CN105112329A - Method for preparing special microbial preparations for homemade 'fruit and vegetable enzymes', products of method and application of products - Google Patents
Method for preparing special microbial preparations for homemade 'fruit and vegetable enzymes', products of method and application of products Download PDFInfo
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Abstract
The invention provides a method for preparing special microbial preparations for homemade 'vegetable and fruit enzymes', products manufactured according to the method and application of the products. The microbial preparations include solid microbial preparations and liquid microbial preparations. A process for preparing the solid microbial preparations includes steps of preparing strains; domesticating and cultivating the strains; preparing materials for inoculants; preparing the microbial preparations. A process for preparing the liquid microbial preparations includes steps of preparing strains; domesticating and cultivating the strains; directly manufacturing the liquid microbial preparations. The method, the products and the application have the advantages that the diversified probiotics strains are domesticated and cultivated to obtain the special microbial preparations for the homemade 'fruit and vegetable enzymes', accordingly, the 'fruit and vegetable enzymes' can be conveniently manufactured at home, the manufacturing success rate can be increased, opportunity of breeding of harmful germs can be reduced as compared with natural fermentation, the sugar contents can be reduced, the fermentation time can be greatly shortened, and the special microbial preparations and the products contain large quantities of effective bioactive components.
Description
Technical field
The present invention relates to food processing field, particularly relate to the preparation method of the special microbial preparation of a kind of family manufacture " vegetables and fruits ferment ", and the application of the product obtained according to the method and this product.
Background technology
" ferment " is the address that TaiWan, China Japan waits for Chinese " enzyme "." vegetables and fruits ferment " as the term suggests, it is the general designation of the daily edible natural bioactive enzyme contained by plant-veterinary antibiotics, and in the market as " vegetables and fruits ferment " that heath food is sold, be that the plants such as vegetable and fruit are fermented as starting material, thus obtain a kind of food article of multiple ferment.
Vegetables and fruits ferment rises from Japan and Korea S for nearly 2 years, after importing China into, becomes the numerous common people and pursue one of healthy first-selected whole food, and especially " vegetables and fruits ferment " it is said cosmetic result, especially one of beauty food of pursuing of Junior Ladies.But commercially available " vegetables and fruits ferment " heath food is expensive, so oneself make " vegetables and fruits ferment " one recently, mainly " fruit ferment " agitation sweeps across China various places, the effect of " fruit ferment " and way, has become the hot issue that the various network platform is propagated.The method of current family manufacture " fruit ferment " be the ratio of sugar, fruit and water from 1:3:10 ~ 1:1:0 not etc. mixing put into bottle, allow its spontaneous fermentation form.
The first, there is the problem of individual worthy of vigilance to be that the formation of bacterium colony affects very large by initial condition in spontaneous fermentation process here.At occurring in nature, the microorganism composition of fruit surface is very various.And except the yeast or milk-acid bacteria that need except fermenting, also have other microorganisms comprising pathogenic bacterium waiting for a chance to cause trouble.Once make process in careless slightly, miscellaneous bacteria in fermenting process " is occupied a hill to act as a lord ", " the fruit ferment " produced so like this not only can not play a role in health care, also health is threatened.It is a risk that mould contamination in living contaminants, comprise multiple mould, some moulds of aspergillus all can produce neurotoxin; Owing to adopting spontaneous fermentation, bacterial classification is wayward, and the product of various microorganism also just can not be controlled, and is wherein no lack of and has the toxic material such as nitrite, methyl alcohol to produce.Although the content of these materials is also unlikely to a level just allowing people fall ill (except living contaminants), but drink " fruit ferment " to pursue not reliable " health care " effect, thus increasing out and out health risk, this loses more than gain really.If by control to bacterial classification, these generations to the unhelpful even objectionable impurities of human body just may be reduced.
Second, during popular at present " vegetables and fruits ferment " family manufacture, sugaring reaches 50% degree, after having fermented, " fruit ferment " general sugar degree also reaches 10%, concerning be originally pursue cosmetic purpose drink, the sugar of high density like this, become high-octane food, virtually add the absorption of sugar.If by controlling fermented bacterium, add so many sugar to suppress the flourish of miscellaneous bacteria with regard to unnecessary.
