CN107699528A - Lactobacillus paracasei and its strain domestication method and cultural method - Google Patents
Lactobacillus paracasei and its strain domestication method and cultural method Download PDFInfo
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- 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
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- C12R2001/00—Microorganisms ; Processes using microorganisms
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- 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
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- 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
- C12N1/36—Adaptation or attenuation of cells
Abstract
The invention discloses a kind of Lactobacillus paracasei and its strain domestication method and cultural method, the Lactobacillus paracasei both ensure that containing Se form be Organic Selenium, and and can plays the physiological function of probiotics.Its strain domestication method includes：Lactobacillus paracasei strain is inoculated on Lactobacillus paracasei seed culture medium and cultivated, when the OD values of nutrient solution reach 1.2~1.6, incubation time h1 is recorded, and adds sodium selenite solution and make it that the Se content in nutrient solution is 0.2~1 μ g/g, then carries out domestication culture h1 durations.Its cultural method includes：The Lactobacillus paracasei thalline obtained after domestication is inoculated into the potato liquid culture medium containing selenium-rich potato and cultivated, when the OD values of the nutrient solution in potato liquid culture medium reach 1.2~1.6, record incubation time h2, and add sodium selenite solution and make it that the Se content in nutrient solution is 0.6~10 μ g/g, then carry out cultivating h2 durations.
Description
Technical field
The present invention relates to Bacteria Culture technical field, in particular to a kind of Lactobacillus paracasei and its strain domestication
Method and cultural method.
Background technology
Lactobacillus paracasei belongs to lactobacillus, as one kind of probiotics, is resistant to the defense mechanism of organism, its
Include enzyme in oral cavity, bile acid of low pH value and small intestine etc. in gastric juice.Meanwhile Lactobacillus paracasei can suppress yeast and mould
The growth of bacterium, nonspecific resistance of the host to microbial pathogens can be strengthened, accelerate the removing of intestinal pathogens；Can
Enteric flora disturbance and intestinal permeability enhancing are treated, so as to prevent food hypersenstivity and acute diarrhea；Make anti-low-density oxidation system
Antibody and the increase of CD4T- lymphocytes, the phagocytosis of granulocyte are remarkably reinforced, and strengthen human immunity and pre- anti-cancer and suppression
The functions such as tumour growth.With the increase of allergic human population, research to Lactobacillus paracasei and made with its development functionality breast
Product are significant.
Most selenium-rich product on domestic and international market and probiotic products are two major classes at present, have a small number of selenium-enriched probioticses to produce
Product are to the addition of selenium-containing compound and the physical mixed product of probiotics respectively, are not that probiotics contains by selenium-rich culturing
The probiotic products of Organic Selenium.The most and sodium selenite of this kind of product addition, that is, inorganic selenium, it is well known that inorganic
The utilization rate of selenium contains certain toxicity well below Organic Selenium, and antagonism can occur for two kinds of functional components in body
Effect.Therefore, existing product lacks the Lactobacillus paracasei product rich in Organic Selenium.
The content of the invention
First purpose of the present invention is to provide a kind of Lactobacillus paracasei, so that it is rich in Organic Selenium.
Second object of the present invention is a kind of strain domestication method for providing Lactobacillus paracasei, to cause its strain
Selenium-rich culturing can be more easily adapted to conform to, shortens fermentation time.
Third object of the present invention is to provide a kind of cultural method of Lactobacillus paracasei, with the pair rich in Organic Selenium
Lactobacillus casei.
The present invention is solved its technical problem and realized using following technical scheme.
The invention provides a kind of strain domestication method of Lactobacillus paracasei, it comprises the following steps：
Lactobacillus paracasei strain is inoculated on Lactobacillus paracasei seed culture medium and cultivated；
When the OD values of nutrient solution reach 1.2~1.6, incubation time h1 is recorded, and add sodium selenite solution so that training
Se content in nutrient solution is 0.2~1 μ g/g, then carries out domestication culture h1 durations.
