CN104988183A - Preparation method of pleurotus ostreatus fermentation broth and application of fermentation broth in degradation of aflatoxin B1 - Google Patents
Preparation method of pleurotus ostreatus fermentation broth and application of fermentation broth in degradation of aflatoxin B1 Download PDFInfo
- Publication number
- CN104988183A CN104988183A CN201510477282.9A CN201510477282A CN104988183A CN 104988183 A CN104988183 A CN 104988183A CN 201510477282 A CN201510477282 A CN 201510477282A CN 104988183 A CN104988183 A CN 104988183A
- Authority
- CN
- China
- Prior art keywords
- preparation
- seed
- sulfate
- medium
- cap fungus
- 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.)
- Pending
Links
Landscapes
- Fodder In General (AREA)
Abstract
The invention relates to microbial fermentation, in particular to a preparation method of pleurotus ostreatus fermentation broth and application of the fermentation broth in degradation of aflatoxin B1. Pleurotus ostreatus CICC14012 is inoculated in an axenic culture medium containing carbon sources, nitrogen sources, inorganic salt and trace elements to conduct aerated fermentation cultivation to prepare the fermentation broth, and the fermentation broth can be used for degrading the virulence of the aflatoxin B1. By means of the method, the technical problems that an existing aflatoxin B1 removal method needs special conditions and special materials, is high in labor intensity and not complete in detoxification, has an effect on breaking nutritious components in processed fodder, and is severe in required condition, prone to producing chemical residuals and causing secondary pollution and the like are solved. The pleurotus ostreatus fermentation broth prepared through the method has the advantages of being easy and convenient in process, free of environment pollution and high in product safety, and has the advantages of being high in degradation speed, high in degradation rate, small in addition amount and the like when used for degrading the aflatoxin B1.
Description
Technical field
The present invention relates to fermentable, refer to that a kind of preparation method of oyster cap fungus fermented liquid and this fermented liquid are at aflatoxin degradation B especially
1in application.
Background technology
Oyster cap fungus is commonly called as flat mushroom, and be a kind of fungi forming sporophore, its growing environment difference is divided into wild and cultivated two kinds.Because of edible containing abundant nutrition composition in sporophore, be also called edible mushrooms.Learnt by a large amount of determination data, about containing crude protein 20 grams, crude fat 4 grams, 50 grams, carbohydrate, robust fibre 6 grams in every 100 grams of dry mushrooms, also containing mineral substance, multivitamin and various trace elements etc.Its protein content is 2.6 times of egg, 4 times of pork; Carbohydrate content accounts for 65.61% of total amount, and wherein reducing sugar accounts for 54.73%; Amino acid about has 18 kinds, and wherein 8 seed amino acid content of needed by human are higher, account for more than 35% of total amino acid content, and the nutritious prod especially as patient is good.
Traditional Chinese medicine is thought, flat mushroom is put down, and taste is sweet, has qi-restoratives, anticancer effect, can improve human metabolism, build up health, and regulating plant is neural.Flat mushroom contains polysaccharide body, has very strong restraining effect, and have immunological characteristic to tumour cell.The effect that flat mushroom also has wind-evil dispelling and cold-evil expelling, stimulates the circulation of the blood and cause the muscles and joints to relax, lumbago and skelalgia can be controlled, numb in every limb, channels and collaterals discomfort waits disease, in addition, flat mushroom is also to hepatitis, chronic gastritis, stomach and duodenal ulcer, and richets, hypertension etc. are all effective in cure, also there is certain curative effect to reducing blood cholesterol and preventing and treating urethral calculus, can opsonization be played to female dimacteric syndrome.
Produced from Pleurotus ostreatus is can to form sporophore containing growth on nutritious solid substance edible, and also can carry out mycelia deep fermentation, in nutrient solution, secretion has a large amount of polysaccharide, enzyme, amino acid and physiologically active substance.Polysaccharide based on acidic polysaccharose, containing β-1,3-dextran, semi-lactosi, seminose, glucuronic acid etc.Confirm through scientific research, liquid fermenting combination product is 91% to euphorbia egg decoctum inhibiting rate, and the glycoprotein of extraction has stronger restraining effect to external S180.Oyster cap fungus glycopeptide component has promotion Lymphokine to kill and wound (LAK) cell and natural killer (NK) cell killing function of tumor.Also there is reducing blood-fat and prevent and treat atherosclerosis, obviously can alleviate serum and liver cholesterol and triglyceride level.Multiple enzyme in fermented liquid is especially higher with three kinds of phenol oxidase content.These enzymes can be used for food fresh keeping and stable, can the multinomial chemical reaction of catalysis, change original molecule structure synthesis novel substance, change molecular substance chemical property, utilize this feature to carry out sewage disposal protection of the environment.
