inoculum cultured according to the embodiment 1 and the microbial inoculum of a single strain are respectively inoculated according to the addition amount of 4%. And packaging the stirred fermentation substrate by using a sealing bag, and fermenting for 3 days at room temperature of 25-37 ℃. And (3) drying the fermented feed at 80 ℃ for 24h, crushing and sieving with a 40-mesh sieve to be tested. The comparative picture of the fermented product is shown in figure 5, and compared with the feed before fermentation, the fermented feed is brown yellow and emits wine fragrance.
Example 6
Example 6 is the content detection of various indexes of the feed.
The content of the feed protein is determined by adopting a Kjeldahl method according to GB T6432-1994 feed crude protein determination. Digesting a sample by concentrated sulfuric acid in the presence of a catalyst to convert organic nitrogen into inorganic ammonium salt, converting the inorganic ammonium salt into ammonia gas under an alkaline condition, distilling the ammonia gas out along with water vapor and absorbing the ammonia gas by excessive boric acid, titrating the ammonia gas by a standard acid solution, and calculating the nitrogen content in the sample by calculating the volume of the acid solution required by titration. 3.00g of potassium sulfate, 0.200g of copper sulfate, 0.200g of sample and 10mL of concentrated sulfuric acid are added into a digestion tube. Digesting at 200 ℃ for 20min, digesting at 300 ℃ for 20min, digesting at 400 ℃ for 90min, cooling, transferring to a conical flask, reacting in an automatic Kjeldahl apparatus, and titrating the reacted liquid by using a standard acid solution.
V0: titration of the volume of the standard acid solution consumed by the blank distillate
V1: titration of sample distillate consumption of standard acid solution volume
C: concentration of standard acid solution
0.014: millimolar mass of nitrogen
F: coefficient of protein
m: quality of sample
The water content of the feed is measured by referring to GBT6435-2014 'water content measurement in feed'. Weighing 2.00g of sample in a pre-dried and weighed empty aluminum box to 0.0002g, placing the sample in a drying oven at 105 ℃, taking out the sample after 4-6h, cooling the sample to room temperature in a dryer and weighing the sample.
m1: quality of empty aluminium box
m2: mass of sample before drying
m3: quality of empty aluminum boxes and dried samples
The method for measuring the crude fiber of the feed refers to an acid-base washing method of GBT5009.10-2003 'crude cellulose measurement'. Under the action of sulfuric acid, the sugar, starch, pectin and hemicellulose in the sample are hydrolyzed and removed, then are treated by alkali to remove protein and fatty acid, and the residual residue is crude fiber.
X: content of crude fiber in sample
G: mass of residue
m: mass of the sample
Indexes of the solid-state fermented feed prepared from the single strain in the example 5 and the microbial agent compounded according to the example 1 are detected and compared with the indexes shown in the table 1.
TABLE 1 comparison of solid fermentation indexes of single strain and complex microbial inoculum
According to table 1, the effect of the microbial inoculum prepared from a single strain and the effect of the microbial composite microbial inoculum on treating the potato residue mixture are both better than the effect of the microbial inoculum prepared from a single strain on treating the initial potato residue mixture without adding the microbial inoculum, and the effect of the microbial composite microbial inoculum on treating the potato residue mixture is better than the effect of the microbial inoculum prepared from a single strain.
The comparison of amino acid components in the fermentation products of the blank and the experimental group inoculated with the mixed microbial agent in example 5 is shown in Table 2.
TABLE 2 comparison of amino acid composition of fermentation products of blank and Experimental groups
As can be seen from Table 2, the amino acid content of the potato residue feed processed by the solid-state fermentation of the microbial inoculum is higher than that of the blank experimental group before the fermentation, the total amount is 13.4%, and the amino acid content is improved by 197.78%. The test detects 16 kinds of amino acids, the glutamic acid content in various amino acids is the highest, the glutamic acid is one of basic amino acids of nitrogen metabolism in organisms, can be used as carbon and nitrogen nutrition to participate in the organism metabolism, and has higher nutritional value. And secondly, leucine, which has effects including repairing muscles, controlling blood sugar, and supplying energy to body tissues in cooperation with isoleucine and valine, wherein leucine is the most effective one of branched chain amino acids, and is effective in preventing muscle loss because it can be decomposed more rapidly into glucose. Because the content of lysine in the cereal food is extremely low, the lysine is called as first limiting amino acid, and the proper amount of lysine is added into the feed, the utilization rate of the feed can be improved, and a certain inhibition effect on the propagation of mould can be achieved. The second limiting amino acid is methionine which is not only required by nutrition but also can inhibit the growth of various mycotoxins, and the feed containing the methionine has the effects of disease prevention and health care on poultry.
