CN101530234A - Method for preparing natural antioxidative food additive - Google Patents
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- CN101530234A CN101530234A CN200910049503A CN200910049503A CN101530234A CN 101530234 A CN101530234 A CN 101530234A CN 200910049503 A CN200910049503 A CN 200910049503A CN 200910049503 A CN200910049503 A CN 200910049503A CN 101530234 A CN101530234 A CN 101530234A
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Abstract
The invention provides a method for preparing a natural antioxidative food additive. The method is characterized by comprising the following steps that: wine Hanseniaspora uvarum, monascus purpureus, Acetobacter pasteurianus and Lactobacillus bulgaricus are cultured on a respective culture medium by an eggplant shaped flask, and are inoculated into LB culture fluid to be fermented and concentrated to obtain the natural antioxidative food additive. The natural antioxidative food additive does not have organic solvent residual, has complete and stable antioxidation capability.
Description
Technical field
The invention provides a kind of preparation method of natural antioxidative food additive, belong to technical field of food additives.
Background technology
Food contacts with airborne oxygen in storage and transportation, and the oxidation reaction that is taken place can cause going bad of food.The rotten nutrition except reduction food of food, flavor takes place beyond the bad change with look, produces harmful substance, crisis health toward the contact meeting.Therefore in the processed food process, all can in food, add the fresh-keeping and anti-oxidation that antioxidant carries out food.
That but for a long time, food service industry is used always is synthetized oxidation preventive agent such as BHT (dibutyl hydroxy toluene), BHA (tert-butyl group 4-BHA), TBHQ (TBHQ) and PG (gallic acid third lipoprotein) etc.In recent years, health mechanism in various countries' has carried out extensive studies for the safety issue of these artificial synthetized oxidation preventive agents.The World Health Organization (FAO/WHO), European Community's protection of the child tissue (HACSG), Britain biological industry association (BIBRA), Japan, the government of the U.S. and studies show that of tissue; synthetized oxidation preventive agent has many side effects; as liver, spleen, lung adverse effect is arranged all, can bring out malignant tumour etc. human body.So FAO/WHO clearly stipulates the addition of synthetized oxidation preventive agent in the food; FDA (Food and Drug Adminstration) (FDA) suggestion is tried one's best in it is generally acknowledged safe food without BHT; The relevant hygiene department of Japan once made the conclusion that bans use of BHA after deliberation; For TBHQ, though a few countries such as the U.S. have been ratified its use in some grease, European Union, Japan and other countries think that its toxicological test data is not perfect, so approval is not used.In a word, artificial synthetized oxidation preventive agent make consumers when the diet to human toxicity, the ubiquity uneasiness and the feared state of mind.
Therefore having antioxidant from natural food safe, characteristics such as oxidation resistance strong, have no side effect, anti-corrosive fresh-keeping now comes into one's own day by day.The fact proves that also in the edible for a long time food of people, the toxicity of natural composition is well below artificial synthetic antioxidant toxicity, and popular in the world organic food is exactly not have pollutant fully, does not contain any artificial synthetic food.The research of natural is sought from nature in countries in the world in recent years, and exploitation wide spectrum, efficient, safe natural become one of focus in the current food additives research field.But general natural additive for foodstuff is to adopt chemical extraction method to extract its active ingredient from natural materials to form, but extracting used solvent often also is chemical reagent, such as ether, methyl alcohol etc., though through processing such as volatilizations, how many all some residual quantities are level do not wait.
Summary of the invention
The preparation method who the purpose of this invention is to provide the food additives that a kind of organic solvent-free is residual, oxidation resistance is comprehensive and stable.
