CN106754482A - A kind of preparation method of the probiotics leaven for producing functional sugar - Google Patents
A kind of preparation method of the probiotics leaven for producing functional sugar Download PDFInfo
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- CN106754482A CN106754482A CN201611060789.5A CN201611060789A CN106754482A CN 106754482 A CN106754482 A CN 106754482A CN 201611060789 A CN201611060789 A CN 201611060789A CN 106754482 A CN106754482 A CN 106754482A
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- functional sugar
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- probiotics leaven
- fermentation
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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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
Abstract
The present invention provides a kind of preparation method of the probiotics leaven for producing functional sugar, and obtained leavening is suitable as industrial fermentation agent with producing acid, producing the premium properties such as fragrant, the antiphagin pollution of viscous, product.A kind of preparation method of the probiotics leaven for producing functional sugar, comprises the following steps：Using lactobacillus gasseri, lactobacillus reuteri or Lactobacillus plantarum as fermentation strain; by the fermentation strain be linked into proliferated culture medium and stage by stage feed supplement to add carbon source; in 35 ~ 40 DEG C of 8 ~ 16h of anaerobic fermentation culture; then aseptically centrifugation obtains lactobacillus cell; 1 ~ 3h of pre-freeze at the lactobacillus cell and freeze drying protectant are blended in into 55 ~ 45 DEG C, then freeze-drying is obtained leavening solid-state bacterium powder.
Description
Technical field
The present invention relates to the biological technology of preparing in fermented dairy product industry, the prebiotic of functional sugar is produced more particularly, to a kind of
The preparation method of bacterium leavening agent.
Background technology
Nearly 20 years, the dairy industry of China experienced rapid growth period, be developed with the growth rate every year more than 20%.
From the point of view of the kind of dairy products, fermented dairy product (including acidified milk, leben, probiotics fermention breast and probiotics breast drink
Material etc.) production and consumption be at present the maximum product category of increasing degree in China's dairy industry.The year of fermented dairy product
Growth rate reaches more than 20%, and still can keep this growth rate within considerable time from now on.2015, I
State's fermented dairy product industry size alreadys exceed 20,000,000,000 yuans, and yield breaks through 1,600,000 tons.Fermented dairy product not only into
To promote the leading products of China's dairy products industry development, and the important side of China's dairy industry product restructuring is turned into
To with development trend from now on.
Although China's fermented dairy product quickly grows, current each Dairy Enterprise production fermented dairy product of China is adopted
Lactic acid bacteria culturers and leavening is substantially all is monopolized by external enterprise and product, and 90% from several big hairs in Europe
The Danisco A/S BJ Rep Office of Jiao Ji companies, such as Denmark, Hansen Corp. of section, the DSM companies of Holland.But, leavening used is such as
Import is delivered directly strain and can only be intended for single use, and does not allow typically subculture to use, and production cost is domestic provide strain for oneself 1.5 times, and
And for a long time go down that the development of China's lactic acid bacteria fermenting agent and fermented dairy product will be restricted.For these reasons, the present invention is to one
The preparation method for planting the probiotics leaven for producing functional sugar is studied in detail.
The content of the invention
Regarding to the issue above, it is an object of the invention to provide a kind of preparation method of the probiotics leaven for producing functional sugar,
Obtained leavening has product acid, produces and glue, produce the premium properties such as perfume, antiphagin pollution, is suitable as industrial fermentation agent.
To reach above-mentioned purpose, the technical solution adopted by the present invention is as follows：
A kind of preparation method of the probiotics leaven for producing functional sugar, comprises the following steps：By lactobacillus gasseri, Luo Yishi breast bars
Bacterium or Lactobacillus plantarum as fermentation strain, by the fermentation strain be linked into proliferated culture medium and stage by stage feed supplement to add
Carbon source, in 35 ~ 40 DEG C of anaerobic fermentation 8 ~ 16h of culture, then aseptically centrifugation obtains lactobacillus cell, by the lactic acid
Bacterium cell and freeze drying protectant be blended in -55 ~ -45 DEG C at 1 ~ 3h of pre-freeze, then freeze-drying is obtained leavening solid-state bacterium powder.
Preferably, the inoculation part by weight of the fermentation strain is 3 ~ 7%.