Aflatoxin be a class to the extremely strong material of humans and animals toxicity, be the secondary metabolite in flavus and Aspergillus parasiticus bacteria growing process.1993 delimited by the World Health Organization (WHO) cancer mechanism is 1 class carcinogens.Produce the fungi of aflatoxin to be prevalent in natural soil, farm crop and fruit, herbage, feed, especially in soybean, paddy, corn, peanut, in ox Milk and milk products, also often detect its toxin.In annual summer and autumn, temperature is higher, and atmospheric moisture is large, food crop prematuration period be easy to by mould contamination, the especially kind grain of pathology precocity very easily grow mold produce mycotoxin.Can directly life threatening after people is edible, endanger animal life or reduction breeding performonce fo animals after feed intake.The main source that people contacts aflatoxin is the food that meals pollute, and contaminated food has two types, and one is take in by aflatoxin-contaminated plant seed food; Two is that aflatoxin-contaminated feed enters in poultry, fowl body to proceed in milk, egg and meat product again and taken in by people, final harm humans life and health.The people such as Gao Xiufen have spot-check 279 parts, the corn feed sample of Jilin, Henan, Hubei, Sichuan, Guangdong, Guangxi 6 province in 2010, aflatoxin positive rate is 75.63%, mean concns 44.04 μ g/kg, concentration range 0.20-888.30 μ g/kg; Sichuan agricultural university Zhang Ziqiang determines 11 provinces, the whole nation for 2008-2009 years and amounts to 1013 parts of Feed Samples, wherein AFBs
1(AFB
1) recall rate is up to 99.51%, especially higher with cotton dregs, the dish dregs of rice, corn content.
The hazardness of aflatoxin is people and the effect of animal livers disorganization, and hepatitis occurs, and liver cirrhosis, can cause liver cancer even dead time serious.With AFB in the food of natural contamination
1the most common, its toxicity and carinogenicity also the strongest.Learn after deliberation, AFB
1the Lethal Dose 50 male (100g) of rat is 7.2 mgs/kg of body weight, female (150g) is 17.9 mgs/kg of body weight, belongs to the poison range (large 10 times of its toxicity ratio potassium cyanide, larger than arsenic 68 times) of special severe toxicity.Poisoning rear clinical manifestation is had a stomach upset, appetite stimulator, feels sick, vomiting, abdominal distension and hepatic region tenderness etc., and oedema appears in severe patient, stupor, twitches and dead.AFB
1(AFB
1) be the most strong carcinogen found at present, its carcinogenicity is 900 times of butter yellow, higher 75 times than position canceration probabilities such as induced by dimethylnitrosamine stomach, kidney, rectum, mammary gland, ovary and small intestines.AFB
1(AFB
1) and other poisonous mineral compound, toxicity as potassium cyanide is compared, its toxicity mechanism is more complicated, and it is mainly by suppressing nucleic acid and albumen synthesis, interference carbohydrate, lipid metabolism, Immunosuppression, reduce hormonal activity, T suppression cell ATP synthesizes, thus causes the suppression of carbohydrate and lipid metabolism and albumen synthesis, cell is lost activity, causes poisoning.So it is again a kind of cytotoxin.For this reason, AFB
1harm has become the focus that people pay close attention to.
AFB
1(AFB
1) be insoluble in water, be soluble in the organic solvents such as oil, methyl alcohol, acetone and chloroform, high temperature resistant, decomposition temperature is 268 DEG C, and separation, the degraded difficulty of above feature decision aflatoxin are large.The pattern removing aflatoxin harm loses toxicity for being separated removal Sum decomposition chemical structure.The detoxification that people adopt in recent years is divided into physical method, chemical process and biological method.Physical is washing, mineral substance absorption, x ray irradiation x etc., and this method labour intensity is large, and mineral substance adsorption saturation limits the treatment capacity to feed, and irradiation removing toxic substances is not thorough; Chemical method all adopts the chemical reagent such as strong acid, highly basic and strong oxidizer degraded toxin, is characterized in that degraded toxin is thorough, and feed nutrient has and destroys in various degree, all has chemical residual, brings secondary injury (secondary pollution) to poultry, domestic animal; Biological methods is Recent study comparatively multi-method, is also the method that people pay attention to most.The experimental results shows, biological methods is the most most scientific detoxification, is characterized in mild condition, and side effect is little, does not produce secondary pollution, does not destroy nutritive ingredient etc., is more and more subject to people's attention.The many activeconstituentss containing aflatoxin degradation of edible fungal hypha body deep fermentation product are also that people study more bacterial classification.
Microbial fermentation preparation aflatoxin degradation mainly adopts bacterium and fungi, and external great mass of data research display result is that fungi is many but bacterium report is less.
Summary of the invention
The object of the present invention is to provide the preparation method of oyster cap fungus fermented liquid and this fermented liquid at aflatoxin degradation B
1in application.
