CN104161172A - Method for producing probiotic peptides by using two-step solid-state fermentation of peanut meal - Google Patents
Method for producing probiotic peptides by using two-step solid-state fermentation of peanut meal Download PDFInfo
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Abstract
The invention belongs to the technical field of animal feeds and bioengineering, specifically relates to a method for producing probiotic peptides by using the two-step solid-state fermentation of peanut meal. The method comprises the following steps: using the peanut meal as a raw material, killing bacteria after uniformly mixing and churning the peanut meal and water, adding 2%(v/w) aspergillus niger(CGMCC9183) spore liquid to the water solution of the peanut meal and the water according to the weight of the peanut meal after killing the bacteria, culturing the mixture of the spore liquid and the water solution for 24-28 hours at the temperature of 28-32 DEG C, and turning over and mixing the materials once every four hours; and adding 10%(v/w) pediococcus pentosaceus(CGMCC9182) to the mixture of the spore liquid and the water solution according to the weight of the peanut meal after culturing the mixture of the spores liquid and the water solution, uniformly mixing the mixture of the peanut meal, the water, the spore liquid and the pediococcus pentosaceus, anaerobically fermenting the mixture of the peanut meal, the water, the spore liquid and the pediococcus pentosaceus for 36-42 hours at the temperature of 37-46 DEG C, drying and crushing the mixture of the peanut meal, the water, the spore liquid and the pediococcus pentosaceusat the temperature of 60 DEG C after fermenting, and obtaining the probiotic peptides. The polypeptides content of the probiotic peptides achieves more than 25%, the lactic acid content achieves more than 8.0%, the viable bacteria count of lactobacilli is more than 109CFU/g, and acidic proteinases achieve more than 200U/g. The method disclosed by the invention has the advantages that the cost of production is low and the fermenting process is easy to control, so that the quality of products is stable.
Description
Technical field
The invention belongs to animal feed and bionic technical field, be specifically related to a kind of method of utilizing peanut meal two step solid state fermentations to produce prebiotic peptide.
Background technology
Peanut meal is the byproduct of shelled peanut after oil plant is refined in squeezing, containing abundant vegetable protein.The nutritive value of peanut meal is higher, and its metabolizable energy is the highest in dregs of rice class, and gross protein value reaches more than 48%, is the important plant origin protein feed of China, and the annual peanut meal resource of China is about more than 300 ten thousand tons of left and right.Yet, because the protein quality of peanut meal is the same with other plant albumen, all with macromolecular structure, exist, and have ANFs, the digestion of impede protein, absorption, the utilization rate of albumen is relatively on the low side.
Yet; at present also relatively less to the exploitation of peanut meal; the polypeptide that obtains high-load (absolute magnitude is higher than 25%) is all to be undertaken by the method for liquid enzymolysis; product yield is lower; drying cost is too high; and because product application surface is narrower, alternative materials is abundant, be difficult to realize the large-scale production on peanut meal.Recently, solid state fermentation, because water content is few, does not produce discarded object substantially, low in the pollution of the environment, and the easy advantage such as processings, in studied personnel's application and peanut meal high-valued.At present, adopt bacillus and the lactic acid bacteria generation fermented peanut meal that jointly ferments, as feedstuff, obtained larger concern.Yet, because bacillus produces neutral proteinase, under the common fermentation of lactic acid bacteria, fermentation system becomes acidity fast, bacillus and neutral proteinase are difficult to play a role, and bacillus and lactic acid bacteria are respectively aerobic microbiological and anaerobe, both are inconsistent to the demand of oxygen, limited (the product lactic acid bacteria 10 of growth of microorganism
8cFU/g), also cause the useful metabolite content of product limited, if content of peptides is in 8% left and right, lactic acid content 3% left and right.
