CN104232506A - Lactic acid bacteria complex inoculant for Chinese sauerkraut fermentation - Google Patents
Lactic acid bacteria complex inoculant for Chinese sauerkraut fermentation Download PDFInfo
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- CN104232506A CN104232506A CN201410341787.8A CN201410341787A CN104232506A CN 104232506 A CN104232506 A CN 104232506A CN 201410341787 A CN201410341787 A CN 201410341787A CN 104232506 A CN104232506 A CN 104232506A
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Abstract
The invention discloses a lactic acid bacteria complex inoculant for Chinese sauerkraut fermentation and relates to a lactic acid bacteria complex inoculant. The lactic acid bacteria complex inoculant is mainly used for solving the problems that the existing fermentation process cannot achieve short fermentation time, good flavor and high quality at the same time. Active ingredients of the lactic acid bacteria complex inoculant comprise Lactobacillus sunkii, Lactobacillus curvatus, Lactobacillus parabuchneri and Lactobacillus oligofermentans. The inoculant has high synergistic effect and forms a stable ecological system in an autologous manner, so that not only can vegetables quickly enter a fermentation state, but also flavor substances such as lactic acid, acetic acid and alcohol can be produced by quick breeding in a fermentation system, and thus, the flavor of the artificially fermented vegetables is close to that of naturally fermented vegetables, the operation is simple and easy, and the cost is low. And moreover, the lactic acid bacteria complex inoculant can be used for fermenting vegetables such as Chinese cabbages, cabbages, radishes and cucumbers.
Description
Technical field
The present invention relates to a kind of lactic acid bacteria composite fungicide.
Background technology
Traditional zymotic vegetables are one that in China's fruits and vegetables product, output is maximum, utilize the microorganisms such as naturally occurring milk-acid bacteria to carry out ferment making and form.Before about 3000, just there has been the manufacture craft of fermented vegetables in China, as Pickles, Sichuan Style, hot pickled mustard tube, Yangzhou pickles etc.Along with the development of modern food industry and human consumer improving constantly food safety and health requirements; in the urgent need to producing the fermented vegetables of less salt, low nitrite content; realize the stdn of process for processing, mass-producing, artificial fermentation vegetables can meet these requirements of people completely.
The fermented vegetables processing of the developed countries such as the current U.S., Japan, Korea S adopts artificial fermentation more; and accomplish scale production; and the most product of China or spontaneous fermentation; because in fermenting process, micro-flora is complicated and changeable; course of fermentation is caused to be difficult to control; quality product fluctuation is indefinite, the large-scale development of serious restriction vegetable fermentation industry.
Northeast sauerkraut is traditional zymotic vegetable based food, take Chinese cabbage as dominant fermentation raw material, the salty acid of its taste, and mouthfeel is tender and crisp, and appetizing is refreshed oneself, and sobers up greasy, not only can improve a poor appetite, help digest, and can also promote that human body is to the absorption of ferro element.In the past, on China northeast and North China and other places, because winter time is long, fresh vegetable should not be preserved, and in order to be saved for a long time by Chinese cabbage, people have invented the method for pickling sauerkraut.Along with the raising of sauerkraut popularity and the quickening of modern life rhythm; increasing to its demand; specification of quality is also more and more higher, and the method for traditional natural fermentation can not meet people's needs, and the continuous expansion of demand accelerates the mass-producing of sauerkraut production.
But up to now, the processing enterprise of whole northeast sauerkraut still more adopts traditional fermentation vat to carry out spontaneous fermentation, there is the problems such as fermentation period is long, product quality is poor, quality is unstable.Manual fermentation technique has the advantages that fermentation time is short, little by seasonal effect, quality is easy to control, but the sauerkraut not as good as spontaneous fermentation on its local flavor is natural, mellow.Therefore, adopt the mode of human controllable to ferment, make its local flavor progressively access expansion fermentation be the targets that northeast sauerkraut large-scale production needs constantly to make great efforts.
Summary of the invention
The present invention will solve existing zymotechnique cannot take into account the good problem of the short and local flavor of fermentation time, quality, provides a kind of lactic acid bacteria composite fungicide for sauerkraut fermentation.
