CN114058543A - Pediococcus acidilactici DY15 and application thereof, feed and preparation method thereof - Google Patents
Pediococcus acidilactici DY15 and application thereof, feed and preparation method thereof Download PDFInfo
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- CN114058543A CN114058543A CN202111384226.2A CN202111384226A CN114058543A CN 114058543 A CN114058543 A CN 114058543A CN 202111384226 A CN202111384226 A CN 202111384226A CN 114058543 A CN114058543 A CN 114058543A
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- pediococcus acidilactici
- feed
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- fermentation
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- TUNFSRHWOTWDNC-HKGQFRNVSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCC[14C](O)=O TUNFSRHWOTWDNC-HKGQFRNVSA-N 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 239000001393 triammonium citrate Substances 0.000 description 1
- 235000011046 triammonium citrate Nutrition 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
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Abstract
The invention provides pediococcus acidilactici DY15 and application thereof, a feed and a preparation method thereof, and relates to the technical field of biology. The research of the inventor discovers that the Pediococcus acidilactici DY15 provided by the invention has bacteriostatic activity, has good inhibitory action on the growth of escherichia coli, salmonella and staphylococcus aureus, and the fermentation product of the Pediococcus acidilactici DY15 comprises various flavor substances, can be applied to the field of feeds, improves the nutritional value of the feeds, improves the palatability, improves the immunity of poultry and livestock, and establishes a good intestinal microbial system. According to the preparation method of the feed, the palm meal and the soybean hulls are treated in a mode of bacteria-enzyme synergistic fermentation, the method is simple and convenient, the bacteriostatic ability of the palm meal and the soybean hulls and the content of organic acid are greatly improved, the palatability of the feed is improved, and the palm meal and the soybean hulls can become the feed.
Description
Technical Field
The invention relates to the technical field of biology, in particular to pediococcus acidilactici DY15 and application thereof, a feed and a preparation method thereof.
Background
Lactic acid bacteria are a general term for gram-positive, non-spore bacteria, and the main metabolite of the fermented sugars is lactic acid. Lactic acid bacteria, which are living microorganisms, play an important role in human and animal health, and their antibacterial and probiotic properties are applied in various fields such as food fermentation, livestock and poultry husbandry, medical care, and industrial production of lactic acid, and thus lactic acid bacteria have become important research targets. The health effect of pediococcus acidilactici is also used for treating intestinal infection and inflammatory bowel disease, and the addition of lactobacillus in livestock industry replaces the effect with antibiotics, so that the nutrition of the feed is enriched, the balance of intestinal flora is maintained, and the healthy development of animals is promoted.
The palm meal is a byproduct of oil extraction of oil palm fruits, has the characteristics of large yield, low price, no aflatoxin and high contents of fat, crude fiber and amino acid. The soybean hull is a byproduct of soybean processing, and has low lignin content, high fiber content and easy absorption. Due to the limitation of the evaluation of the nutritional ingredients and the feeding value of the unconventional feed raw materials, the maximum adding proportion of the unconventional feed raw materials cannot be accurately determined when the daily ration is prepared. In addition, because the total carbohydrate in the palm meal is mostly non-starch polysaccharide such as mannan which is difficult to digest, the bean skin also contains antinutritional factors such as glycinin which can influence the digestion and absorption of nutrient components, and the content of polypeptide substances is low, thus being not beneficial to the digestion and absorption of animals.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a Pediococcus acidilactici strain DY15 which is preserved in China center for type culture Collection at 10 months and 18 days 2021, wherein the preservation numbers are as follows: CCTCC NO: m20211291.
The second purpose of the invention is to provide a microbial inoculum.
The third purpose of the invention is to provide application of the Pediococcus acidilactici (Pediococcus acidilactici) DY15 or microbial inoculum in preparation of antibacterial products.
The fourth purpose of the invention is to provide the application of the Pediococcus acidilactici DY15 or the microbial inoculum in feed.
The fifth purpose of the invention is to provide a preparation method of the feed.
A sixth object of the present invention is to provide a feed.
In a first aspect, the invention provides a Pediococcus acidilactici strain DY15, wherein the Pediococcus acidilactici strain DY15 is stored in the China center for type culture Collection with the following preservation numbers: CCTCC NO: m20211291.