Overall technology design of the present invention is:
The preparation method of oyster cap fungus fermented liquid is inoculated in by oyster cap fungus (Pleurotus ostreatus) CICC14012 to make fermented liquid containing carbon source, nitrogenous source, inorganic salt and trace element without carrying out aerobic fermentation in bacteria fermentation culture medium;
Fermention medium is made up of the raw material of following mass percentage:
Potato starch 6.0%-7.0%, sucrose 1.8%-2.2%, yeast extract paste 0.18%-0.22%, ammonium sulfate 0.29%-0.33%, potassium primary phosphate 0.25%-0.30%, magnesium sulfate 0.15%-0.18%, vitamins B
10.0001%-0.0003%, trace element solution 5.0%-6.0%, veratryl alcohol 430 μ L/L-470 μ L/L, tween 80 0.1%-0.13%, surplus is sterilized water, pH=4.9-5.3;
Trace element solution is made up of following raw material:
Calcium chloride 1500mg, ferrous sulfate 150mg, zinc sulfate 100mg, copper sulfate 150mg, manganous sulfate 1000mg, rose vitriol 200mg, potassium aluminium sulfate 20mg, boric acid 10mg, Sodium orthomolybdate 20mg, is settled to 1000mL;
The processing condition of aerobic fermentation are as follows:
Inoculum size is volume ratio=6-8: 100, and temperature is 26 DEG C-28 DEG C, and air flow is 1: 0.7-1.2vvm, and pressure is 0.03Mpa-0.05Mpa, and culture cycle is for being not less than 240 hours.
Can it is evident that the control of fermentation termination is except time factor, in conjunction with 722 visible spectrophotometers, protein concn in Coomassie Brilliant Blue monitoring fermented liquid should be adopted, determine fermentation termination.
Oyster cap fungus fermented liquid prepared by employing aforesaid method is at aflatoxin degradation B
1in application.
Concrete technical conceive of the present invention also has:
For the needs of suitability for industrialized production, preferred technical scheme is, described oyster cap fungus bacterial classification access without bacteria fermentation culture medium before through enlarged culturing.
More preferred technical scheme is, described enlarged culturing comprises the preparation of slant strains, the preparation of plate bacterial classification, the preparation of first order seed and the preparation of secondary seed.
The preparation of slant strains adopts potato culture, and the preparation of plate bacterial classification adopts dextrose culture-medium, and temperature is 25 DEG C-27 DEG C, and culture cycle is 4-5 days.
In the preparation of first order seed and the preparation of secondary seed, seed culture medium adopts the raw material of following mass percentage to form:
Murphy juice 18%-21%, glucose 1.8%-2.2%, yeast extract paste 0.25%-0.30%, ammonium sulfate 0.29%-0.33%, potassium primary phosphate 0.25%-0.30%, magnesium sulfate 0.15%-0.18%, vitamins B
10.0001%-0.0003%, trace element solution 3.5%-4.5%, surplus is sterilized water, pH=5.0-5.5;
Trace element solution is made up of the raw material of following quality:
Calcium chloride 1500mg, ferrous sulfate 150mg, zinc sulfate 100mg, copper sulfate 150mg, manganous sulfate 1000mg, rose vitriol 200mg, potassium aluminium sulfate 20mg, boric acid 10mg, Sodium orthomolybdate 20mg, is settled to 1000mL.
The preparation of first order seed adopts following processing step:
Select the plate bacterial classification that growth is vigorous, healthy and strong, diameter 7mm-13mm disk is cut at the vigorous position of mycelial growth, being inoculated into by every bottle of 3-5 sheet inoculum size is equipped with in primary-seed medium triangular flask, within shaking culture 85-96 hour under temperature is 26 DEG C of-28 DEG C of conditions, makes first order seed; Loading amount is that the triangular flask of 1000ml loads 160-200ml primary-seed medium, and have a certain amount of mycelium suspended to estimate in primary-seed medium, uniformity sturdy with microscopy mycelial growth cultivates terminal for first order seed.
The preparation of secondary seed adopts following processing step:
According to the ratio of volume ratio=9-11: 100 of first order seed and secondary seed medium, be equipped with after first order seed being accessed sterilizing in the fermentor tank of secondary seed medium, loading amount is volume ratio=60-70:100, it is 26 DEG C-28 DEG C in temperature, air flow is 1: 0.5-0.7vvm, pressure is that the aerated culture carrying out being not less than 72 hours under the condition of 0.03Mpa-0.05Mpa makes secondary seed, have a large amount of mycelium suspended to estimate in liquid nutrient medium, uniformity sturdy with microscopy mycelial growth cultivates terminal for secondary seed.
Adopt the fermented liquid that method of the present invention obtains, the liquid obtained after carrying out solid-liquid separation is according to a conventional method oyster cap fungus fermentation liquor preparation, the method of solid-liquid separation, including, but not limited to adopting the prior aries such as sheet frame separation, whizzer separation, does not all depart from technical spirit of the present invention.
Substantive distinguishing features acquired by the present invention and significant technical progress are:
1, present invention process is easy, ripe, only needs submerged fermentation equipment, and namely the liquid especially after solid-liquid separation can be used for removing toxic substances, the application capable of using as feed of the mycelium after solid-liquid separation, preparation process three-waste free pollution.
2, the Produced from Pleurotus ostreatus used in the present invention, not at pathogenic species scope (edible oyster mushrooms), safe and reliable.
3, in the present invention, the main raw material of substratum is potato starch and edible sucrose, has safe, nontoxic, green feature.
