CN104719695B - A kind of acetic acid type sauerkraut composite ferment and the preparation method and application thereof - Google Patents
A kind of acetic acid type sauerkraut composite ferment and the preparation method and application thereof Download PDFInfo
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Abstract
A kind of acetic acid type sauerkraut composite ferment disclosed by the invention and the preparation method and application thereof, belongs to fruits and vegetables technical field.Sauerkraut type composite ferment provided by the present invention is mainly made of bacterium powder and calcium chloride.Wherein bacterium powder is made of Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus.Leavening provided by the present invention has the characteristics that vigor height, raciness, easy to use, of fine qualities.Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and the viable count of lactobacillus acidophilus reach 10 in leavening provided by the present invention10cfu/g.Using, not only containing the soft lactic acid of mouthfeel, the flavor substances such as acetic acid also felt well containing mouthfeel acid are consistent with the sauerkraut taste of traditional natural fermentation after acetic acid type composite ferment Fermented Cabbage provided by the present invention.
Description
Technical field
The present invention relates to a kind of acetic acid type sauerkraut composite ferments and the preparation method and application thereof, belong to fruits and vegetables technology
Field.
Background technology
Sauerkraut is the salt by adding low concentration into Chinese cabbage, and undergoing microbial fermentation forms and flavor product is with breast
A kind of fermented vegetable product based on acid.Due to that can greatly prolong the holding time after Chinese cabbage harvests, and flavor also obtains
Very big improvement, therefore sauerkraut is liked by northeast, North China people.Unique flavor, nutrient health, mouthfeel acid is refreshing, quality
Tender and crisp, with rich flavor, with fragrance striking the nose, bright etc. are the most apparent and most popular features of sauerkraut.With people's life water
Flat raising, and to the understanding of healthy diet, using lactic acid bacteria as dominant microflora, fermented supplemented by saccharomycete, acetic acid bacteria etc.,
And containing there are many sauerkraut of the prebiotic factor, is gradually liked and eaten by more and more people.
The marinated of sauerkraut originates from traditional salting pickled cabbages at home, which belongs to the spontaneous fermentation of Chinese cabbage.East
The sauerkraut in backlands area refers in the fall after Chinese cabbage harvest, using Chinese cabbage as raw material, by adding salt, in winter spontaneous fermentation 1-
Fermented vegetables products made of 2 months or so.In traditional Chinese cabbage curing process, since the microorganism of a large amount of distincts is attached
It in Chinese cabbage surface, wherein there are a variety of harmful microorganisms, such as mould, saccharomycete and Escherichia coli, it can be common with lactic acid bacteria
Growth and presence, when cause a hidden trouble to people's safe diet, second is that limitation lactic acid bacteria Rapid Fermentation, third, to the normal of sauerkraut
Quality holding damages.Therefore, the spontaneous fermentation of traditional-family's formula is with the production cycle is long, miscellaneous bacteria is more, product quality is not easy
Control, the drawbacks such as product quality is poor, it is impossible to be used in standardization and industrialized production.
Lactic acid bacteria and other strain fermentation Chinese cabbages are put into the form of leavening, are sauerkraut production industrial expansion trend.
Research at present about sauerkraut leavening is although more, but really can apply to the seldom of actual production.Sauerkraut hair therein
Ferment agent is mostly composite ferment, and species and other compositions are had nothing in common with each other, and such as patent CN 101961048A, is disclosed by pair
The leavening of 4 kinds of freeze-drying bacterium powder compositions such as Lactobacillus casei, lactobacillus buchneri, lactobacillus plantarum and bacillus coagulans, simultaneously
Also contain the auxiliary elements such as natamycin and calcium chloride；Leavening disclosed in patent CN 101317646 is lactobacillus plantarum, breast
The freeze-drying bacterium powder of 5 plants of bacterium such as sour piece coccus, Leuconostoc mesenteroides, Lactococcus lactis, Lactobacillus brevis；Patent CN 102212477A
Disclosed leavening is lactobacillus plantarum, lactobacillus acidophilus, leuconostoc pseudomesenteroides, Lactococcus lactis breast subspecies and rhamnose breast
The freeze-drying bacterium powder of 5 plants of bacterium such as bacillus, the leavening can at 0-25 DEG C by fermentation time reduction be 15-20 days.
Existing sauerkraut leavening can greatly shorten fermentation period, and containing the specific lactic acid bacteria or probiotics of inoculation, but
All it is substantially lactic acid in its tunning, and traditional sauerkraut contains the flavor substances such as the acetic acid generated in fermentation process, therefore it is existing
There is the refreshing flavor of the acid of lactobacillus inoculum sauerkraut and traditional sauerkraut inconsistent.Meanwhile existing lactic acid bacteria fermenting agent is inoculated into Chinese cabbage
Later, it is difficult to which the growth for effectively inhibiting saccharomycete in aerobic environment in the early stage causes yeast in Chinese cabbage fermentation process largely to be deposited
And continue to ferment, or even sauerkraut is being caused to rise packet and the problems such as rot after processing and packaging.