The invention further relates to a kind of cultural method of Lactobacillus paracasei, it comprises the following steps：
The Lactobacillus paracasei thalline obtained after the strain domestication method of above-mentioned Lactobacillus paracasei is tamed connects
Plant into the potato liquid culture medium containing selenium-rich potato and cultivated；
When the OD values of the nutrient solution in potato liquid culture medium reach 1.2~1.6, incubation time h2 is recorded, and add
Add sodium selenite solution make it that the Se content in nutrient solution is 0.6~10 μ g/g, then carry out cultivating h2 durations.
The invention further relates to a kind of Lactobacillus paracasei, and it is obtained by the cultural method culture of above-mentioned Lactobacillus paracasei.
The Lactobacillus paracasei obtained by the cultural method culture of the present invention and the product based on the probiotics had both been protected
It is Organic Selenium to have demonstrate,proved containing Se form, can reduce selenium toxicity, improves the physiologically active and absorptivity of selenium, and and can plays the life of probiotics
Manage function, defensive ability/resistance ability of the enhancing body to disease.
Embodiment
, below will be to embodiment party of the present invention to make the purpose, technical scheme and advantage of embodiment of the present invention clearer
Technical scheme in formula is clearly and completely described.Unreceipted actual conditions person in embodiment or embodiment, according to routine
The condition that condition or manufacturer suggest is carried out.Agents useful for same or the unreceipted production firm person of instrument, being can be by commercially available purchase
Buy the conventional products of acquisition.
The Lactobacillus paracasei to embodiment of the present invention and its strain domestication method and cultural method have below
Body explanation.
Some embodiments of the present invention provide a kind of strain domestication method of Lactobacillus paracasei, and it includes following step
Suddenly：
Lactobacillus paracasei strain is inoculated on Lactobacillus paracasei seed culture medium and cultivated；
When the OD values of nutrient solution reach 1.2~1.6, incubation time h1 is recorded, and add sodium selenite solution so that training
Se content in nutrient solution is 0.2~1 μ g/g, preferably 0.4~0.8 μ g/g, more preferably 0.4~0.6 μ g/g, then carry out domestication culture
H1 durations.
By cultivating Lactobacillus paracasei strain, after its fermentation, afterwards to add less inorganic selenium molten to a certain extent
Liquid is sodium selenite solution so that the Lactobacillus paracasei strain can be enlarged culture before can be to the environment containing selenium
Adapted to, to shorten fermentation time during the selenium-rich culturing in later stage, and Se form will be contained and be converted into Organic Selenium, dropped
Low selenium toxicity, the physiologically active and absorptivity of selenium are improved, and can plays the physiological function of probiotics, and enhancing body is to disease
Defensive ability/resistance ability.
OD is the abbreviation of optical density (optical density), represents the optical density that detected material sponges, OD values are light
Density value, it can reflect the concentration of bacterium, reach 1.2~1.6 in the OD values for the nutrient solution that Lactobacillus paracasei is fermented
Add sodium selenite solution so that Lactobacillus paracasei can better adapt to selenium, and then complete to tame and docile Lactobacillus paracasei
Change.
Wherein, the content of the selenium of addition has large effect to domestication result, the too low effect for not reaching domestication of its content,
Too high levels can influence the fermentation process of Lactobacillus paracasei on the contrary.Therefore, selection make it that the content of the selenium in nutrient solution is 0.2
~1 μ g/g can reach optimal domestication effect.
According to some embodiments, the inoculum concentration of Lactobacillus paracasei strain is the body of Lactobacillus paracasei seed culture medium
Long-pending 1~5%, for example, selection volume ratio is 2%, or 3%, or 4% etc..Carrying out inoculated and cultured by above-mentioned inoculum concentration can
Enable culture medium fully to meet the growth of strain, complete domestication process.
