CN111748504A - Technical method for producing enterococcus faecium by using white spirit vinasse - Google Patents
Technical method for producing enterococcus faecium by using white spirit vinasse Download PDFInfo
- Publication number
- CN111748504A CN111748504A CN202010847873.1A CN202010847873A CN111748504A CN 111748504 A CN111748504 A CN 111748504A CN 202010847873 A CN202010847873 A CN 202010847873A CN 111748504 A CN111748504 A CN 111748504A
- Authority
- CN
- China
- Prior art keywords
- parts
- enterococcus faecium
- distiller
- culture
- grains
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
- A23K10/38—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material from distillers' or brewers' waste
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Animal Husbandry (AREA)
- Food Science & Technology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Mycology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Botany (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a technical method for producing enterococcus faecium by utilizing white spirit vinasse, which comprises the steps of purifying and rejuvenating freeze-dried powder of the enterococcus faecium, inoculating the freeze-dried powder of the enterococcus faecium into a primary seed culture solution, and performing shake culture to obtain the primary seed solution; then further inoculating the seed into a secondary seed culture solution to culture to obtain a secondary seed solution; inoculating the prepared secondary seed liquid into a solid fermentation culture medium containing the pretreated distiller's grains, and performing fermentation culture; and after the fermentation is finished, drying and crushing the fermentation product to obtain the enterococcus faecium solid fermentation product. In the obtained solid fermentation product, the number of strains per gram of dry matter reaches more than 180 hundred million, and the viable bacteria rate is still more than 80 percent after the solid fermentation product is stored for 12 months at low temperature.
Description
Technical Field
The invention relates to the technical field of microbial culture, in particular to a technical method for producing enterococcus faecium by using white spirit vinasse.
Background
Enterococcus faecium is one of 16 directly feedable feed microorganism varieties allowed by the Ministry of agriculture in China. Enterococcus faecium is one of lactobacillus family members, has good effects of improving weight gain rate of young livestock and poultry, improving animal immunity, regulating intestinal microecological balance, improving nutrient absorption, reducing diarrhea rate, reducing death rate, and the like, and has good tolerance to partial antibiotics. Therefore, the lactobacillus is a kind of lactobacillus product that is popular among many manufacturers in the current microecological preparation industry, especially in feed additive enterprises. Common enterococcus faecium in the market is obtained by liquid state fermentation, although the viable count of the product is high, the product has higher price, easy inactivation and harsh preservation conditions, thereby greatly restricting the popularization and application of the product in the market.
The vinasse lost yield generated in the white spirit brewing industry of China every year is more than 3500 million tons, the fresh vinasse has high acidity and water content of more than 60 percent, and contains about 10 percent of crude fat, about 17 percent of crude protein, about 25 percent of crude fiber, B vitamins, phosphorus, potassium and other inorganic elements. In addition, distillers grains contain trace amounts of effective components such as amino acids, ribonucleic acids, and purines produced by microbial cells, which are not comparable to grains. If the treatment is not carried out in time, the vinasse is easy to decay, which not only causes a great deal of waste of resources, but also seriously pollutes the surrounding environment.
Disclosure of Invention
Based on the technical problems, the invention provides a technical method for producing enterococcus faecium by using distiller's grains. The pretreated white spirit vinasse is used as a solid culture medium of the enterococcus faecium, and the solid fermentation culture process of the enterococcus faecium is provided, so that the technical problem of preparing the enterococcus faecium by liquid fermentation in the prior art is solved.