In a second aspect, the invention provides a microbial inoculum, which comprises the Pediococcus acidilactici (Pediococcus aciliacticii) DY 15.
In a third aspect, the invention provides an application of the Pediococcus acidilactici (Pediococcus acidilactici) DY15 or a microbial inoculum in preparation of an antibacterial product.
As a further aspect, the inhibiting includes inhibiting at least one of staphylococcus aureus, salmonella, or escherichia coli.
In a fourth aspect, the invention provides an application of the Pediococcus acidilactici (Pediococcus acidilactici) DY15 or a microbial inoculum in any one of the following a-c:
a. preparing a feed;
b. improving the palatability of the feed;
c. improving the nutritive value of the feed.
In a fifth aspect, the invention provides a preparation method of a feed, comprising the following steps: mixing a substrate with alkaline protease, mannanase and the Pediococcus acidilactici DY15 or a microbial inoculum, and fermenting to prepare the feed;
the substrate comprises palm meal and soybean hull.
According to a further technical scheme, the mass ratio of the palm meal to the soybean hulls is 6: 2-6: 6, preferably 6: 4;
preferably, the substrate further comprises molasses; the mass percentage of the molasses in the substrate is 1% -5%, and the preferential percentage is 3%.
As a further technical scheme, after mixing, the concentration of the Pediococcus acidilactici DY15 is 1 x 107CFU/g is above;
preferably, the dosage of the alkaline protease is 1000-1500U/g substrate, preferably 1200U/g substrate;
the dosage of the mannase is 2000-3000U/g substrate, preferably 2500U/g substrate.
As a further technical scheme, the water content of the substrate is 45-65% by mass, and the water content is preferably 55%;
preferably, the fermentation temperature is 36-38 ℃, and preferably 37 ℃;
preferably, the fermentation time is 36-60 h, preferably 48 h;
preferably, the fermentation mode is sealed fermentation.
In a sixth aspect, the invention provides a feed prepared by the preparation method.
Compared with the prior art, the invention has the following beneficial effects:
the research of the inventor discovers that the Pediococcus acidilactici DY15 provided by the invention has bacteriostatic activity, has good inhibitory action on the growth of escherichia coli, salmonella and staphylococcus aureus, and the fermentation product of the Pediococcus acidilactici DY15 comprises various flavor substances, can be applied to the field of feeds, improves the nutritional value of the feeds, improves the palatability, improves the immunity of poultry and livestock, and establishes a good intestinal microbial system.
The preparation method of the feed provided by the invention takes the palm meal and the soybean hulls as substrates, takes Pediococcus acidilactici (DY 15) as zymocyte, simultaneously adds alkaline protease and mannase, and adopts a mode of bacteria-enzyme synergistic fermentation to treat the palm meal and the soybean hulls, the method is simple and convenient, the bacteriostatic ability of the palm meal and the soybean hulls and the content of organic acid are greatly improved, the palatability of the feed is also improved, and the palm meal and the soybean hulls become the feed possibly; after the fermentation is finished, the content of lactic acid is increased by 671.93 times compared with that before the fermentation, the total acid content is increased by 6.34 times compared with that before the fermentation, the acid production capability is moderate, and the excessive acidification of the palm meal and beans cannot be caused; the content of small peptide is improved by 28.03 percent compared with that before fermentation, the added value of the raw material is improved, and the waste is favorably recycled; after fermentation, detecting flavor substances with different contents, such as ethyl caprylate, methyl laurate, acetoin, methyl myristate, acetic acid, isoamyl caprylate, phenylacetaldehyde and the like, wherein a plurality of the flavor substances are common materials for preparing flavors and fragrances, and the palatability of the fermented palm meal and the bean curd skin is improved.
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 description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 shows the morphology of Pediococcus acidilactici DY15 in a plate medium;
FIG. 2 is a growth curve of Pediococcus acidilactici DY15 in MRS medium.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to embodiments and examples, but those skilled in the art will understand that the following embodiments and examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. Those who do not specify the conditions are performed according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
In a first aspect, the invention provides a Pediococcus acidilactici (Pediococcus acidilactici) DY15, which is classified and named as: pediococcus acidilactici DY15, latin literature name: pediococcus acitilacticii DY15, deposited in China center for type culture Collection, with the deposition address: wuhan university, preservation date: 18/10/2021, accession number: CCTCC NO: m20211291.