The activeconstituents that the present invention is used for the lactic acid bacteria composite fungicide of sauerkraut fermentation comprises Lactobacillus sunkii, Lactobacillus curvatus, Lactobacillus parabuchneri and Lactobacillus oligofermentans.
In described lactic acid bacteria composite fungicide, the colony forming unit number of Lactobacillus sunkii, Lactobacillus curvatus, Lactobacillusparabnchneri and Lactobacillus oligofermentans is than being (1 ~ 3): (4 ~ 6): (4 ~ 6): (1 ~ 3).
Described Lactobacillus sunkii specifically can be Lactobacillus sunkii DSM19904; Described Lactobacillus curvatus specifically can be Lactobacillus curvatus DSM20019; Described Lactobacillus parabuchneri specifically can be Lactobacillus parabuchneri DSM20173; Described Lactobacillus oligofermentans specifically can be Lactobacillus oligofermentans DSM15707.
In described lactic acid bacteria composite fungicide, Lactobacillus sunkii, Lactobacillus curvatus, Lactobacillus parabuchneri, Lactobacillus oligofermentans can independently pack, and also can mix.
Lactic acid bacteria composite fungicide of the present invention can be liquid preparation, comprises activeconstituents, glycerine and EDTA, and the proportioning of described activeconstituents, glycerine and EDTA is (1 ~ 1.2) × 10
11cFU: (1 ~ 5) g glycerine: 1gEDTA.
Lactic acid bacteria composite fungicide of the present invention can be solid preparation; the production of microbial inoculum adopts lyophilization; centrifugal by cultured bacterium liquid; the centrifugal 10min of rotating speed 5800g; abandon supernatant, added by lyophilized vaccine, additional proportion is 5: 1 by bacterium liquid and protectant volume ratio; in-20 DEG C of freezing 10h, afterwards pressure be 3 ~ 5Pa, temperature be-50 DEG C of conditions under lyophilize 24h obtain the bacterium powder of freeze-drying.
Microbial inoculum of the present invention can Fermented Cabbage be sauerkraut.
Microbial inoculum of the present invention is preferential but be not limited only to Fermented Cabbage, and its fermentation object also comprises the vegetables such as Caulis et Folium Brassicae capitatae, radish, cucumber.Described microbial inoculum has the synergy of height, the ecosystem that autologous formation is stable, vegetables not only can be made to enter fermentation state rapidly, and can in fermentation system the flavour substances such as Fast-propagation lactic acid producing, acetic acid, ethanol, make artificial fermentation vegetables access expansion fermented flavour, and operate simple and easy, with low cost.
Embodiment
Technical solution of the present invention is not limited to following cited embodiment, also comprises the arbitrary combination between each embodiment.
Embodiment one: the activeconstituents that present embodiment is used for the lactic acid bacteria composite fungicide of sauerkraut fermentation comprises Lactobacillus sunkii, Lactobacillus curvatus, Lactobacillus parabuchneri and Lactobacillus oligofermentans.
Embodiment two: present embodiment and embodiment one unlike: in described lactic acid bacteria composite fungicide, the colony forming unit number of Lactobacillus sunkii, Lactobacillus curvatus, Lactobacillus parabuchneri and Lactobacillus oligofermentans is than being (1 ~ 3): (4 ~ 6): (4 ~ 6): (1 ~ 3).Other is identical with embodiment one.
Embodiment three: present embodiment and embodiment two unlike: in described lactic acid bacteria composite fungicide, the colony forming unit number ratio of Lactobacillus sunkii, Lactobacillus curvatus, Lactobacillus parabuchneri and Lactobacillus oligofermentans is 1: 4: 4: 1 or 1: 2: 2: 1.Other is identical with embodiment two.
Embodiment four: one of present embodiment and embodiment one to three unlike: described Lactobacillus sunkii is Lactobacillus sunkii DSM19904; Described Lactobacillus curvatus is Lactobacillus curvatus DSM20019; Described Lactobacillus parabuchneri is Lactobacillus parabuchneri DSM20173; Described Lactobacillus oligofermentans is Lactobacillus oligofermentans DSM15707.Other is identical with one of embodiment one to three.