Some embodiments of the present invention provide a kind of strain domestication method of Lactobacillus paracasei, and it includes following step
Suddenly：
Lactobacillus paracasei strain is inoculated on Lactobacillus paracasei seed culture medium and cultivated；
When the OD values of nutrient solution reach 1.2~1.6, incubation time h1 is recorded, and add sodium selenite solution so that training
Se content in nutrient solution is 0.2~1 μ g/g, then carries out domestication culture h1 durations；
The nutrient solution obtained after cultivating domestication is centrifuged, and the Lactobacillus paracasei thalline obtained after centrifugation is freezed
Preserve.
According to some embodiments, Lactobacillus paracasei seed culture medium count in parts by weight including：9~11 parts of junket peptone,
7~9 parts of powdered beef, 3~5 parts of dusty yeast, 19~21 parts of glucose, 0.1~0.3 part of magnesium sulfate, 4~6 parts of sodium acetate, citric acid
Three 1~3 part of ammoniums, 1~3 part of dipotassium hydrogen phosphate, 0.03~0.06 part of manganese sulfate, 0.8~1.2 part of Tween 80 and distilled water 900
~1100 parts, and the pH value of the Lactobacillus paracasei seed culture medium is 6~6.4.Lactobacillus paracasei seed culture medium is
It is mixed to get according to the various raw materials of above-mentioned weight.Above-mentioned Lactobacillus paracasei seed culture medium it is various into
The fermented and cultured of Lactobacillus paracasei strain can fully be met by dividing, and reach the purpose tamed to strain.
According to some embodiments, Lactobacillus paracasei seed culture is based on sterilizing in high-pressure sterilizing pot before inoculation,
In some embodiments, sterilising temp be 115~125 DEG C, for example, 115 DEG C, 116 DEG C, 117 DEG C, 118 DEG C, 119 DEG C, 120 DEG C,
121 DEG C or 122 DEG C etc., preferably 121 DEG C, sterilization time are 18~25min, preferably 20min.
According to some embodiments, storage temperature is -19~-17 DEG C, for example, storage temperature is -17 DEG C, -18 DEG C, or -
19℃。
Some embodiments of the present invention provide a kind of cultural method of Lactobacillus paracasei, and it comprises the following steps：
The Lactobacillus paracasei thalline that will be obtained after the strain domestication method of above-mentioned Lactobacillus paracasei is tamed
It is inoculated into the potato liquid culture medium containing selenium-rich potato and is cultivated；
When the OD values of the nutrient solution in potato liquid culture medium reach 1.2~1.6, incubation time h2 is recorded, and add
Sodium selenite solution is added to cause the Se content in nutrient solution to be 0.6~10 μ g/g, preferably 2~8 μ g/g, more preferably 3~6 μ g/g,
Carry out cultivating h2 durations again.
The Lactobacillus paracasei thalline that is obtained after domestication is directly fermented, kind of a liquid preparation time is reduced, directly enters
Enter to produce fermentation in enormous quantities.Selenium-rich training after the Lactobacillus paracasei strain by sodium selenite domestication is more easily adapted to conform to simultaneously
Support, that is, shorten the laundering period of kind of liquid, so as to shorten fermentation time.
According to some embodiments, potato liquid culture medium is counted 190~220 parts of selenium-rich potatos in parts by weight
25~35min is boiled after being mixed with 900~1100 parts, after filtering plus water mend to 900~1100 parts, then with 1~2 part of magnesium sulfate
0.8~1.2 part of ammonium sulfate and 1.8~2.2 parts of dipotassium hydrogen phosphates are mixed to get.Pass through the potato specifically for selenium-rich culturing
The configuration of liquid culture medium so that it can complete the fermentation of Lactobacillus paracasei, and cause inorganic selenium to be converted into Organic Selenium.
According to some embodiments, high-temperature sterilization is carried out before inoculation to potato liquid culture medium, sterilising temp is 115~
125 DEG C, for example, 115 DEG C, 116 DEG C, 117 DEG C, 118 DEG C, 119 DEG C, 120 DEG C, 121 DEG C or 122 DEG C etc., preferably 121 DEG C, sterilizing
Time is 18~25min, preferably 20min.