The Pediococcus acidilactici DY15 provided by the invention is separated from the pickled Chinese cabbage purchased from the tin-free European and Shang supermarket, the thallus of the Pediococcus acidilactici DY15 is medium in size, the colony on the culture medium is milky white, the surface is smooth and moist and is raised, and the pickled Chinese cabbage has a peculiar sour and fragrant taste.
The research of the inventor discovers that the Pediococcus acidilactici DY15 provided by the invention has bacteriostatic activity, has good inhibitory action on the growth of escherichia coli, salmonella and staphylococcus aureus, and the fermentation product of the Pediococcus acidilactici DY15 comprises various flavor substances, can be applied to the field of feeds, improves the nutritional value of the feeds, improves the palatability, improves the immunity of poultry and livestock, and establishes a good intestinal microbial system.
In a second aspect, the invention provides a microbial inoculum, which comprises the Pediococcus acidilactici (Pediococcus acidilactici) DY15, or further comprises other strains or auxiliary materials well known to those skilled in the art, and the microbial inoculum contains the Pediococcus acidilactici (Pediococcus acidilactici) DY15, so the microbial inoculum has all the beneficial effects of the strain.
In a third aspect, the invention provides an application of the Pediococcus acidilactici (Pediococcus acidilactici) DY15 or a microbial inoculum in preparation of an antibacterial product.
The research of the inventor finds that the Pediococcus acidilactici DY15 provided by the invention has bacteriostatic activity and has a good inhibitory effect on the growth of escherichia coli, salmonella and staphylococcus aureus, so that the Pediococcus acidilactici DY15 can be used for preparing bacteriostatic products.
In a fourth aspect, the invention provides an application of the Pediococcus acidilactici (Pediococcus acidilactici) DY15 or a microbial inoculum in any one of the following a-c:
a. preparing a feed;
b. improving the palatability of the feed;
c. improving the nutritive value of the feed.
The research of the inventor discovers that the Pediococcus acidilactici DY15 provided by the invention has bacteriostatic activity, has good inhibitory action on the growth of escherichia coli, salmonella and staphylococcus aureus, and the fermentation product of the Pediococcus acidilactici DY15 comprises various flavor substances, can be applied to the field of feeds, improves the nutritional value of the feeds, improves the palatability, improves the immunity of poultry and livestock, and establishes a good intestinal microbial system. Therefore, the Pediococcus acidilactici DY15 provided by the invention can be applied to preparing feed, improving the palatability of the feed and improving the nutritional value of the feed.
In a fifth aspect, the invention provides a preparation method of a feed, comprising the following steps: mixing a substrate with alkaline protease, mannanase and the Pediococcus acidilactici DY15 or a microbial inoculum, and fermenting to prepare the feed;
the substrate comprises palm meal and soybean hull.
The preparation method provided by the invention is simple and convenient, the palm meal and the soybean hulls are treated by adopting a mode of bacteria-enzyme synergistic fermentation, the bacteriostatic ability of the palm meal and the soybean hulls and the content of organic acid are improved, the colonization of pathogenic bacteria in animal intestinal tracts can be prevented, the disease resistance of animals is enhanced, anti-nutritional factors in raw materials are reduced, the utilization rate of nutrient substances in the feed and the digestion capacity of the animals are improved, and the preparation method has great significance for improving the feeding value of the palm meal and the soybean hulls and utilizing unconventional feed.
As a further technical solution, the mass ratio of the palm meal to the soybean hull may be, but is not limited to, 6:2, 6:3, 6:4, 6:5 or 6:6, preferably 6: 4;
preferably, the substrate further comprises molasses; the mass percentage of molasses in the substrate may be, for example, but not limited to, 1%, 2%, 3%, 4% or 5%, preferably 3%.
As a further technical scheme, after mixing, the concentration of the Pediococcus acidilactici DY15 is 1 x 107CFU/g is above;
preferably, the alkaline protease may be used, for example, but not limited to, 1000U/g substrate, 1100U/g substrate, 1200U/g substrate, 1300U/g substrate, 1400U/g substrate or 1500U/g substrate, preferably 1200U/g substrate;
the mannanase may be used, for example, but not limited to, 2000U/g substrate, 2200U/g substrate, 2400U/g substrate, 2600U/g substrate, 2800U/g substrate or 3000U/g substrate, preferably 2500U/g substrate.
