CN103734310B - A kind of novel Yoghourt fermentation mix bacterium agent - Google Patents
A kind of novel Yoghourt fermentation mix bacterium agent Download PDFInfo
- Publication number
- CN103734310B CN103734310B CN201310741754.8A CN201310741754A CN103734310B CN 103734310 B CN103734310 B CN 103734310B CN 201310741754 A CN201310741754 A CN 201310741754A CN 103734310 B CN103734310 B CN 103734310B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus
- lactobacillus plantarum
- yoghourt
- fermentation
- mix bacterium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 235000013618 yogurt Nutrition 0.000 title claims abstract description 41
- 238000000855 fermentation Methods 0.000 title claims abstract description 36
- 230000004151 fermentation Effects 0.000 title claims abstract description 36
- 241000894006 Bacteria Species 0.000 title claims abstract description 34
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 34
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 34
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 34
- 239000000203 mixture Substances 0.000 claims abstract description 24
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 claims abstract description 15
- 241001465754 Metazoa Species 0.000 claims abstract description 15
- 229940004208 lactobacillus bulgaricus Drugs 0.000 claims abstract description 15
- 241000186000 Bifidobacterium Species 0.000 claims abstract description 14
- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 claims abstract 8
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 10
- 241000194020 Streptococcus thermophilus Species 0.000 abstract description 5
- 238000002360 preparation method Methods 0.000 abstract description 5
- 238000012545 processing Methods 0.000 abstract description 4
- 241000186660 Lactobacillus Species 0.000 abstract description 3
- 229940039696 lactobacillus Drugs 0.000 abstract description 3
- 235000013365 dairy product Nutrition 0.000 abstract description 2
- 230000001580 bacterial effect Effects 0.000 description 23
- 239000003795 chemical substances by application Substances 0.000 description 17
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 14
- 229920001221 xylan Polymers 0.000 description 13
- 239000002994 raw material Substances 0.000 description 9
- 239000007788 liquid Substances 0.000 description 8
- 150000004823 xylans Chemical class 0.000 description 8
- 244000199885 Lactobacillus bulgaricus Species 0.000 description 7
- 239000001963 growth medium Substances 0.000 description 7
- 239000004310 lactic acid Substances 0.000 description 7
- 235000014655 lactic acid Nutrition 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- IDCPFAYURAQKDZ-UHFFFAOYSA-N 1-nitroguanidine Chemical compound NC(=N)N[N+]([O-])=O IDCPFAYURAQKDZ-UHFFFAOYSA-N 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 239000000796 flavoring agent Substances 0.000 description 6
- 235000019634 flavors Nutrition 0.000 description 6
- 238000011081 inoculation Methods 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 5
- -1 MRS xylan Chemical class 0.000 description 5
- 210000000481 breast Anatomy 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 235000013336 milk Nutrition 0.000 description 5
- 239000008267 milk Substances 0.000 description 5
- 210000004080 milk Anatomy 0.000 description 5
- 231100000350 mutagenesis Toxicity 0.000 description 5
- 238000002703 mutagenesis Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 230000001954 sterilising effect Effects 0.000 description 5
- 238000004659 sterilization and disinfection Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 230000002068 genetic effect Effects 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 235000020122 reconstituted milk Nutrition 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 241001478240 Coccus Species 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 229960003692 gamma aminobutyric acid Drugs 0.000 description 3
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 3
- 235000020256 human milk Nutrition 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 235000011194 food seasoning agent Nutrition 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 229940099596 manganese sulfate Drugs 0.000 description 2
- 239000011702 manganese sulphate Substances 0.000 description 2
- 235000007079 manganese sulphate Nutrition 0.000 description 2
- BZDIAFGKSAYYFC-UHFFFAOYSA-N manganese;hydrate Chemical compound O.[Mn] BZDIAFGKSAYYFC-UHFFFAOYSA-N 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 238000009928 pasteurization Methods 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 235000021262 sour milk Nutrition 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- WTLKTXIHIHFSGU-UHFFFAOYSA-N 2-nitrosoguanidine Chemical compound NC(N)=NN=O WTLKTXIHIHFSGU-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 201000004384 Alopecia Diseases 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 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 description 1
- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 239000005862 Whey Substances 0.000 description 1
- 102000007544 Whey Proteins Human genes 0.000 description 1