4, this Produced from Pleurotus ostreatus fermentation liquor preparation is used for aflatoxin degradation B
1, degradation speed is fast, and degradation rate is high, mild condition, and selectivity is good, stable performance.For corn feed (AFB contaminated under natural condition of degrading
1content≤97.7 μ g/kg) and bean-dregs feed (AFB
1content≤93.5 μ g/kg) middle AFB
1, add in feed relative≤9 ‰ ratios, contratoxin degradation rate>=73.29%.
5, the product aflatoxin degradation B adopting the present invention to prepare
1, using method is simple, and have good operability, detoxification processes can not cause secondary to endanger.
Embodiment
Below in conjunction with embodiment, the invention will be further described; but should not be construed as limitation of the invention; the content that protection scope of the present invention is recorded with claim is as the criterion, any according to the equivalent technical elements replacement done by specification sheets, does not all depart from protection scope of the present invention.
Embodiment 1
The preparation method of oyster cap fungus fermented liquid, is inoculated in oyster cap fungus (Pleurotus ostreatus) CICC14012 and carries out aerobic fermentation in the aseptic culture medium containing carbon source, nitrogenous source, inorganic salt and trace element and make fermented liquid;
Fermention medium is made up of the raw material of following mass percent:
Potato starch 6.5%, sucrose 2.0%, yeast extract paste 0.18%, ammonium sulfate 0.31%, potassium primary phosphate 0.28%, magnesium sulfate 0.17%, vitamins B
10.0001%, trace element solution 6.0%, veratryl alcohol 450 μ L/L, tween 80 0.13%, surplus is sterilized water, pH=5.0;
Trace element solution: calcium chloride 1500mg, ferrous sulfate 150mg, zinc sulfate 100mg, copper sulfate 150mg, manganous sulfate 1000mg, rose vitriol 200mg, potassium aluminium sulfate 20mg, boric acid 10mg, Sodium orthomolybdate 20mg, is settled to 1000mL.
The processing condition of aerobic fermentation are as follows:
Inoculum size is volume ratio=7: 100, temperature 26 DEG C, air flow 1: 0.8vvm, and pressure is 0.03Mpa, and culture cycle is for being not less than 250 hours.
The control of fermentation termination, except time factor, in conjunction with 722 visible spectrophotometers, should adopt protein concn in Coomassie Brilliant Blue monitoring fermented liquid, determines fermentation termination.
Described oyster cap fungus bacterial classification access without bacteria fermentation culture medium before through enlarged culturing.
Described enlarged culturing comprises the preparation of slant strains, the preparation of plate bacterial classification, the preparation of first order seed and secondary seed.
The preparation of described slant strains adopts potato culture, and the preparation of plate bacterial classification adopts dextrose culture-medium, and temperature is 26 DEG C, culture cycle 4.5 days.
The preparation of described first order seed comprises following processing step:
Slant strains is inoculated on the solid medium in plate, cultivate 4.5 days under 26 DEG C of conditions, select the plate bacterial classification of robust growth, 10mm disk is cut at the vigorous position of mycelial growth, being inoculated into by every bottle of 3 inoculum sizes is equipped with in primary-seed medium triangular flask, under 27 DEG C of conditions, shaking culture makes first order seed in 90 hours, loading amount is that the triangular flask of 1000ml loads 200ml primary-seed medium, have a certain amount of mycelium suspended to estimate in primary-seed medium, uniformity sturdy with microscopy mycelial growth cultivates terminal for first order seed.
The preparation of described secondary seed comprises following processing step:
Volume ratio=10 according to first order seed and secondary seed medium: the ratio of 100, be equipped with after first order seed being accessed sterilizing in the fermentor tank of secondary seed medium, loading amount is volume ratio=70:100, it is 27 DEG C in temperature, air flow is 1: 0.7vvm, pressure is that the aerated culture carrying out being not less than 72 hours under the condition of 0.05Mpa makes secondary seed, have a large amount of mycelium suspended to estimate in liquid nutrient medium, uniformity sturdy with microscopy mycelial growth cultivates terminal for secondary seed.
Described primary-seed medium and secondary seed medium adopt the raw material of following mass percentage to form:
Murphy juice 21%, glucose 1.9%, yeast extract paste 0.28%, ammonium sulfate 0.29%, potassium primary phosphate 0.25%, magnesium sulfate 0.17%, vitamins B
10.0002%, trace element solution 4.0%, surplus is sterilized water, pH=5.3;
Trace element solution: calcium chloride 1500mg, ferrous sulfate 150mg, zinc sulfate 100mg, copper sulfate 150mg, manganous sulfate 1000mg, rose vitriol 200mg, potassium aluminium sulfate 20mg, boric acid 10mg, Sodium orthomolybdate 20mg, is settled to 1000mL.
The fermented liquid adopting the method for the present embodiment to obtain carries out solid-liquid separation according to a conventional method and namely can be made into fermentation liquor preparation, and solid-liquid separation is including, but not limited to adopting sheet frame separation, whizzer separation etc.
