CN104719695A - Acetic acid type pickled Chinese cabbage composite leavening agent, preparation method thereof and application - Google Patents
Acetic acid type pickled Chinese cabbage composite leavening agent, preparation method thereof and application Download PDFInfo
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Abstract
The invention discloses an acetic acid type pickled Chinese cabbage composite leavening agent, a preparation method thereof and an application, and belongs to the technical field of vegetable processing. The acetic acid type pickled Chinese cabbage composite leavening agent mainly comprises bacterial powder and calcium chloride; and the bacterial powder consists of acetobacter pasteurianus, leuconostoc mesenteroides, lactobacillus plantarum, lactobacillus brevis, lactobacillus buchneri and lactobacillus acidophilus. The leavening agent has the advantages of high activity, good taste, convenience in use and reliable quality. The viable count of the acetobacter pasteurianus, the leuconostoc mesenteroides, the lactobacillus plantarum, the lactobacillus brevis, the lactobacillus buchneri and the lactobacillus acidophilus reaches 1010cfu/g. After being fermented by the acetic acid type pickled Chinese cabbage composite leavening agent, Chinese cabbages contain lactic acid which tastes soft, and also contains flavor substances such as acetic acid tasting sour, and the taste of the Chinese cabbages which are fermented by the acetic acid type pickled Chinese cabbage composite leavening agent is the same with that of pickled Chinese cabbages which are fermented naturally by the traditional method.
Description
Technical field
The present invention relates to a kind of acetic acid type sauerkraut composite ferment and preparation method thereof and application, belong to fruits and vegetables technical field.
Background technology
Sauerkraut is the salt by adding low concentration in Chinese cabbage, undergoes microbial fermentation and forms and a kind of fermented vegetable product of local flavor product based on lactic acid.Owing to greatly can extend the holding time after Chinese cabbage results, and local flavor is also greatly improved, and therefore sauerkraut is extensively by northeast, the liking of North China people.The acid of unique flavor, nutrient health, mouthfeel is refreshing, quality is tender and crisp, with rich flavor, with fragrance striking the nose, bright etc., is the obvious and the most most popular feature of sauerkraut.Along with the raising of people's living standard, and the understanding to health diet, taking lactic acid bacteria as dominant microflora, is that auxiliary fermentation forms with saccharomycete, acetic acid bacteria etc., and the sauerkraut containing the multiple prebiotic factor, liked by increasing people gradually and eat.
Pickling of sauerkraut originates from traditional domestic curing sauerkraut, and this curing process belongs to the spontaneous fermentation of Chinese cabbage.The sauerkraut of the Northeast refers in the fall after Chinese cabbage results, take Chinese cabbage as raw material, by adding salt, and the fermented vegetables products of about spontaneous fermentation 1-2 month in the winter time.In traditional Chinese cabbage curing process, because the microbial adhesion of a large amount of distinct is in Chinese cabbage surface, wherein there is multiple harmful microorganism, as mould, saccharomycete and Escherichia coli etc., can with lactic acid bacteria syntrophism and existence, one is cause a hidden trouble to people's safe diet, and two is restriction lactic acid bacteria Rapid Fermentations, and three is keep damaging to the normal quality of sauerkraut.Therefore, traditional-family's formula spontaneous fermentation has production cycle long, the drawback such as miscellaneous bacteria is more, product quality is wayward, product quality is poor, can not be used for standardization and suitability for industrialized production.
Dropping into lactic acid bacteria and other strain fermentation Chinese cabbages with leavening form, is the development trend of sauerkraut manufacture.Although more about the research of sauerkraut leavening at present, really the little of actual production can be applied to.Sauerkraut leavening mostly wherein is composite ferment, species and other compositions are had nothing in common with each other, as patent CN 101961048A, disclose the leavening be made up of 4 kinds of freeze-dried vaccine powder such as lactobacillus paraceasi, Bu Shi lactobacillus, Lactobacillus plantarum and bacillus coagulanses, simultaneously also containing the auxiliary element such as natamycin and calcium chloride; Leavening disclosed in patent CN 101317646 is the freeze-dried vaccine powder of the 5 strain bacterium such as Lactobacillus plantarum, Pediococcus acidilactici, Leuconostoc mesenteroides, Lactococcus lactis, Lactobacillus brevis; Leavening disclosed in patent CN 102212477A is Lactobacillus plantarum, lactobacillus acidophilus, leuconostoc pseudomesenteroides, Lactococcus lactis breast subspecies and Lactobacillus rhamnosus etc. 5 freeze-dried vaccine powder of strain bacterium, and fermentation time reduction can be 15-20 days by this leavening at 0-25 DEG C.