- 108010046377 Whey Proteins Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 235000015140 cultured milk Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000000280 densification Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- 150000002475 indoles Chemical class 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 230000037323 metabolic rate Effects 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 230000000394 mitotic effect Effects 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000001009 osteoporotic effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 210000004243 sweat Anatomy 0.000 description 1
- 235000012976 tarts Nutrition 0.000 description 1
- 230000035922 thirst Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Dairy Products (AREA)
Abstract
The invention belongs to dairy produce processing and manufacturing field, relate to a kind of preparation method of Yoghourt, described mixed culture fermentation agent is Lactobacillus plantarum, lactobacillus bulgaricus and animal bifidobacteria composition; Described mix bacterium agent: the mass ratio of Lactobacillus plantarum, lactobacillus bulgaricus and animal bifidobacteria is 1:0.2-0.5:0.1-0.2; Lactobacillus plantarum (Lactobacillus? plantarum) Li-2013-01, does is deposit number CGMCC? No.7928, the present invention adopts Lactobacillus plantarum, streptococcus thermophilus, three kinds of bacterium of lactobacillus bulgaricus to carry out the cooperative fermentation of Yoghourt, and the yoghurt of producing also has the feature that viscosity is high, retentiveness is strong, rear acidifying is good.
Description
Technical field
The invention belongs to dairy produce processing and manufacturing field, relate in particular to a kind of preparation method of Yoghourt.
Background technology
Yoghourt is taking fresh breast or reconstituted milk as raw material, adds wherein lactobacillus-fermented after pasteurizeAgent, about 40 DEG C fermentations, the fermented milk goods that are prepared from. Yoghourt be divided into again pure yogurt, seasoning yogurt,Fruit is expected yogurt. Pure yogurt refers to taking breast or reconstituted milk as raw material, the product of making through inoculation fermentation; Seasoning yogurtTaking the breast of 65-80% or reconstituted milk as raw material, through inoculation fermentation or after add other raw material systems such as flavor enhancementThe product becoming; And fruit expects that yogurt refers to that more than 50-80% breast or reconstituted milk are raw material, add before or after inoculation fermentationThe product made from other raw materials such as flavor enhancement, fruits and vegetables, cereal. All kinds of sour milk products are with its nutrition, comfortableColor and luster, tasty and refreshing mouthfeel of quenching one's thirst and enrich tempting local flavor, extremely liking of consumers in general, exists in recent yearsChina's development is very fast. In addition, at present in vast samll cities and towns of China and rural area, the production and consumption of sour milk productsGrow out of nothing, accepted by people gradually.
Yoghourt not only mouthfeel is good, and its nutritive value and health care more merit attention. It is easy that Yoghourt has the appetite of raisingIn digesting, fall end cholesterol, prevention and treatment constipation, anti-cancer, the osteoporotic effect of control. Such as:The patent of invention that application number is " 201210009973.2 " discloses a kind of preparation method of the GABA of being rich in Yoghourt,After being activated, expand cultivation, the bacterial classification of high yield GABA and conventional bacterial strain be mixed for fermented yoghourt,And a kind of safe, new type of health type Yoghourt that GABA content is high of production department; Application number isThe patent of invention of " 200910223090.X " discloses a kind of Yoghourt and production method thereof containing profitable probliotics, logicalCross lactobacillus acidophilus, Bifidobacterium and basic bacterial classification at a lower temperature, jointly fermenting and producing obtains, and hasConventionally enteron aisle, improve flora ecological environment, promote the effect that nutritional labeling is digested and assimilated.
Generally, when the raw materials for production of yoghurt and apparatus and process fixing after, the quality of strain quality just becomesDetermine the leading factor of yogurt quality, and the bacillus that the quality of bacterial classification depends primarily on leavening in Yoghourt withThe quantitative proportion of coccus, yogurt fermentation adopts the multiculture of lactobacillus bulgaricus and streptococcus thermophilus conventionally,Going down to posterity in preservation process of bacterial classification, unstable due to bacterial classification, often easily encounters bacillus and coccus ratio and losesThe problem of weighing apparatus, curdled appearance is poor in process of production, mouthfeel is sour and astringent, produce fragrant deficiency or acidity deficiency to cause YoghourtNot outstanding with yoghurt typical case local flavor.