Embodiment 2
The difference of the present embodiment and embodiment 1 is:
Fermention medium is made up of the raw material of following mass percentage:
Potato starch 7.0%, sucrose 1.8%, yeast extract paste 0.20%, ammonium sulfate 0.29%, potassium primary phosphate 0.25%, magnesium sulfate 0.18%, vitamins B
10.0002%, trace element solution 5.0%, veratryl alcohol 430 μ L/L, tween 80 0.10%, surplus is sterilized water, pH=4.9.
In aerobic fermentation process, inoculum size is volume ratio=6: 100, temperature 28 DEG C, air flow 1: 0.7vvm, and pressure is 0.04Mpa, and culture cycle is for being not less than 265 hours.
In the preparation of slant strains and plate bacterial classification, culture temperature is 25 DEG C, culture cycle 5 days.
The preparation of first order seed comprises following processing step:
Slant strains is inoculated on the solid medium in plate, cultivate 5 days under 25 DEG C of conditions, select the plate bacterial classification of robust growth, 13mm disk is cut at the vigorous position of mycelial growth, being inoculated into by every bottle of 4 inoculum sizes is equipped with in primary-seed medium triangular flask, under 28 DEG C of conditions, shaking culture makes first order seed in 85 hours, and loading amount is that the triangular flask of 1000ml loads 180ml primary-seed medium.
The preparation of secondary seed comprises following processing step:
Volume ratio=9 according to first order seed and secondary seed medium: the ratio of 100, be equipped with after first order seed being accessed sterilizing in the fermentor tank of secondary seed medium, loading amount is volume ratio=65:100, it is 26 DEG C in temperature, air flow is 1: 0.6vvm, and pressure is that the aerated culture carrying out being not less than 72 hours under the condition of 0.03Mpa makes secondary seed.
Described primary-seed medium and secondary seed medium adopt the raw material of following mass percentage to form:
Murphy juice 18%, glucose 2.2%, yeast extract paste 0.30%, ammonium sulfate 0.31%, potassium primary phosphate 0.28%, magnesium sulfate 0.15%, vitamins B
10.0001%, trace element solution 4.5%, surplus is sterilized water, pH=5.5.
Embodiment 3
The difference of the present embodiment and embodiment 1 is:
Fermention medium is made up of the raw material of following mass percentage:
Potato starch 6.0%, sucrose 2.2%, yeast extract paste 0.22%, ammonium sulfate 0.33%, potassium primary phosphate 0.30%, magnesium sulfate 0.15%, vitamins B
10.0003%, trace element solution 5.5%, veratryl alcohol 470 μ L/L, tween 80 0.12%, surplus is sterilized water, pH=5.3.
In aerobic fermentation process, inoculum size is volume ratio=8: 100, temperature 27 DEG C, air flow 1: 1.2vvm, and pressure is 0.05Mpa, and culture cycle is for being not less than 280 hours.
In the preparation of slant strains and plate bacterial classification, culture temperature is 27 DEG C, culture cycle 4 days.
The preparation of first order seed comprises following processing step:
Slant strains is inoculated on the solid medium in plate, cultivate 4 days under 27 DEG C of conditions, select the plate bacterial classification of robust growth, 7mm disk is cut at the vigorous position of mycelial growth, being inoculated into by every bottle of 5 inoculum sizes is equipped with in primary-seed medium triangular flask, under 26 DEG C of conditions, shaking culture makes first order seed in 96 hours, and loading amount is that the triangular flask of 1000ml loads 160ml primary-seed medium.
The preparation of secondary liquid seed comprises following processing step:
Volume ratio=11 according to first order seed and secondary seed medium: the ratio of 100, be equipped with after first order seed being accessed sterilizing in the fermentor tank of secondary seed medium, loading amount is volume ratio=60:100, it is 28 DEG C in temperature, air flow is 1: 0.5vvm, and pressure is that the aerated culture carrying out being not less than 72 hours under the condition of 0.04Mpa makes secondary seed.
Primary-seed medium and secondary seed medium adopt the raw material of following mass percentage to form:
Murphy juice 19%, glucose 1.8%, yeast extract paste 0.25%, ammonium sulfate 0.33%, potassium primary phosphate 0.30%, magnesium sulfate 0.18%, vitamins B
10.0003%, trace element solution 3.5%, surplus is sterilized water, pH=5.0.Application test
The oyster cap fungus fermentation liquor preparation that applicant prepares with regard to above-described embodiment is respectively used to containing AFB
1the aqueous solution, corn feed and bean-dregs feed, carry out aflatoxin degradation B
1ability and addition test.Test-results is as follows:
Detoxification experiment 1
Utilizing the oyster cap fungus fermentation liquor preparation that the method for embodiment 1-3 obtains, is that 3 ‰, 6 ‰, 9 ‰ ratios add different concns AFB respectively in mass ratio
1fully mix in the aqueous solution of (content is at 5-100 μ g/L), insulation degraded toxin more than 72 hours under 37 DEG C of conditions, according to AFB in GB/17480-2008 feed
1measuring method-enzyme-linked immunosorbent assay, measure degraded after AFB
1content.Degraded toxicity test result is as table 1.