In order to achieve the above object, technical scheme of the present invention provides a kind of preparation method of natural antioxidative food additive, it is characterized in that, concrete steps are:
The first step: grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus were cultivated 45～50 hours at 30～40 ℃ on culture medium separately with the eggplant bottle, standby 2～6 ℃ of preservations;
Second step: first step gained grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus are inoculated into respectively in the LB nutrient solution, the weight ratio of grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter, lactobacillus bulgaricus and LB nutrient solution is 25～30:5～10:10～15:20～30:100, cultivates 6-10 hour at 30-40 ℃ in shaking bottle;
Second step: with the fermentation tank temperature is the steam sterilizing 25-35min that 120-125 ℃ of pressure is 0.142-0.152Mpa, add the fermentation base-material, with pasteurization 70 ℃～80 ℃ the sterilization 10-20min, described fermentation base-material is made up of following raw material in weight portion: catsup leftover bits and pieces 1-3 part, Rosa roxburghii Normal juice leftover bits and pieces 2-4 part, kelp 0.5-2.0 part, sunflower seed shell 2-5 part, dregs of beans 1-3 part, 35 parts of murphy juices, honey 5-7 part and water 75-90 part, be cooled to 25～40 ℃ after the sterilization, the weight ratio of the second step gained culture by 0.1～1:100 is inoculated in the described fermentation base-material, semi-hermetic fermentation 7-15 days finishes fermentation when the pH=4 of zymotic fluid;
The 4th the step: with zymotic fluid with the centrifugal 3～8min of speed 1500-2500r/min, getting supernatant is that 20-25 ℃, vacuum are that under the condition of 0.001-0.005 MPa filtrate to be concentrated to water content be 20-30wt% in temperature, at 85-95 ℃ of sterilization 20-30min, obtain the water-soluble natural antioxidative food additive; Described water-soluble natural antioxidative food additive is mixed with weight ratio 100:5-10:15-25 with lecithin and soybean lecithin,, obtain fat-soluble natural antioxidative food additive at 55-65 ℃ of emulsification 16-25 hour.
Further, the culture medium of the grape wine spore Hansenula yeast in the described first step is made up of following raw material in weight portion: glucose 30-50 part, yeast extract 4-8 part, peptone 4-8 part, agar 10-25 part and distilled water 500-800 part.
Monascus purpureus culture medium in the described first step is made up of following raw material in weight portion: pol is brewer's wort 80-120 part and agar 1-3 part of 8-20Brix.
Pasteur's acetobacter culture medium in the described first step is made up of following raw material in weight portion: glucose 80-120 part, yeast extract 8-15 part, calcium carbonate 15-30 part, agar 15-30 part and distilled water 700-1200 part.
Lactobacillus bulgaricus culture medium in the described first step is made up of following raw material in weight portion: yeast extract 5-10 part, peptone 5-10 part, glucose 8-15 part, potassium dihydrogen phosphate 0.1-0.4 part, Tomato juice (50)-(150) part, Tween 80 0.3-0.8 part, distilled water 500-1000 part and agar powder 10-30 part.
The LB nutrient solution in described second step is made up of following raw material in weight portion: yeast extract 3-10 part, peptone 5-20 part, sodium chloride 3-10 part, agar 10-30 part and water 800-1200 part.
The present invention has selected for use grape wine that spore Hansenula yeast (Hanseniaspora uvarum is arranged, ACCC20310), and monascus purpureus (Monascus purpureus, ACCC30140), Pasteur's acetobacter (Acetobac terpas tuerianu5, ACCC10112), and lactobacillus bulgaricus (Lactobacillus bulgaricus, ACCC10638), after screening taming the cultivation, worked in coordination with the utilization of composite fermentation technology, multistage good anaerobic fermentation has obtained various biological metabolism active material.The natural antioxidative food additive of gained is a kind of yellowish liquid formulation to brown, tart flavour is arranged, and organic solvent-free is residual, can resist the oxidation of multiple radical ion to food, therefore its oxidation resistance is comprehensive and stable, and TAC reaches 800-1000U/rml.
The specific embodiment
The invention will be further described below in conjunction with embodiment.
Embodiment 1
A kind of preparation method of natural antioxidative food additive, concrete steps are:
The first step: grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus were cultivated 45 hours at 30 ℃ on culture medium separately with the eggplant bottle, standby 2 ℃ of preservations;
Second step: first step gained grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus are inoculated into respectively in the LB nutrient solution, the weight ratio of grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter, lactobacillus bulgaricus and LB nutrient solution is 25:5:10:20:100, cultivates 6 hours at 30 ℃ in shaking bottle;
Second step: with the fermentation tank temperature is the steam sterilizing 25min that 120 ℃ of pressure are 0.142Mpa, add the fermentation base-material, with pasteurization 70 ℃ the sterilization 10min, described fermentation base-material is made up of following raw material in weight portion: 1 part in catsup leftover bits and pieces, 2 parts in Rosa roxburghii Normal juice leftover bits and pieces, 0.5 part of kelp, 2 parts of sunflower seed shells, 1 part of dregs of beans, 3 parts of murphy juices, 75 parts in 5 parts of honey and water, be cooled to 25 ℃ after the sterilization, the weight ratio of the second step gained culture by 0.1:100 is inoculated in the described fermentation base-material, semi-hermetic fermentation 7 days finishes fermentation when the pH=4 of zymotic fluid;
The 4th the step: with zymotic fluid with the centrifugal 3min of speed 1500r/min, getting supernatant is that 20 ℃, vacuum are that under the condition of 0.001 MPa filtrate to be concentrated to water content be 30wt% in temperature, at 85 ℃ of sterilization 20min, obtain the water-soluble natural antioxidative food additive; Described water-soluble natural antioxidative food additive is mixed with weight ratio 100:5:15 with lecithin and soybean lecithin,, obtain fat-soluble natural antioxidative food additive 55 ℃ of emulsifications 16 hours.