Existing sauerkraut leavening can shorten fermentation period greatly, and containing the specific lactic acid bacteria inoculated or probio, but be substantially all lactic acid in its tunning, and traditional sauerkraut contains the flavor substances such as the acetic acid produced in sweat, therefore the acid of existing lactobacillus inoculum sauerkraut and traditional sauerkraut local flavor of feeling well is inconsistent.Simultaneously, after existing lactic acid bacteria fermenting agent is inoculated in Chinese cabbage, to be difficult in aerobic environment in the early stage effectively suppress saccharomycetic growth, to cause yeast in Chinese cabbage sweat to exist in a large number and continue fermentation, after processing and packaging, the problem such as even cause sauerkraut to rise to wrap and rot.
Summary of the invention
For solving the problem, the invention provides a kind of sauerkraut composite ferment, the technical scheme taked is as follows:
One object of the present invention is to provide a kind of acetic acid type sauerkraut composite ferment, and this composite ferment is primarily of bacterium powder and calcium chloride composition; Wherein bacterium powder is made up of Pasteur's acetobacter Acetobacter pasteurianus, Leuconostoc mesenteroides Leuconostocmesenteroides, Lactobacillus plantarum Lactobacillus plantarum, Lactobacillus brevis Lactobacillus brevis, Bu Shi lactobacillus Lactobacillus buchneri and lactobacillus acidophilus Lactobacillus acidophilus.
Preferably, described Pasteur's acetobacter is Pasteur's acetobacter CICC 23561; Described Leuconostoc mesenteroides is Leuconostoc mesenteroides CICC 22246; Described Lactobacillus plantarum is Lactobacillus plantarum CICC 23138; Described Lactobacillus brevis is Lactobacillus brevis CICC 6239; Described Bu Shi lactobacillus is CICC 20015; Described lactobacillus acidophilus is lactobacillus acidophilus CICC 6082.
Preferably, described bacterium powder, viable count is than being 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; Described calcium chloride and bacterium powder, calcium chloride: the mass ratio of bacterium powder is 1 ~ 8:1.
More preferably, described bacterium powder, viable count is than being Pasteur's acetobacter: Leuconostoc mesenteroides: Lactobacillus plantarum: Lactobacillus brevis: Bu Shi lactobacillus: lactobacillus acidophilus=1:20:30:10:10:10, calcium chloride: the mass ratio of bacterium powder is 4:1.
Described arbitrary composite ferment is applied to fermenting and producing sauerkraut.
The step of described application is as follows:
1) Chinese cabbage is removed root, extract surperficial stale dish leaf, clean with water purification and drain Chinese cabbage surface moisture;
2) Chinese cabbage is placed in 5-6% saline solution pickled dehydration 10-15h;
3) pickled complete Chinese cabbage is taken out, inject the saline solution of 1.5-3% wherein, and the composite ferment mixed containing 0.1g/kg Chinese cabbage in saline solution, by Chinese cabbage compacting, Chinese cabbage is immersed in water;
4) in 0-40 DEG C of fermentation 3-30 days, and processing or edible can be taken out.
Another object of the present invention is to the preparation method providing a kind of described composite ferment, the method is bacterium colony activation culture in acetic acid bacteria fluid nutrient medium of picking Pasteur acetobacter, distinguish picking Leuconostoc mesenteroides again, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus bacterium colony carry out activation culture in MRS fluid nutrient medium, recycling acetic acid bacteria fluid nutrient medium is to Pasteur's acetobacter, utilize MRS fluid nutrient medium to Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus carry out 3 high density respectively and amplify cultivation, add freeze drying protectant freeze-drying after collected by centrifugation thalline respectively again, 6 kinds of freeze-dried vaccine powder are mixed in proportion rear acquisition bacterium powder, finally bacterium powder and calcium chloride are carried out composite rear acquisition composite bacteria agent capable.
Preferably, Pasteur's acetobacter described in described method, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, the bacterium colony unit numbers ratio of picking is 1-5:15-25:25-35:5-15:5-15:5-15; Described 6 kinds of freeze-dried vaccine powder to be mixed in proportion, be according to viable count than being Pasteur's acetobacter: Leuconostoc mesenteroides: Lactobacillus plantarum: Lactobacillus brevis: Bu Shi lactobacillus: the ratio mixing of lactobacillus acidophilus=1:20:30:10:10:10.
