CN110804569A - Quadruple bacteria direct vat type pickle starter culture and preparation method and application thereof - Google Patents
Quadruple bacteria direct vat type pickle starter culture and preparation method and application thereof Download PDFInfo
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- 241000894006 Bacteria Species 0.000 title claims abstract description 46
- 239000007858 starting material Substances 0.000 title claims abstract description 42
- 235000021110 pickles Nutrition 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims description 24
- 238000012136 culture method Methods 0.000 title description 2
- 241000193749 Bacillus coagulans Species 0.000 claims abstract description 33
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 33
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 33
- 229940054340 bacillus coagulans Drugs 0.000 claims abstract description 32
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 30
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 30
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 30
- 235000014897 Streptococcus lactis Nutrition 0.000 claims abstract description 29
- 230000001580 bacterial effect Effects 0.000 claims abstract description 22
- 230000003993 interaction Effects 0.000 claims abstract description 7
- 241000194035 Lactococcus lactis Species 0.000 claims abstract 7
- 239000001963 growth medium Substances 0.000 claims description 41
- 229960000892 attapulgite Drugs 0.000 claims description 27
- 229910052625 palygorskite Inorganic materials 0.000 claims description 27
- 239000000725 suspension Substances 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 238000002156 mixing Methods 0.000 claims description 15
- 230000003068 static effect Effects 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 13
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 230000001954 sterilising effect Effects 0.000 claims description 12
- 235000021108 sauerkraut Nutrition 0.000 claims description 11
- 241001052560 Thallis Species 0.000 claims description 10
- 238000001179 sorption measurement Methods 0.000 claims description 10
- 239000006041 probiotic Substances 0.000 claims description 9
- 235000018291 probiotics Nutrition 0.000 claims description 9
- 230000010355 oscillation Effects 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 7
- 102000004169 proteins and genes Human genes 0.000 claims description 7
- 108090000623 proteins and genes Proteins 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 235000015278 beef Nutrition 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 6
- 230000001105 regulatory effect Effects 0.000 claims description 6
- 239000007836 KH2PO4 Substances 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 5
- 238000007710 freezing Methods 0.000 claims description 5
- 230000008014 freezing Effects 0.000 claims description 5
- 235000021109 kimchi Nutrition 0.000 claims description 5
- 238000009630 liquid culture Methods 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 229940099596 manganese sulfate Drugs 0.000 claims description 5
- 239000011702 manganese sulphate Substances 0.000 claims description 5
- 235000007079 manganese sulphate Nutrition 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 5
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 5
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 5
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000002504 physiological saline solution Substances 0.000 claims 1
- 238000000855 fermentation Methods 0.000 abstract description 27
- 230000004151 fermentation Effects 0.000 abstract description 27
- 239000000796 flavoring agent Substances 0.000 abstract description 8
- 235000019634 flavors Nutrition 0.000 abstract description 8
- 235000013305 food Nutrition 0.000 abstract description 4
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 abstract description 3
- 230000036528 appetite Effects 0.000 abstract description 3
- 235000019789 appetite Nutrition 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 230000004060 metabolic process Effects 0.000 abstract description 3
- 230000007413 intestinal health Effects 0.000 abstract description 2
- 230000004936 stimulating effect Effects 0.000 abstract description 2
- 244000057717 Streptococcus lactis Species 0.000 description 22
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 12
- 210000001035 gastrointestinal tract Anatomy 0.000 description 6
- 239000004310 lactic acid Substances 0.000 description 6
- 235000014655 lactic acid Nutrition 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 235000014633 carbohydrates Nutrition 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 239000004382 Amylase Substances 0.000 description 2
- 102000013142 Amylases Human genes 0.000 description 2
- 108010065511 Amylases Proteins 0.000 description 2
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 2
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 235000019418 amylase Nutrition 0.000 description 2
- 239000003114 blood coagulation factor Substances 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 102000038379 digestive enzymes Human genes 0.000 description 2
- 108091007734 digestive enzymes Proteins 0.000 description 2
- 210000004347 intestinal mucosa Anatomy 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000194032 Enterococcus faecalis Species 0.000 description 1
- 240000001929 Lactobacillus brevis Species 0.000 description 1
- 235000013957 Lactobacillus brevis Nutrition 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 241000186684 Lactobacillus pentosus Species 0.000 description 1
- 241000577554 Lactobacillus zeae Species 0.000 description 1
- 241000194036 Lactococcus Species 0.000 description 1
- 241001468192 Leuconostoc citreum Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000235342 Saccharomycetes Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940032049 enterococcus faecalis Drugs 0.000 description 1
- 235000010855 food raising agent Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000003871 intestinal function Effects 0.000 description 1
- 229940089442 lacticare Drugs 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 235000021049 nutrient content Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
- A23L19/20—Products from fruits or vegetables; Preparation or treatment thereof by pickling, e.g. sauerkraut or pickles
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/231—Lactis
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
Abstract
The invention discloses a quadruple bacteria direct vat set pickle starter which is prepared by the interaction of four bacterial strains of lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto, wherein the viable count content of the lactobacillus plantarum, the lactococcus lactis, the bacillus coagulans and the bacillus natto in the quadruple bacteria direct vat set pickle starter is 109~1011CFU/g. The direct-vat-set pickle starter culture prepared by the invention can be directly used, and has high viable bacteria contentTotal viable count of 109~1011CFU/g, can shorten the fermentation period of the pickle to 1/3-1/2 of natural fermentation, is easy to control the production process, has stable product quality, can improve the fermentation flavor quality, reduce the nitrite content, improve the safety of the pickled food, and improve the performances of the pickle in the aspects of maintaining the intestinal health environment, helping physiological metabolism, stimulating appetite and the like.