AFB in table 1 oyster cap fungus fermentation liquor preparation degradation water solution
1result
Note: Biao Zhong result data unit is μ g/kg.
Illustrated, at AFB by test-results in table 1
1content is in 10.9 μ g/kg solution, adds 3 ‰ oyster cap fungus fermentation liquor preparations, to AFB
1degradation rate>=85.32%; AFB
1content is in 50.6 μ g/kg solution, adds 6 ‰ oyster cap fungus fermentation liquor preparations, to AFB
1degradation rate>=83.8%; AFB
1content is in 98.9 μ g/kg solution, adds 9 ‰ oyster cap fungus fermentation liquor preparations, to AFB
1degradation rate>=81.5%.Summary sheet 1 test-results, AFB in the aqueous solution
1during the μ g/kg of content≤98.9, add≤9 ‰ oyster cap fungus fermentation liquor preparations respectively, contratoxin degradation rate can reach more than 81.5%.
Detoxification experiment 2
Equally, get 3 ‰ of corn feed respectively, 6 ‰, 9 ‰ ratios (W:W) oyster cap fungus fermentation liquor preparation, after adding the dilution of certain water gaging, more respectively with containing 11.0 μ g/kg, 54.3 μ g/kg, 97.7 μ g/kg AFBs
1corn feed mixing, with corn feed surface wettability as well (with certain globule), under being enclosed in 37 DEG C of conditions, insulation degraded more than 72 hours, according to AFB in GB/17480-2008 feed
1measuring method-enzyme-linked immunosorbent assay, to measure after degraded AFB in corn feed
1content, measurement result is as table 2.
AFB in table 2 oyster cap fungus fermentation liquor preparation degrading maize feed
1result
Note: Biao Zhong result data unit is μ g/kg.
Drawn by table 2 result: adopt oyster cap fungus fermentation liquor preparation, AFB in degrading maize feed
1content, when 11.0 μ g/kg, adds oyster cap fungus fermentation liquor preparation amount 3 ‰, its removing toxic substances rate>=80.00%; AFB
1content, when 54.3 μ g/kg, adds oyster cap fungus fermentation liquor preparation amount 6 ‰, its removing toxic substances rate>=68.32%; AFB
1content, when 97.7 μ g/kg, adds oyster cap fungus fermentation liquor preparation amount 9 ‰, its removing toxic substances rate>=73.29%.
Detoxification experiment 3
Get 3 ‰ of bean-dregs feed respectively, 6 ‰, 9 ‰ ratios (W:W) oyster cap fungus fermentation liquor preparation, after adding water dilution, weight is identical with dregs of beans weight, more respectively with contain 13.3 μ g/kg, 44.1 μ g/kg, 93.5 μ g/kg AFBs
1bean-dregs feed mixing, under being enclosed in 37 DEG C of conditions, insulation removing toxic substances more than 72 hours, according to AFB in GB/17480-2008 feed
1measuring method-enzyme-linked immunosorbent assay, to measure after degraded AFB in bean-dregs feed
1content, measurement result is in table 3.
AFB in table 3 oyster cap fungus fermentation liquor preparation degraded bean-dregs feed
1result
Note: Biao Zhong result data unit is μ g/kg.
Result shows, and oyster cap fungus fermentation liquor preparation is for AFB in bean-dregs feed of degrading
1, after increasing the humidity of dregs of beans, aflatoxin content 13.4 μ g/kg, adds 3 ‰ fermentation preparations, degradation rate>=81.34%; AFB
1content 44.1 μ g/kg, adds 6 ‰ ratios, degradation rate>=71.43%; AFB
1content 93.5 μ g/kg, adds 9 ‰ ratios, degradation rate>=75.40%.Relatively Degrading experiment 2 and 3 experimental result, for AFB in solid feed
1degraded, increases content of water in system and is conducive to fermentation liquor preparation aflatoxin degradation B
1action effect.
Drawn by detoxification experiment 1,2,3 result, adopt oyster cap fungus fermentation liquor preparation aflatoxin degradation B
1toxicity, detoxifying effect is better than in corn in solids, bean-dregs feed in aqueous, is more conducive to catalyzed reaction and carries out in water solution system.AFB in solid feed
1contamination level≤15 μ g/kg, this fermentation preparation adds by 3 ‰, removing toxic substances rate>=80%; Contamination level≤54.3 μ g/kg, adds 6 ‰ fermentation liquor preparations, removing toxic substances rate>=68.32%; Contamination level≤97.7 μ g/kg, adds 9 ‰ fermentation liquor preparations, removing toxic substances rate>=73.29%.In the light of actual conditions, AFB in general feeds
1pollute≤54.3 μ g/kg, add oyster cap fungus fermentation liquor preparation in>=6 ‰ ratios, all meet country after degraded to AFB in feed
1limitation requires (≤20 μ g/kg).Increase fermentation preparation adding proportion, detoxifying effect is better.
