CN103734310A - Novel mixed fungicide for fermenting yogurt - Google Patents
Novel mixed fungicide for fermenting yogurt Download PDFInfo
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- CN103734310A CN103734310A CN201310741754.8A CN201310741754A CN103734310A CN 103734310 A CN103734310 A CN 103734310A CN 201310741754 A CN201310741754 A CN 201310741754A CN 103734310 A CN103734310 A CN 103734310A
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- lactobacillus plantarum
- lactobacillus
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- yogurt
- yoghourt
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- 235000013618 yogurt Nutrition 0.000 title claims abstract description 44
- 230000000855 fungicidal effect Effects 0.000 title abstract 3
- 239000000417 fungicide Substances 0.000 title abstract 3
- 238000000855 fermentation Methods 0.000 claims abstract description 38
- 230000004151 fermentation Effects 0.000 claims abstract description 38
- 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
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 claims abstract description 21
- 229940004208 lactobacillus bulgaricus Drugs 0.000 claims abstract description 21
- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 claims abstract 8
- 241000894006 Bacteria Species 0.000 claims description 30
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- 238000003754 machining Methods 0.000 abstract 1
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- 239000003795 chemical substances by application Substances 0.000 description 18
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- 239000001963 growth medium Substances 0.000 description 7
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- 238000001816 cooling Methods 0.000 description 5
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- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 4
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- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 3
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- 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
- 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 2
- 239000001888 Peptone Substances 0.000 description 2
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- 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
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
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Abstract
The invention belongs to the field of machining and manufacturing of a dairy product, and relates to a preparation method of yogurt. A mixed fungicide is prepared from lactobacillus plantarum, lactobacillus bulgaricus and bifidobacterium animalis, wherein the mass ratio of the lactobacillus plantarum to the lactobacillus bulgaricus to the bifidobacterium animalis in the mixed fungicide is 1:(0.2-0.5):(0.1-0.2); the lactobacillus plantarum is Li-2013-01 with the preservation number being CGMCC No.7928. Cooperative fermentation of the yogurt is carried out by adopting the lactobacillus plantarum, streptococcus thermophilus and the lactobacillus bulgaricus, and the generated yogurt also has the characteristics of being high in viscosity, strong in water binding capacity and good in post-acidification.
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 take fresh breast or reconstituted milk as raw material, adds wherein lactic acid bacteria fermenting agent, about 40 ℃ fermentations, the fermented milk goods that are prepared from after pasteurize.Yoghourt is divided into again pure yogurt, seasoning yogurt, fruit is expected yogurt.Pure yogurt refers to take breast or reconstituted milk as raw material, the product of making through inoculation fermentation; Seasoning yogurt is take the breast of 65-80% or reconstituted milk as raw material, through inoculation fermentation or after add the product that other raw materials such as flavor enhancement are made; And fruit expects that yogurt refers to that more than 50-80% breast or reconstituted milk are raw material, the product that other raw materials such as the front or rear interpolation of inoculation fermentation flavor enhancement, fruits and vegetables, cereal are made.All kinds of sour milk products are with its nutrition, comfortable color and luster, tasty and refreshing mouthfeel of quenching one's thirst and enrich tempting local flavor, and extremely liking of consumers in general, very fast in China's development in recent years.In addition, at present in the vast samll cities and towns of China and rural area, the production and consumption of sour milk products is grown out of nothing, and is accepted gradually by people.
Yoghourt not only mouthfeel is good, and its nutritive value and health care more merit attention.Yoghourt has the appetite of raising and is easy to digest, falls 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; The patent of invention that application number is " 200910223090.X " discloses a kind of Yoghourt and production method thereof containing profitable probliotics, by lactobacillus acidophilus, Bifidobacterium and basic bacterial classification at a lower temperature, common fermentation and producing obtains, and has common enteron aisle, improves flora ecological environment, promotes 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 becomes the leading factor that determines yogurt quality, and the quality of bacterial classification depends primarily on the bacillus of leavening and the quantitative proportion of coccus in Yoghourt, 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 encounter the problem of bacillus and coccus proportional imbalance, curdled appearance is poor in process of production to cause Yoghourt, mouthfeel is sour and astringent, producing the not enough and yoghurt typical case local flavor of fragrant deficiency or acidity does not give prominence to.