In order to realize the technical purpose, the invention provides a technical method for producing enterococcus faecium by utilizing white spirit vinasse, which comprises the following steps:
(1) placing enterococcus faecium freeze-dried powder in an improved MRS culture medium, performing streak culture, selecting 3-5 typical enterococcus faecium single colonies, continuously purifying, subculturing and rejuvenating for 3 times, and selecting an optimal enterococcus faecium single colony as a first-level seed liquid strain;
(2) selecting single colony of screened enterococcus faecium to inoculate in the primary seed culture solution by aseptic operation, and performing shake culture to obtain primary seed solution; then further inoculating the seed into a secondary seed culture solution to culture to obtain a secondary seed solution;
(3) inoculating the prepared secondary seed liquid into a solid fermentation culture medium containing the pretreated distiller's grains, and performing fermentation culture;
(4) and after the fermentation is finished, drying and crushing the fermentation product to obtain the enterococcus faecium solid fermentation product.
Preferably, in the step (1), the MRS medium comprises the following components in parts by mass: 1-2 parts of glucose, 0.5-1.5 parts of peptone, 0.5-1.5 parts of beef extract, 0.2-1 part of yeast extract powder, 0.2-0.8 part of sodium chloride, 1-3 parts of agar powder, 0.1-3 parts of calcium carbonate, 0.1-1 part of diammonium citrate, 0.08-0.1 part of tween, 0.02-0.1 part of magnesium sulfate, 0.01-0.05 part of manganese sulfate and 85-96 parts of purified water; the culture conditions were 35-37 ℃.
Preferably, the primary culture medium in the step (2) comprises the following components in parts by mass: 1-2 parts of sucrose, 1-2 parts of glucose, 0.5-1.5 parts of peptone, 1-5 parts of calcium carbonate, 0.2-1 part of yeast extract powder, 0.2-0.8 part of sodium chloride, 0.5-1.5 parts of beef extract, 0-0.8 part of magnesium sulfate, 0.1-0.8 part of manganese sulfate, 0.8-3.5 parts of dipotassium hydrogen phosphate and 86-95 parts of purified water;
the secondary medium comprises the following components: 2-3 parts of cane sugar, 0-1.5 parts of glucose, 0.5-1.5 parts of peptone, 1-5 parts of calcium carbonate, 0.2-1 part of yeast extract powder, 0.2-0.8 part of sodium chloride, 0.5-1.5 parts of beef extract, 0-0.8 part of magnesium sulfate, 0.1-0.8 part of manganese sulfate, 0.8-3.5 parts of dipotassium hydrogen phosphate and 86-95 parts of purified water;
the inoculation amount of the first-stage seed liquid inoculated to the second-stage culture medium is 5-10%;
the culture conditions are 30-40 deg.C, the culture time for preparing the first-stage seed solution is 8-12h, and the culture time for preparing the second-stage seed solution is 12-24 h.
Preferably, the solid state fermentation medium containing the pretreated distiller's grains in the step (3) contains the following components in parts by mass: 30-60 parts of pretreated distiller's grains, 0.1-2 parts of yeast extract powder, 0-1 part of magnesium sulfate, 0-1 part of manganese sulfate, 30-40 parts of water, and sterilizing at high temperature, wherein the pH value is 6-7.5; the inoculation amount is 5-10%.
Preferably, the preparation method of the pretreated distiller's grains comprises the following steps:
firstly, screening rice hulls from fresh distiller's grains, then placing the rice hulls in water, adjusting the pH value to 6-8, drying after soaking treatment, crushing and screening to obtain distiller's grain powder;
② adding water into the obtained distiller's grains powder and corn starch, uniformly mixing, then carrying out puffing treatment, crushing and sieving to obtain the puffed distiller's grains/corn starch mixture.
Preferably, in the step I, the soaking time is 12-48h, the soaking temperature is 60-80 ℃, and the mixture is crushed and sieved by a 40-mesh sieve.
Preferably, in the second step, the mixing ratio of the distiller's grains and the corn flour is (6-8) to 2, and the water adding amount is 30-50% of the total amount of the mixture of the distiller's grains flour and the corn starch.
Preferably, in the second step, the expansion temperature is 100-150 ℃, and the powder is crushed and sieved by a 60-mesh sieve after the expansion treatment.