Invention content
To solve the above problems, the present invention provides a kind of sauerkraut composite ferment, the technical solution taken is as follows：
It is an object of the present invention to provide a kind of acetic acid type sauerkraut composite ferments, and the composite ferment is mainly by bacterium
Powder and calcium chloride composition；Wherein bacterium powder is by Pasteur's acetobacter Acetobacter pasteurianus, Leuconostoc mesenteroides
Leuconostoc mesenteroides, lactobacillus plantarum Lactobacillus plantarum, Lactobacillus brevis
Lactobacillus brevis, lactobacillus buchneri Lactobacillus buchneri and lactobacillus acidophilus
Lactobacillus acidophilus compositions.
Preferably, Pasteur's acetobacter is Pasteur's acetobacter CICC 23561；The Leuconostoc mesenteroides is goldbeater's skin
Leukonid CICC 22246；The lactobacillus plantarum is lactobacillus plantarum CICC 23138；The Lactobacillus brevis is short breast
Bacillus CICC 6239；The lactobacillus buchneri is CICC 20015；The lactobacillus acidophilus is lactobacillus acidophilus CICC
6082。
Preferably, the bacterium powder, viable count ratio are Pasteur's acetobacter：Leuconostoc mesenteroides：Lactobacillus plantarum：Short breast bar
Bacterium：Lactobacillus buchneri：Lactobacillus acidophilus=1：15-25：25-35：5-15：5-15：5-15；The calcium chloride and bacterium powder, chlorination
Calcium：The mass ratio of bacterium powder is 1~8：1.
It is highly preferred that the bacterium powder, viable count ratio is Pasteur's acetobacter：Leuconostoc mesenteroides：Lactobacillus plantarum：Short breast bar
Bacterium：Lactobacillus buchneri：Lactobacillus acidophilus=1：20：30：10：10：10, calcium chloride：The mass ratio of bacterium powder is 4：1.
Any composite ferment is applied to fermenting and producing sauerkraut.
The step of application, is as follows：
1) Chinese cabbage is removed into root, extracts the stale dish leaf in surface, is cleaned with water purification and drain Chinese cabbage surface moisture；
2) Chinese cabbage is placed in pickling dehydration 10-15h in 5-6% saline solutions；
3) Chinese cabbage being pickled is taken out, injects the saline solution of 1.5-3% thereto, and contain 0.1g/kg in saline solution
The uniformly mixed composite ferment of Chinese cabbage, Chinese cabbage is compacted, and Chinese cabbage is submerged in water；
4) it ferments 3-30 days in 0-40 DEG C, and can be taken off processing or eat.
Another object of the present invention is to provide a kind of preparation methods of the composite ferment, and this method is picking bar
The bacterium colony of family name's acetobacter activation culture in acetic acid bacteria fluid nutrient medium, then respectively picking Leuconostoc mesenteroides, lactobacillus plantarum,
Lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus bacterium colony carry out activation culture in MRS fluid nutrient mediums, recycle acetic acid bacteria
Fluid nutrient medium is to Pasteur's acetobacter, using MRS fluid nutrient mediums to Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, cloth
Family name's lactobacillus and lactobacillus acidophilus carry out 3 high density amplification cultures respectively, then freeze-drying is added after thalline were collected by centrifugation respectively and protects
Agent freeze-drying is protected, bacterium powder is obtained after 6 kinds of freeze-drying bacterium powders are mixed in proportion, is answered after finally being compounded bacterium powder and calcium chloride
Combined bacteria agent.
Preferably, Pasteur's acetobacter described in the method, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, Bu Shi
The bacterium colony units mesh ratio of lactobacillus and lactobacillus acidophilus, picking is 1-5：15-25：25-35：5-15：5-15：5-15；Institute
State by 6 kinds freeze-drying bacterium powders mix in proportion, be according to viable count ratio be Pasteur's acetobacter：Leuconostoc mesenteroides：Lactobacillus plantarum：
Lactobacillus brevis：Lactobacillus buchneri：Lactobacillus acidophilus=1：20：30：10：10：10 ratio mixing.