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, 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;
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 ℃, adds mixed fermenting agent, the 2%-4% that addition is material.
Stir fermentation:.
Cooling, broken curdled milk, filling; Cooling, after-ripening.
Beneficial effect:
The present invention carries out on the basis of Yoghourt fermentation having inherited traditional streptococcus thermophilus and lactobacillus bulgaricus, increased and produced the Lactobacillus plantarum that acid amount is high, stability is strong, because Lactobacillus plantarum can be through continuous passage ten times, heredity is better, itself also has the function that maintains colony balance, therefore collaborative streptococcus thermophilus and lactobacillus bulgaricus, can improve the stability of overall bacterial classification, effectively prevent the problem of bacillus and coccus proportional imbalance, thereby well solved the quality problems such as Yoghourt curdled appearance is poor, mouthfeel is sour and astringent; Lactobacillus plantarum can also produce a large amount of aldehydes and Ester in production sweat, makes Yoghourt tart flavour suitable tasty and refreshing, and has 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, shortens the production cycle, saves 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 cooperative fermentation of Yoghourt, and 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 (Lactobacillus plantarum) Li-2013-01, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.7928, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, 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, and atrichia does not produce gemma; On solid medium, bacterium colony is white, smooth surface, and densification, form is circular, edge is more neat.Physicochemical characteristics is: catalase (-), and gelatin liquefaction (-), indoles experiment (+), motility (-), fermentation gas (-), nitrite reduction (-), produces hydrogen sulfide gas (-), growth (+) in pH4.5MRS culture medium.
Lactobacillus plantarum of the present invention adopts following flow process to carry out seed selection:
Original sieve again → mitotic stability of bacterial classification → test tube activation → dithyl sulfate (DES) mutagenesis → dull and stereotyped primary dcreening operation → nitrosoguanidine (NTG) mutagenesis → dull and stereotyped primary dcreening operation → shaking flask test of setting out.
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.In order to improve its lactic acid production, adopt successively DES and NTG to carry out mutagenesis to this bacterial classification, mutagenesis adopts MRS calcium carbonate flat board to carry out primary dcreening operation, then adopt 500mL shake flask fermentation, 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 CGMCC No.7928 genetic stability result shows: through continuous passage ten times, property indices is all more stable, and heredity is better, the object bacterial strain therefore bacterial strain CGMCC No.7928 being obtained as seed selection.
Object bacterial strain CGMCC No.7928 is done to the experiment of 10L fermentation tank, and result shows: after fermentation 72h, take xylan as carbon source, the lactic acid concn of Lactobacillus plantarum CGMCC No.7928 can reach 57g/L, has improved 356% compared with starting strain.
Object bacterial strain CGMCC No.7928 is done to the experiment of 10L fermentation tank, and result shows: after fermentation 72h, take glucose as carbon source, the lactic acid concn of Lactobacillus plantarum CGMCC No.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 CICC 21711 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, access is equipped with in the 250mL triangular flask of 50mL liquid MRS xylan culture medium, and 200rpm cultivates 12h left and right for 40 ℃, makes thalline in logarithmic growth in earlier stage.
(2) get 5mL bacterium liquid, the centrifugal 10min of 5000rpm collects thalline, with physiological saline washing 2 times.
(3) with pH7.0 phosphate buffer, be diluted to 107/mL bacteria suspension.
(4) kaliumphosphate buffer, 8mL bacteria suspension, 0.4mLDES of getting 32mLpH7.0 fully mixes at the 150mL triangular flask of putting in advance rotor, and making DES ultimate density is 1%(v/v).
(5) 150rpm reaction 30min in 30 ℃ of shaking tables, gets 1mL mixed liquor, adds 0.5mL25%Na
2s
2o
3solution stopped reaction.
(6) dilution spread is in the MRS xylan screening solid medium plate containing 90g/L xylan.At the bacterial strain of 40 ℃ of cultivations picking transparent circle/colony diameter maximum after 2-3 days, label is DES bacterium.