Summary of the invention
The object of the invention is, in order to overcome above-mentioned defect, provides a kind of Yoghourt fermentation mix bacterium agent.
For achieving the above object, the technical solution used in the present invention is as follows:
Yoghourt fermentation is composed as follows with mix bacterium agent: described mixed culture fermentation agent is Lactobacillus plantarum, bulgarian milkBacillus and animal bifidobacteria composition;
Described mix bacterium agent: the mass ratio of Lactobacillus plantarum, lactobacillus bulgaricus and animal bifidobacteriaFor 1:0.2-0.5:0.1-0.2;
A preparation method who adopts mix bacterium agent fermented yoghourt, comprises the steps:
Raw material processing: the examination of former milk, sterilization, storage, standardization, batching, degassed, homogeneous;
Re-pasteurization:
Cooling, inoculation: the material after sterilization is cooled to 43-45 DEG C, adds mixed fermenting agent, and addition isThe 2%-4% of material.
Stir fermentation:.
Cooling, broken curdled milk, filling; Cooling, after-ripening.
Beneficial effect:
The present invention is on the basis of having inherited traditional streptococcus thermophilus and lactobacillus bulgaricus and carry out Yoghourt fermentationUpper, increase and produced the Lactobacillus plantarum that acid amount is high, stability is strong, because Lactobacillus plantarum can be through passing continuouslyDai Shici, heredity is better, and itself also has the function that maintains colony balance, therefore collaborative streptococcus thermophilusWith lactobacillus bulgaricus, can improve the stability of overall bacterial classification, effectively prevent bacillus and the mistake of coccus ratioThe problem of weighing apparatus, thus the quality problems such as Yoghourt curdled appearance is poor, mouthfeel is sour and astringent well solved; Plant breast barBacterium can also produce a large amount of aldehydes and Ester in production sweat, makes Yoghourt tart flavour suitable refreshingMouthful, and there is the outstanding yoghurt local flavor of typical case; Meanwhile, the Lactobacillus plantarum sour power of strong product during the fermentation,Can accelerate the fermenting speed of yoghurt, shorten the production cycle, save the cost of the aspects such as manpower, energy consumption, equipment.
The present invention adopts Lactobacillus plantarum, streptococcus thermophilus, three kinds of bacterium of lactobacillus bulgaricus to carry out the association of YoghourtWith fermentation, the yoghurt of producing also has the feature that viscosity is high, retentiveness is strong, rear acidifying is good.
Specific implementation method
The present invention will be described further by following examples.
Lactobacillus plantarum provided by the present invention (Lactobacillusplantarum) Li-2013-01, this bacteriumStrain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCCNo.7928, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Chinese Academy of Sciences microorganism is groundStudy carefully institute, postcode 100101, preservation date on July 15th, 2013.
This bacterial strain feature is as follows: examine under a microscope, this bacterial strain is rod-short, and Gram's staining is positive,Atrichia, does not produce gemma; On solid medium, bacterium colony is white, smooth surface, and densification, form is circleShape, edge is more neat. Physicochemical characteristics is: catalase (-), and gelatin liquefaction (-), indoles experiment (+),Motility (-), fermentation gas (-), nitrite reduction (-), produces hydrogen sulfide gas (-), pH4.5Growth (+) in MRS culture medium.
Lactobacillus plantarum of the present invention adopts following flow process to carry out seed selection:
Original bacterial classification → test tube activation → dithyl sulfate (DES) mutagenesis → dull and stereotyped primary dcreening operation → Asia of setting outThe test of sieve again → mitotic stability of nitroguanidine (NTG) mutagenesis → dull and stereotyped primary dcreening operation → shaking flask.
The original bacterial classification that sets out is CICC20242, is purchased from Chinese industrial microorganism fungus kind preservation administrative center.