"1200U/g substrate" means that 1200U of the enzyme was added to 1g of the substrate.
As a further embodiment, the substrate may have a moisture content of, for example, but not limited to, 45%, 49%, 53%, 57%, 61%, or 65%, preferably 55%;
preferably, the temperature of the fermentation may be, for example, but not limited to, 36 ℃, 37 ℃ or 38 ℃, preferably 37 ℃;
preferably, the fermentation time may be, for example, but not limited to, 36h, 40h, 44h, 48h, 52h, 56h or 60h, preferably 48 h;
preferably, the fermentation mode is sealed fermentation.
By further optimizing and adjusting the fermentation process, the fermentation of the palm meal and the soybean hulls is better realized, and the palatability and the nutritional value of the feed are improved.
In a sixth aspect, the invention provides a feed prepared by the preparation method.
The feed provided by the invention has high nutritive value, is easy to digest and good in palatability, and can enhance the disease resistance of livestock.
The invention is further illustrated by the following specific examples and comparative examples, but it should be understood that these examples are for purposes of illustration only and are not to be construed as limiting the invention in any way.
EXAMPLE 1 screening of strains
1, culture medium: MRS medium (g/L): 10.0 parts of peptone, 8.0 parts of beef extract, 4.0 parts of yeast powder, 20.0 parts of glucose, 2.0 parts of dipotassium phosphate, 2.0 parts of triammonium citrate, 5.0 parts of sodium acetate, 0.58 part of magnesium sulfate heptahydrate, 0.25 part of manganese sulfate tetrahydrate, 801 mL of Tween and 1L of distilled water, and sterilizing at 115 ℃ for 20 minutes.
2, screening: the strain is separated from pickled Chinese cabbage purchased from Wuxi European and Shang supermarket, the pickled Chinese cabbage is firstly cultured in MRS culture medium at 37 ℃ for 24 hours, the culture solution is diluted by 100 times and 1000 times respectively by sterile water, 100 mu L of the culture solution is taken and coated on MRS solid culture medium, the culture solution is cultured at 37 ℃ for 24 hours, and the strain is separated by streaking to obtain a lactic acid strain (shown in figure 1). The microbiological characteristics are gram-positive typical staining, medium thallus, milk white bacterial colony on the culture medium, smooth and moist surface, and raised appearance, and has a peculiar sour and fragrant taste. The strain was cultured in MRS medium and the growth curve is shown in fig. 2.
3: identification of the strains: the selected strains are coated on an MRS plate, a single colony is selected to be amplified by using universal primers 1492R (GGTTACCTTGTTACGACTT (SEQ ID NO.1)) and 27F (AGAGTTTGATCCTGGCTCAG (SEQ ID NO.2)), an amplification product is handed to a biological engineering (Shanghai) company Limited to carry out 16SrRNA sequencing, and the obtained sequence result is subjected to homology comparison with a model strain in Genbank through Nucleotide BLAST of NCBI. The comparison result shows that the strain has 99 percent of similarity I with the 16sRNA of the related model strain in the Genbank, and the strain is determined to be the Pediococcus acidilactici and is named as the Pediococcus acidilactici DY 15.
Example 2 identification of the physiological and biochemical characteristics of Pediococcus acidilactici DY15
(1) Litmus milk experiment: the milk mainly contains lactose and casein, and litmus is added as an indicator and a redox indicator, so that when the litmus is neutral, is in light purple acidity, is in pink alkalinity and is in blue reduction, the milk is faded and reduced to white from top to bottom. Lactic acid bacteria ferment lactose to produce acid, litmus turns red, and when the acidity is very high, milk can be solidified. Observing the experimental results of the litmus milk after inoculation and culture, and observing the acid production and coagulation reaction of the litmus milk after one day of culture at 37 ℃.
(2) Starch hydrolysis experiments: inoculating the strain in a basal culture medium containing soluble starch, culturing at 37 deg.C for 24 hr, taking a little culture solution in a colorimetric disc, taking the culture solution not inoculated as a control, and adding Lugol iodine solution therein respectively. The absence of color indicates hydrolysis of the starch, and the presence of bluish black or bluish violet indicates that the starch is not hydrolyzed or incompletely hydrolyzed.