For solving the defect of the feeding quality of peanut meal, in conjunction with current conventional polypeptide products, probio, probiotic and the enzyme preparation of field of fodder, by biotechnology processing procedure, the function of these benefit-additives is rolled into one, prepared prebiotic peptide.First, sweat can be regarded nutriment predigestion process in vitro as, the ANFs removal that is decomposed in peanut meal, macro-molecular protein is broken down into polypeptide and the little peptide of part, the absorption of peptide has that transport speed is fast, low, the carrier of consuming energy is difficult for transporting uncontested property and inhibition between the advantage such as saturated and various peptide, animal tissue can also directly utilize the amino acid synthetic tissue albumen in peptide, reduces energy consumption; Secondly, young animal gastrointestinal physiology and microbiota are grown also not sound, stomachial secretion hydrochloric acid ability is poor, after Achlorhydria, cause in stomach pH value to raise and suppress pepsin activity and along with the interpolation in feed, make the digestibility of nutriment lower, a large amount of of the external source digestive ferment of sweat accumulation and useful metabolin supplement, and contribute to improve intestinal microflora balance, strengthen animal body immunity, significantly improve nutriment digestibility and utilization efficiency in animal body.Meanwhile, because food security more and more receives publicity, European Union completely forbade and in feed, adds antibiotic in 2006, and the work of finding Substitutes For Antibiotic has also obtained certain progress.Research is found, in feed, add the microbial environment that probio (lactic acid bacteria) or its metabolite lactic acid can effectively improve animal stomach, effectively prevent disease of digestive tract, excitating organism immunologic function, improve body disease-resistant, anti-stress ability, improve utilization rate and the breeding performonce fo animals of feed.
Therefore, on the basis of current Animal nutrition theoretical system, for the deficiency that has at present technology, by aerobic microbiological and anaerobe two-step fermentation, realize the external degradation of peanut meal albumen, accumulate useful metabolite polypeptide and digestive ferment, the probio that Sync enrichment is a large amount of, prepares prebiotic peptide, to improving utilization rate and the breeding performonce fo animals of feed, alleviate abuse of antibiotics and there is important effect, the healthy and sustainable development of feed industry and aquaculture is there is to the meaning of the property changed.
Summary of the invention
Technical problem to be solved by this invention is to utilize peanut meal two step solid state fermentations to produce prebiotic peptide, and the content of peptides of prebiotic peptide reaches more than 25%, and lactic acid content reaches more than 8.0%, and viable count of lactobacillus reaches 10
9more than CFU/g, more than acid protease reaches 200U/g.Two step solid state fermentations are respectively aerobic fermentation and anaerobic fermentation, the microorganism that adopts meets < < feed addictive kind catalogue (2013) > >, is respectively aspergillus niger (CGMCC No.9183) and the Pediococcus pentosaceus (CGMCC No.9182) high temperature resistant, high-yield quick lactic acid of high yield acid protease.
Pediococcus pentosaceus, circular bacterium colony, milky, smooth, projection, neat in edge, opaque, Gram-positive, coccus.Utilize glucose, wood sugar, arabinose, galactolipin, mannose, the multiple sugar such as Glucosamine and cellobiose produces acid, and acidproof, bile tolerance, has good enteron aisle colonization ability.
Aspergillus niger, pitchy, top capsule is large, spherical, and stigma is double-deck, and conidium is spherical, darkly, pitchy, level and smooth or coarse.The flourishing multi-branched of mycelia, has the many cells fungi every multinuclear.Produce acid protease, phytase, cellulase and zytase etc.
The incubation step of Pediococcus pentosaceus is:
37 ℃ of the activated inclined-plane of Pediococcus pentosaceus access MRS culture mediums are cultivated 36h, then with the 5% inoculum concentration 12h that spreads cultivation step by step, until enough solid state fermentation consumptions.
MRS culture medium (gL-1): glucose 20, beef extract 10, tryptone 10, yeast extract 5, K
2hPO
42, Triammonium citrate 2, sodium acetate 5, MgSO
40.1, MnSO
4h
2o 0.05, Tween-80 1, pH nature, 121 ℃ of sterilizing 20 min.
The incubation step of aspergillus niger is:
Behind the activated inclined-plane of aspergillus niger, the dull and stereotyped 30 ℃ of cultivation 5d of line potato culture, dull and stereotyped with aseptic water washing, make spore suspension.
Potato culture: potato 20g, sucrose 2g, running water 100mL, agar 2g, pH nature (approximately 6.0) 121 ℃ of 20min that go out.
Potato processing method: peeling potatoes, be cut into piece and add water, boil 30min, by filtered through gauze, filtrate sugaring, supplies water to 100ml, packs triangular flask into.