Original strain of the present invention is in xylan culture medium, and the output of lactic acid is 12.5g/L. ForImprove its lactic acid production, adopt successively DES and NTG to carry out mutagenesis to this bacterial classification, mutagenesis adopts MRS calcium carbonate flatPlate carries out primary dcreening operation, then adopts 500mL shake flask fermentation, and biosensor analysis instrument carries out multiple sieve to Producing Strain,The lactobacillus plantarum strain that seed selection is good, the experiment of then going down to posterity, evaluates its genetic stability.
Bacterial strain CGMCCNo.7928 genetic stability result shows: through continuous passage ten times, properties refers toMark all more stablely, heredity is better, the object bacterium therefore bacterial strain CGMCCNo.7928 being obtained as seed selectionStrain.
Object bacterial strain CGMCCNo.7928 is done to the experiment of 10L fermentation tank, and result shows: after fermentation 72h, with woodGlycan is carbon source, and the lactic acid concn of Lactobacillus plantarum CGMCCNo.7928 can reach 57g/L, with starting strainCompare and improved 356%.
Object bacterial strain CGMCCNo.7928 is done to the experiment of 10L fermentation tank, and result shows: after fermentation 72h, with PortugalGrape sugar is carbon source, and the lactic acid concn of Lactobacillus plantarum CGMCCNo.7928 can reach 68g/L.
Lactobacillus bulgaricus of the present invention and streptococcus thermophilus are bacterium powder or the microbial inoculum on market, sold.
Animal bifidobacteria CICC21711 is purchased from Chinese industrial microorganism fungus kind preservation center.
Embodiment 1
The concrete Breeding Process of described Lactobacillus plantarum is as follows:
1. dithyl sulfate (DES) mutagenic and breeding
(1) on super-clean bench, get Lactobacillus plantarum one ring on test tube slant, 50mL liquid MRS is equipped with in accessIn the 250mL triangular flask of xylan culture medium, 200rpm, cultivates 12h left and right, makes thalline in right for 40 DEG CNumber growth in earlier stage.
(2) get 5mL bacterium liquid, the centrifugal 10min of 5000rpm collects thalline, with physiological saline washing 2 times.
(3) be diluted to 107/mL bacteria suspension with pH7.0 phosphate buffer.
(4) kaliumphosphate buffer, 8mL bacteria suspension, the 0.4mLDES that get 32mLpH7.0 are putting in advanceIn the 150mL triangular flask of rotor, fully mix, making DES ultimate density is 1%(v/v).
(5) 150rpm reaction 30min in 30 DEG C of shaking tables, gets 1mL mixed liquor, adds 0.5mL25%Na2S2O3Solution stopped reaction.
(6) dilution spread is in the MRS xylan screening solid medium plate containing 90g/L xylan. ?Cultivate the bacterial strain of picking transparent circle/colony diameter maximum after 2-3 days for 40 DEG C, label is DES bacterium.
2. nitrosoguanidine mutagenesis
(1) on super-clean bench, get Lactobacillus plantarum DES mono-ring on test tube slant, 50mL liquid is equipped with in accessIn the 250mL triangular flask of MRS xylan culture medium, 200rpm, cultivates about 12h for 40 DEG C, make thalline inLogarithmic growth in earlier stage.
(2) get the centrifugal 10min of 5mL bacterium liquid 5000rpm and collect thalline, with physiological saline washing 2 times.
(3) be diluted to 10 with pH6.0 phosphate buffer7Individual/mL bacteria suspension.
(4) get 10mL bacteria suspension and be transferred in 100mL triangular flask, add the NTG of 10mg, be mixed with endConcentration is the NTG solution of 10mg/mL, and adds 4-5 to drip acetone, is beneficial to NTG and dissolves.
(5) 200rpm oscillating reactions 30min at 30 DEG C, the centrifugal 10min of 5000rpm collects thalline,By SPSS washing several, stopped reaction.
(6) by after 5 times of bacterium liquid dilutions, get last dilution bacterium liquid 0.2mL, dilution spread is in containing 90g/LIn the MRS xylan screening solid medium plate of xylan. 40 DEG C cultivate picking transparent circle after 2-3 days/150 of the larger bacterial strains of colony diameter.