Preferably, the step of described method is as follows:
1) picking Pasteur acetobacter colony inoculation is in 5mL acetic acid bacteria fluid nutrient medium, 24-48h is cultivated at 25-37 DEG C, distinguish the single bacterium colony of picking Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus again, be seeded in the small test tube filling 5mL MRS fluid nutrient medium respectively, cultivate 24-48h in 30-40 DEG C, obtain activated strains;
The composition of described acetic acid bacteria fluid nutrient medium is glucose 100.0g, yeast extract 10.0g, CaCO
320.0g, agar 15.0g, distilled water 1.0L, pH 6.8;
Described Pasteur's acetobacter, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, the bacterium colony unit numbers ratio of picking is 1-5:15-25:25-35:5-15:5-15:5-15;
2) acetic acid bacteria fluid nutrient medium is utilized to step 1) Pasteur's acetobacter of gained, utilizing MRS fluid nutrient medium to step 1) Leuconostoc mesenteroides of gained, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus carry out 3 high density respectively and amplify and cultivate, condition of culture is: Pasteur's acetobacter inoculum concentration 1-3%, cultivation temperature: 25-37 DEG C, incubation time: 24-48h; The cultivation temperature of Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus: 30-40 DEG C, incubation time: 24-48h; In 3 high density amplification cultivations, the volume of acetic acid bacteria fluid nutrient medium and MRS fluid nutrient medium is followed successively by 100mL, 2000mL and 40L; High Density Cultivation terminates rear acquisition 6 kinds of bacterium liquid;
3) by step 2) the bacterium liquid of gained respectively under 5000-10000r/min, centrifugal 5-15min, centrifuging temperature is 4-10 DEG C, retains precipitation;
4) by step 2) the long-pending 50%-100% of the bacteria liquid of gained is to step 3) add freeze drying protectant mixing in gained precipitation after, in-20 DEG C to-80 DEG C after pre-freeze 10-15h, utilize freeze drier freeze-drying, acquisition freeze-dried vaccine powder;
5) by step 4) 6 kinds of freeze-dried vaccine powder of gained according to viable count than being Pasteur's acetobacter: Leuconostoc mesenteroides: Lactobacillus plantarum: Lactobacillus brevis: Bu Shi lactobacillus: obtain bacterium powder after the ratio mixing of lactobacillus acidophilus=1:20:30:10:10:10, then by calcium chloride and bacterium powder according to calcium chloride: the mass ratio of bacterium powder is the composite acquisition composite bacteria agent capable of ratio of 1 ~ 8:1.
Preferably, step 4 in described method) described freeze drying protectant comprise the skimmed milk of percentage by weight 10%, the trehalose of 3% and 1.5% sodium glutamate.
The present invention achieves following beneficial effect:
1. leavening provided by the present invention has that vigor is high, raciness, feature easy to use, of fine qualities.The viable count of Pasteur's acetobacter in leavening provided by the present invention, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus reaches 10
10cfu/g.Leavening, with packed packaged, can use after taking apart.
2. leavening provided by the present invention is acetic acid type sauerkraut composite ferment, wherein containing acetic acid bacteria (Pasteur's acetobacter) and lactic acid bacteria (Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus), not only containing the lactic acid that mouthfeel is soft after Fermented Cabbage, also containing flavor substances such as the acetic acid making the acid of sauerkraut mouthfeel refreshing, consistent with the sauerkraut taste that traditional natural ferments.
3. leavening provided by the present invention can effectively suppress saccharomycetic breeding, and what prevent saccharomycete continuation breeding in sweat from causing is putrid and deteriorated, thus extends the shelf time of sauerkraut.
4. leavening provided by the present invention is except containing except multiple bacterial classification, also containing calcium chloride, rapidly dissolvable after dropping into sauerkraut, and effectively can keep the vivid color and luster of Chinese cabbage and the tender and crisp feature of quality.