Technical scheme of the present invention is:
A method of utilizing peanut meal two step solid state fermentations to produce prebiotic peptide, step is as follows:
Take peanut meal as raw material, by material water weight ratio, 1:0.8-1:1.1 mixes rear 115-121 ℃ of sterilizing 10-20min with running water, after sterilizing, press the 2%(v/w of peanut meal weight) access aspergillus niger spore liquid, after mixing, at 28 ℃ ~ 32 ℃, cultivate 24 ~ 28h, every 4h stirring is once; After cultivation by 3 batches of mixing, by peanut meal weight access 10%(v/w) Pediococcus pentosaceus, after mixing at 37 ℃ ~ 46 ℃ anaerobic fermentation 36 ~ 42h, after fermentation ends 60 ℃ dry, pulverize, make prebiotic peptide; Described aspergillus niger (Aspergillus niger) preservation is to the Chinese common micro-organisms culture presevation administrative center that is positioned at No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and on May 20 2014 preservation time, its deposit number is CGMCC No.9183; Pediococcus pentosaceus (Pediococcus pentosaceus) preservation is to the Chinese common micro-organisms culture presevation administrative center that is positioned at No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and on May 20 2014 preservation time, its deposit number is CGMCC No.9182.
Beneficial effect of the present invention is:
The present invention is by the flora of screening effect excellence, and by optimizing preparation technology and parameters, a kind of method of the new prebiotic peptide of production is provided, be specially and utilize peanut meal two step solid state fermentations to produce prebiotic peptide, the content of peptides of the prebiotic peptide obtaining reaches more than 25%, lactic acid content reaches more than 8.0%, and viable count of lactobacillus reaches 10
9more than CFU/g, more than acid protease reaches 200U/g.The external predigestion of high molecular weight protein in this Realization of Product peanut meal, and accumulate polypeptide and the acid protease of useful metabolite lactic acid, absorption easy to digest, the lactic acid bacteria that enrichment is useful.
The prebiotic peptide product that the present invention obtains adds 5% left and right in feed formula, can improve the abilities of digestive and absorption of animal, regulates animal alimentary canal microecological balance, suppresses the pathogenic microorganisms growths such as Escherichia coli, salmonella, strengthens the disease resistance ability of animal.
In a word, preparation method of the present invention is easy to control, the prebiotic peptide product steady quality obtaining, and performance is good, and the method possesses outstanding substantive distinguishing features and progressive significantly.
contrast effect:
The fermented peanut meal that adopts one-step fermentation to prepare, content of peptides is 8%, and lactic acid content is 3.0%, and beneficial bacterium number of viable is 10
8cFU/g, the prebiotic peptide prepared far below this method.
test example:
The basal feed that adds 6% fish meal of take is test control group, the fish meal that substitutes respectively 2%, 4%, 6% in feed with prebiotic peptide (is equivalent to 1/3 of fish meal total amount, 2/3,3/3), be made into etc. nitrogen, etc. can feed, in each fish meal alternate sets, supplement crystal lysine, methionine and threonine simultaneously, make it reach the level consistent with control group.Feedstuff is all pulverized with pulverizer, crosses 40 mesh sieves, is crushed to sinking pellet diet (85 ℃ of pelleting temperatures) after mixing, and dry at ventilation lucifugal place, plastic bag sealing.Carry out 8 weeks raising experiments:
the prepared prebiotic peptide of table 1 the present invention replaces the impact of fish meal on Growth Op Tilapia
Note: the different alphabetical persons of colleague's data tool, represent significant difference (P<0.05)
the prebiotic peptide of table 2 replaces the impact of fish meal on Tilapia mossambica body index
Note: the different alphabetical persons of colleague's data tool, represent significant difference (P<0.05)
By table 1, table 2 is known, in the tilapia feed that is 6% in fish meal content, under the condition of lysine supplement, methionine and threonine, the prepared prebiotic peptide of the present invention can replace 2/3 (4%) of fish meal consumption, and test group Growth Op Tilapia index and body index and control group are quite even better than control group.In the feeding process of the prebiotic peptide Substitute For Partial of product of the present invention fish meal, improve acid-base balance in Tilapia mossambica body, improved the rate of digesting and absorbing, improved immunity, reduced the water pollution bringing in breeding process.