Compared with the prior art, the invention has the following beneficial effects:
(1) the purified and rejuvenated dominant enterococcus faecium has stronger adaptability and reproductive capacity, and can ensure that the dominant enterococcus faecium still has high reproductive activity in a solid culture medium containing distiller's grains;
(2) the white spirit vinasse is placed in the alkaline solution to be soaked at a certain temperature, so that the degradation rate of cellulose and hemicellulose in the white spirit vinasse can be improved, and meanwhile, when the white spirit vinasse is mixed and puffed with corn flour in the subsequent process, the swelling reaction of the white spirit vinasse and the corn flour can be promoted, so that richer nutrient substances are provided for enterococcus faecium, the vinasse is easier to absorb when being used as an enterococcus faecium culture medium, and a foundation is provided for the subsequent process of being used as a;
(3) the distiller's grains are rich in crude protein and can provide sufficient nitrogen source for the thalli, but the carbon source in the distiller's grains is basically and completely utilized, so that the carbon source is insufficient, the corn flour contains high sugar and provides sufficient carbon source for the thalli, and the corn flour and the thalli have synergistic effect to promote the enterococcus faecium to keep high growth activity in the fermentation tank. Simultaneously, the pretreated white lees and the corn flour are uniformly mixed and then are puffed, in the puffing process, on one hand, starch in the corn flour is gelatinized, hydrogen bonds among starch molecules are broken to form gelatinized starch, part of the starch is degraded into reducing sugar, and the digestion utilization rate of the corn starch is obviously improved;
(4) because the pretreated distiller's grains exist in a mixture with corn flour, the pretreated distiller's grains are rich in amino compounds, the corn flour is high in sugar content, free amino acids in the amino compounds and free carboxyl groups of carbonyl compounds can be promoted to be condensed to form imine derivatives in the puffing process, Maillard reaction occurs, Maillard reaction products can show a strong inhibition effect on staphylococcus aureus, escherichia coli and the like, and therefore the inhibition effect of impurity bacteria on enterococcus faecium in the solid culture process can be avoided. Meanwhile, special flavor substances are given to the enterococcus faecium fermentation product, so that the enterococcus faecium fermentation product does not influence the mouthfeel of the feed when being used as an additive for livestock and poultry feeds, and the palatability of the feed is improved;
(5) the pretreated white lees and the corn flour are subjected to puffing reaction in a mixture form, so that the white lees and the corn flour can be more fully mixed and combined for reaction, and the supply of nutrient substances in all aspects can be further ensured when the enterococcus faecium is attached to the puffed mixture, so that the nutrient substance requirements in all aspects of the enterococcus faecium can be met on the premise that other raw materials are not required to be added into the solid culture medium;
(6) after the fermentation is finished, the fermentation product is dried and crushed to obtain the enterococcus faecium solid fermentation finished product, the strain quantity of each gram of dry matter in the obtained solid fermentation product reaches more than 180 hundred million, and the microorganisms at the moment are still attached to the solid culture medium with a pore structure, so that necessary nutrient substance supply can be still provided for the enterococcus faecium in the storage process, the survival rate of the enterococcus faecium in the storage process can be ensured, the storage life of the strain product is prolonged, and the viable bacteria rate is still more than 80 percent after the enterococcus faecium is stored for 12 months at low temperature.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
Example 1