5. starter bacteria powder provided by the present invention is made up of a strain acetic acid bacteria and five strains of lactic acid bacteria, and due in sauerkraut spontaneous fermentation process, what the starting stage grew is aerobic acetic acid bacteria and amphimicrobian saccharomycete, and acetic acid bacteria consumes oxygen and carbohydrate produces acetic acid and CO
2, saccharomycete then produces CO
2and ethanol, generate CO by consuming oxygen
2for oxygen free condition is created in the growth of lactic acid bacteria, and acetic acid and ethanol are the important flavor substances in sauerkraut, wherein acetic acid can give sauerkraut salubrious tart flavour, and ethanol can generate the dulcet ester of tool with lactic acid, make the local flavor that sauerkraut is exclusive, because acetic acid bacteria creates acetic acid, can effectively suppress saccharomycete in the early stage after survival and growth, can prevent the microbial variable color of black yeast, produce the phenomenons such as smelly and aerogenesis, the quality being conducive to controlling sauerkraut keeps; In five kinds of lactic acid bacterias, Leuconostoc mesenteroides, Lactobacillus brevis and Bu Shi lactobacillus are heterolactic fermentation bacterium, except generation lactic acid, ethanol and acetic acid can also be produced, strengthen the acid of sauerkraut to feel well local flavor and aromatic character, and Lactobacillus plantarum and lactobacillus acidophilus are homofermentative lactic bacterium, acetic acid bacteria, heterolactic fermentation bacterium and homofermentative lactic bacterium can produce acid by Rapid Fermentation, and form local flavor specific to sauerkraut.
Accompanying drawing explanation
In Fig. 1 sauerkraut that to be embodiment 1 ferment with the composite ferment utilizing embodiment 4-6 to make, lactic acid and acetic acid content contrast.
Fig. 2 is that embodiment 1 produces saccharomycete and acetic acid bacteria variation tendency in sauerkraut with the composite ferment utilizing embodiment 6 to prepare.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention will be further described, but the present invention is not by the restriction of embodiment.
Material therefor of the present invention, reagent, instruments and methods, without specified otherwise, be this area conventional material, reagent, instruments and methods, all can be obtained by commercial channel.
Embodiment 1
Present embodiments provide a kind of preparation method utilizing the ferment-fermented sauerkraut of commercially available lactic acid type, concrete steps are as follows:
(1), after being dug up the roots by Chinese cabbage and removing rotten leaf, airing is placed under sunlight one to three days, remove portion moisture.
(2) clear water is used to clean.
(3) the Chinese cabbage dress cylinder will cleaned, often put one deck Chinese cabbage and spread one deck salt, total salt consumption is about 10-15g/kg Chinese cabbage.
(4) use stone or weight to be pushed down by Chinese cabbage, add commercially available lactic acid type leavening according to the ratio of 0.1g/Kg Chinese cabbage and in Chinese cabbage, add clear water again, the water surface did not have Chinese cabbage.
(5) terminate after fermenting 3 days.
Embodiment 2
Present embodiments provide a kind of fermentation culture method of required bacterial classification, concrete steps are as follows:
(1) picking Pasteur acetobacter colony inoculation is in 5mL acetic acid bacteria fluid nutrient medium, and at 30 DEG C, 24h is cultivated in concussion.Wherein, the composition of acetic acid bacteria fluid nutrient medium is glucose 100.0g, yeast extract 10.0g, CaCO
320.0g, agar 15.0g, distilled water 1.0L, pH 6.8.
(2) the single bacterium colony of difference picking Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, is seeded in the small test tube filling 5mLMRS fluid nutrient medium, cultivates 24h in 40 DEG C, obtain activated strains.
Wherein, step 1) and step 2) in Pasteur's acetobacter, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus bacterium colony unit numbers ratio be 1:25:35:15:15:15
(3) the Pasteur's acetobacter activated in small test tube is seeded in 100mL acetic acid bacteria fluid nutrient medium with 2% ratio, 24h is cultivated in 30 DEG C of concussions, by activate in small test tube Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, be seeded in 100mLMRS fluid nutrient medium with 2% ratio, cultivate 24h in 40 DEG C.
(4) the Pasteur's acetobacter activated by 100mL is seeded in 2000mL acetic acid bacteria fluid nutrient medium with 2% ratio, 24h is cultivated in 30 DEG C of concussions, by activate in small test tube Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, be seeded in 2000mLMRS fluid nutrient medium with 2% ratio, cultivate 24h in 40 DEG C.
Embodiment 3
Present embodiments provide a kind of fermentation culture method of required bacterial classification, concrete steps are as follows:
1) picking Pasteur acetobacter colony inoculation is in 5mL acetic acid bacteria fluid nutrient medium, and at 25 DEG C, 48h is cultivated in concussion.Wherein, the composition of acetic acid bacteria fluid nutrient medium is glucose 100.0g, yeast extract 10.0g, CaCO
320.0g, agar 15.0g, distilled water 1.0L, pH 6.8.
2) the single bacterium colony of difference picking Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, is seeded in the small test tube filling 5mLMRS fluid nutrient medium, cultivates 48h in 35 DEG C, obtain activated strains.