The specific embodiment
Below by specific embodiment, technical scheme of the present invention is described in detail.
embodiment 1
Take peanut meal as raw material, by material-water ratio 1:0.8, mix rear 121 ℃ of sterilizing 20min with running water, after sterilizing, press the 2%(v/w of peanut meal weight) access aspergillus niger (CGMCC No.9183) spore liquid, after mixing, at 28 ℃ ~ 32 ℃, cultivate 28h, every 4h stirring is once; After cultivation by 3 batches of mixing, by peanut meal weight access 10%(v/w) Pediococcus pentosaceus (CGMCC No.9182), after mixing at 37 ℃ ~ 46 ℃ anaerobic fermentation 42h, dry after 60 ℃ of fermentation ends, pulverize, make prebiotic peptide.The content of peptides of prebiotic peptide reaches 28.4%, and lactic acid content reaches 8.8%, and viable count of lactobacillus reaches 1.2 * 10
9cFU/g, acid protease reaches 228U/g.
embodiment 2
Take peanut meal as raw material, by material-water ratio 1:0.8, mix rear 121 ℃ of sterilizing 20min with running water, after sterilizing, press the 2%(v/w of peanut meal weight) access aspergillus niger (CGMCC No.9183) spore liquid, after mixing, at 28 ℃ ~ 32 ℃, cultivate 26h, every 4h stirring is once; After cultivation by 3 batches of mixing, by peanut meal weight access 10%(v/w) Pediococcus pentosaceus (CGMCC No.9182), after mixing at 37 ℃ ~ 46 ℃ anaerobic fermentation 36h, dry after 60 ℃ of fermentation ends, pulverize, make prebiotic peptide.The content of peptides of prebiotic peptide reaches 26.2%, and lactic acid content reaches 8.3%, and viable count of lactobacillus reaches 1.0 * 10
9cFU/g, acid protease reaches 212U/g.
embodiment 3
Take peanut meal as raw material, by material-water ratio 1:1.1, mix rear 120 ℃ of sterilizing 15min with running water, after sterilizing, press the 2%(v/w of peanut meal weight) access aspergillus niger (CGMCC No.9183) spore liquid, after mixing, at 28 ℃ ~ 32 ℃, cultivate 28h, every 4h stirring is once; After cultivation by 3 batches of mixing, by peanut meal weight access 10%(v/w) Pediococcus pentosaceus (CGMCC No.9182), after mixing at 37 ℃ ~ 46 ℃ anaerobic fermentation 36h, dry after 60 ℃ of fermentation ends, pulverize, make prebiotic peptide.The content of peptides of prebiotic peptide reaches 25.4%, and lactic acid content reaches 8.2%, and viable count of lactobacillus reaches 1.0 * 10
9cFU/g, acid protease reaches 201U/g.
More than shown and described basic principle of the present invention and principal character and advantage of the present invention.The technical staff of the industry should understand; the present invention is not restricted to the described embodiments; description in above-described embodiment and description principle of the present invention is just described; without departing from the spirit and scope of the present invention; the present invention also has various changes and modifications; these changes and improvements all fall in the claimed scope of the invention, and in the claimed scope of the present invention, appending claims and equivalent thereof define.
Claims (8)
1. a method of utilizing peanut meal two step solid state fermentations to produce prebiotic peptide, step is as follows:
Take peanut meal as raw material, by material water weight ratio, 1:0.8-1:1.1 mixes rear 115-121 ℃ of sterilizing 10-20min with running water, after sterilizing, press the 2%(v/w of peanut meal weight) access aspergillus niger spore liquid, after mixing, at 28 ℃ ~ 32 ℃, cultivate 24 ~ 28h, every 4h stirring is once; After cultivation by 3 batches of mixing, by peanut meal weight access 10%(v/w) Pediococcus pentosaceus, after mixing at 37 ℃ ~ 46 ℃ anaerobic fermentation 36 ~ 42h, after fermentation ends 60 ℃ dry, pulverize, make prebiotic peptide.
2. the method for utilizing peanut meal two step solid state fermentations to produce prebiotic peptide according to claim 1, it is characterized in that, described aspergillus niger is the aspergillus niger of high yield acid protease, the extremely Chinese common micro-organisms culture presevation of preservation administrative center, and its deposit number is CGMCC9183.