Description
Technical Field
The invention belongs to the technical field of direct vat set starter, and particularly relates to a quadruple bacteria direct vat set pickle starter, and a preparation method and application thereof.
Background
The pickle is a food of vegetables fermented by various microorganisms taking lactic acid bacteria as the leading factor, and has special flavor, nutritive value and certain probiotic function. The research on the pickle microorganism at home and abroad mainly focuses on the aspect of lactic acid bacteria, and the research mainly comprises lactobacillus plantarum, lactobacillus brevis, leuconostoc citreum, lactobacillus pentosus, lactobacillus zeae, lactobacillus casei, lactococcus lactis, enterococcus faecalis and the like, and in addition, acetic acid bacteria, saccharomycetes, butyric acid bacteria, mold and the like. The fermentation strain identification and application of the pickle, the preservation of the nutrient content of the pickle and the like are researched in the United states and Japan; korea has been studied more on the nutrient components and health functions of kimchi; the research of China on the aspects of screening and identification of strains, flavor substances, nutrient components and the like is developed, the productivity level is obviously improved particularly in the aspects of pretreatment, production automation, mechanization, packaging, sterilization and the like of pickle raw materials, and the large-scale industrial development of pickle is promoted particularly by the popularization of a direct-vat-set lactic acid bacteria microbial inoculum.
The direct-vat-set starter is dry starter strain which is regulated, concentrated and standardized, can be directly used for fermentation in the process of making pickles, and has high viable count, strong and stable fermentation capacity. However, the performance of the direct vat set starter in the prior art in improving the functions of the pickle such as maintaining the healthy environment of the intestinal tract, helping the physiological metabolism and stimulating the appetite is still to be further improved. Lactic acid bacteria are gram-positive probiotics, can utilize fermentable carbohydrates to produce a large amount of lactic acid, and are common leavening agents for pickles. However, no studies have been made on the use of Bacillus coagulans and Bacillus natto for kimchi. The bacillus coagulans is gram-positive probiotics and can decompose macromolecular substances to generate substances such as L-lactic acid, amylase, protease, coagulation factors and the like, so that beneficial bacteria in intestinal tracts can grow and reproduce better, and the digestion and absorption of the intestinal tracts are facilitated. The bacillus natto is gram-positive probiotics and can ferment and decompose carbohydrate, protein and the like to form various substances such as organic acid, amino acid and the like which are easy to be absorbed and digested by a human body, and also can form various digestive enzymes and vitamins to promote the proliferation of small intestinal mucosa cells and maintain the intestinal function.
Disclosure of Invention
The invention provides a quadruple bacteria direct vat set pickle starter and a preparation method and application thereof aiming at the defects in the background technology, and the invention can effectively solve the problems of long pickle fermentation period, complex operation, unstable product quality and the like, and simultaneously can shorten the pickle fermentation period, improve the fermentation flavor quality, reduce the nitrite content, improve the safety of pickled food, and improve the performances of the pickle in the aspects of maintaining intestinal health environment, helping physiological metabolism, promoting appetite and the like.