Wherein, step 1) and step 2) in Pasteur's acetobacter, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus bacterium colony unit numbers ratio be 1:15:25:5:5:5.
3) the Pasteur's acetobacter activated in small test tube is seeded in 100mL acetic acid bacteria fluid nutrient medium with 3% ratio, 48h is cultivated in 25 DEG C of concussions, by activate in small test tube Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, be seeded in 100mLMRS fluid nutrient medium with 3% ratio, cultivate 48h in 35 DEG C.
4) the Pasteur's acetobacter activated by 100mL is seeded in 2000mL acetic acid bacteria fluid nutrient medium with 3% ratio, 48h is cultivated in 25 DEG C, by activate in small test tube Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, be seeded in 2000mLMRS fluid nutrient medium with 3% ratio, cultivate 48h in 35 DEG C.
Embodiment 4
Present embodiments provide a kind of preparation method of acetic acid type compound sauerkraut leavening, concrete steps are as follows:
(1) bacterial classification is cultivated according to the method (except the picking ratio of bacterium colony unit) of embodiment 3, obtain the bacterium liquid being amplified to 2000mL, the Pasteur's acetobacter activated by 2000mL is again seeded in 40L acetic acid bacteria fluid nutrient medium with 1% ratio, 10h is cultivated in 37 DEG C, the Leuconostoc mesenteroides that 2000mL is activated, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, be seeded in 40L acetic acid bacteria fluid nutrient medium with 1% ratio, cultivate 10h in 40 DEG C.
Wherein, the bacterium colony unit numbers ratio of Pasteur's acetobacter of picking before activation, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus is 1:15:25:5:5:5.
(2) getting six kinds of bacterium liquid respectively carries out centrifugal; rotating speed is 5000r/min; time is 15min, and temperature controls, at 4-10 DEG C, to abandon supernatant; collect thalline; the freeze drying protectant of original bacteria liquid 50%-100% volume is joined thalline, and mixes, be placed in-20 DEG C to-80 DEG C pre-freeze 15h; re-use freeze drier and carry out freeze-drying, obtain bacterium powder.Wherein, freeze drying protectant contains the sodium glutamate of the skimmed milk of 10% (W/W), the trehalose and 1.5% (W/W) of 3% (W/W).
(3) by step 2) 6 kinds of freeze-dried vaccine powder of gained according to viable count than being Pasteur's acetobacter: Leuconostoc mesenteroides: Lactobacillus plantarum: Lactobacillus brevis: Bu Shi lactobacillus: obtain bacterium powder after the ratio mixing of lactobacillus acidophilus=1:20:30:10:10:10, again the mass ratio 1:1 of calcium chloride and bacterium powder is carried out composite, obtain acetic acid type sauerkraut composite ferment.
Embodiment 5
Present embodiments provide a kind of preparation method of acetic acid type compound sauerkraut leavening, concrete steps are as follows:
1) bacterial classification is cultivated according to the method (except the picking ratio of bacterium colony unit) of embodiment 3, obtain the bacterium liquid being amplified to 2000mL, the Pasteur's acetobacter activated by 2000mL is again seeded in 40L acetic acid bacteria fluid nutrient medium with 2% ratio, 11h is cultivated in 30 DEG C, the Leuconostoc mesenteroides that 2000mL is activated, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, be seeded in 40L acetic acid bacteria fluid nutrient medium with 2% ratio, cultivate 11h in 35 DEG C.
Wherein, the bacterium colony unit numbers ratio of Pasteur's acetobacter of picking before activation, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus is 1:25:35:15:15:15.
2) getting six kinds of bacterium liquid respectively carries out centrifugal; rotating speed is 8000r/min; time is 10min, and temperature controls, at 4-10 DEG C, to abandon supernatant; collect thalline; the freeze drying protectant of original bacteria liquid 50%-100% volume is joined thalline, and mixes, be placed in-20 DEG C to-80 DEG C pre-freeze 12h; re-use freeze drier and carry out freeze-drying, obtain bacterium powder.Wherein, freeze drying protectant contains the sodium glutamate of the skimmed milk of 10% (W/W), the trehalose and 1.5% (W/W) of 3% (W/W).
3) by step 2) 6 kinds of freeze-dried vaccine powder of gained according to viable count than being Pasteur's acetobacter: Leuconostoc mesenteroides: Lactobacillus plantarum: Lactobacillus brevis: Bu Shi lactobacillus: obtain bacterium powder after the ratio mixing of lactobacillus acidophilus=1:20:30:10:10:10, the mass ratio 8:1 of calcium chloride and bacterium powder carries out composite again, obtains acetic acid type sauerkraut composite ferment.