Preferably, the proliferated culture medium includes the component of following weight portion：5 ~ 15 parts of sucrose, 5 ~ 15 parts of glucose, ferment
1 ~ 2 part of female powder, 1 ~ 2 part of tryptone, K2HPO43 ~ 5 parts, MgSO4•7H20.2 ~ 0.8 part of O, CaCl20.01 ~ 0.03 part, tell
80 0.5 ~ 1.5 parts of temperature.
It is highly preferred that the pH of the proliferated culture medium is 5 ~ 6.
Preferably, stream plus glucose after the 3 ~ 6h that ferments.
It is highly preferred that the concentration for flowing the glucose for adding is 3 ~ 7g/L.
Preferably, cell number is 1.0 × 10 in the proliferated culture medium9~ 6.5×109 cfu/ml。
Preferably, the freeze drying protectant includes the component of following weight portion：90 ~ 110 parts of skimmed milk power, sucrose 90 ~ 110
Part, 30 ~ 45 parts of glycerine, 45 ~ 55 parts of maltodextrin, 90 ~ 110 parts of trehalose, 8 ~ 12 parts of L-sodium, Cys 3 ~ 7
Part.
Preferably, the lactobacillus cell and freeze drying protectant are with 1：（0.5~1.5）Ratio mixing.
Preferably, the viable count in the leavening solid-state bacterium powder is 2.1 × 1010More than cfu/g.
The present invention has the following advantages that compared to existing technology：
（1）Exploitation leavening production technology and product, fill up the blank in the field, break the technology barriers of foreign countries and the ridge of product
It is disconnected；
（2）With reference to lactic acid bacteria characteristic in itself, exploitation high-efficient culture, preparation and active holding technology obtain the benefit of high-biomass
Raw bacterium and can to greatest extent keep probiotic active；
（3）Product prepared by the present invention can be applied to the multiple fields such as food, nutrient and healthcare products.
Specific embodiment
Presently preferred embodiments of the present invention is described in detail below so that advantages and features of the invention can be easier to by
It will be understood by those skilled in the art that.
Embodiment 1
Using Lactobacillus plantarum as fermentation strain, by 5% proliferated culture medium for being inoculated into pH5.5.Each component in proliferated culture medium
Concentration is as follows：The g/L of sucrose 10, the g/L of glucose 10, the g/L of dusty yeast 1.5, tryptone 1.5 g/L, K2HPO44.0 g/L,
MgSO4·7H2O 0.5 g/L, CaCl2 0.02 g/L, Tween 80 1mL.In 37 °C of anaerobic fermentation culture 12h, wherein in fermentation
Stream plus glucose after 6h, control concentration of glucose is in 5g/L.Cell number reaches 1.4 × 10 in final culture medium9Cfu/ml, will
Zymocyte liquid is aseptically centrifuged acquisition lactobacillus cell.Then 1 is pressed:1 ratio adds freeze drying protectant, freeze drying protectant
The concentration of middle each component is as follows：Skimmed milk power 100g/L, sucrose 100g/L, glycerine 30ml/L, maltodextrin 50g/L, trehalose
100g/L, L-sodium 10g/L, Cys 5g/L.- 50 DEG C of pre-freeze 1h are placed in after well mixed, then carry out freezing and done
Dry acquisition leavening solid-state bacterium powder, viable count 4.5 × 1010 cfu/g。
Embodiment 2
With lactobacillus gasseri as fermentation strain, by 5% proliferated culture medium for being inoculated into pH5.5.Each component is dense in proliferated culture medium
Degree is as follows：The g/L of sucrose 10, the g/L of glucose 10, the g/L of dusty yeast 1.5, tryptone 1.5 g/L, K2HPO44.0 g/L,
MgSO4·7H2O 0.5 g/L, CaCl2 0.02 g/L, Tween 80 1mL.In 37 °C of anaerobic fermentation culture 12h, wherein in fermentation
Stream plus glucose after 3h, control concentration of glucose is in 5g/L.Cell number reaches 4.1 × 10 in final culture medium9Cfu/ml, will
Zymocyte liquid is aseptically centrifuged acquisition lactobacillus cell.Then 1 is pressed:1 ratio adds freeze drying protectant, freeze drying protectant
The concentration of middle each component is as follows：Skimmed milk power 100g/L, sucrose 100g/L, glycerine 30ml/L, maltodextrin 50g/L, trehalose