According to some embodiments, after being first diluted to Lactobacillus paracasei thalline with physiological saline or phosphate buffer
Again inoculation fermentation culture is carried out in potato liquid culture medium.Preferably, the body of the physiological saline of dilution or phosphate buffer
Product is equal to the volume of the nutrient solution after carrying out above-mentioned domestication culture.
According to some embodiments, the inoculum concentration of Lactobacillus paracasei thalline is the volume of the potato liquid culture medium
5~10%.For example, volume ratio is 6%, 7%, or 8% etc..Inoculated and cultured is carried out by above-mentioned inoculum concentration so that unobvious
On the premise of spinning out fermentation time, the inoculum concentration of required minimum magnitude, so as to reduce kind of a liquid cost, the effect for expanding culture is improved
Rate.
Some embodiments of the present invention provide a kind of cultural method of Lactobacillus paracasei, and it includes：
S1. Lactobacillus paracasei seed culture medium is prepared：In parts by weight, by 9~11 parts of junket peptone, powdered beef 7~9
Part, 3~5 parts of dusty yeast, 19~21 parts of glucose, 0.1~0.3 part of magnesium sulfate, 4~6 parts of sodium acetate, Triammonium citrate 1~3
Part, 1~3 part of dipotassium hydrogen phosphate, 0.03~0.06 part of manganese sulfate, 0.8~1.2 part of Tween 80 and 900~1100 parts of distilled water
Mixing, the pH value of regulation Lactobacillus paracasei seed culture medium is 6~6.4, and in autoclaving under 115~125 DEG C of high temperature
18~25min of sterilizing, can be used in pot, by volume 1%~5% inoculation Lactobacillus paracasei.
Plus selenium S2.：Each two hour detects zymotic fluid OD values, when reaching 1.2~1.6, records total fermentation time h1, and
Addition sodium selenite solution make it that the Se content of zymotic fluid is 0.2~1 μ g/g.
S3. centrifuge：After adding selenium, then the h1 durations that ferment, zymotic fluid is centrifuged afterwards.
S4. preserve：The Lactobacillus paracasei thalline by sodium selenite domestication of precipitation is protected in -19~-17 DEG C of freezings
Deposit.
S5. potato fluid nutrient medium is prepared：Count in parts by weight and 190~220 parts of selenium-rich potatos are cleaned into peeling, cut
Into after fritter, 25~35min is boiled after being mixed with 900~1100 parts, after filtering plus water is mended to 900~1100 parts, then with 1~2
Part 0.8~1.2 part of ammonium sulfate of magnesium sulfate and 1.8~2.2 parts of dipotassium hydrogen phosphates are mixed to get and under 115~125 DEG C of high temperature
Sterilize 18~25min, can use.
S6. it is inoculated with：When Lactobacillus paracasei is produced in enormous quantities, directly with the physiology with zymotic fluid equal volume before centrifugation
Salt solution or phosphate buffer dilute above-mentioned freezing thalline, are inoculated into potato fluid nutrient medium by 5%-10% volume ratio
Row fermented and cultured.
S7. centrifuge：Each two hour monitors zymotic fluid OD values, when reaching 1.2~1.6, records total fermentation time h2, and
Addition sodium selenite solution make it that zymotic fluid Se content is 0.6 μ g/g-10 μ g/g.After adding selenium, then the h2 durations that ferment, centrifuge afterwards
Zymotic fluid, that is, obtain selenium-rich Lactobacillus paracasei thalline.
The invention further relates to a kind of Lactobacillus paracasei, and it is obtained by the cultural method culture of above-mentioned Lactobacillus paracasei
Arrive.
The feature and performance of the present invention are described in further detail with reference to embodiments.