Embodiment 6
Present embodiments provide a kind of preparation method of acetic acid type compound sauerkraut leavening, concrete steps are as follows:
1) bacterial classification is cultivated according to the method (except the picking ratio of bacterium colony unit) of embodiment 3, obtain the bacterium liquid being amplified to 2000mL, the Pasteur's acetobacter activated by 2000mL is again seeded in 40L acetic acid bacteria fluid nutrient medium with 3% ratio, 10h is cultivated in 25 DEG C, the Leuconostoc mesenteroides that 2000mL is activated, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, be seeded in 40L acetic acid bacteria fluid nutrient medium with 3% ratio, cultivate 10h in 30 DEG C.
Wherein, the bacterium colony unit numbers ratio of Pasteur's acetobacter of picking before activation, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus is 1:20:30:10:10:10.
2) getting six kinds of bacterium liquid respectively carries out centrifugal; rotating speed is 10000r/min; time is 5min, and temperature controls, at 4-10 DEG C, to abandon supernatant; collect thalline; the freeze drying protectant of original bacteria liquid 50%-100% volume is joined thalline, and mixes, be placed in-20 DEG C to-80 DEG C pre-freeze 10h; re-use freeze drier and carry out freeze-drying, obtain bacterium powder.Wherein, freeze drying protectant contains the sodium glutamate of the skimmed milk of 10% (W/W), the trehalose and 1.5% (W/W) of 3% (W/W).
3) by step 2) 6 kinds of freeze-dried vaccine powder of gained according to viable count than being Pasteur's acetobacter: Leuconostoc mesenteroides: Lactobacillus plantarum: Lactobacillus brevis: Bu Shi lactobacillus: obtain bacterium powder after the ratio mixing of lactobacillus acidophilus=1:20:30:10:10:10, to be that 4:1 carries out composite for the ratio of calcium chloride and bacterium powder again, obtains acetic acid type sauerkraut composite ferment.
Embodiment 7
Present embodiments provide a kind of method utilizing the composite ferment prepared by embodiment 4,5 and 6 to prepare sauerkraut, concrete steps are as follows:
1) Chinese cabbage is removed root, extract surperficial stale dish leaf, clean with water purification and drain Chinese cabbage surface moisture;
2) Chinese cabbage is placed in 5% saline solution pickled dehydration 12h;
3) prepare a certain amount of saline solution according to Chinese cabbage weight, its concentration is 2% (W/W), dissolves mixing.
4) according to Chinese cabbage weight, drop into the composite ferment prepared by embodiment 4,5 and 6 in saline solution respectively, input amount is 0.1g/kg Chinese cabbage, mixing.
5) by Chinese cabbage compacting, in Chinese cabbage, inject the saline solution containing leavening, make the water surface exceed Chinese cabbage;
6) in 20 DEG C of fermentations 3 days, and processing or edible can be taken out.
Embodiment 8
The effects such as the local flavor of the present embodiment to the sauerkraut prepared by embodiment 1,4,5 and 6, the inhibitory action to yeast measure.Method and result as follows:
(1) local flavor
To embodiment 1, 4, 5 and 6 pH producing sauerkraut measure, wherein embodiment 1 pickles acid, pH value is 3.08, embodiment 4, 5 and 6 produce sauerkraut pH value reaches 3.01 respectively, 3.02 with 2.93, by lactic acid and acetic acid content in high effective liquid chromatography for measuring sauerkraut, as shown in Figure 1, embodiment 4, 5 and 6 produce lactic acid content in sauerkraut is respectively 19.12, 18.66 with 20.35g/kg Chinese cabbage, and be 18.73g/kg Chinese cabbage in embodiment 1 pickling sauerkraut, embodiment 4, 5 and 6 produce acetic acid content in sauerkraut is respectively 3.04, 2.94 and 3.07g/kg Chinese cabbage, and in embodiment 1 pickling sauerkraut, be only 1.42g/kg Chinese cabbage, it can thus be appreciated that, embodiment 4, 5 and 6 produce sauerkraut and embodiment 1 to make the lactic acid content of sauerkraut basically identical, and acetic acid content is 214.1% of embodiment 1 respectively, 207.0% and 216.2%, its product special flavour obviously improves, more close to the sauerkraut that the daily life of a family is pickled.