(1) Aseptic operation, placing a small amount of enterococcus faecium freeze-dried powder (purchased from China center for type culture Collection in Wuhan university with the preservation number of CCTCC M: 2017191) in a culture medium containing 2 parts of glucose, 1.5 parts of peptone, 0.5 part of beef extract, 0.2 part of yeast extract powder, 0.2 part of sodium chloride, 1 part of agar powder, 3 parts of calcium carbonate, 0.5 part of diammonium citrate, 0.08 part of Tween, 0.1 part of magnesium sulfate, 0.05 part of manganese sulfate and 96 parts of purified water, marking in the culture medium, culturing at 37 ℃ for 24h, selecting 5 typical enterococcus faecium single colonies, respectively marking in an MRS culture medium, continuously purifying and carrying out subculture rejuvenation for 3 times, selecting the rejuvenated enterococcus faecium for physiological and biochemical identification, and displaying that the enterococcus faecium is enterococcus faecium, selecting the optimal enterococcus faecium single colony as a seed strain as a primary seed strain liquid;
(2) selecting screened single enterococcus faecium colonies to inoculate into 2 parts of cane sugar, 2 parts of glucose, 1.5 parts of peptone, 5 parts of calcium carbonate, 1 part of yeast extract powder, 0.8 part of sodium chloride, 1.5 parts of beef extract, 0.8 part of magnesium sulfate, 0.8 part of manganese sulfate, 3.5 parts of dipotassium hydrogen phosphate and 95 parts of purified water, and culturing for 12 hours in a shaking table at the temperature of 35 ℃ and the rotating speed of 180r/min to obtain a primary seed solution;
(3) inoculating the prepared primary seed liquid into 2 parts of glucose, 1.5 parts of peptone, 5 parts of calcium carbonate, 1 part of yeast extract powder, 0.8 part of sodium chloride, 1.5 parts of beef extract, 0.8 part of magnesium sulfate, 0.8 part of manganese sulfate, 3.5 parts of dipotassium hydrogen phosphate and 95 parts of purified water at the temperature of 35 ℃ in a shaking table at the rotating speed of 240r/min in a sterile environment at the inoculation rate of 10% for 24 hours to obtain secondary seed liquid.
(4) Preparing a solid culture medium:
sieving fresh distiller's grains to remove rice hull, placing in water, adjusting pH to 8, soaking at 80 deg.C for 24 hr, drying, pulverizing, and sieving with 40 mesh sieve to obtain distiller's grain powder;
mixing the obtained distiller's grains powder and corn starch at a ratio of 8: 2, adding 40% of water, blending uniformly, performing puffing treatment at 120 ℃, pulverizing, and sieving with 60 mesh sieve to obtain puffed distiller's grains/corn starch mixture;
taking 60 parts of a mixture of the expanded distiller's grains and the corn starch, 2 parts of yeast extract powder, 1 part of magnesium sulfate, 1 part of manganese sulfate and 40 parts of water, adjusting the pH value to 7, and sterilizing at high temperature to obtain the solid culture medium.
(5) Inoculating the secondary seed liquid into the prepared solid culture medium by 10 percent of inoculation amount, and performing fermentation culture for 72 hours at 35 ℃ in a sterile environment; and after the fermentation is finished, drying the fermentation product at 30 ℃, and crushing to 50 meshes to obtain the enterococcus faecium solid fermentation product.
Example 2
The difference from the example 1 is that in the step (4), the rice hull of the fresh distiller's grains is separated and put into water, the pH value is adjusted to 8, the fresh distiller's grains are soaked for 24 hours at 80 ℃, and then are frozen and dried at 40 ℃ below zero, crushed and sieved by a 40-mesh sieve to obtain distiller's grain powder, and the distiller's grain powder and the corn starch are mixed according to the proportion of 8: 2 and then are directly used as the raw material of the solid culture medium without being expanded.
Example 3
The difference from the example 1 is that the rice hull of the fresh spirit vinasse is separated by screening in the step (4), and then the fresh spirit vinasse is not soaked in alkaline water solution, and is directly dried, crushed and sieved by a 40-mesh sieve to obtain spirit vinasse powder, and the spirit vinasse powder is mixed with corn starch for puffing treatment.
Example 4
The difference from example 1 is that the second-stage seed solution was obtained by direct scale-up culture without the strain purification and rejuvenation step (1).