(3) Gelatin liquefaction experiment: the experimental strains were inoculated in a basal medium containing gelatin and cultured at 37 ℃ for 24h, and an uninoculated test tube was used as a control. And placing the inoculated control tube and the non-inoculated control tube in a refrigerator at 4 ℃, recording the experimental result after the control tube is solidified, and repeatedly observing and comparing for many times. If the control tube is solidified, the inoculation tube is liquefied into a positive reaction, and the solidification is a negative reaction.
(4) V-P experiment: the experimental strain is inoculated in PYG culture medium, cultured for 24h at 37 ℃, the culture solution is taken, added with the theaphenol and the potassium hydroxide, placed in a test tube without a cover for mixing, and shaken for 30 minutes, and the red color is shown as positive reaction.
(5) Dextran assay: fresh cultures were inoculated on medium slants containing sucrose and cultured at 37 ℃ for 24 h. The culture on the slant formed a thick lawn indicating glucan production as a positive reaction, otherwise as a negative reaction.
(6) Hydrogen sulfide test: inoculating fresh culture into a culture medium containing cysteine or cystine, clamping a lead acetate filter paper strip by using a sterile forceps, suspending the lead acetate filter paper strip in an inoculation test tube, wherein the lower end of the lead acetate filter paper strip is close to the surface of the culture medium and does not contact the liquid level, the upper end of the lead acetate filter paper strip is tightly plugged by using a cotton plug, a blank control is arranged in the experiment, the lead acetate filter paper strip is suspended on the non-inoculated test tube culture medium, culturing is carried out at 37 ℃ for 24h, and observation and comparison are carried out, so that the paper strip turns black to be a positive reaction.
(7) Sugar fermentation experiment: carbohydrates such as sugar or alcohols to be measured are added to the basal medium and separately dispensed into 5mL test tubes. Shaking and culturing at 37 deg.C for 24 h. During detection, a small amount of culture solution is taken and placed in a colorimetric disc, meanwhile, culture solution without carbohydrate is taken as a control, BTB-MR reagent is dripped to compare the change of color, and the strength of acid production is recorded.
The physiological and biochemical characteristics of the DY15 strain are shown in tables 1 and 2, and the results show that litmus milk and V-P experiments are positive, starch hydrolysis and hydrogen sulfide experiments and glucan and gelatin liquefaction are negative, and the strain can produce acid by fermenting D-galactose, maltose, rhamnose, sucrose, fructose, lactose, glucose and D-mannose.
TABLE 1 physiological Properties of Pediococcus acidilactici DY15
Note: "-" indicates negative, and "+" indicates positive.
TABLE 2 Pediococcus acidilactici DY15 sugar fermentation results
Note: "+" indicates strong fermentation and "d" indicates slight fermentation.
Example 3 method for measuring bacteriostatic diameter of bacterial strain
Preparing an indicator bacterium liquid: three indicator bacteria of escherichia coli, salmonella and staphylococcus aureus are inoculated in an LB liquid culture medium and cultured for 24h at 37 ℃.
Adopting an oxford cup method: taking a flat plate with the diameter of about 90mm, respectively pouring 15-20mL of the nutrient agar culture medium which is melted by heating, uniformly spreading the nutrient agar culture medium in the flat plate, and placing the flat plate on a horizontal table surface to solidify the nutrient agar culture medium to be used as a bottom layer. Heating and melting semisolid nutrient agar culture medium (agar content is 1%), cooling to 48-50 deg.C, adding 0.1-0.2mL indicator bacteria suspension (bacteria concentration is 10) per 50-100mL culture medium8CFU/mL), 5mL was added to each 1 plate to spread evenly on the bottom layer as a bacterial layer. 4 Oxford cups are uniformly arranged in equal distance in each 1 plate for standby, 200 mu L of lactobacillus supernatant is respectively dripped into the Oxford cup in each double-layer plate, after the culture is carried out for 10 to 13 hours at 37 ℃, the diameter (or the area) of each inhibition zone is measured, and the result is shown in Table 3.