(2) acetic acid bacteria is to the suppression of yeast
Complete in initial 0 day of fermentation and fermentation respectively and sauerkraut was produced to embodiment 7 in 3 days and carry out saccharomycete and acetic acid bacteria and measure, respectively initial 0 day of fermentation and after ferment to embodiment 1 and embodiment 7 in utilize embodiment 6 composite ferment to prepare sauerkraut carry out saccharomycete and acetic acid bacteria mensuration, as shown in Figure 2, embodiment 7 is produced in sauerkraut process, and the initial bacterium number of saccharomycete is 10
2cfu/mL, because below 10cfu/mL is down to by the suppression saccharomycete by acetic acid bacteria when having fermented, and 10 when embodiment 1 acetic acid bacteria has initial
4cfu/mL, the by fermentation oxygen consumption of process, viable count is also down to below 10cfu/mL thereupon; Comparatively speaking, embodiment 1 makes in sauerkraut process, although due to oxygen consumption formed anaerobic environment, acetic acid bacteria viable count is down to below 10cfu/mL, but saccharomycete viable count has almost no change, and thus sauerkraut is after being processed into product, still have and continue the rotten possibility of fermentation gas.
(3) sauerkraut storage period
The sauerkraut utilizing embodiment 6 composite ferment to prepare in embodiment 7 and embodiment 1 are made sauerkraut, the vacuum packaging when not adding anticorrisive agent, under being positioned over room temperature (22 DEG C) environment, result shows, the sauerkraut that embodiment 7 is produced can store two months, and embodiment 1 makes sauerkraut period of storage instability, started to 15 days rise bag or variable color.
(4) Pasteur's acetobacter adds control experiment
For the effect of Pasteur's acetobacter in evaluation leavening, use the leavening that with the addition of Pasteur's acetobacter respectively, and do not add the leavening of Pasteur's acetobacter, carry out sauerkraut and produce contrast experiment, test sample is taken out when fermentation to pH value reaches 3.0, as shown in table 1, being added to of Pasteur's acetobacter impacts the maturation time of sauerkraut, and the fermentation deadline is all 3 days; But do not add saccharomycete viable count in the sauerkraut of Pasteur's acetobacter far above the sauerkraut adding Pasteur's acetobacter, it can thus be appreciated that Pasteur's acetobacter effectively can suppress saccharomycetic growth and survival in sauerkraut.
Table 1 adds and the fermentation index of not adding Pasteur's acetobacter
Although the present invention with preferred embodiment openly as above; but it is also not used to limit the present invention, any person skilled in the art, without departing from the spirit and scope of the present invention; can do various change and modification, what therefore protection scope of the present invention should define with claims is as the criterion.
Claims (10)
1. an acetic acid type sauerkraut composite ferment, is characterized in that, primarily of bacterium powder and calcium chloride composition; Wherein bacterium powder is made up of Pasteur's acetobacter Acetobacter pasteurianus, Leuconostoc mesenteroides Leuconostoc mesenteroides, Lactobacillus plantarum Lactobacillus plantarum, Lactobacillus brevis Lactobacillus brevis, Bu Shi lactobacillus Lactobacillus buchneri and lactobacillus acidophilus Lactobacillus acidophilus.
2. composite ferment described in claim 1, is characterized in that, described Pasteur's acetobacter, is Pasteur's acetobacter CICC 23561; Described Leuconostoc mesenteroides is Leuconostoc mesenteroides CICC 22246; Described Lactobacillus plantarum is Lactobacillus plantarum CICC23138; Described Lactobacillus brevis is Lactobacillus brevis CICC 6239; Described Bu Shi lactobacillus is CICC 20015; Described lactobacillus acidophilus is lactobacillus acidophilus CICC 6082.
3. composite ferment described in claim 1, it is characterized in that, described bacterium powder, viable count is than being 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; Described calcium chloride and bacterium powder, calcium chloride: the mass ratio of bacterium powder is 1 ~ 8:1.
4. composite ferment described in claim 3, it is characterized in that, described bacterium powder, viable count is than being Pasteur's acetobacter: Leuconostoc mesenteroides: Lactobacillus plantarum: Lactobacillus brevis: Bu Shi lactobacillus: lactobacillus acidophilus=1:20:30:10:10:10, calcium chloride: the mass ratio of bacterium powder is 4:1.
5. arbitrary composite ferment described in claim 1-4, is characterized in that, is applied to fermenting and producing sauerkraut.