Claims (8)
1. the preparation method of oyster cap fungus fermented liquid is inoculated in by oyster cap fungus (Pleurotus ostreatus) CICC 14012 to make fermented liquid containing carbon source, nitrogenous source, inorganic salt and trace element without carrying out aerobic fermentation in bacteria fermentation culture medium; It is characterized in that:
Fermention medium is made up of the raw material of following mass percentage:
Potato starch 6.0%-7.0%, sucrose 1.8%-2.2%, yeast extract paste 0.18%-0.22%, ammonium sulfate 0.29%-0.33%, potassium primary phosphate 0.25%-0.30%, magnesium sulfate 0.15%-0.18%, vitamins B
10.0001%-0.0003%, trace element solution 5.0%-6.0%, veratryl alcohol 430 μ L/L-470 μ L/L, tween 80 0.1%-0.13%, surplus is sterilized water, pH=4.9-5.3;
Trace element solution is made up of following raw material:
Calcium chloride 1500mg, ferrous sulfate 150mg, zinc sulfate 100mg, copper sulfate 150mg, manganous sulfate 1000mg, rose vitriol 200mg, potassium aluminium sulfate 20mg, boric acid 10mg, Sodium orthomolybdate 20mg, is settled to 1000mL;
The processing condition of aerobic fermentation are as follows:
Inoculum size is volume ratio=6-8: 100, and temperature is 26 DEG C-28 DEG C, and air flow is 1: 0.7-1.2vvm, and pressure is 0.03Mpa-0.05Mpa, and culture cycle is for being not less than 240 hours.
2. the preparation method of oyster cap fungus fermented liquid according to claim 1, is characterized in that described oyster cap fungus bacterial classification before access is without bacteria fermentation culture medium through enlarged culturing.
3. the preparation method of oyster cap fungus fermented liquid according to claim 2, is characterized in that described enlarged culturing comprises the preparation of slant strains, the preparation of plate bacterial classification, the preparation of first order seed and secondary seed.
4. the preparation method of oyster cap fungus fermented liquid according to claim 3, it is characterized in that the preparation of described slant strains adopts potato culture, the preparation of plate bacterial classification adopts dextrose culture-medium, and temperature is 25 DEG C-27 DEG C, and culture cycle is 4-5 days.
5. the preparation method of oyster cap fungus fermented liquid according to claim 3, is characterized in that in the preparation of described first order seed and the preparation of secondary seed, seed culture medium adopts the raw material of following mass percentage to form:
Murphy juice 18%-21%, glucose 1.8%-2.2%, yeast extract paste 0.25%-0.30%, ammonium sulfate 0.29%-0.33%, potassium primary phosphate 0.25%-0.30%, magnesium sulfate 0.15%-0.18%, vitamins B
10.0001%-0.0003%, trace element solution 3.5%-4.5%, surplus is sterilized water, pH=5.0-5.5;
Trace element solution is made up of the raw material of following quality:
Calcium chloride 1500mg, ferrous sulfate 150mg, zinc sulfate 100mg, copper sulfate 150mg, manganous sulfate 1000mg, rose vitriol 200mg, potassium aluminium sulfate 20mg, boric acid 10mg, Sodium orthomolybdate 20mg, is settled to 1000mL.
6. the preparation method of the oyster cap fungus fermented liquid according to claim 3 or 5, is characterized in that the preparation of described first order seed adopts following processing step:
Select the plate bacterial classification that growth is vigorous, healthy and strong, diameter 7mm-13mm disk is cut at the vigorous position of mycelial growth, being inoculated into by every bottle of 3-5 sheet inoculum size is equipped with in primary-seed medium triangular flask, within shaking culture 85-96 hour under temperature is 26 DEG C of-28 DEG C of conditions, makes first order seed; Loading amount is that the triangular flask of 1000ml loads 160-200ml primary-seed medium, and have a certain amount of mycelium suspended to estimate in primary-seed medium, uniformity sturdy with microscopy mycelial growth cultivates terminal for first order seed.
7. the preparation method of the oyster cap fungus fermented liquid according to claim 3 or 5, is characterized in that the preparation of described secondary seed adopts following processing step:
According to the ratio of volume ratio=9-11: 100 of first order seed and secondary seed medium, be equipped with after first order seed being accessed sterilizing in the fermentor tank of secondary seed medium, loading amount is volume ratio=60-70:100, it is 26 DEG C-28 DEG C in temperature, air flow is 1: 0.5-0.7vvm, pressure is that the aerated culture carrying out being not less than 72 hours under the condition of 0.03Mpa-0.05Mpa makes secondary seed, have a large amount of mycelium suspended to estimate in liquid nutrient medium, uniformity sturdy with microscopy mycelial growth cultivates terminal for secondary seed.