Example 5
The difference from example 1 is that step (4) is omitted, and the solid medium used in step (5) is 10 parts of soybean meal, 2 parts of rice bran, 10 parts of wheat bran, 1 part of calcium carbonate, 0.1 part of yeast extract powder and 78 parts of purified water.
The enterococcus faecium solid state fermentation products prepared in examples 1 to 5 were subjected to effective viable count measurement respectively for products stored at-4 ℃ for 6 months and-20 ℃ for 12 months, and the detection results are shown in table 1;
TABLE 1
| 0 month | Low temperature storage at-4 deg.C for 6 months | Low temperature storage at-20 deg.C for 12 months | |
| Example 1 | 182 hundred million/g | 165 hundred million/g | 155 hundred million/g |
| Example 2 | 150 hundred million/g | 100 hundred million/g | 85 hundred million/g |
| Example 3 | 135 hundred million/g | 102 hundred million/g | 94 hundred million/g |
| Example 4 | 126 hundred million/g | 105 hundred million/g | 112 hundred million/g |
| Example 5 | 105 hundred million/g | 45 hundred million/g | 37 hundred million/g |
As can be seen from Table 1, the purified and rejuvenated enterococcus faecium can be more suitable for the solid culture medium containing distiller's grains prepared by the invention, and the mixture after the swelling treatment can provide necessary nutrient substance guarantee for the growth of the enterococcus faecium, so that the viable count and the activity after storage are obviously improved.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included therein.
Claims (8)
1. A technical method for producing enterococcus faecium by using white spirit vinasse is characterized by comprising the following steps:
(1) placing enterococcus faecium freeze-dried powder in an improved MRS culture medium, performing streak culture, selecting 3-5 typical enterococcus faecium single colonies, continuously purifying, subculturing and rejuvenating for 3 times, and selecting an optimal enterococcus faecium single colony as a first-level seed liquid strain;
(2) selecting single colony of screened enterococcus faecium to inoculate in the primary seed culture solution by aseptic operation, and performing shake culture to obtain primary seed solution; then further inoculating the seed into a secondary seed culture solution to culture to obtain a secondary seed solution;
(3) inoculating the prepared secondary seed liquid into a solid fermentation culture medium containing the pretreated distiller's grains, and performing fermentation culture;
(4) and after the fermentation is finished, drying and crushing the fermentation product to obtain the enterococcus faecium solid fermentation product.
2. The technical method for producing enterococcus faecium by using distiller's grains according to claim 1, wherein in the step (1), the MRS culture medium comprises the following components in parts by mass: 1-2 parts of glucose, 0.5-1.5 parts of peptone, 0.5-1.5 parts of beef extract, 0.2-1 part of yeast extract powder, 0.2-0.8 part of sodium chloride, 1-3 parts of agar powder, 0.1-3 parts of calcium carbonate, 0.1-1 part of diammonium citrate, 0.08-0.1 part of tween, 0.02-0.1 part of magnesium sulfate, 0.01-0.05 part of manganese sulfate and 85-96 parts of purified water; the culture conditions were 35-37 ℃.
3. The technical method for producing enterococcus faecium by using the distiller's grains according to claim 1, wherein the primary culture medium in the step (2) comprises the following components in parts by mass: 1-2 parts of sucrose, 1-2 parts of glucose, 0.5-1.5 parts of peptone, 1-5 parts of calcium carbonate, 0.2-1 part of yeast extract powder, 0.2-0.8 part of sodium chloride, 0.5-1.5 parts of beef extract, 0-0.8 part of magnesium sulfate, 0.1-0.8 part of manganese sulfate, 0.8-3.5 parts of dipotassium hydrogen phosphate and 86-95 parts of purified water;
the secondary medium comprises the following components: 2-3 parts of cane sugar, 0-1.5 parts of glucose, 0.5-1.5 parts of peptone, 1-5 parts of calcium carbonate, 0.2-1 part of yeast extract powder, 0.2-0.8 part of sodium chloride, 0.5-1.5 parts of beef extract, 0-0.8 part of magnesium sulfate, 0.1-0.8 part of manganese sulfate, 0.8-3.5 parts of dipotassium hydrogen phosphate and 86-95 parts of purified water;
the inoculation amount of the first-stage seed liquid inoculated to the second-stage culture medium is 5-10%;
the culture conditions are 30-40 deg.C, the culture time for preparing the first-stage seed solution is 8-12h, and the culture time for preparing the second-stage seed solution is 12-24 h.