The parameter indexes and heavy metal ion contents of the fermentation products of the blank group and the experimental group added with the mixed microbial agent in the embodiment example 5 are compared in tables 3 and 4.
TABLE 3 comparison of fermentation product parameters for blank and Experimental groups
TABLE 4 comparison of heavy metal ion content in fermentation products of blank and experimental groups
Although specific embodiments of the invention have been described herein, it will be understood by those skilled in the art that the specific embodiments described are illustrative only and are not limiting to the scope of the invention, and that equivalent modifications and variations can be made by those skilled in the art without departing from the spirit of the invention, which is to be limited only by the appended claims.
Claims (10)
1. Bacillus subtilis BCSW-21 with the preservation number of CGMCC 23931 is preserved in the China general microbiological culture Collection center at the preservation address of No. 3 Xilu No. 1 Beijing, Chaoyang, with the preservation date of 2021 year, 11 months and 18 days.
2. A microbial agent for producing protein feed by solid state fermentation of potato residues is characterized by comprising Bacillus subtilis BCSW-21 with the preservation number of CGMCC 23931, Saccharomyces cerevisiae BCGQ2107 with the preservation number of CGMCC 23129 and alteromonas G1-3 with the preservation number of CGMCC 14457.
3. The microbial agent for producing protein feed by solid state fermentation of potato residues according to claim 2, wherein the microbial agent comprises 1-3 parts by volume of saccharomyces cerevisiae, 1-3 parts by volume of bacillus subtilis and 1-2 parts by volume of alteromonas.
4. The microbial agent for producing protein feed by solid state fermentation of potato residues as claimed in claim 1, wherein the microbial agent can inhibit Escherichia coli, Staphylococcus aureus and Streptomyces scabies.
5. A method for producing protein feed by solid state fermentation of potato dregs based on the microbial inoculant as defined in any one of claims 1 to 4, comprising the steps of: (1) respectively culturing and optimally expanding saccharomyces cerevisiae, bacillus subtilis and alteromonas, and mixing to prepare a microbial agent; (2) mixing the microbial agent with a fermentation substrate, and sealing for solid state fermentation; oven drying, and pulverizing to obtain protein feed;
the expanding culture medium of the saccharomyces cerevisiae comprises, by mass, 0.3-0.5 part of molasses, 0.3-0.5 part of potassium sulfate, yeast extract and ammonium sulfate, wherein the mass ratio of the yeast extract to the ammonium sulfate is 4:1, and the culture condition is that the saccharomyces cerevisiae is shake-cultured for 30 hours at the temperature of 28-30 ℃ and the rotating speed of 180-200 r/min until OD is achieved6001.5 to 1.7; the expanding culture medium of the bacillus subtilis and the alteromonas comprises molasses 0.1-0.3 part by mass, potassium sulfate 0.3-0.5 part by mass, yeast extract and ammonium chloride 3:1 by mass, and the culture condition is that the expanding culture medium is shake-cultured for 28 hours at the temperature of 35-37 ℃ and the rotating speed of 180-200 r/min until OD is OD6000.8 to 1.0.
6. The method for producing the protein feed by solid state fermentation of the potato residues as claimed in claim 5, wherein the microbial agent is mixed with the fermentation substrate by an addition amount of 4-5% by volume for solid state fermentation to prepare the protein feed.
7. The method for producing the protein feed by solid state fermentation of the potato residues as claimed in claim 5, wherein the fermentation substrate comprises 55-85% of the potato residues, 0-10% of the straws, 0-15% of the bean pulp, 8-20% of the bran, 5-10% of the caragana microphylla powder, 2% of ammonium sulfate and 0.2% of potassium sulfate by mass percentage.
8. The method for producing protein feed by solid state fermentation of potato dregs in claim 7, wherein the fermentation substrate comprises, by mass, 60-80% of potato dregs, 0-10% of straw, 0-15% of soybean meal, 10-15% of bran, 5-10% of corn caragana microphylla powder, 2% of ammonium sulfate and 0.2% of potassium sulfate.
9. The method for producing the protein feed by solid state fermentation of the potato residues in the claim 5, wherein the temperature of the sealed solid state fermentation is 25-40 ℃, preferably 25-37 ℃; the time for sealed fermentation is 2-10 days, preferably 3-5 days; and after fermentation, drying at 50-60 ℃ until the water content is 10% -20%, and storing.
10. The method for producing the protein feed by solid state fermentation of the potato residues in the claim 5, wherein the pH of the fermentation substrate before solid state fermentation is 5.0-7.8, preferably 5.5-6.5; the water content is 45-55 wt.%, and the preferred water content is 50-52.5 wt.%.
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