100g/L, L-sodium 10g/L, Cys 5g/L.- 50 DEG C of pre-freeze 3h are placed in after well mixed, then carry out freezing and done
Dry acquisition leavening solid-state bacterium powder, viable count 2.1 × 1010 cfu/g。
Embodiment 3
Using lactobacillus reuteri as fermentation strain, by 5% proliferated culture medium for being inoculated into pH5.5.Each component in proliferated culture medium
Concentration it is as follows：The g/L of sucrose 10, the g/L of glucose 10, the g/L of dusty yeast 1.5, tryptone 1.5 g/L, K2HPO4 4.0 g/
L, MgSO4·7H2O 0.5 g/L, CaCl2 0.02 g/L, Tween 80 1mL.In 37 °C of anaerobic fermentation culture 12h, wherein in hair
Stream plus glucose after ferment 6h, control concentration of glucose is in 5g/L.Cell number reaches 1.4 × 10 in final culture medium9Cfu/ml,
Zymocyte liquid is aseptically centrifuged acquisition lactobacillus cell.Then 1 is pressed:1 ratio adds freeze drying protectant, frozen-dried protective
The concentration of each component is as follows in agent：Skimmed milk power 100g/L, sucrose 100g/L, glycerine 30ml/L, maltodextrin 50g/L, marine alga
Sugared 100g/L, L-sodium 10g/L, Cys 5g/L.- 50 DEG C of pre-freeze 1h are placed in after well mixed, then are freezed
Dry and obtain leavening solid-state bacterium powder, viable count 3.0 × 1010 cfu/g。
Embodiment 4
Using Lactobacillus plantarum as fermentation strain, by 3% proliferated culture medium for being inoculated into pH5.Each component is dense in proliferated culture medium
Degree is as follows：Sucrose 5g/L, the g/L of glucose 5, the g/L of dusty yeast 2, tryptone 1g/L, K2HPO45.0 g/L, MgSO4·
7H2O 0.8 g/L, CaCl2 0.03 g/L, Tween 80 1.5mL.In 35 °C of anaerobic fermentation culture 8h, wherein being flowed after the 3h that ferments
Plus glucose, control concentration of glucose is in 3g/L.Cell number reaches 1.0 × 10 in final culture medium9Cfu/ml, by zymophyte
Liquid is aseptically centrifuged acquisition lactobacillus cell.Then 1 is pressed:0.5 ratio adds freeze drying protectant, each in freeze drying protectant
The concentration of component is as follows：Skimmed milk power 110g/L, sucrose 110g/L, glycerine 23ml/L, maltodextrin 45g/L, trehalose 90g/
L, L-sodium 8g/L, Cys 7g/L.- 55 DEG C of pre-freeze 3h are placed in after well mixed, then carry out freeze-drying acquisition
Leavening solid-state bacterium powder, viable count 2.3 × 1010 cfu/g。
Embodiment 5
Using Lactobacillus plantarum as fermentation strain, by 7% proliferated culture medium for being inoculated into pH6.Each component is dense in proliferated culture medium
Degree is as follows：Sucrose 15g/L, glucose 15 g/L, dusty yeast 1g/L, tryptone 2g/L, K2HPO43.0 g/L, MgSO4·
7H2O 0.2 g/L, CaCl2 0.01 g/L, Tween 80 0.5mL.In 40 °C of anaerobic fermentation culture 16h, wherein after the 5h that ferments
Stream plus glucose, control concentration of glucose is in 7g/L.Cell number reaches 6.5 × 10 in final culture medium9Cfu/ml, will ferment
Bacterium solution is aseptically centrifuged acquisition lactobacillus cell.Then 1 is pressed:1.5 ratios add freeze drying protectant, in freeze drying protectant
The concentration of each component is as follows：Skimmed milk power 90g/L, sucrose 90g/L, glycerine 35ml/L, maltodextrin 55g/L, trehalose 110g/
L, L-sodium 12g/L, Cys 3g/L.- 45 DEG C of pre-freeze 2h are placed in after well mixed, then carry out freeze-drying acquisition
Leavening solid-state bacterium powder, viable count 3.7 × 1010 cfu/g。
The above embodiments merely illustrate the technical concept and features of the present invention, is a kind of preferred embodiment, its object is to allow
Person skilled in the art will appreciate that present disclosure and implements according to this, can not limit protection model of the invention with this
Enclose.The equivalent change or modification that all Spirit Essences of the invention are made, should all be included within the scope of the present invention.