The culture medium of the grape wine spore Hansenula yeast in the described first step is made up of following raw material in weight portion: 500 parts of 30 parts of glucose, 4 parts of yeast extracts, 4 parts of peptones, 10 parts in agar and distilled water.
Monascus purpureus culture medium in the described first step is made up of following raw material in weight portion: pol is 1 part in 80 parts of the brewer's worts of 8Brix and an agar.
Pasteur's acetobacter culture medium in the described first step is made up of following raw material in weight portion: 700 parts of 80 parts of glucose, 8 parts of yeast extracts, 15 parts in calcium carbonate, 15 parts in agar and distilled water.
Lactobacillus bulgaricus culture medium in the described first step is made up of following raw material in weight portion: 10 parts of 5 parts of yeast extracts, 5 parts of peptones, 8 parts of glucose, 0.1 part of potassium dihydrogen phosphate, 50 parts of Tomato juice, 0.3 part of Tween 80,500 parts of distilled water and agar powders.
The LB nutrient solution in described second step is made up of following raw material in weight portion: 800 parts in 3 parts of yeast extracts, 5 parts of peptones, 3 parts in sodium chloride, 10 parts in agar and water.
Embodiment 2
A kind of preparation method of natural antioxidative food additive, concrete steps are:
The first step: grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus were cultivated 50 hours at 40 ℃ on culture medium separately with the eggplant bottle, standby 6 ℃ of preservations;
Second step: first step gained grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus are inoculated into respectively in the LB nutrient solution, the weight ratio of grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter, lactobacillus bulgaricus and LB nutrient solution is 30:10:15:30:100, cultivates 10 hours at 40 ℃ in shaking bottle;
The 3rd step: with the fermentation tank temperature is the steam sterilizing 35min that 125 ℃ of pressure are 0.152Mpa, add the fermentation base-material, with pasteurization 80 ℃ the sterilization 20min, described fermentation base-material is made up of following raw material in weight portion: 2 parts in catsup leftover bits and pieces, 3 parts in Rosa roxburghii Normal juice leftover bits and pieces, 1 part of kelp, 4 parts of sunflower seed shells, 2 parts of dregs of beans, 4 parts of murphy juices, 80 parts in 6 parts of honey and water, be cooled to 40 ℃ after the sterilization, the weight ratio of the second step gained culture by 1:100 is inoculated in the described fermentation base-material, semi-hermetic fermentation 15 days finishes fermentation when the pH=4 of zymotic fluid;
The 4th the step: with zymotic fluid with the centrifugal 8min of speed 2500r/min, getting supernatant is that 25 ℃, vacuum are that under the condition of 0.005 MPa filtrate to be concentrated to water content be 25wt% in temperature, at 95 ℃ of sterilization 30min, obtain the water-soluble natural antioxidative food additive; Described water-soluble natural antioxidative food additive is mixed with weight ratio 100:10:25 with lecithin and soybean lecithin,, obtain fat-soluble natural antioxidative food additive 65 ℃ of emulsifications 20 hours.
The culture medium of the grape wine spore Hansenula yeast in the described first step is made up of following raw material in weight portion: 800 parts of 30 parts of glucose, 4 parts of yeast extracts, 4 parts of peptones, 25 parts in agar and distilled water.
Monascus purpureus culture medium in the described first step is made up of following raw material in weight portion: pol is 3 parts in 120 parts of the brewer's worts of 20Brix and an agar.
Pasteur's acetobacter culture medium in the described first step is made up of following raw material in weight portion: 1200 parts of 120 parts of glucose, 15 parts of yeast extracts, 30 parts in calcium carbonate, 30 parts in agar and distilled water.