3. shaking flask is sieved again
(1) on super-clean bench, get respectively Lactobacillus plantarum one ring on each test tube slant, 50mL is equipped with in accessIn the 250mL triangular flask of liquid MRS xylan culture medium, 200rpm, cultivates 3-4 days for 40 DEG C, and detect every dayConcentration of glucose and Pfansteihl change in concentration. After fermentation ends, relatively the xylan of 150 strain bacterial classifications consumes speedRate and lactic acid produce conversion ratio and the heteroacid content of speed, lactic acid.
(2) selection xylan metabolic rate is fast, lactic acid concn is high, conversion ratio is high and the poor bacterium of heteroacidPlant as final bacterial classification called after Li bacterium.
4. genetic stability test
Li-2013-01 bacterial strain is gone down to posterity for continuous ten times on inclined-plane, and the method for sieving again by shaking flask detects eachFermentation situation after going down to posterity. Experiment is found, on inclined-plane, goes down to posterity for continuous ten times, and this bacterial classification proterties does not obviously becomeChange, property indices is all normal, illustrates that the genetic stability of this bacterial classification is stronger.
Described culture medium consists of: liquid MRS xylan culture medium (beef extract 2g, peptone 10g, yeastCream 5g, xylan 20g, sodium acetate 5g, ammonium citrate 2g, dipotassium hydrogen phosphate 2g, epsom salt 0.2g,Seven water manganese sulfate 0.05g, after dissolving one by one, running water constant volume 1000mL, regulates pH7.1); MRS xylanScreening solid medium (beef extract 2g, peptone 10g, yeast extract 5g, xylan 90g, sodium acetate 5g,Ammonium citrate 2g, dipotassium hydrogen phosphate 2g, epsom salt 0.2g, seven water manganese sulfate 0.05g, dissolve one by oneAfter, running water constant volume 1000mL, regulates pH7.1, adds 20g agar);
Embodiment 2:
Yoghourt fermentation is composed as follows with mix bacterium agent: described mixed culture fermentation agent is Lactobacillus plantarum, bulgarian milkBacillus and animal bifidobacteria composition;
Described mix bacterium agent: the mass ratio of Lactobacillus plantarum, lactobacillus bulgaricus and animal bifidobacteriaFor 1:0.5:0.1;
A preparation method for Yoghourt, comprises the steps:
Raw material processing: the examination of former milk, sterilization, storage, standardization, batching, degassed, homogeneous;
Re-pasteurization: after homogeneous, material carries out sterilization by heat exchanger, condition is 95 DEG C, 300s;
Cooling, inoculation: the material after sterilization is cooled to 43-45 DEG C, adds hybrid working fermenting agent, addsAmount is 2% of material.
Stir fermentation: after inoculation work completes, open and stir, after stirring 15min, stop, fermentation jar temperature is protectedBe held in 45 DEG C, ferment 4 hours, pH is 4.5 o'clock, stops fermentation.
Cooling, broken curdled milk, filling: when the acidity of milk reaches requirement in fermentation tank, fall through plate type heat exchangerTemperature is to 17 DEG C, then by mechanical force destroy fermentation-causing curdling colloid, gelinite particle diameter reaches 0.2mm, carries outFilling, the temperature fall time of every tank material 55 minutes. After filling completing, be cooled to rapidly 9 DEG C of left and right with cold wind,Be placed on and in 0 DEG C of freezer, carry out refrigeration after-ripening in 24 hours, and product is cooled to 4 DEG C at warehouse-in in latter 8 hours.
This product shelf-life under 10 DEG C of refrigerated conditions is 23 days. Within the shelf-life, viscosity is stabilized in 14200cp,Titratable acidity is stabilized in 86 ° of T, separates out without whey.
Embodiment 3: Yoghourt fermentation is composed as follows with mix bacterium agent: described mixed culture fermentation agent is Lactobacillus plantarum, guarantorAdd Leah lactobacillus and animal bifidobacteria composition; Described mix bacterium agent: Lactobacillus plantarum, bulgarian milkThe mass ratio of bacillus and animal bifidobacteria is 1:0.2:0.1.