2. nitrosoguanidine mutagenesis
(1) on super-clean bench, get Lactobacillus plantarum DES mono-ring on test tube slant, access is equipped with in the 250mL triangular flask of 50mL liquid MRS xylan culture medium, and 200rpm cultivates 12h left and right for 40 ℃, makes thalline in logarithmic growth in earlier stage.
(2) get the centrifugal 10min of 5mL bacterium liquid 5000rpm and collect thalline, with physiological saline washing 2 times.
(3) with pH6.0 phosphate buffer, be diluted to 10
7individual/mL bacteria suspension.
(4) get 10mL bacteria suspension and be transferred in 100mL triangular flask, add the NTG of 10mg, being mixed with final concentration 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 ℃, 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 the MRS xylan screening solid medium plate containing 90g/L xylan.150 of 40 ℃ of cultivation bacterial strains that after 2-3 days, picking transparent circle/colony diameter is larger.
3. shaking flask is sieved again
(1) on super-clean bench, get respectively Lactobacillus plantarum one ring on each test tube slant, access is equipped with in the 250mL triangular flask of 50mL liquid MRS xylan culture medium, and 200rpm cultivates 3-4 days, detects concentration of glucose and Pfansteihl change in concentration every day for 40 ℃.After fermentation ends, relatively the xylan wear rate of 150 strain bacterial classifications 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 bacterial classification of heteroacid is 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 the fermentation situation after at every turn going down to posterity.Experiment discovery is gone down to posterity for continuous ten times on inclined-plane, and this bacterial classification proterties does not have significant change, and 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, yeast extract 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 xylan screening 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, after dissolving one by one, running water constant volume 1000mL, regulate pH7.1, add 20g agar);
Embodiment 2:
Yoghourt fermentation is composed as follows with mix bacterium agent: 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 1:0.5:0.1;
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 ℃, 300s;
Cooling, inoculation: the material after sterilization is cooled to 43-45 ℃, adds hybrid working fermenting agent, addition is material 2%.
Stir fermentation: after inoculation work completes, open and stir, after stirring 15min, stop, fermentation jar temperature remains on 45 ℃, ferments 4 hours, and pH is, stops fermentation at 4.5 o'clock.
Cooling, broken curdled milk, filling: when the acidity of milk reaches requirement in fermentation tank, through plate type heat exchanger, be cooled to 17 ℃, then by mechanical force destroy fermentation-causing curdling colloid, gelinite particle diameter reaches 0.2mm, carry out filling, the temperature fall time of every tank material 55 minutes.After filling completing, be cooled to rapidly 9 ℃ of left and right with cold wind, be placed on and in 0 ℃ of freezer, carry out refrigeration after-ripening in 24 hours, and product is cooled to 4 ℃ at warehouse-in in latter 8 hours.
This product shelf-life under 10 ℃ of refrigerated conditions is 23 days.Within the shelf-life, viscosity is stabilized in 14200cp, and titratable acidity is stabilized in 86 ° of T, without whey, separates out.
Embodiment 3: Yoghourt fermentation is composed as follows with mix bacterium agent: 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 1:0.2:0.1.
Embodiment 4: Yoghourt fermentation is composed as follows with mix bacterium agent: 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 1:0.5:0.2.
Claims (4)
1. a Yoghourt fermentation mix bacterium agent, described mixed culture fermentation agent is comprised of Lactobacillus plantarum, lactobacillus bulgaricus and animal bifidobacteria; 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, deposit number is CGMCCNo.7928.
2. a kind of Yoghourt fermentation mix bacterium agent according to claim 1, is characterized in that: the mass ratio of Lactobacillus plantarum, lactobacillus bulgaricus and animal bifidobacteria is 1:0.5:0.1.
3. a kind of Yoghourt fermentation mix bacterium agent according to claim 1, is characterized in that: the mass ratio of described Lactobacillus plantarum, 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: the mass ratio of described Lactobacillus plantarum, lactobacillus bulgaricus and animal bifidobacteria is 1:0.5:0.2.
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| 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 |
| CN107509811A (en) * | 2016-06-17 | 2017-12-26 | 扬州市扬大康源乳业有限公司 | A kind of Yogurt and preparation method thereof |
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