In order to achieve the purpose, the technical solution of the invention is as follows:
commercial Lactobacillus plantarum (A) used in the present inventionLactobacillus.plantarum，CCTCC AB 2013128), Bacillus coagulans (B.coagulans)Bacillus .coagulansCCTCC AB 92066), Bacillus natto (Bacillus natto)Bacillus subtilis nattoCCTCCM 2014405) are all purchased from China center for type culture Collection, lactococcus lactis (C.))Lactococcus.lactisCGMCC 1.62) was purchased from China general microbiological culture Collection center.
A direct vat set pickle starter is prepared by the interaction of four strains of lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto.
The viable count contents of Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto in the quadruple bacteria direct vat set pickle starter are all 109~1011CFU/g。
A preparation method of a quadruple bacteria direct vat set pickle starter comprises the following steps:
(1) preparing a bacterial suspension: culturing Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto in corresponding liquid culture medium to late logarithmic phase, collecting thallus, washing with sterile normal saline to remove culture medium, and regulating thallus concentration to 109~1010CFU/ml is reserved;
(2) mixing the bacterial suspension: uniformly mixing the lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto bacterial suspensions according to the ratio of 1:1-5:1-5:1-5 for later use;
(3) adsorbing thalli by attapulgite: mixing the mixed bacterial suspension in the step (2) with attapulgite according to the proportion of 1L: adsorbing at the ratio of 10-100g, wherein the pH value of an adsorption system is 3-6, the temperature is 20-35 ℃, and the adsorption time is 20-50 minutes;
(4) vacuum freeze drying: pre-freezing the attapulgite adsorbing the thalli in the step (3) at-40 to-75 ℃ for 2-3 hours, quickly transferring the attapulgite into a freeze dryer for freeze drying for 20-28 hours to ensure that the water content of the preparation is less than or equal to 2 percent to obtain a direct vat set starter of tetragenous bacteria, and detecting that the number of viable bacteria in the starter is 109~1011CFU/g。
Preferably, the culture conditions of lactobacillus plantarum in the step (1) are as follows: adopting MRS culture medium to perform static culture for 16-20h at 37-39 ℃.
Preferably, the culture conditions of lactococcus lactis in step (1) are as follows: adopting MRS culture medium to perform static culture for 22-26h at 37-39 ℃.
Preferably, the culture conditions of the bacillus coagulans in the step (1) are as follows: adopting a culture medium A to perform static culture for 22-26h at 37-39 ℃; the preparation method of the culture medium A comprises the following steps: adding 30-40g bran, 12-18g beef extract, 0.3-0.8g magnesium sulfate, 0.3-0.8g manganese sulfate and 0.3-0.8g sodium dihydrogen phosphate into each liter of water, adjusting pH to 6.8-7.2, and sterilizing at 121 deg.C for 20 min.
Preferably, the culture conditions of the bacillus natto in the step (1) are as follows: shake culturing at 37-39 deg.C for 46-50h with culture medium B; the preparation method of the culture medium B comprises the following steps: each liter of water is added with 18-22 g of glucose, 4-6g of protein, 50-60 g of NaCI and 4-6g K2HPO4And 1.5-2.5 g KH2PO4Adjusting pH to 6.8-7.2, and sterilizing at 121 deg.C for 20 min.
Preferably, when the attapulgite adsorbs the bacteria in the step (3), a vibration method is adopted to increase the contact between the attapulgite and the probiotics, and the vibration speed is 90-180 rpm.
The application of fermentation agent for directly throwing pickle of tetragenous bacteria is characterized by that the fermentation agent is added according to the total quantity of 103~105The CFU/g (total weight of sauerkraut) can be directly added, so that the fermentation time of sauerkraut can be shortened to 1/3-1/2 of natural fermentation, the flavor is superior to that of natural fermentation and pure bacteria fermentation, and after fermentation is finished, 10% can be maintained4~108Viable count and spore count of CFU/g.
Compared with the prior art, the invention has the following beneficial effects:
(1) the direct-vat-set pickle starter prepared by the invention can be directly used, the viable bacteria content is high, and the total viable bacteria number is 109~1011CFU/g, can shorten the fermentation period of the pickle to 1/3-1/2 of natural fermentation, is easy to control the production process, has stable product quality, can improve the fermentation flavor quality, reduce the nitrite content and improve the safety of the pickled food.