Embodiment 1
(1) Lactobacillus paracasei seed culture medium is prepared：By junket peptone 10.0g, powdered beef 8.0g, dusty yeast 4.0g, grape
Sugared 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, Triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.05g, tween
80 1.0g and distilled water 1000mL mixing, regulation pH value is 6.2, and is sterilized under 121 DEG C of high temperature in high-pressure sterilizing pot
20min, Lactobacillus paracasei strain is inoculated in Lactobacillus paracasei seed culture medium according to volume ratio 3% and cultivated.
(2) selenium is added：Each two hour detects zymotic fluid OD values, when reaching 1.2~1.6, and records total fermentation time h1,
And add sodium selenite solution and make it that the Se content of zymotic fluid is 0.4 μ g/g.
(3) centrifuge：After adding selenium, then the h1 durations that ferment, zymotic fluid is centrifuged afterwards.
(4) preserve：By the Lactobacillus paracasei thalline by sodium selenite domestication of precipitation in -18 DEG C of freezen protectives.
(5) potato fluid nutrient medium is prepared：By 200g selenium-rich potatos, peeling is cleaned, is cut into small pieces, adds water
1000ml, boil 30min, after filtering plus water complements to 1000mL, then with 1.5g magnesium sulfate, 1g ammonium sulfate and 2.0g phosphoric acid hydrogen
Dipotassium mixes, and the 20min that sterilized under 121 DEG C of high temperature.
(5) it is inoculated with：When Lactobacillus paracasei is produced in enormous quantities, directly with the physiology with zymotic fluid equal volume before centrifugation
The Lactobacillus paracasei thalline of salt solution or phosphate buffer dilution refrigeration, is inoculated into fluid nutrient medium by 8% volume ratio.
(6) centrifuge：Each two hour monitors zymotic fluid OD values, when reaching 1.2~1.6, records total fermentation time h2, and
Addition sodium selenite solution make it that zymotic fluid Se content is 0.6 μ g/g.After adding selenium, then the h2 durations that ferment, zymotic fluid is centrifuged afterwards,
The selenium-rich Lactobacillus paracasei thalline that Se content is 259.96 μ g/g is obtained, and dry weight yield is 1.89g/L, selenium high conversion rate
Up to 63.70%.
Embodiment 2
(1) Lactobacillus paracasei seed culture medium is prepared：By junket peptone 10.0g, powdered beef 8.0g, dusty yeast 4.0g, grape
Sugared 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, Triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.05g, tween
80 1.0g and distilled water 1000mL mixing, regulation pH value is 6.2, and is sterilized under 121 DEG C of high temperature in high-pressure sterilizing pot
20min, Lactobacillus paracasei strain is inoculated in Lactobacillus paracasei seed culture medium according to volume ratio 3% and cultivated.
(2) selenium is added：Each two hour detects zymotic fluid OD values, when reaching 1.2~1.6, and records total fermentation time h1,
And add sodium selenite solution and make it that the Se content of zymotic fluid is 0.4 μ g/g.
(3) centrifuge：After adding selenium, then the h1 durations that ferment, zymotic fluid is centrifuged afterwards.
(4) preserve：By the Lactobacillus paracasei thalline by sodium selenite domestication of precipitation in -18 DEG C of freezen protectives.
(5) potato fluid nutrient medium is prepared：By 200g selenium-rich potatos, peeling is cleaned, is cut into small pieces, adds water
1000ml, boil 30min, after filtering plus water complements to 1000mL, then with 1.5g magnesium sulfate, 1g ammonium sulfate and 2.0g phosphoric acid hydrogen
Dipotassium mixes, and the 20min that sterilized under 121 DEG C of high temperature.
(5) it is inoculated with：When Lactobacillus paracasei is produced in enormous quantities, directly with the physiology with zymotic fluid equal volume before centrifugation
The Lactobacillus paracasei thalline of salt solution or phosphate buffer dilution refrigeration, is inoculated into fluid nutrient medium by 8% volume ratio.