3rd, present family manufacture " vegetables and fruits ferment " needs spontaneous fermentation for up to 1 ~ 3 month not etc., and Production Time is long, and fermentation ends point is wayward, causes fermented quality at every turn different, can not control.If by the control to fermented bacterium, just fermentation time can be shortened, ensure fermented quality.
Therefore, seek safe, effective, the simple fermentation process of a kind of family manufacture " vegetables and fruits ferment ", to solve the food safety potential problem of family manufacture " vegetables and fruits ferment ", what just become that family manufacture " vegetables and fruits ferment " is subject to that everybody extremely payes attention to has technical problem to be solved.
Summary of the invention
In order to above-mentioned problems of the prior art can be overcome, for solving the food safety potential problem of family manufacture " vegetables and fruits ferment ", for obtaining for family manufacture " vegetables and fruits ferment " safer, effective, simple fermentation process, a first aspect of the present invention, provide the preparation method of the special microbial preparation of a kind of family manufacture " vegetables and fruits ferment ", it is characterized in that, comprise the following steps:
(1) bacterial classification preparation
First, bacterial classification plant lactobacillus (Lactobacillusplantarum is bought from Chinese agriculture Microbiological Culture Collection administrative center, ACCC11118), Lactobacterium acidophilum (L.acidophilus, ACCC10637), long bifidus bacillus (Bifidobacteriumlongum, ACCC03961), kluyveromyces (Kluyveromyceslactis, ACCC21191), bacterial classification bifidobacterium adolescentis (Bifidobacteriumadolescentis, CICC6070) is bought from Chinese Research for Industrial Microbial Germ preservation administrative center.
Plant lactobacillus and Lactobacterium acidophilum are transferred in the Erlenmeyer flask of BST liquid nutrient medium, 180-220rpm shaking culture 48-72 hour under 35-39 DEG C of anaerobic state, takes out after substratum muddiness, then puts into 2-4 DEG C of Refrigerator store stand-by;
Long bifidus bacillus and bifidobacterium adolescentis are transferred in the Erlenmeyer flask of BBL liquid nutrient medium, 180-220rpm shaking culture 60-80 hour under 35-40 DEG C of anaerobic state, takes out after substratum muddiness, then puts into 2-4 DEG C of Refrigerator store stand-by;
Kluyveromyces is transferred in the Erlenmeyer flask of potato liquid nutrient medium, at 28-32 DEG C, 180-220rpm shaking culture 48-72 hour, take out after substratum muddiness, then put into 2-4 DEG C of Refrigerator store stand-by;
(2) the tame and docile conjunction of bacterial classification is cultivated
Bacterial classification step (1) cultivated matches well by following weight percent and gets: plant lactobacillus 25-40%, Lactobacterium acidophilum 25-40%, long bifidus bacillus 10-25%, bifidobacterium adolescentis 10-25%, kluyveromyces 5-15%; Then:
The bacterial classification carrying out liquid bacterial agent is tamed and dociled conjunction and is spread cultivation: will join the bacterial classification got in step (2) and to put in 1:80-120 ratio the stoste of following proportioning: romote antiquity brown sugar 2-5%, honey 2-5%, magnesium sulfate 2-10%, sodium-chlor 0.5-5%, Cys 0.05-0.1%, pure water 85-98%, the ammoniacal liquor of 14% is used to regulate pH at about 7.0-7.5, put into fermentor tank, 150-200rpm stirs, culture temperature 24-32 DEG C, semi closed cultivates 0.5-2.0 days, then at 28-35 DEG C, airtight cultivation 2-5 days, pH=3.5-5.5, obtain liquid bacterial agent, directly obtained liquid microbe preparation, it is yellowish to filbert.
Or the bacterial classification carrying out solid fungicide tame and docile conjunction spread cultivation for: the bacterial classification got will be joined in step (2) to put in 1:80-120 ratio the mixed-culture medium of 60-70%BST liquid nutrient medium, 25-40%BBL liquid nutrient medium, 5-10% potato liquid nutrient medium, 150-200rpm stirs, culture temperature 24-32 DEG C, semi closed cultivates 0.5-2.0 days, then at 28-35 DEG C, airtight cultivation 3-5 days; Use the ammoniacal liquor of 14% to regulate pH in cultivation, make pH remain on 6.0-6.5, obtain bacterium liquid, its nutrient solution is milky white to faint yellow.