(2) The direct vat set starter is prepared by utilizing the interaction of four bacterial strains, wherein, the lactobacillus plantarum and the lactococcus lactis promote the fermentation of the pickle in the fermentation process and form flavor substances, and the bacillus coagulans can decompose macromolecular substances to generate L-lactic acid, amylase, protease, coagulation factors and other substances, so that beneficial bacteria in intestinal tracts can better grow and reproduce, and the digestion and absorption of the intestinal tracts are facilitated; the bacillus natto can ferment and decompose carbohydrate, protein and the like to form various substances such as organic acid, amino acid and the like which are easily absorbed and digested by human bodies, and can also form various digestive enzymes and vitamins to promote the proliferation of small intestinal mucosa cells and maintain the functions of intestinal tracts.
Detailed Description
In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further described with the specific embodiments.
The preparation method of the MRS culture medium used in the invention comprises the following steps: adding 10g peptone, 10g beef extract, 5g yeast powder, 20g glucose, 2g K into each liter of water2HPO42g of diammonium citrate, 2g of sodium acetate, 1 mL of Tween 80 and 0.5g of MgSO4•7H2O and 0.25 g MnSO4•4H2And O, adjusting the pH value to 6.2-6.4, and sterilizing for 20min at 115 ℃.
Example 1
A direct vat set pickle starter is prepared by the interaction of four strains of lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto. Wherein the viable count content of Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto is 109CFU/g。
A preparation method of a quadruple bacteria direct vat set pickle starter comprises the following steps:
(1) preparing a bacterial suspension: culturing Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto in corresponding liquid culture medium to late logarithmic phase, collecting thallus, washing with sterile normal saline to remove culture medium, and regulating thallus concentration to 109CFU/ml is reserved; wherein, the culture conditions of the lactobacillus plantarum are as follows: performing static culture for 18h at 37 ℃ by adopting MRS culture medium; the culture conditions of lactococcus lactis are as follows: performing static culture for 20h at 38 ℃ by adopting MRS culture medium; the culture conditions of the bacillus coagulans are as follows: adopting a culture medium A to perform static culture for 22h at 37 ℃, wherein the preparation method of the culture medium A comprises the following steps: adding 30 g of bran, 12g of beef extract, 0.3g of magnesium sulfate, 0.3g of manganese sulfate and 0.3g of sodium dihydrogen phosphate into each liter of water, adjusting the pH value to 6.8, and sterilizing at 121 ℃ for 20 min; the culture conditions of the bacillus natto are as follows: adopting a culture medium B to perform shake cultivation for 40h at the constant temperature of 37 ℃, wherein the preparation method of the culture medium B comprises the following steps: 18g of glucose, 4g of protein, 50g of NaCI and 4g K are added into each liter of water2HPO4And 1.5g KH2PO4Adjusting pH to 6.8, and sterilizing at 121 deg.C for 20 min.
(2) Mixing the bacterial suspension: uniformly mixing the lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto bacterial suspensions according to the ratio of 1:1:1:1 for later use.
(3) Adsorbing thalli by attapulgite: mixing the mixed bacterial suspension in the step (2) with attapulgite according to the proportion of 1L: adsorbing at the ratio of 10g, wherein the pH of an adsorption system is 3, the temperature is 20 ℃, the adsorption time is 20 minutes, and the contact between the attapulgite and the probiotics is increased by adopting an oscillation method, wherein the oscillation speed is 90 rpm.
(4) Vacuum freeze drying: pre-freezing the attapulgite adsorbing the thalli in the step (3) at-40 ℃ for 2 hours, quickly transferring the attapulgite into a freeze dryer for freeze drying for 20 hours to ensure that the water content of the preparation is less than or equal to 2 percent to obtain the tetragenous bacteria direct vat set starter, and detecting that the number of viable bacteria in the starter is 109CFU/g。
Example 2
A direct-vat-set starter for preparing pickled vegetables is prepared from lactobacillus plantarum and lactic acid lactosphereThe bacillus coagulans strain is prepared by the interaction of four strains of bacillus coagulans and bacillus natto. Wherein the viable count content of Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto is 1010CFU/g。
A preparation method of a quadruple bacteria direct vat set pickle starter comprises the following steps:
(1) preparing a bacterial suspension: culturing Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto in corresponding liquid culture medium to late logarithmic phase, collecting thallus, washing with sterile normal saline to remove culture medium, and regulating thallus concentration to 1010CFU/ml is reserved; wherein, the culture conditions of the lactobacillus plantarum are as follows: performing static culture for 20h at 38 ℃ by adopting MRS culture medium; the culture conditions of lactococcus lactis are as follows: performing static culture for 24 hours at 37 ℃ by adopting MRS culture medium; the culture conditions of the bacillus coagulans are as follows: adopting a culture medium A to perform static culture for 24 hours at 37 ℃, wherein the preparation method of the culture medium A comprises the following steps: adding 35 g of bran, 15g of beef extract, 0.5g of magnesium sulfate, 0.5g of manganese sulfate and 0.5g of sodium dihydrogen phosphate into each liter of water, adjusting the pH value to 7.0, and sterilizing at 121 ℃ for 20 min; the culture conditions of the bacillus natto are as follows: adopting a culture medium B to perform shake cultivation for 48 hours at a constant temperature of 39 ℃, wherein the preparation method of the culture medium B comprises the following steps: 20g of glucose, 5g of protein, 55 g of NaCI and 5g K are added into each liter of water2HPO4And 2g KH2PO4Adjusting pH to 7.0, and sterilizing at 121 deg.C for 20 min.