Embodiment five: one of present embodiment and embodiment one to four unlike: described lactic acid bacteria composite fungicide comprises activeconstituents, glycerine and EDTA, and the proportioning of described activeconstituents, glycerine and EDTA is (1 ~ 1.2) × 10
11cFU: (1 ~ 5) g glycerine: 1g EDTA.Other is identical with one of embodiment one to four.
Embodiment six: one of present embodiment and embodiment one to four unlike: described lactic acid bacteria composite fungicide comprises activeconstituents and skimmed milk powder, and the proportioning of described activeconstituents and skimmed milk powder is (1-1.2) × 10
8cFU: (1 ~ 6) g skimmed milk powder.Other is identical with one of embodiment one to four.
Embodiment seven: one of present embodiment and embodiment one to six are unlike the purposes of described lactic acid bacteria composite fungicide in Fermented Cabbage, Caulis et Folium Brassicae capitatae, radish or cucumber.Other is identical with one of embodiment one to six.
Beneficial effect of the present invention is verified by following examples:
The screening of activeconstituents in embodiment 1, fermenting agent.
The composition of MRS liquid nutrient medium: peptone 10g, extractum carnis 10g, yeast extract 5.0g, glucose 20g, K
2hPO
42g, dibasic ammonium citrate 2g, anhydrous sodium acetate 5g, tween 80 ml, MgSO
47H
2o0.58g, MnSO
44H
2o0.25g, pH6.2 ~ 64, distilled water 1000ml.
Getting three, northeast area traditional sauerkraut fermented liquid 5ml is linked in 95ml MRS substratum respectively, Anaerobic culturel 10d in 5 DEG C of refrigerators.
After 10d, get above-mentioned three kinds of each 5ml of nutrient solution respectively, or mutually combination cumulative volume is in the MRS substratum that the culture access 95ml of 5ml is fresh, every 4d switching once, continuous succeeding transfer culture.Measure the output of the pH value of nutrient solution, lactic acid, acetic acid and ethanol.PH value is selected to decline rapidly, the more and metastable culture of environmental change to external world of lactic acid producing, acetic acid, ethanol, screening activeconstituents wherein.
By denaturing gradient gel electrophoresis (DGGE) and 16S rDNA clone library, the culture to above-mentioned acquisition is studied, result shows, the composition bacterium in above-mentioned culture is mainly Lactobacillus sunkii, Lactobacillus curvatus, Lactobacillus parabuchneri, Lactobacillus oligofermentans.
Embodiment 2, prepare liquid lactic acid bacteria composite fungicide, ferment for Chinese cabbage.
One, the preparation of microbial inoculum
Bacterial strain activates: receive on MRS solid medium respectively by Lactobacillus sunkii DSM19904, Lactobacillus curvatus DSM20019, Lactobacillus parabuchneri DSM20173, Lactobacillus oligofermentans DSM15707, Lactobacillus sunkii DSM19904, Lactobacillus curvatus DSM20019, Lactobacillus parabuchneri DSM20173 cultivate 2d, Lactobacillus oligofermentans DSM15707 in 30 DEG C and cultivate 2d in 25 DEG C.
The composition of MRS solid medium: peptone 10g, extractum carnis 10g, yeast extract 5.0g, glucose 20g, K
2hPO
42g, dibasic ammonium citrate 2g, anhydrous sodium acetate 5g, tween 80 ml, MgSO
47H
2o0.58g, MnSO
44H
2o0.25g, agar 15g, pH6.2 ~ 6.4, distilled water 1000ml.
Lactobacillus sunkii DSM19904, Lactobacillus curvatus DSM20019, Lactobacillus parabuchneri DSM20173, Lactobacillus oligofermentans DSM15707 are all purchased from German Culture Collection.
Activated bacterial strain list bacterium colony is inoculated in 50mlMRS liquid nutrient medium respectively, under relevant temperature, cultivates 24h.Get Lactobacillus sunkii DSM1990410ml, Lactobacillus curvatus DSM2001940ml, Lactobacillus parabuchneri DSM2017340ml, Lactobacillus oligofermentans DSM1570710ml, the final colony forming unit number ratio making four kinds of bacterium is 1: 4: 4: 1, and the colony forming unit number in this mixed-culture medium is 1 × 10
8cFU/ml.