Then, in order to obtained solid microbe preparation, further comprising the steps of:
1. the material preparation of solid microbial:
By the bacterium liquid of step (2) gained, centrifugal 15-20 minute in rotating speed 8000rpm whizzer, obtains about 20 times of concentrated bacterial culture fluids; Bacterial culture fluid and 20% syrup will be concentrated with the mixing of 30-50% ratio, obtain thalline suspension liquid; Select 30-50% skim-milk, 15-30% W-Gum, 15-30% ground rice as embedding base material;
2. the preparation of solid microbe preparation:
Use fluidized bed granulation seed-coating machine to carry out thalline embedding and drying integrated operation to described thalline suspension liquid, finally obtained target product solid microbe preparation, more obtained described solid microbe preparation aluminium foil bag is distributed into packed bacterium powder, preserve.When after the every bag of 5-10g packing of bacterium powder aluminium foil bag, if need long-term preservation, then need freezing depositing, general family uses normal temperature to deposit also passable, is recommended as stored refrigerated.
Wherein, the thalline survival rate of the microbial preparation of gained can calculate according to the following formula:
Usually, the thalline survival rate of the microbial preparation obtained according to method of the present invention can reach 80-95%.
Preferably, described BST liquid nutrient medium comprises: sugar 5.0%, yeast powder 1.0%, K
2hP0
41.5%, ammonium sulfate 0.5%, 5% tween 80 solution 1.0%, inorganic salt solution 0.1%, its pH is 8.0; Wherein, described inorganic salt solution comprises magnesium sulfate 20.0%, manganous sulfate 1.0%, ferric sulfate 1.0%, sodium-chlor 1.0%.
Preferably, described BBL liquid nutrient medium comprises: peptone 15.0g, yeast powder 2.0g, glucose 20.0g, Zulkovsky starch 0.5g, sodium-chlor 5.0g, Cys 0.5g, tomato leaching powder 5.0g, liver leaching powder 2.0g, tween 80 solution 1.0ml, distilled water 1000.0ml, its pH is 7.0.
Preferably, the specific works condition of described embedding and drying integrated operation is: hot blast temperature 45-60 DEG C, hot blast wind speed is 10-16m/sec, pressurized air pressure of air supply 2.5kg/cm
2, thalline suspension liquid flow 5-20ml/min, described thalline suspension liquid and described embedding base material press 1:10-20 and add.
A second aspect of the present invention, provides the microbial preparation that the obtained family manufacture " vegetables and fruits ferment " of aforesaid method is special.
A third aspect of the present invention, provides the special microbial preparation of above-mentioned family manufacture " vegetables and fruits ferment " and is making the application in " vegetables and fruits ferment ", comprise step:
(1) preparation of vegetable and fruit:
By commercially available vegetables and fruit, clean up, be cut into sheet or bulk;
(2) making of vegetables and fruits ferment:
By step (1) ready vegetables and fruits, load in clean container, sugar or honey is put into by individual need, and pure water, the ratio of liquid microbe preparation described in one bag of described solid microbe preparation or 5-10ml is blent in every 1000g vegetables and fruits and water, evenly add described solid microbe preparation or described liquid microbe preparation, covered container lid, 24-32 DEG C of room temperature fermentation after 5-10 days, liquid in container is poured out, obtains described " vegetables and fruits ferment ", be " vegetables and fruits ferment " beverage, put refrigerator cold-storage to preserve, or directly eat.