Preferably, the step of the method is as follows：
1) in picking Pasteur acetobacter colony inoculation to 5mL acetic acid bacteria fluid nutrient mediums, 24- is cultivated at 25-37 DEG C
48h, then picking Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus single bacterium colony respectively,
It is seeded to respectively in the small test tube for filling 5mL MRS fluid nutrient mediums, cultivates 24-48h in 30-40 DEG C, obtain activated strains；
The composition of the acetic acid bacteria fluid nutrient medium is glucose 100.0g, yeast extract 10.0g, CaCO320.0g,
Agar 15.0g, distilled water 1.0L, pH 6.8；
Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and acidophilus breast bar
The bacterium colony units mesh ratio of bacterium, picking is 1-5：15-25：25-35：5-15：5-15：5-15；
2) utilize acetic acid bacteria fluid nutrient medium to Pasteur's acetobacter obtained by step 1), using MRS fluid nutrient mediums to step
It is rapid 1) obtained by Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus carry out respectively 3 times
High density amplification culture, condition of culture are：Pasteur acetobacter inoculum concentration 1-3%, cultivation temperature：25-37 DEG C, incubation time：24-
48h；Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and the cultivation temperature of lactobacillus acidophilus：30-40
DEG C, incubation time：24-48h；In the amplification culture of 3 high density the volume of acetic acid bacteria fluid nutrient medium and MRS fluid nutrient mediums according to
Secondary is 100mL, 2000mL and 40L；6 kinds of bacterium solutions are obtained after High Density Cultivation；
3) bacterium solution obtained by step 2) is centrifuged into 5-15min, centrifuging temperature 4- respectively under 5000-10000r/min
10 DEG C, retain precipitation；
4) freeze drying protectant is added into precipitation obtained by step 3) by the 50%-100% of the bacterium solution volume obtained by step 2)
It after mixing, in -20 DEG C to -80 DEG C after pre-freeze 10-15h, is lyophilized using freeze drier, obtains freeze-drying bacterium powder；
5) it is Pasteur's acetobacter according to viable count ratio by 6 kinds of freeze-drying bacterium powders obtained by step 4)：Leuconostoc mesenteroides：Plant
Lactobacillus：Lactobacillus brevis：Lactobacillus buchneri：Lactobacillus acidophilus=1：20：30：10：10：Bacterium powder is obtained after 10 ratio mixing,
Again by calcium chloride and bacterium powder according to calcium chloride：The mass ratio of bacterium powder is 1~8：1 ratio compounding obtains composite bacteria agent.
Preferably, the step 4) freeze drying protectant includes the skimmed milk of weight percent 10%, 3% in the method
Trehalose and 1.5% sodium glutamate.
The present invention achieves following advantageous effect：
1. leavening provided by the present invention has the characteristics that vigor height, raciness, easy to use, of fine qualities.This hair
Pasteur's acetobacter in bright provided leavening, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and thermophilic
The viable count of Lactobacillus lactis reaches 1010cfu/g.Leavening is packed in the form of packed, can be used after dismantling.
2. leavening provided by the present invention is acetic acid type sauerkraut composite ferment, wherein containing acetic acid bacteria (Pasteur's vinegar bar
Bacterium) and lactic acid bacteria (Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus), Fermented Cabbage
Afterwards not only containing the soft lactic acid of mouthfeel, also contain the flavor substances such as the acetic acid for keeping sauerkraut mouthfeel acid refreshing, ferments with traditional natural
Sauerkraut taste it is consistent.
3. leavening provided by the present invention can effectively inhibit the breeding of saccharomycete, prevent saccharomycete in fermentation process from continuing
It is putrid and deteriorated caused by breeding, to extend the storage phase of sauerkraut.
4. leavening provided by the present invention in addition to containing there are many strain, also contains calcium chloride, can be quick after input sauerkraut
Dissolving, and can effectively keep the vivid color and luster of Chinese cabbage and the tender and crisp feature of quality.
5. leavening bacterium powder provided by the present invention is made of one plant of acetic acid bacteria and five strains of lactic acid bacteria, since sauerkraut is sent out naturally
During ferment, starting stage growth is the saccharomycete of aerobic acetic acid bacteria and amphimicrobian, and acetic acid bacteria consumes oxygen and carbohydrate
Generate acetic acid and CO2, and saccharomycete then generates CO2And ethyl alcohol, generate CO by consuming oxygen2Nothing is created for the growth of lactic acid bacteria
Oxygen condition, and acetic acid and ethyl alcohol are the important flavor substances in sauerkraut, and wherein acetic acid can assign sauerkraut salubrious tart flavour, and ethyl alcohol
It can be generated with lactic acid and have dulcet ester, made the flavor that sauerkraut is exclusive, since acetic acid bacteria produces acetic acid, can effectively inhibit yeast
Bacterium in the early stage after survival and growth, can prevent black yeast it is microbial discoloration, produce phenomena such as smelly and aerogenesis, be conducive to
The quality for controlling sauerkraut is kept；In five kinds of lactic acid bacterias, Leuconostoc mesenteroides, Lactobacillus brevis and lactobacillus buchneri are that special-shaped lactic acid is sent out
Yeast-like fungi can also generate ethyl alcohol and acetic acid in addition to generating lactic acid, enhance the refreshing flavor of acid and aromatic character of sauerkraut, and plant is newborn
Bacillus and lactobacillus acidophilus are homofermentative lactic bacterium, and acetic acid bacteria, heterolactic fermentation bacterium and homofermentative lactic bacterium can be quick
Fermentation and acid, and form flavor specific to sauerkraut.
Description of the drawings
Fig. 1 is embodiment 1 and lactic acid and acetic acid content in the sauerkraut of the composite ferment fermentation made using embodiment 4-6
Comparison.