By above-mentioned mixed fermentation liquid, glycerine and EDTA according to 1 × 10
3the proportioning mixing of ml: 5g: 1g, obtains microbial inoculum A.
Two, for Fermented Cabbage
Autumn Chinese cabbage results after, airing 2d repairs, and after cleaning, code enters pithos layer by layer, and pithos volume is 750L, add salt, salt and fresh vegetable anharmonic ratio are 1: 100, mixed by 100ml microbial inoculum A, pour in cylinder with water, with the compacting of salted vegetables stone, fermentation medial temperature is 16 DEG C, and fermentation time is 24d.
At the 6d of fermentation, 12d, 18d, 24d sample, and measure milk-acid bacteria and volatile products in sauerkraut fermentation system.
Enumeration (CFUs) adopts plate dilution method, and milk-acid bacteria is cultivated and adopts MRS substratum, cultivates 2d counting for 30 DEG C, and income value is repeat mean value three times.
Adopt the volatile products in HPLC mensuration sauerkraut, chromatographic column is Bio-Rad company of U.S. Aminex87, stratographic analysis organic acid condition is: the sulfuric acid of 5mM is as moving phase, total program time of carrying out is 30min, flow velocity is 0.6mL/min, column temperature 40 DEG C, sample size is 10 μ L, and UV-detector wavelength is 205nm.Replication three times.
Sauerkraut fermentation 18d all salts down, and quality is tender and crisp, in aureus.
Milk-acid bacteria and volatile products analysis in table 1 microbial inoculum A Fermented Cabbage 24d fermentation system
Embodiment 3, prepare solid lactic acid bacterium composite fungus agent, ferment for Chinese cabbage.
One, the preparation of microbial inoculum
Bacterial strain activates: receive on MRS solid medium respectively by Lactobacillus sunkii DSM19904, Lactobacillus curvatus DSM20019, Lactobacillus parabuchneri DSM20173, Lactobacillus oligofermentans DSM15707, Lactobacillus sunkii DSM19904, Lactobacillus curvatus DSM20019, Lactobacillus parabuchneri DSM20173 cultivate 2d, Lactobacillus oligofermentans DSM15707 in 30 DEG C and cultivate 2d in 25 DEG C.
Lactobacillus sunkii DSM19904, Lactobacillus curvatus DSM20019, Lactobacillus parabuchneri DSM20173, Lactobacillus oligofermentans DSM15707 are all purchased from German Culture Collection.
Activated bacterial strain list bacterium colony is inoculated in 50ml liquid MRS substratum respectively, under relevant temperature, cultivates 24h.Get Lactobacillus sunkii DSM1990420ml, Lactobacillus curvatus DSM2001940ml, Lactobacillus parabuchneri DSM2017340ml, Lactobacillus oligofermentans DSM1570720ml, the final colony forming unit number ratio making four kinds of bacterium is 1: 2: 2: 1, and the colony forming unit number in this mixed-culture medium is 1.2 × 10
8cFU/ml.
The skimming milk 100ml of 10% will be added in thalline, as lyophilized vaccine ,-20 DEG C of freezing 10h, afterwards pressure be 3 ~ 5Pa, temperature be-50 DEG C of conditions under lyophilize 24h obtain the bacterium powder of freeze-drying, be microbial inoculum B.
Two, for Fermented Cabbage
Fermentation adopts traditional natural method, namely, after Chinese cabbage results in autumn, airing 2d repairs, and after cleaning, code enters pithos layer by layer, pithos volume is 750L, add salt, salt and fresh vegetable anharmonic ratio are 1: 100, are mixed by 1g microbial inoculum B with water, pour in cylinder, with the compacting of salted vegetables stone, fermentation medial temperature is 16 DEG C, and fermentation time is 24d.
Milk-acid bacteria and volatile products measuring method are with reference to embodiment 2.
Sauerkraut fermentation 18d all salts down, and quality is tender and crisp, in aureus.
Milk-acid bacteria and volatile products analysis in table 2 microbial inoculum B fermentation Chinese cabbage 24d fermentation system
Embodiment 4, prepare solid lactic acid bacterium composite fungus agent, ferment for Caulis et Folium Brassicae capitatae.