(2) Mixing the bacterial suspension: uniformly mixing the lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto bacterial suspensions according to the ratio of 1:2:3:4 for later use.
(3) Adsorbing thalli by attapulgite: mixing the mixed bacterial suspension in the step (2) with attapulgite according to the proportion of 1L: adsorbing at the ratio of 50g, wherein the pH value of an adsorption system is 5, the temperature is 28 ℃, the adsorption time is 35 minutes, and the contact between the attapulgite and the probiotics is increased by adopting an oscillation method, wherein the oscillation speed is 150 rpm.
(4) Vacuum freeze drying: pre-freezing the attapulgite adsorbing the thalli in the step (3) at-60 ℃ for 2.5 hours, and quickly transferring the attapulgite into a freeze dryer for freeze drying for 24 hours to ensure that the water content of the preparation is less than or equal to2 percent to obtain the direct vat set starter of the tetrad bacteria, and the number of the live bacteria in the starter is detected to be 1010CFU/g。
Example 3
A direct vat set pickle starter is prepared by the interaction of four strains of lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto. Wherein the viable count content of Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto is 1011CFU/g。
A preparation method of a quadruple bacteria direct vat set pickle starter comprises the following steps:
(1) preparing a bacterial suspension: culturing Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto in corresponding liquid culture medium to late logarithmic phase, collecting thallus, washing with sterile normal saline to remove culture medium, and regulating thallus concentration to 1011CFU/ml is reserved; wherein, the culture conditions of the lactobacillus plantarum are as follows: performing static culture for 20h at 39 ℃ by adopting MRS culture medium; the culture conditions of lactococcus lactis are as follows: performing static culture for 26h at 39 ℃ by adopting MRS culture medium; the culture conditions of the bacillus coagulans are as follows: adopting a culture medium A to perform static culture for 26 hours at 39 ℃, wherein the preparation method of the culture medium A comprises the following steps: adding 40g of bran, 18g of beef extract, 0.8g of magnesium sulfate, 0.8g of manganese sulfate and 0.8g of sodium dihydrogen phosphate into each liter of water, adjusting the pH value to 7.2, and sterilizing at 121 ℃ for 20 min; the culture conditions of the bacillus natto are as follows: adopting a culture medium B to perform shake cultivation for 50h at a constant temperature of 39 ℃, wherein the preparation method of the culture medium B comprises the following steps: 22g of glucose, 6g of protein, 60 g of NaCI and 6g K are added into each liter of water2HPO4And 2.5g KH2PO4Adjusting pH to 7.2, and sterilizing at 121 deg.C for 20 min.
(2) Mixing the bacterial suspension: uniformly mixing the lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto bacterial suspensions according to the ratio of 1:5:5:5 for later use.
(3) Adsorbing thalli by attapulgite: mixing the mixed bacterial suspension in the step (2) with attapulgite according to the proportion of 1L: adsorbing at the ratio of 10g, wherein the pH value of an adsorption system is 6, the temperature is 35 ℃, the adsorption time is 50 minutes, and the contact between the attapulgite and the probiotics is increased by adopting an oscillation method, wherein the oscillation speed is 180 rpm.
(4) Vacuum freeze drying: pre-freezing the attapulgite adsorbing the thalli in the step (3) at-40 ℃ for 2 hours, quickly transferring the attapulgite into a freeze dryer for freeze drying for 20 hours to ensure that the water content of the preparation is less than or equal to 2 percent to obtain the tetragenous bacteria direct vat set starter, and detecting that the number of viable bacteria in the starter is 1011CFU/g。
The quality analysis of the four-bacterium direct vat set prepared in the above examples 1, 2 and 3 was determined as follows:
the fermentation agent of the four-bacterium direct vat set pickle is added according to the total amount of 103~105The CFU/g (total weight of sauerkraut) can be directly added, so that the fermentation time of sauerkraut can be shortened to 1/3-1/2 of natural fermentation, the flavor is superior to that of natural fermentation and pure bacteria fermentation, and after fermentation is finished, 10% can be maintained4~108Viable count and spore count of CFU/g.