(6) centrifuge：Each two hour monitors zymotic fluid OD values, when reaching 1.2~1.6, records total fermentation time h2, and
Addition sodium selenite solution make it that zymotic fluid Se content is 4 μ g/g.After adding selenium, then the h2 durations that ferment, zymotic fluid is centrifuged afterwards, i.e.,
Obtain the selenium-rich Lactobacillus paracasei thalline that Se content is 2966.12 μ g/g.Dry weight yield is 2.01g/L, and selenium high conversion rate reaches
98.00%.
Embodiment 3
(1) Lactobacillus paracasei seed culture medium is prepared：By junket peptone 10.0g, powdered beef 8.0g, dusty yeast 4.0g, grape
Sugared 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, Triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.05g, tween
80 1.0g and distilled water 1000mL mixing, regulation pH value is 6.2, and is sterilized under 121 DEG C of high temperature in high-pressure sterilizing pot
20min, Lactobacillus paracasei strain is inoculated in Lactobacillus paracasei seed culture medium according to volume ratio 3% and cultivated.
(2) selenium is added：Each two hour detects zymotic fluid OD values, when reaching 1.2~1.6, and records total fermentation time h1,
And add sodium selenite solution and make it that the Se content of zymotic fluid is 0.4 μ g/g.
(3) centrifuge：After adding selenium, then the h1 durations that ferment, zymotic fluid is centrifuged afterwards.
(4) preserve：By the Lactobacillus paracasei thalline by sodium selenite domestication of precipitation in -18 DEG C of freezen protectives.
(5) potato fluid nutrient medium is prepared：By 200g selenium-rich potatos, peeling is cleaned, is cut into small pieces, adds water
1000ml, boil 30min, after filtering plus water complements to 1000mL, then with 1.5g magnesium sulfate, 1g ammonium sulfate and 2.0g phosphoric acid hydrogen
Dipotassium mixes, and the 20min that sterilized under 121 DEG C of high temperature.
(5) it is inoculated with：When Lactobacillus paracasei is produced in enormous quantities, directly with the physiology with zymotic fluid equal volume before centrifugation
The Lactobacillus paracasei thalline of salt solution or phosphate buffer dilution refrigeration, is inoculated into fluid nutrient medium by 8% volume ratio.
(6) centrifuge：Each two hour monitors zymotic fluid OD values, when reaching 1.2~1.6, records total fermentation time h2, and
Addition sodium selenite solution make it that zymotic fluid Se content is 10 μ g/g.After adding selenium, then the h2 durations that ferment, zymotic fluid is centrifuged afterwards,
Obtain the selenium-rich Lactobacillus paracasei thalline that Se content is 6651.12 μ g/g.Dry weight yield is 1.73g/L, selenium high conversion rate
Up to 90.00%.
Embodiment 4
(1) Lactobacillus paracasei seed culture medium is prepared：By junket peptone 9.0g, powdered beef 7.0g, dusty yeast 3.0g, glucose
19.0g, magnesium sulfate 0.1g, sodium acetate 4.0g, Triammonium citrate 1.0g, dipotassium hydrogen phosphate 1.0g, manganese sulfate 0.03g, Tween 80
0.8g and distilled water 900mL mixing, regulation pH value is 6, and the 25min that sterilized under 115 DEG C of high temperature in high-pressure sterilizing pot, is pressed
Lactobacillus paracasei strain is inoculated in into Lactobacillus paracasei seed culture medium according to volume ratio 1% to be cultivated.
(2) selenium is added：Each two hour detects zymotic fluid OD values, when reaching 1.2~1.6, and records total fermentation time h1,
And add sodium selenite solution and make it that the Se content of zymotic fluid is 0.2 μ g/g.
(3) centrifuge：After adding selenium, then the h1 durations that ferment, zymotic fluid is centrifuged afterwards.
(4) preserve：By the Lactobacillus paracasei thalline by sodium selenite domestication of precipitation in -17 DEG C of freezen protectives.
(5) potato fluid nutrient medium is prepared：By 190g selenium-rich potato, peeling is cleaned, is cut into small pieces, adds water 900ml,
25min is boiled, after filtering plus water complements to 900mL, then is mixed with 1g magnesium sulfate, 0.8g ammonium sulfate and 1.8g dipotassium hydrogen phosphates
Close, and the 25min that sterilized under 115 DEG C of high temperature.