Fig. 2 is embodiment 1 and produces saccharomycete and acetic acid bacteria variation in sauerkraut using composite ferment prepared by embodiment 6
Trend.
Specific implementation mode
With reference to specific embodiment, the present invention will be further described, but the present invention should not be limited by the examples.
Material therefor of the present invention, reagent, instrument and method, without specified otherwise, be this field conventional material, reagent,
Instrument and method can be obtained by commercial channel.
Embodiment 1
A kind of production method using the ferment-fermented sauerkraut of commercially available lactic acid type is present embodiments provided, specific steps are such as
Under：
(1) Chinese cabbage is removed into partial moisture except root and except after rotten leaf, being placed under sunlight and drying one to three days.
(2) it is cleaned using clear water.
(3) clean Chinese cabbage is filled into cylinder, often puts one layer of Chinese cabbage and spread one layer of salt, total salt consumption is about 10-15g/kg white
Dish.
(4) Chinese cabbage is pushed down using stone or weight, commercially available lactic acid type, which is added, according to the ratio of 0.1g/Kg Chinese cabbages ferments
Clear water is added in agent into Chinese cabbage again, and the water surface did not had Chinese cabbage.
(5) terminate after fermenting 3 days.
Embodiment 2
A kind of fermentation culture method of required strain is present embodiments provided, is as follows：
(1) in picking Pasteur acetobacter colony inoculation to 5mL acetic acid bacteria fluid nutrient mediums, shake culture is for 24 hours at 30 DEG C.
Wherein, the composition of acetic acid bacteria fluid nutrient medium is glucose 100.0g, yeast extract 10.0g, CaCO320.0g agar
15.0g, distilled water 1.0L, pH 6.8.
(2) picking Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus are single respectively
Bacterium colony is seeded in the small test tube for filling 5mLMRS fluid nutrient mediums, for 24 hours in 40 DEG C of cultures, obtains activated strains.
Wherein, step 1) and Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, Bu Shi in step 2)
The bacterium colony units mesh ratio of lactobacillus and lactobacillus acidophilus is 1:25:35:15:15:15
(3) activated Pasteur's acetobacter in small test tube is seeded to 100mL acetic acid bacteria fluid nutrient mediums with 2% ratio
In, for 24 hours in 30 DEG C of shake cultures, by activated Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, Bu Shi in small test tube
Lactobacillus and lactobacillus acidophilus are seeded to 2% ratio in 100mLMRS fluid nutrient mediums, for 24 hours in 40 DEG C of cultures.
(4) the activated Pasteur's acetobacters of 100mL are seeded to 2% ratio in 2000mL acetic acid bacteria fluid nutrient mediums,
For 24 hours in 30 DEG C of shake cultures, by activated Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, Bu Shi breast bars in small test tube
Bacterium and lactobacillus acidophilus are seeded to 2% ratio in 2000mLMRS fluid nutrient mediums, for 24 hours in 40 DEG C of cultures.
Embodiment 3
A kind of fermentation culture method of required strain is present embodiments provided, is as follows：
1) in picking Pasteur acetobacter colony inoculation to 5mL acetic acid bacteria fluid nutrient mediums, the shake culture 48h at 25 DEG C.
Wherein, the composition of acetic acid bacteria fluid nutrient medium is glucose 100.0g, yeast extract 10.0g, CaCO320.0g agar
15.0g, distilled water 1.0L, pH 6.8.
2) difference picking Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus single bacterium
It falls, is seeded in the small test tube for filling 5mLMRS fluid nutrient mediums, cultivate 48h in 35 DEG C, obtain activated strains.
Wherein, step 1) and Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, Bu Shi in step 2)
The bacterium colony units mesh ratio of lactobacillus and lactobacillus acidophilus is 1:15:25:5:5:5.
3) activated Pasteur's acetobacter in small test tube is seeded to 3% ratio in 100mL acetic acid bacteria fluid nutrient mediums,
In 25 DEG C of shake culture 48h, by activated Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, Bu Shi breast bars in small test tube
Bacterium and lactobacillus acidophilus are seeded to 3% ratio in 100mLMRS fluid nutrient mediums, and 48h is cultivated in 35 DEG C.
4) the activated Pasteur's acetobacters of 100mL are seeded to 3% ratio in 2000mL acetic acid bacteria fluid nutrient mediums, in
25 DEG C of culture 48h, by activated Leuconostoc mesenteroides in small test tube, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and thermophilic
Lactobacillus lactis is seeded to 3% ratio in 2000mLMRS fluid nutrient mediums, and 48h is cultivated in 35 DEG C.
Embodiment 4
A kind of preparation method of the compound sauerkraut leavening of acetic acid type is present embodiments provided, is as follows：
(1) strain is cultivated according to the method for embodiment 3 (except picking ratio of bacterium colony unit), acquisition is amplified to 2000mL
Bacterium solution, then the activated Pasteur's acetobacters of 2000mL are seeded to 1% ratio in 40L acetic acid bacteria fluid nutrient mediums, in 37
DEG C culture 10h, by the activated Leuconostoc mesenteroides of 2000mL, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and acidophilus breast
Bacillus is seeded to 1% ratio in 40L acetic acid bacteria fluid nutrient mediums, and 10h is cultivated in 40 DEG C.