Claims (9)
1. A quadruple bacteria direct vat set pickle starter culture is characterized in that: is prepared by the interaction of four strains of lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto.
2. The direct vat set of kimchi starter of claim 1, wherein: the viable count contents of Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto in the quadruple bacteria direct vat set pickle starter are all 109~1011CFU/g。
3. The method for preparing a starter of kimchi with tetrad bacteria as claimed in claim 1, wherein: the method comprises the following steps:
(1) preparing a bacterial suspension: culturing Lactobacillus plantarum, lactococcus lactis, Bacillus coagulans and Bacillus natto in corresponding liquid culture medium to logarithmic numberCollecting thallus in later growth period, washing with sterile physiological saline to remove culture medium, and regulating thallus concentration to 109~1010CFU/ml is reserved;
(2) mixing the bacterial suspension: uniformly mixing the lactobacillus plantarum, lactococcus lactis, bacillus coagulans and bacillus natto bacterial suspensions according to the ratio of 1:1-5:1-5:1-5 for later use;
(3) adsorbing thalli by attapulgite: mixing the mixed bacterial suspension in the step (2) with attapulgite according to the proportion of 1L: adsorbing at the ratio of 10-100g, wherein the pH value of an adsorption system is 3-6, the temperature is 20-35 ℃, and the adsorption time is 20-50 minutes;
(4) vacuum freeze drying: pre-freezing the attapulgite adsorbing the thalli in the step (3) at-40 to-75 ℃ for 2-3 hours, quickly transferring the attapulgite into a freeze dryer for freeze drying for 20-28 hours to ensure that the water content of the preparation is less than or equal to 2 percent to obtain a direct vat set starter of tetragenous bacteria, and detecting that the number of viable bacteria in the starter is 109~1011CFU/g。
4. The method for preparing a starter of sauerkraut of tetrad bacteria direct vat set as in claim 3, wherein: the culture conditions of the lactobacillus plantarum in the step (1) are as follows: adopting MRS culture medium to perform static culture for 16-20h at 37-39 ℃.
5. The method for preparing a starter of sauerkraut of tetrad bacteria direct vat set as in claim 3, wherein: the culture conditions of the lactococcus lactis in the step (1) are as follows: adopting MRS culture medium to perform static culture for 22-26h at 37-39 ℃.
6. The method for preparing a starter of sauerkraut of tetrad bacteria direct vat set as in claim 3, wherein: the culture conditions of the bacillus coagulans in the step (1) are as follows: adopting a culture medium A to perform static culture for 22-26h at 37-39 ℃; the preparation method of the culture medium A comprises the following steps: adding 30-40g bran, 12-18g beef extract, 0.3-0.8g magnesium sulfate, 0.3-0.8g manganese sulfate and 0.3-0.8g sodium dihydrogen phosphate into each liter of water, adjusting pH to 6.8-7.2, and sterilizing at 121 deg.C for 20 min.
7. The method for preparing a starter of sauerkraut of tetrad bacteria direct vat set as in claim 3, wherein: the culture conditions of the bacillus natto in the step (1) are as follows: shake culturing at 37-39 deg.C for 46-50h with culture medium B; the preparation method of the culture medium B comprises the following steps: each liter of water is added with 18-22 g of glucose, 4-6g of protein, 50-60 g of NaCI and 4-6 gK2HPO4And 1.5-2.5 g KH2PO4Adjusting pH to 6.8-7.2, and sterilizing at 121 deg.C for 20 min.
8. The method for preparing a starter of sauerkraut of tetrad bacteria direct vat set as in claim 3, wherein: and (3) increasing the contact between the attapulgite and the probiotics by adopting an oscillation method when the attapulgite adsorbs the bacteria in the step (3), wherein the oscillation speed is 90-180 rpm.
9. The use of the starter of the quadruple bacteria direct vat set kimchi according to claim 1 or 2, wherein: for preparing sauerkraut, the leaven is 10% of total amount of sauerkraut3~105And directly putting the CFU/g (total pickle mass).
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