8. the oyster cap fungus fermented liquid prepared of method according to claim 1 is at aflatoxin degradation B
1in application.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510477282.9A CN104988183A (en) | 2015-08-06 | 2015-08-06 | Preparation method of pleurotus ostreatus fermentation broth and application of fermentation broth in degradation of aflatoxin B1 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510477282.9A CN104988183A (en) | 2015-08-06 | 2015-08-06 | Preparation method of pleurotus ostreatus fermentation broth and application of fermentation broth in degradation of aflatoxin B1 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104988183A true CN104988183A (en) | 2015-10-21 |
Family
ID=54300077
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510477282.9A Pending CN104988183A (en) | 2015-08-06 | 2015-08-06 | Preparation method of pleurotus ostreatus fermentation broth and application of fermentation broth in degradation of aflatoxin B1 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104988183A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108094712A (en) * | 2017-12-29 | 2018-06-01 | 辽宁工程技术大学 | Aflatoxin B in a kind of distillers ' grains1Biodegrading process |
| CN108419607A (en) * | 2018-03-07 | 2018-08-21 | 安徽农业大学 | A kind of process parameter optimizing method of oyster cap fungus degradation cotton stalk |
| CN109020637A (en) * | 2018-08-03 | 2018-12-18 | 淅川县物华生物科技有限公司 | A method of organic fertilizer is produced using stalk |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101492705A (en) * | 2009-02-19 | 2009-07-29 | 亳州市亳广中药饮片有限公司 | Method of preparing pleurotu ostreatus polysaccharide |
-
2015
- 2015-08-06 CN CN201510477282.9A patent/CN104988183A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101492705A (en) * | 2009-02-19 | 2009-07-29 | 亳州市亳广中药饮片有限公司 | Method of preparing pleurotu ostreatus polysaccharide |
Non-Patent Citations (4)
| Title |
|---|
| 任少亭等: "糙皮侧耳(Pleurotus ostreatus SR—2)产漆酶条件及漆酶性质的研究", 《微生物学杂志》 * |
| 席丹等: "糙皮侧耳菌漆酶的发酵及其对荨麻的应用", 《纺织高校基础科学学报》 * |
| 王会娟等: "高产漆酶平菇的筛选及其在降解黄曲霉毒素B1中的应用", 《核农学报》 * |
| 黄春燕等: "糙皮侧耳液体菌种培养条件的初步研究", 《食用菌学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108094712A (en) * | 2017-12-29 | 2018-06-01 | 辽宁工程技术大学 | Aflatoxin B in a kind of distillers ' grains1Biodegrading process |
| CN108094712B (en) * | 2017-12-29 | 2021-05-04 | 辽宁工程技术大学 | Degradation method of aflatoxin B1 in distiller's grains |
| CN108419607A (en) * | 2018-03-07 | 2018-08-21 | 安徽农业大学 | A kind of process parameter optimizing method of oyster cap fungus degradation cotton stalk |
| CN109020637A (en) * | 2018-08-03 | 2018-12-18 | 淅川县物华生物科技有限公司 | A method of organic fertilizer is produced using stalk |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US8702833B2 (en) | Health food | |
| CN101360829A (en) | Fermentation and culture method, fermented plant extract, composition containing fermented plant extract, method for producing lipopolysaccharide and lipopolysaccharide | |
| CN107593955A (en) | Ferment edible mushroom tea preparation process and fermentation edible mushroom tea | |
| CN105981581B (en) | A kind of artificial culture method of cicada fungus | |
| CN106889306A (en) | A kind of feed fermentation agent and the application on feed fermentation | |
| CN107868778A (en) | A kind of preparation method of Se-enriched yeast | |
| CN104988183A (en) | Preparation method of pleurotus ostreatus fermentation broth and application of fermentation broth in degradation of aflatoxin B1 | |
| Adejumo et al. | Influence of UV light exposure on mineral composition and biomass production of mycomeat produced from different agricultural substrates | |
| CN1005525B (en) | Fast solid culture method for edible fungus mycelium | |
| CN108175040B (en) | Preparation method of millet oat antrodia camphorata mycoplasm health food | |
| KR101082838B1 (en) | Yeast microoganizm material using confectioneries, breads, noodle by-product | |
| KR20030044791A (en) | A preparation method of feed and feed additive comprising mushroom mycelium | |
| CN107027516A (en) | A kind of selenium-rich Periostracum cicadae, its cultural method and application | |
| CN105725163B (en) | A kind of Lenlinus edodes black garlic sauce | |
| KR100557467B1 (en) | Process for feeding fish and livestock a liquid extract of mushroom mycelial culture and product thereof | |
| CN102008012B (en) | Production method of fungal fermentation feed | |
| CN107916229B (en) | One plant of Inonotus obliquus and its application | |
| Michalak et al. | Functional fermented food and feed from seaweed | |
| CN104830701A (en) | Preparation method of trametes versicolor fermentation liquor and application of trametes versicolor fermentation liquor in degrading aflatoxin B1 | |
| CN102283311B (en) | Technology for producing functional biological feed by utilizing edible mushroom fermented sauce residue | |
| CN110358691B (en) | Beer yeast and application thereof | |
| CN1806656A (en) | Elm tree mushroom feedstuff addictive, its preparation method and application | |
| CN102018094B (en) | Product of microorganism fermented forage | |
| KR100422314B1 (en) | Process for feeding ducks using medicinal mushrooms and product thereof | |
| CN112400760B (en) | Method for relieving transportation stress of freshwater fishes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20151021 |
|
| RJ01 | Rejection of invention patent application after publication |