Wherein, Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, the Bu Shi breast bars of picking before activating
The bacterium colony units mesh ratio of bacterium and lactobacillus acidophilus is 1:15:25:5:5:5.
(2) six kinds of bacterium solutions are taken to be centrifuged respectively, rotating speed 5000r/min, time 15min, temperature are controlled in 4-10
DEG C, supernatant is abandoned, thalline is collected, the freeze drying protectant of original bacteria liquid 50%-100% volumes is added to thalline, and mixing, be placed in-
Pre-freeze 15h in 20 DEG C to -80 DEG C, reuses freeze drier and is lyophilized, obtain bacterium powder.Wherein, freeze drying protectant contains 10%
(W/W) sodium glutamate of the trehalose and 1.5% (W/W) of skimmed milk, 3% (W/W).
(3) it is Pasteur's acetobacter according to viable count ratio by 6 kinds of freeze-drying bacterium powders obtained by step 2)：Leuconostoc mesenteroides：It plants
Object lactobacillus：Lactobacillus brevis：Lactobacillus buchneri：Lactobacillus acidophilus=1：20：30：10：10：Bacterium is obtained after 10 ratio mixing
Powder, then by calcium chloride and the mass ratio of bacterium powder 1:1 is compounded, and acetic acid type sauerkraut composite ferment is obtained.
Embodiment 5
A kind of preparation method of the compound sauerkraut leavening of acetic acid type is present embodiments provided, is as follows：
1) strain is cultivated according to the method for embodiment 3 (except picking ratio of bacterium colony unit), acquisition is amplified to 2000mL
Bacterium solution, then the activated Pasteur's acetobacters of 2000mL are seeded to 2% ratio in 40L acetic acid bacteria fluid nutrient mediums, in 30
DEG C culture 11h, by the activated Leuconostoc mesenteroides of 2000mL, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and acidophilus breast
Bacillus is seeded to 2% ratio in 40L acetic acid bacteria fluid nutrient mediums, and 11h is cultivated in 35 DEG C.
Wherein, Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, the Bu Shi breast bars of picking before activating
The bacterium colony units mesh ratio of bacterium and lactobacillus acidophilus is 1:25:35:15:15:15.
2) six kinds of bacterium solutions are taken to be centrifuged respectively, rotating speed 8000r/min, time 10min, temperature are controlled in 4-10
DEG C, supernatant is abandoned, thalline is collected, the freeze drying protectant of original bacteria liquid 50%-100% volumes is added to thalline, and mixing, be placed in-
Pre-freeze 12h in 20 DEG C to -80 DEG C, reuses freeze drier and is lyophilized, obtain bacterium powder.Wherein, freeze drying protectant contains 10%
(W/W) sodium glutamate of the trehalose and 1.5% (W/W) of skimmed milk, 3% (W/W).
3) it is Pasteur's acetobacter according to viable count ratio by 6 kinds of freeze-drying bacterium powders obtained by step 2)：Leuconostoc mesenteroides：Plant
Lactobacillus：Lactobacillus brevis：Lactobacillus buchneri：Lactobacillus acidophilus=1：20：30：10：10：Bacterium powder is obtained after 10 ratio mixing,
Calcium chloride and the mass ratio of bacterium powder 8 again:1 is compounded, and acetic acid type sauerkraut composite ferment is obtained.
Embodiment 6
A kind of preparation method of the compound sauerkraut leavening of acetic acid type is present embodiments provided, is as follows：
1) strain is cultivated according to the method for embodiment 3 (except picking ratio of bacterium colony unit), acquisition is amplified to 2000mL
Bacterium solution, then the activated Pasteur's acetobacters of 2000mL are seeded to 3% ratio in 40L acetic acid bacteria fluid nutrient mediums, in 25
DEG C culture 10h, by the activated Leuconostoc mesenteroides of 2000mL, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and acidophilus breast
Bacillus is seeded to 3% ratio in 40L acetic acid bacteria fluid nutrient mediums, and 10h is cultivated in 30 DEG C.
Wherein, Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, the Bu Shi breast bars of picking before activating
The bacterium colony units mesh ratio of bacterium and lactobacillus acidophilus is 1:20:30:10:10:10.
2) six kinds of bacterium solutions are taken to be centrifuged respectively, rotating speed 10000r/min, time 5min, temperature are controlled in 4-10
DEG C, supernatant is abandoned, thalline is collected, the freeze drying protectant of original bacteria liquid 50%-100% volumes is added to thalline, and mixing, be placed in-
Pre-freeze 10h in 20 DEG C to -80 DEG C, reuses freeze drier and is lyophilized, obtain bacterium powder.Wherein, freeze drying protectant contains 10%
(W/W) sodium glutamate of the trehalose and 1.5% (W/W) of skimmed milk, 3% (W/W).