TABLE 3 Pediococcus acidilactici DY15 bacteriostatic effect
EXAMPLE 4 preparation of Pediococcus acidilactici microbial inoculum
(1) Inoculating the original strain of pediococcus acidilactici to an MRS culture medium according to the inoculation amount of 1%, activating for 24h at 37 ℃, activating for 2-3 generations according to the same manner, and continuously culturing in the MRS culture medium to obtain the bacterial liquid of the pediococcus acidilactici.
(2) Preparing a lactic acid bacteria liquid: and (3) putting the original pediococcus acidilactici strain into an MRS liquid culture medium, activating for 24h at 37 ℃, activating for 2 times according to the steps, and culturing for 24h at 37 ℃ in the MRS liquid culture medium to obtain fermentation liquor.
And (3) counting colonies: taking 1mL of fermentation liquor, diluting with a sterilized 1.5mL centrifuge tube in a clean bench according to a 10-fold dilution method. Sucking 100uL of the suspension by a pipette, injecting the suspension into a centrifugal tube containing 900uL of distilled water, repeatedly blowing and beating the suspension in the liquid for 5 times, and uniformly mixing the suspension by oscillation (paying attention to changing a gun head and fully shaking the suspension to uniformly mix the suspension in each dilution). Diluting step by step according to the above steps, taking 10-7，10-8100 mu L of the two gradiental bacterial suspensions are dripped on a seed culture medium plate for plate coating, and bacterial colonies are counted.
Example 5 Process for fermenting palm meal and bean curd skin by using bacterial enzymes
Using a coarse rod type pulverizer to pulverize raw materials of palm meal and soybean hull according to a mass ratio of 6:4 until the size is 2cm +/-0.5 cm for later use. Adding water, Pediococcus acidilactici DY15, alkaline protease and mannanase into palm pulp and testa Sojae Atricolor, and fermenting; the water content of palm meal and soybean hull was 55% (m/m), the alkaline protease content was 1200U/g substrate, the mannanase content was 2500U/g substrate, and Pediococcus acidilactici DY15 was cultured in the same manner as in example 4, and the bacterial solution was added to the substrate in an inoculation amount of 5% (v/m, mL/g) to a concentration of 1X 107CFU/g。
The content of the small peptide is 62.72mg/g by adopting a Folin-phenol method, which is improved by 28.03%; the total acid content is determined to be 5.07 percent by adopting an acid-base titration method, and is improved by 6.34 times; the number of detected lactic acid bacteria was 2X 107cfu/g, increased by 50%; as a result of measuring the content of organic acids in the fermentation raw material obtained in example 5 by the HPLC method, it was found that the contents of lactic acid, acetic acid and the like were significantly increased as shown in table 4. Wherein the content of lactic acid is increased by 671.93 times after fermentation.
TABLE 4 organic acid content of Pediococcus acidilactici DY15
Note: the control group was the raw material without fermentation treatment.
Example 6: aromatic substance for improving feed palatability before and after fermentation
The feed flavoring agent is also called as phagostimulant and appetite promoter, the action principle of the feed flavoring agent is closely related to the functions of taste, smell, respiratory system, digestive system and the like of animals, and the palatability of the feed can be improved. The headspace test results show that the fermented palm meal and the bean curd skin flavor substances obtained in example 5 have relatively high contents of ethyl caprylate, methyl laurate, acetoin, methyl myristate, acetic acid, isoamyl caprylate, phenylacetaldehyde and the like. The ethyl caprylate is mainly used for preparing food aromatizing agents and spices, has the fragrance similar to brandy and has sweet taste; acetoin is mainly used for preparing edible spices such as cream, dairy products, yoghourt, strawberries and the like, has strong butter and fat fragrance, and has pleasant milk fragrance after being highly diluted; the methyl laurate is mainly used for preparing edible essence and daily chemical essence and has the fragrance of fat, flower fragrance and wine; methyl myristate is commonly used in edible essence such as honey and coconut, daily essence, and organic synthesis. Maltol, ethyl decanoate, myristic acid, delta-dodecalactone, ethyl palmitate and the like are also detected in the fermented feed, and are common materials for preparing flavors and fragrances. In addition, formic acid, ammonium acetate, propionic acid, butyric acid, L-lactic acid, caproic acid, and the like have been detected to inhibit the growth of pathogenic bacteria and undesirable putrefactive microorganisms.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
SEQUENCE LISTING
<110> Shandong Bohai sea practice group Co., Ltd, Shandong Bohai sea grease industry Co., Ltd, Qingdao Bohai sea Kou grain and oil industry Co., Ltd, Guangxi Bohai sea agricultural development Co., Ltd
<120> pediococcus acidilactici DY15 and application thereof, feed and preparation method thereof
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<213> Artificial sequence
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<212> DNA
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Claims (10)
1. The Pediococcus acidilactici (Pediococcus acidilactici) DY15 is characterized in that the Pediococcus acidilactici (Pediococcus acidilactici) DY15 is preserved in China center for type culture Collection with the preservation number: CCTCC NO: m20211291.