Embodiment 4: Yoghourt fermentation is composed as follows with mix bacterium agent: described mixed culture fermentation agent is Lactobacillus plantarum, guarantorAdd Leah lactobacillus and animal bifidobacteria composition; Described mix bacterium agent: Lactobacillus plantarum, bulgarian milkThe mass ratio of bacillus and animal bifidobacteria is 1:0.5:0.2.
Claims (4)
1. a Yoghourt fermentation mix bacterium agent, described mixed culture fermentation agent by Lactobacillus plantarum, lactobacillus bulgaricus andAnimal bifidobacteria composition; Described mix bacterium agent: Lactobacillus plantarum, lactobacillus bulgaricus and animal bifidThe mass ratio of bacillus is 1:0.2-0.5:0.1-0.2; Described Lactobacillus plantarum is Lactobacillus plantarum(Lactobacillusplantarum) Li-2013-01, deposit number is CGMCCNo.7928.
2. a kind of Yoghourt fermentation mix bacterium agent according to claim 1, is characterized in that: Lactobacillus plantarum, lactobacillus bulgaricus and animal bifidobacteria mass ratio be 1:0.5:0.1.
3. a kind of Yoghourt fermentation mix bacterium agent according to claim 1, is characterized in that: described Lactobacillus plantarum,The mass ratio of lactobacillus bulgaricus and animal bifidobacteria is 1:0.2:0.1.
4. a kind of Yoghourt fermentation mix bacterium agent according to claim 1, is characterized in that: described Lactobacillus plantarum,The mass ratio of lactobacillus bulgaricus and animal bifidobacteria is 1:0.5:0.2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310741754.8A CN103734310B (en) | 2013-12-28 | 2013-12-28 | A kind of novel Yoghourt fermentation mix bacterium agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310741754.8A CN103734310B (en) | 2013-12-28 | 2013-12-28 | A kind of novel Yoghourt fermentation mix bacterium agent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103734310A CN103734310A (en) | 2014-04-23 |
| CN103734310B true CN103734310B (en) | 2016-05-25 |
Family
ID=50491447
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310741754.8A Active CN103734310B (en) | 2013-12-28 | 2013-12-28 | A kind of novel Yoghourt fermentation mix bacterium agent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103734310B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105053201B (en) * | 2015-08-25 | 2018-07-13 | 东北农业大学 | A kind of preparation method and high-quality Yoghourt of yoghourt stabilizer |
| CN107509811B (en) * | 2016-06-17 | 2021-01-15 | 扬州市扬大康源乳业有限公司 | Preparation method of yogurt |
| CN106212674A (en) * | 2016-08-05 | 2016-12-14 | 内蒙古普泽生物制品有限责任公司 | A kind of Yoghourt fermentation mix bacterium agent |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009065862A1 (en) * | 2007-11-23 | 2009-05-28 | Dsm Ip Assets B.V. | Improved bioactive peptide production |
| CN102191192A (en) * | 2010-12-21 | 2011-09-21 | 北京博锦元生物科技有限公司 | Animal Bifidobacterium and use method thereof |
| CN102453689A (en) * | 2011-12-06 | 2012-05-16 | 光明乳业股份有限公司 | Lactobacillus plantarum strain producing extracellular polysaccharide, and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5438448B2 (en) * | 2009-09-24 | 2014-03-12 | 大日本スクリーン製造株式会社 | Spatial modulated light generation apparatus and image recording apparatus |
-
2013
- 2013-12-28 CN CN201310741754.8A patent/CN103734310B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009065862A1 (en) * | 2007-11-23 | 2009-05-28 | Dsm Ip Assets B.V. | Improved bioactive peptide production |
| CN102191192A (en) * | 2010-12-21 | 2011-09-21 | 北京博锦元生物科技有限公司 | Animal Bifidobacterium and use method thereof |
| CN102453689A (en) * | 2011-12-06 | 2012-05-16 | 光明乳业股份有限公司 | Lactobacillus plantarum strain producing extracellular polysaccharide, and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103734310A (en) | 2014-04-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104293706B (en) | Pickle production microbial bacterial agent | |
| CN101580809B (en) | Method for preparing direct vat set Lactobacillus casei subsp. rhamnosus freeze-dried powder | |