Lactobacillus bulgaricus culture medium in the described first step is made up of following raw material in weight portion: 30 parts of 10 parts of yeast extracts, 10 parts of peptones, 15 parts of glucose, 0.4 part of potassium dihydrogen phosphate, 150 parts of Tomato juice, 0.8 part of Tween 80,1000 parts of distilled water and agar powders.
The LB nutrient solution in described second step is made up of following raw material in weight portion: 1200 parts in 10 parts of yeast extracts, 20 parts of peptones, 10 parts in sodium chloride, 30 parts in agar and water.
Embodiment 3
A kind of preparation method of natural antioxidative food additive, concrete steps are:
The first step: grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus were cultivated 47 hours at 35 ℃ on culture medium separately with the eggplant bottle, standby 4 ℃ of preservations;
Second step: first step gained grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus are inoculated into respectively in the LB nutrient solution, the weight ratio of grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter, lactobacillus bulgaricus and LB nutrient solution is 27:7:12:25:100, cultivates 8 hours at 35 ℃ in shaking bottle;
The 3rd step: with the fermentation tank temperature is the steam sterilizing 30min that 121 ℃ of pressure are 0.148Mpa, add the fermentation base-material, with pasteurization 75 ℃ the sterilization 15min, described fermentation base-material is made up of following raw material in weight portion: 3 parts in catsup leftover bits and pieces, 4 parts in Rosa roxburghii Normal juice leftover bits and pieces, 2 parts of kelps, 5 parts of sunflower seed shells, 3 parts of dregs of beans, 5 parts of murphy juices, 90 parts in 7 parts of honey and water, be cooled to 35 ℃ after the sterilization, the weight ratio of the second step gained culture by 1:100 is inoculated in the described fermentation base-material, semi-hermetic fermentation 10 days finishes fermentation when the pH=4 of zymotic fluid;
The 4th the step: with zymotic fluid with the centrifugal 5min of speed 2000r/min, getting supernatant is that 22 ℃, vacuum are that under the condition of 0.003 MPa filtrate to be concentrated to water content be 30wt% in temperature, at 90 ℃ of sterilization 25min, obtain the water-soluble natural antioxidative food additive; Described water-soluble natural antioxidative food additive is mixed with weight ratio 100:7:20 with lecithin and soybean lecithin,, obtain fat-soluble natural antioxidative food additive 60 ℃ of emulsifications 25 hours.
The culture medium of the grape wine spore Hansenula yeast in the described first step is made up of following raw material in weight portion: 600 parts of 30 parts of glucose, 4 parts of yeast extracts, 4 parts of peptones, 15 parts in agar and distilled water.
Monascus purpureus culture medium in the described first step is made up of following raw material in weight portion: 90 parts of the brewer's worts of 15Brix, 2 parts in agar.
Pasteur's acetobacter culture medium in the described first step is made up of following raw material in weight portion: 1000 parts of 80 parts of glucose, 10 parts of yeast extracts, 15 parts in calcium carbonate, 20 parts in agar and distilled water.
Lactobacillus bulgaricus culture medium in the described first step is made up of following raw material in weight portion: 20 parts of 5 parts of yeast extracts, 8 parts of peptones, 10 parts of glucose, 0.2 part of potassium dihydrogen phosphate, 50 parts of Tomato juice, 0.3 part of Tween 80,900 parts of distilled water and agar powders.
The LB nutrient solution in described second step is made up of following raw material in weight portion: 800 parts in 5 parts of yeast extracts, 10 parts of peptones, 4 parts in sodium chloride, 10 parts in agar and water.
The using method of gained food additives of the present invention is as follows: directly add in the food, the present invention can not with fall acid additives and use simultaneously; But product normal temperature lucifuge of the present invention is placed and is preserved, and the term of validity is 1 year.
Observe the effect of product of the present invention by following experiment to the food oxydating resistance function:
Getting and newly squeezing rape oil, fresh chicken fat, fresh apple cucumber mud is test meal, and calculating best interpolation concentration according to fat content is 200-1000PPM, directly adds and stirs.After the interpolation behind 80 ℃ of heating 5min, (, carry out quantitative analysis with the assay method of anti peroxidation of lipid ability with optical spectroscopy owing to after lipid peroxidation product MDA (MDA) adds thiobarbituricacid, maximum strong absworption peak is arranged at 530nm.) sample is carried out the detection of peroxidating index, serve as the contrast radix not add this antioxidant, calculate its oxidation resistance.Result such as following table:
Following oxidation resistance detects data and shows that product of the present invention has the ability of comprehensive opposing free radical:
1. remove the mensuration of DPPH free radical ability:
1,1-diphenyl picryl phenylhydrazine (DPPH) is a kind of stable organic free radical, has the last one to absorb at the 517nm place according to DPPH, adopts AAS to carry out quantitative analysis.
The concrete operations step: (1) is mixed 120 μ mmol/L DPPH solution 3.0ml and is shaken up, place 30min in the dark with test liquid 0.1ml; (2) measure the 517nm absorbance; (3) computing formula: clearance rate (%)=[(A
c-(A
1-A
10)/A
c)] * 100%, A in the formula
c: 0.1ml absolute ethyl alcohol and 3.0m] absorbance of DPPH solution mixed liquor; A
1: the absorbance of 0.1ml test liquid and 3.0mlDPPH solution mixed liquor; A
10: the absorbance of 0.1ml absolute ethyl alcohol and 0.1ml test liquid mixed liquor.
Testing result shows the removing DPPH free radical ability 87.35% of weight concentration 10% antioxidative food additive.
2. the mensuration of anti peroxidation of lipid ability:
Because lipid peroxidation product MDA (MDA) behind the interpolation thiobarbituricacid (TBA), has maximum strong absworption peak at 530nm, adopts AAS to carry out quantitative analysis.
The concrete operations step: (1) is that 3% hydrogen peroxide (fresh preparation) 1.0ml, weight concentration are 1.0% TBA solution 5ml and test liquid 0.4ml mixing with oil sample 0.2ml, weight concentration, 37 ℃ of water-baths dissolving 30min, running water cooling; (2) gained solution in (1) is added the distilled water constant volume to 10ml, adding chloroform 1ml, capping plug turned upside down 10 times, 10min leaves standstill clarification (can centrifugal 5min as muddiness); (3) pour gained solution in (2) into cuvette, 530nm (532nm) measures absorbance; (4) calculate A0 in inhibiting rate (%)=[(A0-Ax)/A0)] * 100% formula: the absorbance of contrast solution; Ax: the absorbance of sample solution;
Testing result shows that the anti peroxidation of lipid ability of antioxidative food additive of the present invention reaches 90.20%.
3. remove the mensuration of hydroxy radical (OH) ability:
Hydroxy radical oxidation salicylic acid obtains 2,3 one dihydroxy-benzoic acids, represents what of OH with its light absorption value at the 510nm place.Light absorption value is directly proportional with the amount of OH.Add material in the reaction system and can reduce this light absorption value with removing OH effect.
Concrete operations step: (1) A
0: 9mmol/L salicylic acid (septichen C7H6O3) ethanolic solution 2ml, 9mmol/L FeSO
4Solution 2ml, 8.8mmol/L H
2O
2Solution 2ml and H
2The mixed liquor of O 2ml; A
x: 9mmol/L salicylic acid (septichen C7H6O3) ethanolic solution 2ml, 9mmol/L FeSO
4Solution 2ml, 8.8mmol/L H
2O
2The mixed liquor of solution 2ml and test liquid 2ml; A
X0: 9mmol/L salicylic acid (septichen C7H6O3) ethanolic solution 2ml, test liquid 2ml and H
2The mixed liquor of O 4ml; (2) with three kinds of solution in (1) at 37 ℃ of water-bath 30min; (3) measure the 510nm absorbance of three kinds of solution that (2) are obtained; (4) clearance rate (%)=[(A
0-(A
x-A
X0))/A
0)] * 100%, A in the formula
0: the absorbance of no sample blank contrast liquid; A
x: the absorbance behind the adding sample; A
X0: sample blank contrast liquid absorbance.
Testing result shows that weight concentration of the present invention is that removing hydroxy radical (OH) ability of 20% antioxidative food additive reaches 82.53%.
4. remove ultra-oxygen anion free radical (O
2 -) the mensuration of ability:
Pyrogallol produces O in the rapid autoxidation of alkali condition in the autoxidation process
2 -, O
2 -Quicken pyrogallol autoxidation speed, generate coloured intermediate product simultaneously, the hysteresis that is accumulated in of intermediate product became good linear relationship with the time in the time of 30～45 seconds, generally kept about 4min, slowed down subsequently.Coloured product has strong light absorption at 325nm.
The concrete operations step: (1) with 50mmol/L Tris-HCl (Tri(Hydroxymethyl) Amino Methane Hydrochloride) (pH=8.2) mixture of 4.5ml and 4.2ml distilled water at 25 ℃ of water-baths dissolving 20min; (2) gained solution in (1) is added the 3mmol/L pyrogallol 0.3ml of 25 ℃ of water-bath preheatings; (3) pour gained solution in (2) into cuvette rapidly, every 10s measures absorbance under the 325nm, calculates the increase of per minute absorbance in the range of linearity; (4) 50mmol/L Tris-HC] (pH=8.2) mixed liquor of 4.5ml, distilled water 2.2ml and test liquid 2.0ml, 25 ℃ of water-baths dissolving 20min; (5) gained solution in (4) is added the 3mmol/L pyrogallol solution 0.3ml of 25 ℃ of water-bath preheatings; (6) pour gained solution in (5) into cuvette rapidly, every 10s measures absorbance under the 325nm, calculates the increase of per minute absorbance in the range of linearity; (7) calculate inhibiting rate (%)=[(autoxidation speed A
0Autoxidation speed A behind the-adding sample)/A
0)] * 100%.
Testing result shows that weight concentration of the present invention is the removing ultra-oxygen anion free radical (O of 5% antioxidative food additive
2 -) ability reach 60.01%.
5. the mensuration of TAC:
Adopt the detection method of TAC testing cassete (Nanjing is built up bio-engineering research and produced).
The concrete operations step: placed 10 minutes with the abundant mixing of whirlpool vortex mixer (1), the distilled water zeroing, and the 1cm optical path, the 520nm place surveys respectively manages absorbance; (2) computing formula: extension rate before TAC (U/ milliliter)=(measuring pipe OD value-control tube OD value) ÷ 0.01 ÷ 30 * (reactant liquor cumulative volume ml ÷ sampling amount m1) * sample test.
Testing result shows that the TAC of antioxidative food additive of the present invention is at 921.92-1103.21U/ml.
Claims (6)
1, a kind of preparation method of natural antioxidative food additive, it is characterized in that, concrete steps are: the first step: grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus were cultivated 45～50 hours at 30～40 ℃ on culture medium separately with the eggplant bottle, and standby 2～6 ℃ of preservations;
Second step: first step gained grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter and lactobacillus bulgaricus are inoculated into respectively in the LB nutrient solution, the weight ratio of grape wine spore Hansenula yeast, monascus purpureus, Pasteur's acetobacter, lactobacillus bulgaricus and LB nutrient solution is 25～30:5～10:10～15:20～30:100, cultivates 6-10 hour at 30-40 ℃ in shaking bottle;
The 3rd step: with the fermentation tank temperature is the steam sterilizing 25-35min that 120-125 ℃ of pressure is 0.142-0.152Mpa, add the fermentation base-material, with pasteurization 70 ℃～80 ℃ the sterilization 10-20min, described fermentation base-material is made up of following raw material in weight portion: catsup leftover bits and pieces 1-3 part, Rosa roxburghii Normal juice leftover bits and pieces 2-4 part, kelp 0.5-2.0 part, sunflower seed shell 2-5 part, dregs of beans 1-3 part, murphy juice 3-5 part, honey 5-7 part and water 75-90 part, be cooled to 25～40 ℃ after the sterilization, the weight ratio of the second step gained culture by 0.1～1:100 is inoculated in the described fermentation base-material, semi-hermetic fermentation 7-15 days finishes fermentation when the pH=4 of zymotic fluid;
The 4th the step: with zymotic fluid with the centrifugal 3～8min of speed 1500-2500r/min, getting supernatant is that 20-25 ℃, vacuum are that under the condition of 0.001-0.005 MPa filtrate to be concentrated to water content be 20-30wt% in temperature, at 85-95 ℃ of sterilization 20-30min, obtain the water-soluble natural antioxidative food additive; Described water-soluble natural antioxidative food additive is mixed with weight ratio 100:5-10:15-25 with lecithin and soybean lecithin,, obtain fat-soluble natural antioxidative food additive at 55-65 ℃ of emulsification 16-25 hour.
2, preparation method as claimed in claim 1, it is characterized in that the culture medium of the grape wine spore Hansenula yeast in the described first step is made up of following raw material in weight portion: glucose 30-50 part, yeast extract 4-8 part, peptone 4-8 part, agar 10-25 part and distilled water 500-800 part.
3, preparation method as claimed in claim 1, it is characterized in that the monascus purpureus culture medium in the described first step is made up of following raw material in weight portion: pol is brewer's wort 80-120 part and agar 1-3 part of 8-20Brix.
4, preparation method as claimed in claim 1, it is characterized in that the Pasteur's acetobacter culture medium in the described first step is made up of following raw material in weight portion: glucose 80-120 part, yeast extract 8-15 part, calcium carbonate 15-30 part, agar 15-30 part and distilled water 700-1200 part.
5, preparation method as claimed in claim 1, it is characterized in that the lactobacillus bulgaricus culture medium in the described first step is made up of following raw material in weight portion: yeast extract 5-10 part, peptone 5-10 part, glucose 8-15 part, potassium dihydrogen phosphate 0.1-0.4 part, the 50-150 of Tomato juice part, Tween 80 0.3-0.8 part, distilled water 500-1000 part and agar powder 10-30 part.
6, preparation method as claimed in claim 1, it is characterized in that the LB nutrient solution in described second step is made up of following raw material in weight portion: yeast extract 3-10 part, peptone 5-20 part, sodium chloride 3-10 part, agar 10-30 part and water 800-1200 part.
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| CN103005614A (en) * | 2011-10-18 | 2013-04-03 | 江瀚生物科技（上海）有限公司 | Production method of microorganic antioxidant for foods |
| CN104223318A (en) * | 2014-08-25 | 2014-12-24 | 南通昊友食品添加剂有限公司 | Production method of microorganism food antioxidant |
| CN105875292A (en) * | 2016-04-12 | 2016-08-24 | 江苏大学 | Method for preventing and treating postharvest diseases of citrus reticulata blanco by combining phosphatidylcholine with hanseniaspora uvarum |
| CN108244449A (en) * | 2018-03-13 | 2018-07-06 | 重庆拜月食品科技有限公司 | Anioxidant phytochemicals and its application in food fresh keeping |
| CN108497282A (en) * | 2018-03-13 | 2018-09-07 | 重庆拜月食品科技有限公司 | Triticale steamed bun premixed powder and its manufacture craft |
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Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US5780023A (en) * | 1988-04-04 | 1998-07-14 | The United States Of America As Represented By The Secretary Of Agriculture | Inhibiting plant pathogens with an antagonistic microorganism(s) |
| CN1919216A (en) * | 2005-08-24 | 2007-02-28 | 上海亘卓生物工程有限公司 | Microbiological antioxidant and method for preparing same |
| CN100528326C (en) * | 2007-07-01 | 2009-08-19 | 饶力群 | Method for preparing anti-oxidant bio-microcapsule |
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| CN101971973A (en) * | 2010-10-14 | 2011-02-16 | 上海创博食品技术发展有限公司 | Method for extracting natural food antioxidant from plants |
| CN103005614A (en) * | 2011-10-18 | 2013-04-03 | 江瀚生物科技（上海）有限公司 | Production method of microorganic antioxidant for foods |
| CN104223318A (en) * | 2014-08-25 | 2014-12-24 | 南通昊友食品添加剂有限公司 | Production method of microorganism food antioxidant |
| CN105875292A (en) * | 2016-04-12 | 2016-08-24 | 江苏大学 | Method for preventing and treating postharvest diseases of citrus reticulata blanco by combining phosphatidylcholine with hanseniaspora uvarum |
| CN108244449A (en) * | 2018-03-13 | 2018-07-06 | 重庆拜月食品科技有限公司 | Anioxidant phytochemicals and its application in food fresh keeping |
| CN108497282A (en) * | 2018-03-13 | 2018-09-07 | 重庆拜月食品科技有限公司 | Triticale steamed bun premixed powder and its manufacture craft |
| CN108244449B (en) * | 2018-03-13 | 2021-06-15 | 夏津圣树源农业有限公司 | Plant antioxidant and application thereof in food preservation |
| CN108497282B (en) * | 2018-03-13 | 2021-07-13 | 上海四和食品有限公司 | Premixed black wheat steamed bun flour and preparation process thereof |
| CN115918822A (en) * | 2022-09-23 | 2023-04-07 | 河南省纳普生物技术有限公司 | Preparation method of red pine and antler composite fermentation functional beverage |
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