2. A microbial preparation comprising Pediococcus acidilactici (Pediococcus acililacicici) DY15 according to claim 1.
3. Use of Pediococcus acidilactici (Pediococcus acidilactici) DY15 according to claim 1 or the microbial inoculum according to claim 2 in the preparation of bacteriostatic products.
4. The use of claim 3, wherein the inhibiting bacteria comprises inhibiting at least one of Staphylococcus aureus, Salmonella, or Escherichia coli.
5. Use of Pediococcus acidilactici (Pediococcus acidilactici) DY15 according to claim 1 or the microbial agent according to claim 2 in any one of the following a to c:
a. preparing a feed;
b. improving the palatability of the feed;
c. improving the nutritive value of the feed.
6. The preparation method of the feed is characterized by comprising the following steps: mixing a substrate with alkaline protease, mannanase and Pediococcus acidilactici (Pediococcus acidilactici) DY15 as claimed in claim 1 or microbial inoculum as claimed in claim 2, and fermenting to obtain feed;
the substrate comprises palm meal and soybean hull.
7. The preparation method according to claim 6, wherein the mass ratio of the palm meal to the soybean hull is 6: 2-6: 6, preferably 6: 4;
preferably, the substrate further comprises molasses; the mass percentage of the molasses in the substrate is 1% -5%, and the preferential percentage is 3%.
8. The method according to claim 6, wherein the concentration of Pediococcus acidilactici DY15 is 1X 10 after mixing7CFU/g is above;
preferably, the dosage of the alkaline protease is 1000-1500U/g substrate, preferably 1200U/g substrate;
the dosage of the mannase is 2000-3000U/g substrate, preferably 2500U/g substrate.
9. The preparation method according to claim 6, wherein the substrate contains 45-65% water by mass, preferably 55%;
preferably, the fermentation temperature is 36-38 ℃, and preferably 37 ℃;
preferably, the fermentation time is 36-60 h, preferably 48 h;
preferably, the fermentation mode is sealed fermentation.
10. A feed produced by the production method according to any one of claims 6 to 9.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150201662A1 (en) * | 2012-11-28 | 2015-07-23 | J9 Bio Agricultural Association Corporation | Health supplement food containing pediococcus acidilactici j9, and method for manufacturing same |
| CN105410337A (en) * | 2015-12-11 | 2016-03-23 | 上海邦成生物科技有限公司 | Biological feed additive rich in bioactive peptides and probiotics as well as preparation method of additive |
| CN107034163A (en) * | 2017-05-23 | 2017-08-11 | 北京市农林科学院 | A kind of novel lactic piece coccus and its application |
| CN107227279A (en) * | 2017-07-18 | 2017-10-03 | 邓禹 | One plant of Pediococcus acidilactici and its application |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150201662A1 (en) * | 2012-11-28 | 2015-07-23 | J9 Bio Agricultural Association Corporation | Health supplement food containing pediococcus acidilactici j9, and method for manufacturing same |
| CN105410337A (en) * | 2015-12-11 | 2016-03-23 | 上海邦成生物科技有限公司 | Biological feed additive rich in bioactive peptides and probiotics as well as preparation method of additive |
| CN107034163A (en) * | 2017-05-23 | 2017-08-11 | 北京市农林科学院 | A kind of novel lactic piece coccus and its application |
| CN107227279A (en) * | 2017-07-18 | 2017-10-03 | 邓禹 | One plant of Pediococcus acidilactici and its application |
Non-Patent Citations (1)
| Title |
|---|
| 党建章主编: "发酵工艺教程", 中国轻工业出版社 * |
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