One, the preparation of microbial inoculum
Microbial inoculum B is prepared with reference to embodiment 3.
Two, ferment Caulis et Folium Brassicae capitatae
Fermentation adopts traditional natural method, and cut the wide filament of 0.5cm after being gathered in the crops by Caulis et Folium Brassicae capitatae, load pithos, volume is 750L, add salt, salt and fresh vegetable anharmonic ratio are 1: 100, mixed by 1g microbial inoculum B, pour in cylinder with water, with the compacting of salted vegetables stone, fermentation medial temperature is 20 DEG C, and fermentation time is 10d.
Milk-acid bacteria and volatile products measuring method are with reference to embodiment 2.
Fermentation 5d salts down, and quality is tender and crisp, in sour fragrance.
Table 3 microbial inoculum B fermentation 10d Caulis et Folium Brassicae capitatae milk-acid bacteria and volatile products detected result
Claims (7)
1., for a lactic acid bacteria composite fungicide for sauerkraut fermentation, it is characterized in that the activeconstituents of this lactic acid bacteria composite fungicide comprises Lactobacillus sunkii, Lactobacillus curvatus, Lactobacillus parabuchneri and Lactobacillus oligofermentans.
2. a kind of lactic acid bacteria composite fungicide for sauerkraut fermentation according to claim 1, is characterized in that the colony forming unit number of Lactobacillus sunkii, Lactobacillus curvatus in described lactic acid bacteria composite fungicide, Lactobacillus parabuchneri and Lactobacillus oligofermentans than being (1 ~ 3): (4 ~ 6): (4 ~ 6): (1 ~ 3).
3. a kind of lactic acid bacteria composite fungicide for sauerkraut fermentation according to claim 2, is characterized in that the colony forming unit number ratio of Lactobacillus sunkii, Lactobacillus curvatus in described lactic acid bacteria composite fungicide, Lactobacillus parabuchneri and Lactobacillus oligofermentans is 1: 4: 4: 1 or 1: 2: 2: 1.
4. a kind of lactic acid bacteria composite fungicide for sauerkraut fermentation according to claim 1,2 or 3, is characterized in that described Lactobacillus sunkii is Lactobacillus sunkii DSM19904; Described Lactobacillus curvatus is Lactobacillus curvatus DSM20019; Described Lactobacillus parabuchneri is Lactobacillus parabuchneri DSM20173; Described Lactobacillus oligofermentans is Lactobacillus oligofermentans DSM15707.
5. a kind of lactic acid bacteria composite fungicide for sauerkraut fermentation according to claim 1, it is characterized in that described lactic acid bacteria composite fungicide comprises activeconstituents, glycerine and EDTA, the proportioning of described activeconstituents, glycerine and EDTA is (1 ~ 1.2) × 10
11cFU: (1 ~ 5) g glycerine: 1gEDTA.
6. a kind of lactic acid bacteria composite fungicide for sauerkraut fermentation according to claim 1, it is characterized in that described lactic acid bacteria composite fungicide comprises activeconstituents and skimmed milk powder, the proportioning of described activeconstituents and skimmed milk powder is (1 ~ 1.2) × 10
8cFU: (1 ~ 6) g skimmed milk powder.
7., according to described a kind of lactic acid bacteria composite fungicide for sauerkraut fermentation arbitrary in claim 1 ~ 6, it is characterized in that the purposes of described lactic acid bacteria composite fungicide in Fermented Cabbage, Caulis et Folium Brassicae capitatae, radish or cucumber.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108251343A (en) * | 2018-03-31 | 2018-07-06 | 东北林业大学 | Few L fermentum strain SH-Y15 and its application |
| CN112358989A (en) * | 2020-11-10 | 2021-02-12 | 贵州老高山食品有限公司 | Lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage and preparation method of lactic acid bacteria agent |
| 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 |
-
2014
- 2014-07-17 CN CN201410341787.8A patent/CN104232506A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108251343A (en) * | 2018-03-31 | 2018-07-06 | 东北林业大学 | Few L fermentum strain SH-Y15 and its application |
| CN112358989A (en) * | 2020-11-10 | 2021-02-12 | 贵州老高山食品有限公司 | Lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage and preparation method of lactic acid bacteria agent |
| 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 |
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