3. the method for utilizing peanut meal two step solid state fermentations to produce prebiotic peptide according to claim 1, it is characterized in that, described pentose sheet ball is Pediococcus pentosaceus high temperature resistant, quick lactic acid producing, the extremely Chinese common micro-organisms culture presevation of preservation administrative center, its deposit number is CGMCC9182.
4. the method for utilizing peanut meal two step solid state fermentations to produce prebiotic peptide according to claim 3, is characterized in that, the incubation step of described Pediococcus pentosaceus is:
37 ℃ of the activated inclined-plane of Pediococcus pentosaceus access MRS culture mediums are cultivated 36h, then with the 5% inoculum concentration 12h that spreads cultivation step by step, until enough solid state fermentation consumptions;
MRS culture medium (gL-1): glucose 20, beef extract 10, tryptone 10, yeast extract 5, K
2hPO
42, Triammonium citrate 2, sodium acetate 5, MgSO
40.1, MnSO
4h
2o 0.05, Tween-80 1, pH nature, 121 ℃ of sterilizing 20 min.
5. the method for utilizing peanut meal two step solid state fermentations to produce prebiotic peptide according to claim 2, is characterized in that, the incubation step of described aspergillus niger is:
Behind the activated inclined-plane of aspergillus niger, the dull and stereotyped 30 ℃ of cultivation 5d of line potato culture, dull and stereotyped with aseptic water washing, make spore suspension;
Potato culture: potato 20g, sucrose 2g, running water 100mL, agar 2g, pH nature (approximately 6.0) 121 ℃ of 20min that go out;
Potato processing method: peeling potatoes, be cut into piece and add water, boil 30min, by filtered through gauze, filtrate sugaring, supplies water to 100ml, packs triangular flask into.
6. the prebiotic peptide that prepared by the method as described in claim 1-5 any one.
7. the prebiotic peptide that prepared by the method as described in claim 1-5 any one, content of peptides reaches more than 25%, and lactic acid content reaches more than 8.0%, and viable count of lactobacillus reaches 10
9more than CFU/g, more than acid protease reaches 200U/g.
8. the application of the prebiotic peptide as described in claim 6 or 7 in preparing feed.
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Cited By (5)
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| CN104543374A (en) * | 2015-01-20 | 2015-04-29 | 青岛天祥食品集团有限公司 | High-value utilization method for high-nutrition peanut hot-pressed cakes |
| CN105146050A (en) * | 2015-09-25 | 2015-12-16 | 华南理工大学 | Nutritive matrix containing rich essential amino acid and production method and application thereof |
| CN109258921A (en) * | 2018-11-01 | 2019-01-25 | 青岛农业大学 | A kind of fermentation material of the Maize-Peanut dregs of rice, its fermentation process and application |
| CN110583964A (en) * | 2019-09-23 | 2019-12-20 | 江南大学 | Biological removal method for efficiently removing four aflatoxins in peanut meal |
| CN110818459A (en) * | 2018-08-10 | 2020-02-21 | 中国科学院大连化学物理研究所 | Liquid peanut meal organic fertilizer and preparation method and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104543374A (en) * | 2015-01-20 | 2015-04-29 | 青岛天祥食品集团有限公司 | High-value utilization method for high-nutrition peanut hot-pressed cakes |
| CN105146050A (en) * | 2015-09-25 | 2015-12-16 | 华南理工大学 | Nutritive matrix containing rich essential amino acid and production method and application thereof |
| CN105146050B (en) * | 2015-09-25 | 2019-05-14 | 华南理工大学 | A kind of nutrient base material and its production method and purposes rich in essential amino acid |
| CN110818459A (en) * | 2018-08-10 | 2020-02-21 | 中国科学院大连化学物理研究所 | Liquid peanut meal organic fertilizer and preparation method and application thereof |
| CN110818459B (en) * | 2018-08-10 | 2021-10-22 | 中科绿帅生物科技(广州)有限公司 | Liquid peanut meal organic fertilizer and preparation method and application thereof |
| CN109258921A (en) * | 2018-11-01 | 2019-01-25 | 青岛农业大学 | A kind of fermentation material of the Maize-Peanut dregs of rice, its fermentation process and application |
| CN110583964A (en) * | 2019-09-23 | 2019-12-20 | 江南大学 | Biological removal method for efficiently removing four aflatoxins in peanut meal |
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