3) it is Pasteur's acetobacter according to viable count ratio by 6 kinds of freeze-drying bacterium powders obtained by step 2)：Leuconostoc mesenteroides：Plant
Lactobacillus：Lactobacillus brevis：Lactobacillus buchneri：Lactobacillus acidophilus=1：20：30：10：10：Bacterium powder is obtained after 10 ratio mixing,
The ratio of calcium chloride and bacterium powder is 4 again:1 is compounded, and acetic acid type sauerkraut composite ferment is obtained.
Embodiment 7
A kind of method that the composite ferment using prepared by embodiment 4,5 and 6 prepares sauerkraut is present embodiments provided, is had
Steps are as follows for body：
1) Chinese cabbage is removed into root, extracts the stale dish leaf in surface, is cleaned with water purification and drain Chinese cabbage surface moisture；
2) Chinese cabbage is placed in pickling dehydration 12h in 5% saline solution；
3) a certain amount of saline solution is prepared according to Chinese cabbage weight, a concentration of 2% (W/W) dissolves mixing.
4) it according to Chinese cabbage weight, puts into the composite ferment prepared by embodiment 4,5 and 6 respectively into saline solution, puts into
Amount is 0.1g/kg Chinese cabbages, mixing.
5) Chinese cabbage is compacted, the saline solution containing leavening is injected into Chinese cabbage, it is more than Chinese cabbage to make the water surface；
6) it ferments 3 days in 20 DEG C, and can be taken off processing or eat.
Embodiment 8
The present embodiment is carried out to the flavor of the sauerkraut prepared by embodiment 1,4,5 and 6, to inhibiting effect of yeast and other effects
It measures.Method and result are as follows：
(1) flavor
The pH that sauerkraut is produced to embodiment 1,4,5 and 6 is measured, and wherein the marinated acid of embodiment 1, pH value 3.08 are real
It applies the production of example 4,5 and 6 sauerkraut pH value and respectively reaches 3.01,3.02 and 2.93, pass through newborn in high effective liquid chromatography for measuring sauerkraut
Acid and acetic acid content, as shown in Figure 1, it is respectively 19.12,18.66 and that embodiment 4,5 and 6, which produces lactic acid content in sauerkraut,
20.35g/kg Chinese cabbages, and in 1 pickling sauerkraut of embodiment it is 18.73g/kg Chinese cabbages, embodiment 4,5 and 6 produces acetic acid in sauerkraut and contains
Amount is respectively 3.04,2.94 and 3.07g/kg Chinese cabbages, and in 1 pickling sauerkraut of embodiment is only 1.42g/kg Chinese cabbages, it follows that
The lactic acid content that the produced sauerkraut in embodiment 4,5 and 6 makes sauerkraut with embodiment 1 is almost the same, and acetic acid content is to be respectively
214.1%, 207.0% and the 216.2% of embodiment 1, product special flavour is obviously improved, and is more nearly the marinated sauerkraut of the daily life of a family.
(2) inhibition of the acetic acid bacteria to yeast
It completes to carry out saccharomycete and acetic acid bacteria survey to the production sauerkraut of embodiment 7 in 3 days with fermentation within initial 0 day in fermentation respectively
It is fixed, respectively in initial 0 day of fermentation and after fermentation to embodiment 1 and embodiment 7 in utilize the preparation of 6 composite ferment of embodiment
Sauerkraut carry out saccharomycete and acetic acid bacteria and measure, as shown in Fig. 2, during embodiment 7 produces sauerkraut, the initial bacterium number of saccharomycete is
102Cfu/mL, due to being inhibited saccharomycete to be down to 10cfu/mL by acetic acid bacteria hereinafter, and 1 acetic acid bacteria of embodiment when fermentation is completed
10 when having initial4Cfu/mL, by the oxygen consumption of fermentation process, viable count is also down to 10cfu/mL or less therewith；Comparatively,
During embodiment 1 makes sauerkraut, although since the consumption of oxygen forms anaerobic environment, acetic acid bacteria viable count is down to 10cfu/mL
Hereinafter, but saccharomycete viable count be almost absent variation, thus sauerkraut is after being processed into product, still has and continues fermentation gas and become
The possibility of matter.
(3) sauerkraut storage period
The sauerkraut for utilizing 6 composite ferment of embodiment to prepare in embodiment 7 and embodiment 1 are made into sauerkraut, do not added
It is vacuum-packed, is positioned under room temperature (22 DEG C) environment in the case of preservative, the results showed that, the sauerkraut that embodiment 7 produces can be store
It deposits two months, and the making of embodiment 1 sauerkraut period of storage is unstable, until 15 days start rise bag or discoloration.