4. The technical method for producing enterococcus faecium by using distiller's grains according to claim 1, wherein the solid state fermentation medium containing the pretreated distiller's grains in the step (3) comprises the following components in parts by mass: 30-60 parts of pretreated distiller's grains, 0.1-2 parts of yeast extract powder, 0-1 part of magnesium sulfate, 0-1 part of manganese sulfate, 30-40 parts of water, and sterilizing at high temperature, wherein the pH value is 6-7.5; the inoculation amount is 5-10%.
5. The method of claim 4, wherein the pre-treated distiller's grain is prepared by the steps of:
firstly, screening rice hulls from fresh distiller's grains, then placing the rice hulls in water, adjusting the pH value to 6-8, drying after soaking treatment, crushing and screening to obtain distiller's grain powder;
② adding water into the obtained distiller's grains powder and corn starch, uniformly mixing, then carrying out puffing treatment, crushing and sieving to obtain the puffed distiller's grains/corn starch mixture.
6. The method for producing enterococcus faecium by using distiller's grains according to claim 5, wherein the soaking time is 12-48h, the soaking temperature is 60-80 ℃, and the mixture is crushed and sieved by a 40-mesh sieve.
7. The method of claim 5, wherein the ratio of distiller's grains to corn flour is 6-8: 2, and the amount of water added is 30-50% of the total amount of the mixture of distiller's grain flour and corn starch.
8. The method for producing enterococcus faecium by using distiller's grains of spirit according to claim 5, wherein the swelling temperature is 100 ℃ and 150 ℃, and the powder is crushed and sieved by a 60-mesh sieve after the swelling treatment.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010847873.1A CN111748504B (en) | 2020-08-21 | 2020-08-21 | Technical method for producing enterococcus faecium by using white spirit lees |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010847873.1A CN111748504B (en) | 2020-08-21 | 2020-08-21 | Technical method for producing enterococcus faecium by using white spirit lees |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111748504A true CN111748504A (en) | 2020-10-09 |
| CN111748504B CN111748504B (en) | 2023-06-06 |
Family
ID=72713397
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010847873.1A Active CN111748504B (en) | 2020-08-21 | 2020-08-21 | Technical method for producing enterococcus faecium by using white spirit lees |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111748504B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106978369A (en) * | 2017-04-12 | 2017-07-25 | 福建奥姆龙生物工程有限公司 | A kind of high-purity, resistance to storage at normal temperature VREF solid state fermentation culture process |
| CN110373364A (en) * | 2019-08-23 | 2019-10-25 | 华中农业大学 | A method of bacillus coagulans are produced based on distillers ' grains |
| CN110384179A (en) * | 2019-08-23 | 2019-10-29 | 华中农业大学 | Method, feed addictive and the application of fermentation distillers ' grains are produced using bacillus subtilis |
| CN110384176A (en) * | 2019-08-23 | 2019-10-29 | 华中农业大学 | A method of animal feed is prepared using distillers ' grains and expanded corn powder culture bacillus licheniformis |
| CN110452850A (en) * | 2019-08-23 | 2019-11-15 | 华中农业大学 | A kind of bacillus amyloliquefaciens culture, preparation method and applications |
| CN110447765A (en) * | 2019-08-23 | 2019-11-15 | 华中农业大学 | A kind of method and its application preparing bafillus natto culture using distillers ' grains |
-
2020
- 2020-08-21 CN CN202010847873.1A patent/CN111748504B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106978369A (en) * | 2017-04-12 | 2017-07-25 | 福建奥姆龙生物工程有限公司 | A kind of high-purity, resistance to storage at normal temperature VREF solid state fermentation culture process |