The advantage of microbial preparation of the present invention is: the probiotic bacterium bacterial classification that can be used for food adopting multiple state food drug surveilance general bureau to specify, carry out taming and dociling and close family manufacture " vegetables and fruits ferment " the special microorganism preparation that is just trained, extremely facilitate family manufacture " vegetables and fruits ferment ", improve and be made into power, make it the chance of growing compared with spontaneous fermentation minimizing harmful bacteria, reduce sugar degree, fermentation time shortens dramatically.Simultaneously, family manufacture of the present invention " vegetables and fruits ferment " special microorganism microbial inoculum have employed various prebiotic bacterial classification, these probioticss seize growth vigor in " vegetables and fruits ferment " process made, and what " vegetables and fruits ferment " that therefore this microbiobacterial agent makes made compared with spontaneous fermentation contains more effectively bioactive ingredients.
Embodiment
Below in conjunction with embodiment, the present invention is further elaborated, but the present invention is not limited to following embodiment.
The preparation of embodiment 1 solid microbe preparation
1. bacterial classification preparation
By the plant lactobacillus (Lactobacillusplantarum bought from Chinese agriculture Microbiological Culture Collection administrative center, ACCC11118), Lactobacterium acidophilum (L.acidophilus, ACCC10637) strain transfer enters in the Erlenmeyer flask of BST liquid nutrient medium, 200rpm shaking culture 48 hours under 37 DEG C of anaerobic conditions, take out after substratum muddiness, then put into 4 DEG C of Refrigerator stores stand-by;
By the long bifidus bacillus (Bifidobacteriumlongum bought from Chinese agriculture Microbiological Culture Collection administrative center, ACCC03961) bacterial classification and the bifidobacterium adolescentis (Bifidobacteriumadolescentis from Chinese Research for Industrial Microbial Germ preservation administrative center purchase bacterium, CICC6070) strain transfer enters BBL liquid nutrient medium, 200rpm shaking culture 60 hours under 37 DEG C of anaerobic conditions, take out after substratum muddiness, then put into 4 DEG C of Refrigerator stores stand-by;
By the kluyveromyces (Kluyveromyceslactis bought from Chinese agriculture Microbiological Culture Collection administrative center, ACCC21191) strain transfer enters in the Erlenmeyer flask of potato liquid nutrient medium, at 30 DEG C, 200rpm shaking culture 48 hours, take out after substratum muddiness, then put into 4 DEG C of Refrigerator stores stand-by.
2. the tame and docile conjunction of bacterial classification is cultivated:
(1) bacterial classification step 1 cultivated matches well by following weight percent and gets: plant lactobacillus 30%, Lactobacterium acidophilum 30%, long bifidus bacillus 15%, bifidobacterium adolescentis 15%, kluyveromyces 10%.
(2) bacterial classification is tamed and dociled to close and is cultivated: will join the bacterial classification got above and to put in 1:80 ratio the mixed-culture medium of 65%BST liquid nutrient medium, 30%BBL liquid nutrient medium, 5% potato liquid nutrient medium, 200rpm stirs, culture temperature 30 DEG C, semi closed cultivates 0.5 day, then at 32 DEG C, airtight cultivation, uses the ammoniacal liquor of 14% to regulate pH in cultivation, make pH remain on about 6.5, cultivate 3 days; Obtain bacterium liquid, its nutrient solution is light yellow.
3. the material preparation of microbial inoculum:
(1) close tame and docile the bacterium liquid cultivated, in rotating speed 8,000rpm whizzer centrifugal 15 minutes, obtain about 20 times of concentrated bacterial culture fluids.
(2) bacterial culture fluid and the mixing of 20% syrup 40% ratio will be concentrated, obtain thalline suspension liquid.
(3) embed base material and select 50% skim-milk, 30% W-Gum, 20% ground rice.
4. the preparation of microbial preparation:
(1) fluidized bed granulation seed-coating machine is used to carry out thalline embedding and drying integrated operation to thalline suspension liquid, its working conditions: hot blast temperature 50 DEG C, hot blast wind speed: 15m/sec, pressurized air pressure of air supply 2.5kg/cm
2, thalline suspension liquid flow 12ml/min, thalline suspension liquid and embedding base material press 1:10 add, finally obtain solid microbe preparation; The microbial cells survival rate prepared thus is 90%.