| CN101338283B (en) | Lactobacillus casei and applications thereof in solid-state fermentation | |
| CN103734311B (en) | A kind of preparation method of Yoghourt | |
| JP4772131B2 (en) | Method for producing fermented milk using novel lactic acid bacteria | |
| CN101589829B (en) | Probiotics fermented beverage and production method thereof | |
| CN102604833B (en) | Preparation method and product of multiple lactic acid bacteria fermentation agent for fermented milk | |
| CN105483050B (en) | One lactobacillus plantarum and its application in fermented feed | |
| CN102599245A (en) | Set-style fermented yoghourt rich in probiotics and preparation method of yoghourt | |
| CN104312941A (en) | Vegetable fermentation composite bacterial agent product | |
| CN104719467B (en) | A kind of preparation method of Yoghourt | |
| CN112126599B (en) | High-density culture method of lactobacillus helveticus, preparation of high-activity bacterium powder and application of high-density culture method | |
| CN103120208A (en) | Lactobacillus plantarum fermented milk and preparation method thereof | |
| CN102321550B (en) | Lactococcus lactis fermented at low temperature and method for preparing yoghurt by using Lactococcus lactis | |
| CN101586087B (en) | Acid sensitivity lactobacillus bulgaricus strain and usage thereof | |
| CN103766483B (en) | A kind of preparation method of Yoghourt | |
| CN103734310B (en) | A kind of novel Yoghourt fermentation mix bacterium agent | |
| CN103636779A (en) | Functional fermented milk and preparation method thereof | |
| CN115281247A (en) | Lactobacillus fermentum normal-temperature yogurt and preparation method thereof | |
| CN115136986A (en) | Lactobacillus fermentum solid beverage and preparation method thereof | |
| CN104673692A (en) | Composite microbial preparation for pickle production and preparation method thereof | |
| CN104388332A (en) | Composite bacterial agent product for fermentation of pickled vegetable and preparation method thereof | |
| CN108013134A (en) | A kind of acidified milk rich in lactobacillus plantarum ST-III and preparation method thereof | |
| CN110760458B (en) | Complex function microbial inoculum for fermentation | |
| CN105962337A (en) | Method of preparing probiotic food using produced slurry in production process of bean vermicelli |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| CB03 | Change of inventor or designer information |
Inventor after: Deng Ping Inventor after: Xiao Songyi Inventor after: Cao Wenhai Inventor before: Shao Suying Inventor before: Kong Rixiang |
|
| COR | Change of bibliographic data | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160920 Address after: 410000 room 1402, Jinzhou Avenue venture building, Jing Jing District, Changsha, Hunan, Ningxiang Patentee after: ROYAL GROUP HUNAN USHI DAIRY CO.,LTD. Address before: Tianhe District Tong East Road Guangzhou city Guangdong province 510665 B-101 No. 5, room B-118 Patentee before: GUANGDONG GAOHANG INTELLECTUAL PROPERTY OPERATION Co.,Ltd. Effective date of registration: 20160920 Address after: Tianhe District Tong East Road Guangzhou city Guangdong province 510665 B-101 No. 5, room B-118 Patentee after: GUANGDONG GAOHANG INTELLECTUAL PROPERTY OPERATION Co.,Ltd. Address before: 100079 Beijing City, Fengtai District, No. seven, No. 1603, room 9, No. 44 Patentee before: Shao Suying |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A new mixed bacterial agent for yoghurt fermentation Effective date of registration: 20210917 Granted publication date: 20160525 Pledgee: Ningxiang Hunan rural commercial bank Limited by Share Ltd. Pledgor: ROYAL GROUP HUNAN USHI DAIRY Co.,Ltd. Registration number: Y2021980009485 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20231008 Granted publication date: 20160525 Pledgee: Ningxiang Hunan rural commercial bank Limited by Share Ltd. Pledgor: ROYAL GROUP HUNAN USHI DAIRY CO.,LTD. Registration number: Y2021980009485 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right |