| CN110373364A (en) * | 2019-08-23 | 2019-10-25 | 华中农业大学 | A method of bacillus coagulans are produced based on distillers ' grains |
| CN110384179A (en) * | 2019-08-23 | 2019-10-29 | 华中农业大学 | Method, feed addictive and the application of fermentation distillers ' grains are produced using bacillus subtilis |
| CN110384176A (en) * | 2019-08-23 | 2019-10-29 | 华中农业大学 | A method of animal feed is prepared using distillers ' grains and expanded corn powder culture bacillus licheniformis |
| CN110452850A (en) * | 2019-08-23 | 2019-11-15 | 华中农业大学 | A kind of bacillus amyloliquefaciens culture, preparation method and applications |
| CN110447765A (en) * | 2019-08-23 | 2019-11-15 | 华中农业大学 | A kind of method and its application preparing bafillus natto culture using distillers ' grains |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111748504B (en) | 2023-06-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101940267B (en) | Method for preparing microbial fermentation feed for improving pork quality | |
| CN111436526A (en) | Preparation method and application of fermented rice bran meal with bacterium enzyme for improving growth performance of fattening pigs | |
| CN114601041A (en) | Highland barley vinasse fermented feed for improving yak meat quality and preparation method and application thereof | |
| CN112471325A (en) | Straw biological fermentation feed and preparation method and application thereof | |
| CN110384176B (en) | Method for preparing animal feed by culturing bacillus licheniformis through distiller's grains and puffed corn flour | |
| CN116478867A (en) | Fermented feed with high content of soluble protein as well as preparation method and application thereof | |
| CN106035990B (en) | Method for preparing biological feed by solid-state enzymolysis fermentation of citrus pulp, product and application thereof | |
| CN111296723A (en) | Probiotic agent for multi-strain combined solid state fermentation of rapeseed meal and fermentation method thereof | |
| CN113331318B (en) | Preparation method of feed for enriching yeast, astaxanthin, lactic acid and protease for shrimp and crab culture | |
| CN110558420A (en) | High-dietary-fiber low-antigen-protein fermented soybean hull | |
| CN111748504B (en) | Technical method for producing enterococcus faecium by using white spirit lees | |
| KR101252134B1 (en) | Feed additives for promoting growth of cattle and process for the preparation of feed for breeding cattle using the same | |
| RU2391859C2 (en) | Method of protein-vitamin fodder production | |
| CN110373364B (en) | Method for producing bacillus coagulans based on distiller's grains | |
| RU2243678C1 (en) | Method for preparing protein-vitamin fodder | |
| CN114886008A (en) | Biological fermentation selenium-rich feed and preparation method thereof | |
| CN112586615A (en) | Preparation method of beef cattle feed | |
| CN116391793B (en) | Process for microbial fermentation of feed and application | |
| PL219782B1 (en) | New strain of Lactobacillus buchneri A and multi-agent for preserving high-starch plants | |
| CN109287848B (en) | Fermentation enhancer, preparation method and application thereof | |
| CN116326676A (en) | Preparation method of semi-solid antibiotic-reducing feed by synergistic fermentation of bacterial enzymes and semi-solid antibiotic-reducing feed | |
| CN118266526A (en) | Process for producing feed additive by using lactobacillus plantarum | |
| CN1631202A (en) | Cassava feed with high protein | |
| CN111616262A (en) | Biological feed treatment enzyme preparation and preparation method and application thereof | |
| CN117562215A (en) | Method for preparing rice bran cereal powder by solid-state enzymolysis, fermentation and degreasing of rice bran |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |