CN103005382A - Fungus powder for degrading cholesterol, and compound flavoring paste containing fungus powder for degrading cholesterol as well as applications of compound flavoring paste - Google Patents
Fungus powder for degrading cholesterol, and compound flavoring paste containing fungus powder for degrading cholesterol as well as applications of compound flavoring paste Download PDFInfo
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Abstract
The invention discloses fungus powder for degrading cholesterol and compound flavoring paste containing the fungus powder for degrading the cholesterol as well as applications thereof. The fungus powder for degrading the cholesterol comprises a novel culture LZKDZ2011 Torulopsismogii which is preserved in CCTCC (China Center For Type Culture Collection) in July, 30th, 2012, and the preservation Number is CCTCCM2012303. The compound flavoring paste containing the fungus powder for degrading cholesterol comprises the following raw materials by weight parts of: 550-600 parts of wheat germ, 300-350 parts of salt, 5.6-6.6 parts of fungus powder for degrading the cholesterol, 75-120 parts of spice powder and 500-600 parts of water, and the compound flavoring paste containing the fungus powder for degrading the cholesterol can lower the intake of the cholesterol for people, and can protect the health of people. Because the fungus powder for degrading the cholesterol can effectively lower the cholesterol content in meat products, the fungus powder can be applied to the preparation of various compounds for degrading the cholesterol.
Description
Technical field
The present invention relates to field of food, be specifically related to the preparation of norcholesterol bacterium powder and the preparation of application and norcholesterol composite seasoning sauce.
Background technology
Cholesterol is human and animal's necessary nutritional labeling of surviving, and has important physiological function, but also can work the mischief when excessive in vivo.Along with the development of science and technology and the raising of people's living standard, with cardiovascular and cerebrovascular diseases such as the excessive relevant atherosclerotic that causes of cholesterol, coronary heart disease, headstrokes, human health in serious threat now.Investigation shows, cholesterol every increase by one unit (1mmol/L) in the human body of Asia, and angiocardiopathy death risk probability just increases by 35%, and the apoplexy probability relevant with blood vessel also can increase by 25%.Known health knowledge shows: the metabolism of lipid and cholesterol intake in the dietary restriction is necessary.But along with the raising of people's living standard and the increase of diet nutritional, people too much take in higher fatty acid, high cholesterol food often because of unreasonable meals, cause the blood cholesterol levels content overproof.In order to prevent the generation of cardiovascular and cerebrovascular disease, many people dietary restriction of having to, this pursues growth in the living standard with people and contradicts.So, study dietary restriction not, do not disturb the human homergy, do not affect flavour of food products, food and the processing method thereof of economical and practical minimizing cholesterol absorption, become the important development direction of food-processing industry.
Reduce the method for the cholesterol in the food, at present, mainly contain 3 kinds of methods: (1) removes cholesterol in the raw-food material with physical method; (2) with the cholesterol in the chemical method removal raw-food material; (3) with the cholesterol in microorganism or the microbial enzyme conversion food, namely utilize biotransformation method to process food.
Utilize physics, chemical method to process food, the method that reduces cholesterol level in the raw-food material mainly contains at present: b-cyclodextrine absorption inclusion, edible oil and solvent extraction, molecular clock, vacuum distillation, electrodialysis, CO
2Supercritical extract, succinyl oxide or saponin chemical treatment etc.There is respectively following shortcoming in these methods: cyclodextrin is not legal food additives in many countries, must decompose remaining cyclodextrin with enzyme, and this method cost is higher; The organic solvent method solvent for use can remain in the oil; Steaming process is too high to the requirement of equipment; CO
2The investment of supercritical extraction is too large; Molecular clock is difficult to realize industrialization etc.And these methods can affect flavour of food products or nutritional labeling when shifting cholesterol.
It is a kind of both economical and effectively reduce the method for Food Cholesterol relying on microorganism and enzyme thereof or active material to act as main biological method, its efficient is high, cost is lower and better the advantages such as the original nutritional labeling of retaining food product and local flavor just obtaining more and more researchers' favors.At present, utilizing microbial degradation cholesterol field, scientific workers had done a lot of valuable researchs.Can reduce the content of the serum cholesterol of human body such as many food of lactobacillus-fermented that studies show that.Richardson (1977) and Mann studies show that: contain a kind of 3-hydroxyl-3-methyl glucoside acid and orotic acid and can suppress from the factor of acetic acid synthetic cholesterol in yogurt, a large amount of absorption yogurts can reduce the cholesterol in the blood in meals.This perseverance of Chinese scholar Guo health care about lactic acid bacteria in functional dairy product points out, edible sour milk can reduce the function of human serum cholesterol levels.
All contain cholesterol oxidase in many microorganisms, thereby the cholesterol oxidation Decomposition can be made it to remove from food.But, the enzyme activity of the cholesterol oxidase of separating from present microorganism is lower, roughly at 60～80 μ mol/(minL) between, oxidation Decomposition cholesterol indifferent, the biological function of the product courage Gona-4-en-3-one of cholesterol oxidase and the impact of health is still waiting further research simultaneously, the cholesterol that therefore cholesterol oxidase is used for removing food is also well used.
Summary of the invention
Purpose of the present invention just is for the deficiencies in the prior art, and a kind of norcholesterol bacterium powder is provided, this norcholesterol bacterium powder contain parent shrimp (
Torulopsis mogii) novel bacterial LZKDZ 2011, the enzyme activity of the cholesterol oxidase that parent shrimp LZKDZ 2011 separates is high, and it can effectively reduce the cholesterol level in the meat products, thereby reduces the absorption of people's cholesterol, and the protection people's is healthy.Simultaneously the present invention also provides norcholesterol composite seasoning sauce that contains norcholesterol bacterium powder and preparation method thereof, it is take natural materials as raw material, adopt biofermentation technique, be prepared into the norcholesterol composite seasoning sauce, not only can effectively reduce the cholesterol level in the meat products, do not affect flavour of food products but also have, characteristics easy to use, economical and practical.
To achieve these goals, the technical solution used in the present invention is such:
Norcholesterol bacterium powder of the present invention contain parent shrimp (
Torulopsis mogii) novel bacterial LZKDZ 2011, described parent shrimp novel bacterial LZKDZ 2011 is preserved in Wuhan City, Hubei Province Wuhan University Chinese Typical Representative culture collection center (CCTCC) on July 30th, 2012, and preserving number is CCTCC M 2012303.
The preparation method of norcholesterol bacterium powder of the present invention may further comprise the steps:
(1) preparation liquid enriched medium: form according to weight portion: 10 parts of peptones, 3 parts of beef extracts, 5 parts in sodium chloride, 1000 parts of distilled water take by weighing each raw material, the peptone, beef extract and the sodium chloride that take by weighing are dissolved in the distilled water, proofreading and correct pH value is 7 ± 0.2, and at 98KPa, autoclaving 15min obtains the liquid enriched medium under 121 ℃ the condition;
(2) enrichment culture: from the vegetables and fruits of spontaneous fermentation, carry out aseptic sampling, taking by weighing sample 1g adding fills in the conical flask of bead and 100ml sterilized water, concussion 20min, in 10% inoculum concentration access 100ml liquid enriched medium, at 30 ℃, shaking table is cultivated and was obtained enrichment culture liquid in 20 hours under the condition of 200rpm;
(3) dilution enrichment culture liquid: respectively measure enrichment culture liquid 10ml, add respectively and make 4～8 dilution enrichment culture dilutions of difference in 90ml, the liquid enriched medium of 900ml equal-volume by 10 times of amplifications;
(4) preparation solid screening and culturing base is dull and stereotyped: form according to weight portion: 2 parts in cholesterol, 1.6 parts of potassium dihydrogen phosphates, 0.2 part in magnesium sulfate, 0.2 part of manganese sulfate, 0.2 part in ammonium nitrate, 5 parts in sodium chloride, 1 part in surfactant, 20 parts in agar take by weighing each raw material, all are dissolved in the distilled water raw material that takes by weighing except agar, proofreading and correct pH value is 7 ± 0.2, add the agar take by weighing, during the shape that is heated to that agar fully dissolves, liquid is translucent, be sub-packed in the conical flask, at 98KPa, autoclaving 15min obtains solid screening and culturing base flat board under 121 ℃ the condition;
(5) separation and purification single strain: 4～8 dilution enrichment culture dilutions of difference are applied to respectively on each solid screening and culturing base flat board, after being inverted cultivation 18h under 30 ℃ of temperature, the bacterial strain of choosing different colonial morphologies carries out plate streaking to be separated, and obtains the single strain of purifying;
(6) bacterium: the active height of the cholesterol oxidase that the single strain that mensuration obtains from the separation and purification of 4～8 dilution enrichment culture dilutions of difference produces filters out the active high bacterial strain of cholesterol oxidase;
(7) preservation strain: the preparation method according to solid screening and culturing base flat board prepares the solid slant medium in 15 * 150 test tube, and the inoculation that filters out on the solid slant medium, is treated that strain growth to clump count is 10
15～10
16Individual/cm
2, the closed test tube opening is with test tube freezing under 2～8 ℃ temperature conditions;
(8) preparation liquid seeds activation culture liquid: picking one articulating enters in the 100mL liquid enriched medium from the test tube of preservation strain, and at 30 ℃, shaking table is cultivated and obtained bacterial strain clump count 10 in 24 hours under the condition of 200rpm
8～10
9The liquid seeds activation culture liquid of individual/ml;
(9) preparation fermentation medium: form according to weight portion: 300 parts of malt flours, 1000 parts of distilled water take by weighing each raw material, to be heated to 38 ℃ of insulation 2h in the malt flour adding distilled water that take by weighing, be warming up to again 45 ℃ of insulation 30min, bring up to again 50 ℃ of insulation 30min, rise to again 60 ℃, saccharification 1～1.5h obtains saccharified liquid, after saccharification is complete saccharified liquid is heated to 100 ℃ and boils half an hour, the saccharified liquid after then will heating filters and obtains clarified solution, and this clarified solution namely is fermentation medium;
(10) zymotic fluid is cultivated: the inoculum concentration by 5% is with in the liquid seeds activation culture liquid access fermentation medium, and at 30 ℃, shaking table is cultivated 48h and obtained zymotic fluid under the condition of 200rpm;
(11) preparation norcholesterol bacterium powder: zymotic fluid was placed under-18 ± 4 ℃ the temperature freezing 18～24 hours, put into the vacuum drying refrigerator after freezing, under the vacuum state of 1.3～13 handkerchiefs, freeze drying 3～4 hours 90% the abundant drying of moisture to the zymotic fluid obtains the piece that ferments under-25～-40 ℃ temperature conditions, and the fermentation piece is pulverized and namely got norcholesterol bacterium powder.
As preferably, the spontaneous fermentation vegetables and fruits of aseptic sampling are one or more in Fermented Cabbage, acid pepper, apple, pineapple, the orange in preparation method's (2) step enrichment culture of norcholesterol bacterium powder of the present invention.
The inventor by a large amount of experiment find the norcholesterol bacterium powder of the present invention's preparation contain parent shrimp (
Torulopsis mogii) novel bacterial LZKDZ 2011, the enzyme activity of the cholesterol oxidase that parent shrimp LZKDZ 2011 separates is high.
Cholesterol oxidase enzyme activity determination method is as follows:
The enzyme activity definition: at 37 ℃, under the condition of pH7.5, catalysis 1 μ mol cholesterol is oxidized to the needed enzyme of courage Gona-4-en-3-one in the 1min.
Assay method: get the 0.2mL zymotic fluid, the Triton X-100 solution of adding 0.05%, use pH7.5,0.1mol/ the kaliumphosphate buffer of L is measured with ultraviolet specrophotometer at the 241nm place after diluting 10 times, then above-mentioned mixed solution is incubated 3min 37 ℃ times, 1% cholesterol of adding equality of temperature preheating-ethanolic solution 100 L, 37 ℃ of lower vibrations, accurately react 15 min, add immediately 1mol/L NaOH-ethanolic solution and 5mL cyclohexane, vibration 30s, leave standstill extraction 2h, draw the 1mL upper layer of extraction liquid and in the scale test tube, be settled to 5mL with absolute ethyl alcohol, measure with ultraviolet specrophotometer at the 241nm place.
Computing formula is: enzyme activity unit (μ mol/ (minL))=(O.D.E-O.D.O) ÷ 0.03548 * 125 ÷ 384 ÷ 15 * 1000
In the formula: O.D.E---ultraviolet absorption value after the reaction;
O.D.O---ultraviolet absorption value before the reaction;
0.03548---courage Gona-4-en-3-one UV absorption slope of standard curve;
125---extension rate;
384---courage Gona-4-en-3-one relative molecular mass;
15---the reaction time (min);
1000---reaction volume (ml).
After measured, the oxidasic enzyme activity of norcholesterol that parent shrimp novel bacterial LZKDZ 2011 separates in the norcholesterol bacterium powder of the present invention is 90～180 μ mol/(minL).
The norcholesterol composite seasoning sauce that contains norcholesterol bacterium powder of the present invention comprises the raw material that following weight parts forms: wheat malt 550-600 part, salt 300-350 part, norcholesterol bacterium powder 5.6-6.6 part, condiment powder 75-120 part and water 500-600 part.
The preparation method of norcholesterol composite seasoning sauce of the present invention comprises following production stage:
(1) preparation wheat malt:
A. select full wheat, soaked 48～72 hours with clear water, after the abundant imbibition of wheat, taking-up drains away the water;
B. the wheat that drains is placed shallow bottom tray, cover clean gauze, 30 ℃ of warm water of every 8h spray 1 time stir simultaneously slightly;
Day c.2～4 after, observe wheat and grow root and plumule situation, treat that 80～90% Fructus Hordei Germinatus grow, remove overcover, the wheat that grows Fructus Hordei Germinatus namely is wheat malt;
(2) take by weighing each raw material according to the parts by weight of raw materials composition;
The wheat malt that (3) will take by weighing steams 30～40min under 80-100 ℃ temperature, the water of weighing 250-300 part, 2.25 parts salt are poured in the watering can, be sprayed on behind the norcholesterol bacterium powder that adding takes by weighing in the watering can on the wheat malt after steaming, then wheat malt put into incubator and cultivated 24 hours;
(4) water of weighing 250-300 part, 2.25 parts salt join in the wheat malt after the cultivation and mix fragmentation, place the liquid fermentation tank fermentation to obtain fermentate in 24-72 hour;
(5) salt of remainder and condiment powder are joined in the fermentate mix, place the ceramic jar sealing to shine 30-60 processed days;
(6) material that shines in the rear ceramic jar of system obtains the norcholesterol composite seasoning sauce through high-temperature sterilization, can.
As preferably, the condition of culture of described incubator is 37 ± 1 ℃ of temperature, relative humidity 60%-75%; The fermentation condition of described liquid fermentation tank is 37 ± 1 ℃ of temperature.
Because norcholesterol bacterium powder of the present invention can effectively reduce the cholesterol level in the meat products, thereby reduce the absorption of people's cholesterol, therefore bacterium powder of the present invention can be applied to prepare the compound of various norcholesterols, such as tartar sauce, health food etc.
The invention has the advantages that:
(1) norcholesterol bacterium powder of the present invention is the live body solids that extracts the parent shrimp novel bacterial LZKDZ 2011 with cholesterol degradation ability for preparing from the spontaneous fermentation vegetables and fruits, its mechanism of degradation is the fermentation through cholesterol bacterium powder, parent shrimp novel bacterial LZKDZ 2011 produces the norcholesterol oxidizing ferment, the oxidase catalyzed cholesterol degradation reaction of norcholesterol, thereby the cholesterol level in the reduction food.The oxidasic enzyme activity of norcholesterol that parent shrimp novel bacterial LZKDZ 2011 separates in the norcholesterol bacterium powder of the present invention is 90～180 μ mol/(minL), the cholesterol in the meat products of effectively degrading, the cholesterol level of reduction food;
(2) the norcholesterol oxidizing ferment belongs to small protein, and heat resistance is not strong, and 60-80 ℃ gets final product deactivation, and easily by pepsin and trypsin degradation, human body is not produced harm;
(3) norcholesterol oxydasis CHF only produces single product: the courage Gona-4-en-3-one is conducive to food security.There is at present a large amount of courage Gona-4-en-3-ones that studies show that to have the effect of reducing blood lipid, fat-reducing, therefore norcholesterol bacterium powder is applied to prepare the norcholesterol compound, can increase the health care of norcholesterol compound;
(4) contain the preparation method of norcholesterol composite seasoning sauce of norcholesterol bacterium powder simple; easy to use, join in the meat products according to usage ratio and to get final product, it does not only affect the original local flavor of food; but also can reduce cholesterol level in the food, the protection people's is healthy.
The specific embodiment
In order more clearly to understand purpose of the present invention, technical scheme and beneficial effect, below the present invention is described further, but protection scope of the present invention is not limited in following examples.
One, preparation wheat malt:
(1) select full wheat, soaked 48～72 hours with clear water, after the abundant imbibition of wheat, taking-up drains away the water;
(2) wheat that drains is placed shallow bottom tray, cover clean gauze, 30 ℃ of warm water of every 8h spray 1 time stir simultaneously slightly;
After (3) 2～4 days, observe wheat and grow root and plumule situation, treat that 80～90% Fructus Hordei Germinatus grow, remove overcover, the wheat that grows Fructus Hordei Germinatus namely is wheat malt;
Two, preparation norcholesterol bacterium powder:
(1) preparation liquid enriched medium: form according to weight portion: 10 parts of peptones, 3 parts of beef extracts, 5 parts in sodium chloride, 1000 parts of distilled water take by weighing each raw material, the peptone, beef extract and the sodium chloride that take by weighing are dissolved in the distilled water, proofreading and correct pH value is 7 ± 0.2, and at 98KPa, autoclaving 15min obtains the liquid enriched medium under 121 ℃ the condition;
(2) enrichment culture: from the vegetables and fruits of spontaneous fermentation, carry out aseptic sampling, taking by weighing sample 1g adding fills in the conical flask of bead and 100ml sterilized water, concussion 20min, in 10% inoculum concentration access 100ml liquid enriched medium, at 30 ℃, shaking table is cultivated and was obtained enrichment culture liquid in 20 hours under the condition of 200rpm; As preferably, the spontaneous fermentation vegetables and fruits are one or more in Fermented Cabbage, acid pepper, apple, pineapple, the orange.
(3) dilution enrichment culture liquid: respectively measure enrichment culture liquid 10ml, add respectively and make 4～8 dilution enrichment culture dilutions of difference in 90ml, the liquid enriched medium of 900ml equal-volume by 10 times of amplifications;
(4) preparation solid screening and culturing base is dull and stereotyped: form according to weight portion: 2 parts in cholesterol, 1.6 parts of potassium dihydrogen phosphates, 0.2 part in magnesium sulfate, 0.2 part of manganese sulfate, 0.2 part in ammonium nitrate, 5 parts in sodium chloride, 1 part in surfactant, 20 parts in agar take by weighing each raw material, all are dissolved in the distilled water raw material that takes by weighing except agar, proofreading and correct pH value is 7 ± 0.2, add the agar take by weighing, during the shape that is heated to that agar fully dissolves, liquid is translucent, be sub-packed in the conical flask, at 98KPa, autoclaving 15min obtains solid screening and culturing base flat board under 121 ℃ the condition;
(5) separation and purification single strain: 4～8 dilution enrichment culture dilutions of difference are applied to respectively on each solid screening and culturing base flat board, after being inverted cultivation 18h under 30 ℃ of temperature, the bacterial strain of choosing different colonial morphologies carries out plate streaking to be separated, and obtains the single strain of purifying;
(6) bacterium: the active height of the cholesterol oxidase that the single strain that mensuration obtains from the separation and purification of 4～8 dilution enrichment culture dilutions of difference produces filters out the active high bacterial strain of cholesterol oxidase;
(7) preservation strain: the preparation method according to solid screening and culturing base flat board prepares the solid slant medium in 15 * 150 test tube, and the inoculation that filters out on the solid slant medium, is treated that strain growth to clump count is 10
15～10
16Individual/cm
2, the closed test tube opening is with test tube freezing under 2～8 ℃ temperature conditions;
(8) preparation liquid seeds activation culture liquid: picking one articulating enters in the 100mL liquid enriched medium from the test tube of preservation strain, and at 30 ℃, shaking table is cultivated and obtained bacterial strain clump count 10 in 24 hours under the condition of 200rpm
8～10
9The liquid seeds activation culture liquid of individual/ml;
(9) preparation fermentation medium: form according to weight portion: 300 parts of malt flours, 1000 parts of distilled water take by weighing each raw material, to be heated to 38 ℃ of insulation 2h in the malt flour adding distilled water that take by weighing, be warming up to again 45 ℃ of insulation 30min, bring up to again 50 ℃ of insulation 30min, rise to again 60 ℃, saccharification 1～1.5h obtains saccharified liquid, after saccharification is complete saccharified liquid is heated to 100 ℃ and boils half an hour, the saccharified liquid after then will heating filters and obtains clarified solution, and this clarified solution namely is fermentation medium;
(10) zymotic fluid is cultivated: the inoculum concentration by 5% is with in the liquid seeds activation culture liquid access fermentation medium, and at 30 ℃, shaking table is cultivated 48h and obtained zymotic fluid under the condition of 200rpm;
(11) preparation norcholesterol bacterium powder: zymotic fluid was placed under-18 ± 4 ℃ the temperature freezing 18～24 hours, put into the vacuum drying refrigerator after freezing, under the vacuum state of 1.3～13 handkerchiefs, freeze drying 3～4 hours 90% the abundant drying of moisture to the zymotic fluid obtains the piece that ferments under-25～-40 ℃ temperature conditions, and the fermentation piece is pulverized and namely got norcholesterol bacterium powder.
According to cholesterol oxidase enzyme activity determination method, recording the oxidasic enzyme activity of norcholesterol of separating in the norcholesterol bacterium powder is 90～180 μ mol/(minL).
Embodiment one: preparation norcholesterol composite seasoning sauce
Average 15～20 ℃ of outdoor temperature
1, takes by weighing 550 parts of wheat malts, 300 parts of salt, 5.6 parts in norcholesterol bacterium powder, 500 parts in 75 parts of condiment powders and water;
The wheat malt that 2, will take by weighing steams 30min under 80 ℃ temperature, the water that weighing is 250 parts, 2.25 parts salt are poured in the watering can, be sprayed on behind the norcholesterol bacterium powder that adding takes by weighing in the watering can on the wheat malt after steaming, then wheat malt being put into incubator cultivated 24 hours, wherein the condition of culture of incubator is 37 ± 1 ℃ of temperature, relative humidity 60%-75%.；
3, the water of 250 parts of weighings, 2.25 parts salt join in the wheat malt after the cultivation and mix fragmentation, place the liquid fermentation tank fermentation to obtain fermentate in 24 hours, and wherein the fermentation condition of liquid fermentation tank is 37 ± 1 ℃ of temperature.；
4, the salt of remainder and condiment powder are joined in the fermentate mix, place under the ceramic jar sealing sunlight and shone system 45 days;
5, after the solarization system material in the ceramic jar is taken out in the steam-jacked kettle, while stirring heating keeps sterilization in 15 minutes at 90 ℃ ± 5 ℃ constant temperature;
6, pour bottle placer into, at high temperature, the heat seal packing.
Embodiment two: preparation norcholesterol composite seasoning sauce
Average 20～30 ℃ of outdoor temperature
1, takes by weighing 600 parts of wheat malts, 350 parts of salt, 6.6 parts in norcholesterol bacterium powder, 600 parts in 120 parts of condiment powders and water;
The wheat malt that 2, will take by weighing steams 40min under 100 ℃ temperature, the water that weighing is 300 parts, 2.25 parts salt are poured in the watering can, be sprayed on behind the norcholesterol bacterium powder that adding takes by weighing in the watering can on the wheat malt after steaming, then wheat malt being put into incubator cultivated 24 hours, wherein the condition of culture of incubator is 37 ± 1 ℃ of temperature, relative humidity 60%-75%.；
3, the water of 300 parts of weighings, 2.25 parts salt join in the wheat malt after the cultivation and mix fragmentation, place the liquid fermentation tank fermentation to obtain fermentate in 24-72 hour, and wherein the fermentation condition of liquid fermentation tank is 37 ± 1 ℃ of temperature.；
4, the salt of remainder and condiment powder are joined in the fermentate mix, place the ceramic jar sealing to shine system 30 days;
5, after the solarization system material in the ceramic jar is taken out in the steam-jacked kettle, while stirring heating keeps sterilization in 20 minutes at 90 ℃ ± 5 ℃ constant temperature;
6, pour bottle placer into, at high temperature, the heat seal packing.
Embodiment three: preparation norcholesterol composite seasoning sauce
Average 4～15 ℃ of outdoor temperature
1, takes by weighing 575 parts of wheat malts, 325 parts of salt, 6 parts in norcholesterol bacterium powder, 560 parts in 100 parts of condiment powders and water;
The wheat malt that 2, will take by weighing steams 35min under 90 ℃ temperature, the water that weighing is 280 parts, 2.25 parts salt are poured in the watering can, be sprayed on behind the norcholesterol bacterium powder that adding takes by weighing in the watering can on the wheat malt after steaming, then wheat malt being put into incubator cultivated 24 hours, wherein the condition of culture of incubator is 37 ± 1 ℃ of temperature, relative humidity 60%-75%.；
3, the water of 280 parts of weighings, 2.25 parts salt join in the wheat malt after the cultivation and mix fragmentation, place the liquid fermentation tank fermentation to obtain fermentate in 24-72 hour, and wherein the fermentation condition of liquid fermentation tank is 37 ± 1 ℃ of temperature.；
4, the salt of remainder and condiment powder are joined in the fermentate mix, place the ceramic jar sealing to shine system 60 days;
5, after the solarization system material in the ceramic jar is taken out in the steam-jacked kettle, while stirring heating keeps sterilization in 30 minutes at 90 ℃ ± 5 ℃ constant temperature;
6, pour bottle placer into, at high temperature, the heat seal packing.
Embodiment four: contrast experiment's (preparation does not add the composite seasoning sauce of norcholesterol bacterium powder)
Average 20～30 ℃ of outdoor temperature
1, takes by weighing 600 parts of wheat malts, 350 parts of salt, 600 parts in 120 parts of condiment powders and water;
The wheat malt that 2, will take by weighing steams 40min under 100 ℃ temperature, the water that weighing is 300 parts, 2.25 parts salt are poured on the wheat malt that is sprayed in the watering can after steaming, then wheat malt being put into incubator cultivated 24 hours, wherein the condition of culture of incubator is 37 ± 1 ℃ of temperature, relative humidity 60%-75%.；
3, the water of 300 parts of weighings, 2.25 parts salt join in the wheat malt after the cultivation and mix fragmentation, place the liquid fermentation tank fermentation to obtain fermentate in 24-72 hour, and wherein the fermentation condition of liquid fermentation tank is 37 ± 1 ℃ of temperature.；
4, the salt of remainder and condiment powder are joined in the fermentate mix, place the ceramic jar sealing to shine system 30 days;
5, after the solarization system material in the ceramic jar is taken out in the steam-jacked kettle, while stirring heating keeps sterilization in 20 minutes at 90 ℃ ± 5 ℃ constant temperature;
6, pour bottle placer into, at high temperature, the heat seal packing.
Detect according to the performance indications of following index to the tartar sauce of embodiment one to four preparation:
Testing result sees the following form shown in 1:
Take by weighing respectively pig, the beef or mutton of 100g, detect its cholesterol level, then the norcholesterol composite seasoning sauce that embodiment one to three is made, the composite seasoning sauce that embodiment four makes joins respectively in the above-mentioned meat products according to 1: 10 ratio, pickled after stirring 6-8 hour, detect the cholesterol level of meat products, testing result sees Table two.
Can find out from above each table, the property indices that contains the norcholesterol composite seasoning sauce of norcholesterol bacterium powder of the present invention all reaches requirement, and it is easy to use, joins in the meat products according to usage ratio to get final product.It not only can effectively reduce the cholesterol level in the meat products, does not affect flavour of food products but also have, characteristics easy to use, economical and practical.
Norcholesterol bacterium powder of the present invention can effectively reduce the cholesterol level in the meat products simultaneously, thereby reduces the absorption of people's cholesterol, therefore bacterium powder of the present invention can be applied to prepare the compound of various norcholesterols, such as tartar sauce, health food etc.
The preservation explanation
The parent shrimp of mentioning in the specification (
Torulopsis mogii) novel bacterial LZKDZ 2011 is being positioned at Chinese Typical Representative culture collection center (CCTCC) preservation of Wuhan City, Hubei Province Wuhan University on July 30th, 2012, preserving number is CCTCC M 2012303.
Claims (8)
1. norcholesterol bacterium powder is characterized in that: described norcholesterol bacterium powder contain parent shrimp (
Torulopsis mogii) novel bacterial LZKDZ 2011, described parent shrimp novel bacterial LZKDZ 2011 is preserved in Chinese Typical Representative culture collection center (CCTCC) on July 30th, 2012, and preserving number is CCTCC M 2012303.
2. the preparation method of a kind of norcholesterol bacterium powder according to claim 1, it is characterized in that: described preparation method comprises the steps:
(1) preparation liquid enriched medium: form according to weight portion: 10 parts of peptones, 3 parts of beef extracts, 5 parts in sodium chloride, 1000 parts of distilled water take by weighing each raw material, the peptone, beef extract and the sodium chloride that take by weighing are dissolved in the distilled water, proofreading and correct pH value is 7 ± 0.2, and at 98KPa, autoclaving 15min obtains the liquid enriched medium under 121 ℃ the condition;
(2) enrichment culture: from the vegetables and fruits of spontaneous fermentation, carry out aseptic sampling, taking by weighing sample 1g adding fills in the conical flask of bead and 100ml sterilized water, concussion 20min, in 10% inoculum concentration access 100ml liquid enriched medium, at 30 ℃, shaking table is cultivated and was obtained enrichment culture liquid in 20 hours under the condition of 200rpm;
(3) dilution enrichment culture liquid: respectively measure enrichment culture liquid 10ml, add respectively and make 4～8 dilution enrichment culture dilutions of difference in 90ml, the liquid enriched medium of 900ml equal-volume by 10 times of amplifications;
(4) preparation solid screening and culturing base is dull and stereotyped: form according to weight portion: 2 parts in cholesterol, 1.6 parts of potassium dihydrogen phosphates, 0.2 part in magnesium sulfate, 0.2 part of manganese sulfate, 0.2 part in ammonium nitrate, 5 parts in sodium chloride, 1 part in surfactant, 20 parts in agar take by weighing each raw material, all are dissolved in the distilled water raw material that takes by weighing except agar, proofreading and correct pH value is 7 ± 0.2, add the agar take by weighing, during the shape that is heated to that agar fully dissolves, liquid is translucent, be sub-packed in the conical flask, at 98KPa, autoclaving 15min obtains solid screening and culturing base flat board under 121 ℃ the condition;
(5) separation and purification single strain: 4～8 dilution enrichment culture dilutions of difference are applied to respectively on each solid screening and culturing base flat board, after being inverted cultivation 18h under 30 ℃ of temperature, the bacterial strain of choosing different colonial morphologies carries out plate streaking to be separated, and obtains the single strain of purifying;
(6) bacterium: the active height of the cholesterol oxidase that the single strain that mensuration obtains from the separation and purification of 4～8 dilution enrichment culture dilutions of difference produces filters out the active high bacterial strain of cholesterol oxidase;
(7) preservation strain: the preparation method according to solid screening and culturing base flat board prepares the solid slant medium in 15 * 150 test tube, and the inoculation that filters out on the solid slant medium, is treated that strain growth to clump count is 10
15～10
16Individual/cm
2, the closed test tube opening is with test tube freezing under 2～8 ℃ temperature conditions;
(8) preparation liquid seeds activation culture liquid: picking one articulating enters in the 100mL liquid enriched medium from the test tube of preservation strain, and at 30 ℃, shaking table is cultivated and obtained bacterial strain clump count 10 in 24 hours under the condition of 200rpm
8～10
9The liquid seeds activation culture liquid of individual/ml;
(9) preparation fermentation medium: form according to weight portion: 300 parts of malt flours, 1000 parts of distilled water take by weighing each raw material, to be heated to 38 ℃ of insulation 2h in the malt flour adding distilled water that take by weighing, be warming up to again 45 ℃ of insulation 30min, bring up to again 50 ℃ of insulation 30min, rise to again 60 ℃, saccharification 1～1.5h obtains saccharified liquid, after saccharification is complete saccharified liquid is heated to 100 ℃ and boils half an hour, the saccharified liquid after then will heating filters and obtains clarified solution, and this clarified solution namely is fermentation medium;
(10) zymotic fluid is cultivated: the inoculum concentration by 5% is with in the liquid seeds activation culture liquid access fermentation medium, and at 30 ℃, shaking table is cultivated 48h and obtained zymotic fluid under the condition of 200rpm;
(11) preparation norcholesterol bacterium powder: zymotic fluid was placed under-18 ± 4 ℃ the temperature freezing 18～24 hours, put into the vacuum drying refrigerator after freezing, under the vacuum state of 1.3～13 handkerchiefs, freeze drying 3～4 hours 90% the abundant drying of moisture to the zymotic fluid obtains the piece that ferments under-25～-40 ℃ temperature conditions, and the fermentation piece is pulverized and namely got norcholesterol bacterium powder.
3. the preparation method of a kind of norcholesterol bacterium powder according to claim 2, it is characterized in that: the spontaneous fermentation vegetables and fruits of aseptic sampling are one or more in Fermented Cabbage, acid pepper, apple, pineapple, the orange in described (2) the step enrichment culture.
4. norcholesterol composite seasoning sauce that contains norcholesterol bacterium powder claimed in claim 1 is characterized in that: comprise the raw material that following weight parts forms:
Wheat malt 550-600 part, salt 300-350 part, norcholesterol bacterium powder 5.6-6.6 part, condiment powder 75-120 part and water 500-600 part.
5. the preparation method of a kind of norcholesterol composite seasoning sauce according to claim 4 is characterized in that: comprise following production stage:
(1) preparation wheat malt:
A. select full wheat, soaked 48～72 hours with clear water, after the abundant imbibition of wheat, taking-up drains away the water;
B. the wheat that drains is placed shallow bottom tray, cover clean gauze, 30 ℃ of warm water of every 8h spray 1 time stir simultaneously slightly;
Day c.2～4 after, observe wheat and grow root and plumule situation, treat that 80～90% Fructus Hordei Germinatus grow, remove overcover, the wheat that grows Fructus Hordei Germinatus namely is wheat malt;
(2) take by weighing each raw material according to weight portion composition claimed in claim 4;
The wheat malt that (3) will take by weighing steams 30～40min under 80-100 ℃ temperature, the water of weighing 250-300 part, 2.25 parts salt are poured in the watering can, be sprayed on behind the norcholesterol bacterium powder that adding takes by weighing in the watering can on the wheat malt after steaming, then wheat malt put into incubator and cultivated 24 hours;
(4) water of weighing 250-300 part, 2.25 parts salt join in the wheat malt after the cultivation and mix fragmentation, place the liquid fermentation tank fermentation to obtain fermentate in 24-72 hour;
(5) salt of remainder and condiment powder are joined in the fermentate mix, place the ceramic jar sealing to shine 30-60 processed days;
(6) material that shines in the rear ceramic jar of system obtains the norcholesterol composite seasoning sauce through high-temperature sterilization, can.
6. the preparation method of a kind of norcholesterol composite seasoning sauce according to claim 5, it is characterized in that: the condition of culture of described incubator is 37 ± 1 ℃ of temperature, relative humidity 60%-75%.
7. the preparation method of a kind of norcholesterol composite seasoning sauce according to claim 5, it is characterized in that: the fermentation condition of described liquid fermentation tank is 37 ± 1 ℃ of temperature.
8. the application of norcholesterol bacterium powder claimed in claim 1 in preparation norcholesterol compound.
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| CN107581458A (en) * | 2017-08-24 | 2018-01-16 | 宁夏红山河食品股份有限公司 | A kind of special barbecue sauce of Pan-Fried Beef Steak with Black Pepper and preparation method thereof |
| CN107581582A (en) * | 2017-08-24 | 2018-01-16 | 宁夏红山河食品股份有限公司 | A kind of fragrant spicy barbecue sauce and preparation method thereof |
| CN114698826A (en) * | 2022-04-18 | 2022-07-05 | 江南大学 | Low-cholesterol chicken liver sauce and preparation method thereof |
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| CN107495300A (en) * | 2017-08-24 | 2017-12-22 | 宁夏红山河食品股份有限公司 | A kind of spicy barbecue sauce and preparation method thereof |
| CN107495275A (en) * | 2017-08-24 | 2017-12-22 | 宁夏红山河食品股份有限公司 | It is a kind of effectively to prevent chafing dish bottom flavorings of Oxidation of Fat and Oils and preparation method thereof |
| CN107581458A (en) * | 2017-08-24 | 2018-01-16 | 宁夏红山河食品股份有限公司 | A kind of special barbecue sauce of Pan-Fried Beef Steak with Black Pepper and preparation method thereof |
| CN107581582A (en) * | 2017-08-24 | 2018-01-16 | 宁夏红山河食品股份有限公司 | A kind of fragrant spicy barbecue sauce and preparation method thereof |
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| CN107581582B (en) * | 2017-08-24 | 2020-12-04 | 宁夏红山河食品股份有限公司 | Spicy barbecue sauce and preparation method thereof |
| CN114698826A (en) * | 2022-04-18 | 2022-07-05 | 江南大学 | Low-cholesterol chicken liver sauce and preparation method thereof |
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