Claims (10)
1. it is a kind of produce functional sugar probiotics leaven preparation method, it is characterised in that comprise the following steps：By grignard breast bar
The fermentation strain is linked into proliferated culture medium and divided by bacterium, lactobacillus reuteri or Lactobacillus plantarum as fermentation strain
To add carbon source, in 35 ~ 40 DEG C of anaerobic fermentation 8 ~ 16h of culture, then aseptically centrifugation obtains lactic acid bacteria for stage feed supplement
Cell, 1 ~ 3h of pre-freeze at the lactobacillus cell and freeze drying protectant are blended in into -55 ~ -45 DEG C, then freeze-drying are obtained fermentation
Agent solid-state bacterium powder.
2. it is according to claim 1 produce functional sugar probiotics leaven preparation method, it is characterised in that the fermentation
The inoculation part by weight of bacterial strain is 3 ~ 7%.
3. it is according to claim 1 produce functional sugar probiotics leaven preparation method, it is characterised in that the propagation
Culture medium includes the component of following weight portion：5 ~ 15 parts of sucrose, 5 ~ 15 parts of glucose, 1 ~ 2 part of dusty yeast, 1 ~ 2 part of tryptone,
K2HPO43 ~ 5 parts, MgSO4•7H20.2 ~ 0.8 part of O, CaCl20.01 ~ 0.03 part, 0.5 ~ 1.5 part of Tween 80.
4. it is according to claim 3 produce functional sugar probiotics leaven preparation method, it is characterised in that the propagation
The pH of culture medium is 5 ~ 6.
5. it is according to claim 1 produce functional sugar probiotics leaven preparation method, it is characterised in that fermentation 3 ~
Stream plus glucose after 6h.
6. it is according to claim 5 produce functional sugar probiotics leaven preparation method, it is characterised in that stream plus Portugal
The concentration of grape sugar is 3 ~ 7g/L.
7. it is according to claim 1 produce functional sugar probiotics leaven preparation method, it is characterised in that the propagation
Cell number is 1.0 × 10 in culture medium9~ 6.5×109 cfu/ml。
8. the preparation method of the probiotics leaven for producing functional sugar according to claim 1, it is characterised in that described lyophilized
Protective agent includes the component of following weight portion：90 ~ 110 parts of skimmed milk power, 90 ~ 110 parts of sucrose, 30 ~ 45 parts of glycerine, maltodextrin
45 ~ 55 parts, 90 ~ 110 parts of trehalose, 8 ~ 12 parts of L-sodium, 3 ~ 7 parts of Cys.
9. it is according to claim 1 produce functional sugar probiotics leaven preparation method, it is characterised in that the lactic acid
Bacterium cell and freeze drying protectant are with 1：（0.5~1.5）Ratio mixing.
10. it is according to claim 1 produce functional sugar probiotics leaven preparation method, it is characterised in that the hair
Viable count in ferment agent solid-state bacterium powder is 2.1 × 1010More than cfu/g.
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Cited By (5)
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| CN107723260A (en) * | 2017-10-19 | 2018-02-23 | 江苏中通生物科技有限公司 | A kind of preparation method of probiotics leaven |
| WO2019027017A1 (en) * | 2017-08-04 | 2019-02-07 | 株式会社明治 | Method for culturing lactic acid bacteria |
| CN111820349A (en) * | 2019-04-17 | 2020-10-27 | 吉林省绿色食品工程研究院 | Direct-throwing fruit and vegetable juice lactobacillus accelerant |
| CN111961631A (en) * | 2020-09-04 | 2020-11-20 | 国科智盛(沁阳市)生物科技有限公司 | Fermentation production process of lactobacillus fermentum |
| CN112063547A (en) * | 2020-08-14 | 2020-12-11 | 润盈生物工程（上海）有限公司 | Method for high-density fermentation and bacterial powder quality improvement of lactobacillus gasseri LG-G12 |
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Cited By (6)
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| CN111820349A (en) * | 2019-04-17 | 2020-10-27 | 吉林省绿色食品工程研究院 | Direct-throwing fruit and vegetable juice lactobacillus accelerant |
| CN112063547A (en) * | 2020-08-14 | 2020-12-11 | 润盈生物工程（上海）有限公司 | Method for high-density fermentation and bacterial powder quality improvement of lactobacillus gasseri LG-G12 |
| CN111961631A (en) * | 2020-09-04 | 2020-11-20 | 国科智盛(沁阳市)生物科技有限公司 | Fermentation production process of lactobacillus fermentum |
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