6. apply described in claim 5, it is characterized in that, step is as follows:
1) Chinese cabbage is removed root, extract surperficial stale dish leaf, clean with water purification and drain Chinese cabbage surface moisture;
2) Chinese cabbage is placed in 5-6% saline solution pickled dehydration 10-15h;
3) pickled complete Chinese cabbage is taken out, inject the saline solution of 1.5-3% wherein, and the composite ferment mixed containing 0.1g/kg Chinese cabbage in saline solution, by Chinese cabbage compacting, Chinese cabbage is immersed in water;
4) in 0-40 DEG C of fermentation 3-30 days, and processing or edible can be taken out.
7. the preparation method of composite ferment described in a claim 1, it is characterized in that, bacterium colony activation culture in acetic acid bacteria fluid nutrient medium of picking Pasteur acetobacter, distinguish picking Leuconostoc mesenteroides again, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus bacterium colony carry out activation culture in MRS fluid nutrient medium, recycling acetic acid bacteria fluid nutrient medium is to Pasteur's acetobacter, utilize MRS fluid nutrient medium to Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus carry out 3 high density respectively and amplify cultivation, add freeze drying protectant freeze-drying after collected by centrifugation thalline respectively again, 6 kinds of freeze-dried vaccine powder are mixed in proportion rear acquisition bacterium powder, finally bacterium powder and calcium chloride are carried out composite rear acquisition composite bacteria agent capable.
8. method described in claim 7, it is characterized in that, described Pasteur's acetobacter, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, the bacterium colony unit numbers ratio of picking is 1-5:15-25:25-35:5-15:5-15:5-15; Described 6 kinds of freeze-dried vaccine powder to be mixed in proportion, be according to viable count than being Pasteur's acetobacter: Leuconostoc mesenteroides: Lactobacillus plantarum: Lactobacillus brevis: Bu Shi lactobacillus: the ratio mixing of lactobacillus acidophilus=1:20:30:10:10:10.
9. method described in claim 7, is characterized in that, step is as follows:
1) picking Pasteur acetobacter colony inoculation is in 5mL acetic acid bacteria fluid nutrient medium, 24-48h is cultivated at 25-37 DEG C, distinguish the single bacterium colony of picking Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus again, be seeded in the small test tube filling 5mL MRS fluid nutrient medium respectively, cultivate 24-48h in 30-40 DEG C, obtain activated strains; The composition of described acetic acid bacteria fluid nutrient medium is glucose 100.0g, yeast extract 10.0g, CaCO
320.0g, agar 15.0g, distilled water 1.0L, pH 6.8;
Described Pasteur's acetobacter, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus, the bacterium colony unit numbers ratio of picking is 1-5:15-25:25-35:5-15:5-15:5-15;
2) acetic acid bacteria fluid nutrient medium is utilized to step 1) Pasteur's acetobacter of gained, utilizing MRS fluid nutrient medium to step 1) Leuconostoc mesenteroides of gained, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus carry out 3 high density respectively and amplify and cultivate, condition of culture is: Pasteur's acetobacter inoculum concentration 1-3%, cultivation temperature: 25-37 DEG C, incubation time: 24-48h; The cultivation temperature of Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus brevis, Bu Shi lactobacillus and lactobacillus acidophilus: 30-40 DEG C, incubation time: 24-48h; In 3 high density amplification cultivations, the volume of acetic acid bacteria fluid nutrient medium and MRS fluid nutrient medium is followed successively by 100mL, 2000mL and 40L; High Density Cultivation terminates rear acquisition 6 kinds of bacterium liquid;
3) by step 2) the bacterium liquid of gained respectively under 5000-10000r/min, centrifugal 5-15min, centrifuging temperature is 4-10 DEG C, retains precipitation;
4) by step 2) the long-pending 50%-100% of the bacteria liquid of gained is to step 3) add freeze drying protectant mixing in gained precipitation after, in-20 DEG C to-80 DEG C after pre-freeze 10-15h, utilize freeze drier freeze-drying, acquisition freeze-dried vaccine powder;
5) by step 4) 6 kinds of freeze-dried vaccine powder of gained according to viable count than being Pasteur's acetobacter: Leuconostoc mesenteroides: Lactobacillus plantarum: Lactobacillus brevis: Bu Shi lactobacillus: obtain bacterium powder after the ratio mixing of lactobacillus acidophilus=1:20:30:10:10:10, then by calcium chloride and bacterium powder according to calcium chloride: the mass ratio of bacterium powder is the composite acquisition composite bacteria agent capable of ratio of 1 ~ 8:1.
10. method described in claim 9, is characterized in that, step 4) described freeze drying protectant comprise the skimmed milk of percentage by weight 10%, the trehalose of 3% and 1.5% sodium glutamate.
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