The preparation of embodiment 2 liquid microbe preparation
1. bacterial classification preparation: with the step 1 of embodiment 1;
2. the tame and docile conjunction of bacterial classification is cultivated:
(1) bacterial classification step 1 cultivated matches well by following weight percent and gets: plant lactobacillus 35%, Lactobacterium acidophilum 35%, long bifidus bacillus 10%, bifidobacterium adolescentis 10%, kluyveromyces 10%.
(2) bacterial classification is tamed and dociled to close and is cultivated: will join the bacterial classification got above and to put in 1:100 ratio the stoste of following proportioning: romote antiquity brown sugar 5%, honey 5%, magnesium sulfate 5%, sodium-chlor 2%, Cys 0.05%, pure water 83%, the ammoniacal liquor of 14% is used to regulate pH about 7.2, put into fermentor tank, stir with speed 150rpm, culture temperature 27 DEG C, semi closed cultivates 1.0 days, then at 32 DEG C, airtight cultivation 5 days, during pH<5, obtain liquid bacterial agent, directly obtained liquid microbe preparation, it is yellowish to filbert.
The application of embodiment 3 solid microbe preparation
(1) preparation of vegetable and fruit:
By commercially available celery 100g, onion 150g, Radix Dauci Sativae 100g, blueberry 100g, apple 200g, lemon 50g, clean up, be cut into small pieces shape.
(2) preparation of vegetables and fruits ferment:
By ready to (1) vegetables and fruits, load in clean container, bacterium powder of the present invention for 3g (solid microbe preparation) is added 20ml pure water furnishing bacterium liquid and pour container into, then add pure water 280ml, covered container lid, ferments 6 days 28-30 DEG C of room temperature.Fermented liquid in container-" vegetables and fruits ferment " is poured out, puts refrigerator cold-storage and preserve, add a little honey and water every day in right amount and eat.
Use the Application comparison of " the vegetables and fruits ferment " that microbial preparation of the present invention ferments and prepared by spontaneous fermentation
| Working method | Color | Taste | Energy | Be applicable to crowd |
| Microbial preparation ferments | Vandyke brown | Band point tart flavour | Low | Be applicable to any crowd |
| Spontaneous fermentation | Light brown | Very sweet | High | Diabetic subject should not drink |
Table 1
Above specific embodiments of the invention have been described in detail, but it is just as example, and the present invention is not restricted to specific embodiment described above.To those skilled in the art, any equivalent modifications that the present invention is carried out and substituting also all among category of the present invention.Therefore, equalization conversion done without departing from the spirit and scope of the invention and amendment, all should contain within the scope of the invention.
Claims (6)
1. a preparation method for the microbial preparation that family manufacture " vegetables and fruits ferment " is special, is characterized in that, comprise the following steps:
(1) bacterial classification preparation
Plant lactobacillus and Lactobacterium acidophilum are transferred in the Erlenmeyer flask of BST liquid nutrient medium, 180-220rpm shaking culture 48-72 hour under 35-39 DEG C of anaerobic state, takes out after substratum muddiness, then puts into 2-4 DEG C of Refrigerator store stand-by;
Long bifidus bacillus and bifidobacterium adolescentis are transferred in the Erlenmeyer flask of BBL liquid nutrient medium, 180-220rpm shaking culture 60-80 hour under 35-40 DEG C of anaerobic state, takes out after substratum muddiness, then puts into 2-4 DEG C of Refrigerator store stand-by;
Kluyveromyces is transferred in the Erlenmeyer flask of potato liquid nutrient medium, at 28-32 DEG C, 180-220rpm shaking culture 48-72 hour, take out after substratum muddiness, then put into 2-4 DEG C of Refrigerator store stand-by;
(2) the tame and docile conjunction of bacterial classification is cultivated
Bacterial classification step (1) cultivated matches well by following weight percent and gets: plant lactobacillus 25-40%, Lactobacterium acidophilum 25-40%, long bifidus bacillus 10-25%, bifidobacterium adolescentis 10-25%, kluyveromyces 5-15%;
Then:
The bacterial classification carrying out liquid bacterial agent is tamed and dociled conjunction and is spread cultivation: will join the bacterial classification got in step (2) and to put in 1:80-120 ratio the stoste of following proportioning: romote antiquity brown sugar 2-5%, honey 2-5%, magnesium sulfate 2-10%, sodium-chlor 0.5-5%, Cys 0.05-0.1%, pure water 85-98%, the ammoniacal liquor of 14% is used to regulate pH at about 7.0-7.5, put into fermentor tank, 150-200rpm stirs, culture temperature 24-32 DEG C, semi closed cultivates 0.5-2.0 days, then at 28-35 DEG C, airtight cultivation 2-5 days, pH=3.5-5.5, obtain liquid bacterial agent, directly obtained liquid microbe preparation,
Or the bacterial classification carrying out solid fungicide tame and docile conjunction spread cultivation for: the bacterial classification got will be joined in step (2) to put in 1:80-120 ratio the mixed-culture medium of 60-70%BST liquid nutrient medium, 25-40%BBL liquid nutrient medium, 5-10% potato liquid nutrient medium, 150-200rpm stirs, culture temperature 24-32 DEG C, semi closed cultivates 0.5-2.0 days, then at 28-35 DEG C, airtight cultivation 3-5 days; Use the ammoniacal liquor of 14% to regulate pH in cultivation, make pH remain on 6.0-6.5, obtain bacterium liquid;
Then, in order to obtained solid microbe preparation, further comprising the steps of:
1. the material preparation of solid microbial:
By the bacterium liquid of step (2) gained, centrifugal 15-20 minute in rotating speed 8000rpm whizzer, obtains about 20 times of concentrated bacterial culture fluids; Bacterial culture fluid and 20% syrup will be concentrated with the mixing of 30-50% ratio, obtain thalline suspension liquid; Select 30-50% skim-milk, 15-30% W-Gum, 15-30% ground rice as embedding base material;
2. the preparation of solid microbe preparation:
Use fluidized bed granulation seed-coating machine to carry out thalline embedding and drying integrated operation to described thalline suspension liquid, finally obtained target product solid microbe preparation, more obtained described solid microbe preparation aluminium foil bag is distributed into packed bacterium powder, preserve.
2. preparation method according to claim 1, is characterized in that, described BST liquid nutrient medium comprises: sugar 5.0%, yeast powder 1.0%, K
2hP0
41.5%, ammonium sulfate 0.5%, 5% tween 80 solution 1.0%, inorganic salt solution 0.1%, its pH is 8.0; Wherein, described inorganic salt solution comprises magnesium sulfate 20.0%, manganous sulfate 1.0%, ferric sulfate 1.0%, sodium-chlor 1.0%.
3. preparation method according to claim 1, it is characterized in that, described BBL liquid nutrient medium comprises: peptone 15.0g, yeast powder 2.0g, glucose 20.0g, Zulkovsky starch 0.5g, sodium-chlor 5.0g, Cys 0.5g, tomato leaching powder 5.0g, liver leaching powder 2.0g, tween 80 solution 1.0ml, distilled water 1000.0ml, its pH is 7.0.
4. preparation method according to claim 1, is characterized in that, the specific works condition of described embedding and drying integrated operation is: hot blast temperature 45-60 DEG C, hot blast wind speed is 10-16m/sec, pressurized air pressure of air supply 2.5kg/cm
2, thalline suspension liquid flow 5-20ml/min, described thalline suspension liquid and described embedding base material press 1:10-20 and add.
5. the microbial preparation that the family manufacture " vegetables and fruits ferment " that the method according to any one of the claims is obtained is special.
6. microbial preparation according to claim 5 is making the application in " vegetables and fruits ferment ", comprises step:
(1) preparation of vegetable and fruit:
By commercially available vegetables and fruit, clean up, be cut into sheet or bulk;
(2) making of vegetables and fruits ferment:
By step (1) ready vegetables and fruits, load in clean container, sugar or honey is put into by individual need, and pure water, the ratio of liquid microbe preparation described in one bag of described solid microbe preparation or 5-10ml is blent in every 1000g vegetables and fruits and water, evenly add described solid microbe preparation or described liquid microbe preparation, covered container lid, 24-32 DEG C of room temperature fermentation after 5-10 days, liquid in container is poured out, obtain described " vegetables and fruits ferment ", put refrigerator cold-storage and preserve, or directly eat.
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