(4) Pasteur's acetobacter adds control experiment
To evaluate the effect of Pasteur's acetobacter in leavening, respectively using the leavening for being added to Pasteur's acetobacter, and not
The leavening of Pasteur's acetobacter is added, sauerkraut is carried out and produces contrast experiment, fermentation takes out test sample when reaching 3.0 to pH value, such as table 1
Shown, being added to for Pasteur's acetobacter impacts the maturation time of sauerkraut, and the fermentation deadline is all 3 days；But it is not added with
Saccharomycete viable count is far above the sauerkraut for adding Pasteur's acetobacter in the sauerkraut of Pasteur's acetobacter, it follows that Pasteur's acetobacter
The growth and survival of saccharomycete in sauerkraut can effectively be inhibited.
Table 1 adds and is not added with the fermentation index of Pasteur's acetobacter
Although the present invention is disclosed as above with preferred embodiment, it is not limited to the present invention, any to be familiar with this
The people of technology can do various changes and modification, therefore the protection of the present invention without departing from the spirit and scope of the present invention
Range should be subject to what claims were defined.
Claims (10)
1. a kind of acetic acid type sauerkraut composite ferment, which is characterized in that be mainly made of bacterium powder and calcium chloride；Wherein bacterium powder by bar
Family name's acetobacter Acetobacter pasteurianus, Leuconostoc mesenteroides Leuconostoc mesenteroides, plant breast
Bacillus Lactobacillus plantarum, Lactobacillus brevis Lactobacillus brevis, lactobacillus buchneri
Lactobacillus buchneri and lactobacillus acidophilus Lactobacillus acidophilus compositions
Wherein, the bacterium powder, viable count ratio are Pasteur's acetobacter：Leuconostoc mesenteroides：Lactobacillus plantarum：Lactobacillus brevis：Bu Shi
Lactobacillus：Lactobacillus acidophilus=1：15-25：25-35：5-15：5-15：5-15.
2. composite ferment described in claim 1, which is characterized in that Pasteur's acetobacter is Pasteur's acetobacter CICC
23561；The Leuconostoc mesenteroides is Leuconostoc mesenteroides CICC 22246；The lactobacillus plantarum is lactobacillus plantarum
CICC 23138；The Lactobacillus brevis is Lactobacillus brevis CICC 6239；The lactobacillus buchneri is CICC 20015；It is described
Lactobacillus acidophilus is lactobacillus acidophilus CICC 6082.
3. composite ferment described in claim 1, which is characterized in that the calcium chloride and bacterium powder, calcium chloride：The mass ratio of bacterium powder
It is 1~8：1.
4. composite ferment described in claim 3, which is characterized in that the bacterium powder, viable count ratio are Pasteur's acetobacter：Goldbeater's skin is bright
Beading bacterium：Lactobacillus plantarum：Lactobacillus brevis：Lactobacillus buchneri：Lactobacillus acidophilus=1：20：30：10：10：10, calcium chloride：Bacterium
The mass ratio of powder is 4：1.
5. the application of any composite ferments of claim 1-4, which is characterized in that be applied to fermenting and producing sauerkraut.
6. the application of composite ferment described in claim 5, which is characterized in that steps are as follows：
1) Chinese cabbage is removed into root, extracts the stale dish leaf in surface, is cleaned with water purification and drain Chinese cabbage surface moisture；
2) Chinese cabbage is placed in pickling dehydration 10-15h in 5-6% saline solutions；
3) Chinese cabbage being pickled is taken out, injects the saline solution of 1.5-3% thereto, and contain 0.1g/kg Chinese cabbages in saline solution
Uniformly mixed composite ferment, Chinese cabbage is compacted, Chinese cabbage is submerged in water；
4) it ferments 3-30 days in 0-40 DEG C, you can take out processing or edible.
7. the preparation method of composite ferment described in a kind of claim 1, which is characterized in that the bacterium colony of picking Pasteur's acetobacter exists
Activation culture in acetic acid bacteria fluid nutrient medium, then picking Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, Bu Shi breasts respectively
Bacillus and lactobacillus acidophilus bacterium colony carry out activation culture in MRS fluid nutrient mediums, recycle acetic acid bacteria fluid nutrient medium to bar
Family name's acetobacter, using MRS fluid nutrient mediums to Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and acidophilus
Lactobacillus carries out 3 high density amplification cultures respectively, then freeze drying protectant freeze-drying is added after thalline were collected by centrifugation respectively, by 6 kinds
Freeze-drying bacterium powder obtains bacterium powder after mixing in proportion, and composite bacteria agent is obtained after finally being compounded bacterium powder and calcium chloride.
8. claim 7 the method, which is characterized in that Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, short
The bacterium colony units mesh ratio of lactobacillus, lactobacillus buchneri and lactobacillus acidophilus, picking is 1-5：15-25：25-35：5-15：5-
15：5-15；It is described by 6 kinds freeze-drying bacterium powders mix in proportion, be according to viable count ratio be Pasteur's acetobacter：Leuconostoc mesenteroides：
Lactobacillus plantarum：Lactobacillus brevis：Lactobacillus buchneri：Lactobacillus acidophilus=1：20：30：10：10：10 ratio mixing.
9. claim 7 the method, which is characterized in that steps are as follows：
1) in picking Pasteur acetobacter colony inoculation to 5mL acetic acid bacteria fluid nutrient mediums, 24-48h is cultivated at 25-37 DEG C, then
Picking Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus single bacterium colony respectively, connect respectively
In kind to the small test tube for filling 5mLMRS fluid nutrient mediums, 24-48h is cultivated in 30-40 DEG C, obtains activated strains；
The composition of the acetic acid bacteria fluid nutrient medium is glucose 100.0g, yeast extract 10.0g, CaCO320.0g agar
15.0g distilled water 1.0L, pH 6.8；
Pasteur's acetobacter, Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus, choose
The bacterium colony units mesh ratio taken is 1-5：15-25：25-35：5-15：5-15：5-15；
2) utilize acetic acid bacteria fluid nutrient medium to Pasteur's acetobacter obtained by step 1), using MRS fluid nutrient mediums to step 1)
Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and the lactobacillus acidophilus of gained carry out respectively 3 times it is highly dense
Degree amplification culture, condition of culture are：Pasteur acetobacter inoculum concentration 1-3%, cultivation temperature：25-37 DEG C, incubation time：24-48h；
Leuconostoc mesenteroides, lactobacillus plantarum, Lactobacillus brevis, lactobacillus buchneri and the cultivation temperature of lactobacillus acidophilus：30-40 DEG C, training
Support the time：24-48h；The volume of acetic acid bacteria fluid nutrient medium and MRS fluid nutrient mediums is followed successively by 3 high density amplification cultures
100mL, 2000mL and 40L；6 kinds of bacterium solutions are obtained after High Density Cultivation；
3) it is 4-10 DEG C respectively under 5000-10000r/min, centrifuging 5-15min, centrifuging temperature by the bacterium solution obtained by step 2),
Retain precipitation；
4) freeze drying protectant mixing is added into precipitation obtained by step 3) by the 50%-100% for pressing the bacterium solution volume obtained by step 2)
Afterwards, it is lyophilized using freeze drier after pre-freeze 10-15h in -20 DEG C to -80 DEG C, obtains freeze-drying bacterium powder；
5) it is Pasteur's acetobacter according to viable count ratio by 6 kinds of freeze-drying bacterium powders obtained by step 4)：Leuconostoc mesenteroides：Plant breast bar
Bacterium：Lactobacillus brevis：Lactobacillus buchneri：Lactobacillus acidophilus=1：20：30：10：10：Bacterium powder is obtained after 10 ratio mixing, then will
Calcium chloride and bacterium powder are according to calcium chloride：The mass ratio of bacterium powder is 1~8：1 ratio compounding obtains composite bacteria agent.
10. claim 9 the method, which is characterized in that the step 4) freeze drying protectant includes weight percent 10%
Skimmed milk, 3% trehalose and 1.5% sodium glutamate.
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| CN105394669A (en) * | 2016-01-05 | 2016-03-16 | 四川东坡中国泡菜产业技术研究院 | Manufacturing method of novel semisolid quick-fermentation pickle |
| CN106942623A (en) * | 2017-04-12 | 2017-07-14 | 成都大学 | A kind of salted vegetables type composite fermentation local flavor meat products and its processing method |
| CN107183732B (en) * | 2017-06-13 | 2020-09-04 | 贵州大学 | Method for improving comprehensive quality of roxburgh rose pomace through microbial fermentation |
| CN107518378A (en) * | 2017-10-23 | 2017-12-29 | 江苏棉海园林工程有限公司 | A kind of sauerkraut leavening |
| CN108618034B (en) * | 2018-05-14 | 2021-06-18 | 湖北工业大学 | Preparation method of rapid compound fermented cowpea |
| CN109393440B (en) * | 2018-11-06 | 2022-04-15 | 吉林大学 | Preparation method of direct vat set composite lactobacillus starter for pickling pickled Chinese cabbages |
| CN110583871A (en) * | 2019-09-30 | 2019-12-20 | 中国农业科学院兰州畜牧与兽药研究所 | Biological fermentation product of traditional Chinese medicine residue of Maiwei rehmannia pills and preparation method and application thereof |
| CN111329019B (en) * | 2020-04-07 | 2023-03-21 | 佳木斯大学 | Preparation method of honey-flavored instant pickled Chinese cabbage |
| CN113149776A (en) * | 2021-03-31 | 2021-07-23 | 海南自贸区沃佳实业有限公司 | Selenium-rich organic fertilizer and preparation method thereof |
| CN114304564A (en) * | 2021-12-30 | 2022-04-12 | 哈尔滨果垒生物科技有限公司 | Microbial fermentation composition and application thereof |
| CN114668125B (en) * | 2022-03-30 | 2023-09-12 | 广西来宾恒荣农业科技有限公司 | Composite lactic acid bacteria for fermenting pickled Chinese cabbage and preparation method and application thereof |
| CN115109716B (en) * | 2022-05-20 | 2023-11-10 | 华东师范大学 | Pickle starter and preparation method and application thereof |
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