(5) it is inoculated with：When Lactobacillus paracasei is produced in enormous quantities, directly with the physiology with zymotic fluid equal volume before centrifugation
The Lactobacillus paracasei thalline of salt solution or phosphate buffer dilution refrigeration, is inoculated into fluid nutrient medium by 5% volume ratio.
(6) centrifuge：Each two hour monitors zymotic fluid OD values, when reaching 1.2~1.6, records total fermentation time h2, and
Addition sodium selenite solution make it that zymotic fluid Se content is 0.4 μ g/g.After adding selenium, then the h2 durations that ferment, zymotic fluid is centrifuged afterwards,
The selenium-rich Lactobacillus paracasei thalline that Se content is 128.75 μ g/g is obtained, and dry weight yield is 1.82g/L, selenium high conversion rate
Up to 51.50%.
Embodiment 5
(1) Lactobacillus paracasei seed culture medium is prepared：By junket peptone 10.0g, powdered beef 8.0g, dusty yeast 5.0g, grape
Sugared 21.0g, magnesium sulfate 0.3g, sodium acetate 6.0g, Triammonium citrate 3.0g, dipotassium hydrogen phosphate 3.0g, manganese sulfate 0.06g, tween
80 1.2g and distilled water 1100mL mixing, regulation pH value is 6.4, and is sterilized under 125 DEG C of high temperature in high-pressure sterilizing pot
18min, Lactobacillus paracasei strain is inoculated in Lactobacillus paracasei seed culture medium according to volume ratio 5% and cultivated.
(2) selenium is added：Each two hour detects zymotic fluid OD values, when reaching 1.2~1.6, and records total fermentation time h1,
And add sodium selenite solution and make it that the Se content of zymotic fluid is 1 μ g/g.
(3) centrifuge：After adding selenium, then the h1 durations that ferment, zymotic fluid is centrifuged afterwards.
(4) preserve：By the Lactobacillus paracasei thalline by sodium selenite domestication of precipitation in -19 DEG C of freezen protectives.
(5) potato fluid nutrient medium is prepared：By 220g selenium-rich potatos, peeling is cleaned, is cut into small pieces, adds water
1100ml, boil 35min, after filtering plus water complements to 1100mL, then with 2g magnesium sulfate, 1.2g ammonium sulfate and 2.2g phosphoric acid hydrogen
Dipotassium mixes, and the 18min that sterilized under 125 DEG C of high temperature.
(5) it is inoculated with：When Lactobacillus paracasei is produced in enormous quantities, directly with the physiology with zymotic fluid equal volume before centrifugation
The Lactobacillus paracasei thalline of salt solution or phosphate buffer dilution refrigeration, is inoculated into fluid nutrient medium by 10% volume ratio.
(6) centrifuge：Each two hour monitors zymotic fluid OD values, when reaching 1.2~1.6, records total fermentation time h2, and
Addition sodium selenite solution make it that zymotic fluid Se content is 0.5 μ g/g.After adding selenium, then the h2 durations that ferment, zymotic fluid is centrifuged afterwards,
The selenium-rich Lactobacillus paracasei thalline that Se content is 242.96 μ g/g is obtained, and dry weight yield is 1.83g/L, selenium high conversion rate
Up to 63.31%.
Embodiment 6
(1) Lactobacillus paracasei seed culture medium is prepared：By junket peptone 9.5g, powdered beef 8.5g, dusty yeast 4.5g, glucose
20.5g, magnesium sulfate 0.25g, sodium acetate 5.5g, Triammonium citrate 2.5g, dipotassium hydrogen phosphate 2.5g, manganese sulfate 0.04g, Tween 80
1.5g and distilled water 1050mL mixing, regulation pH value is 6.3, and is sterilized under 122 DEG C of high temperature in high-pressure sterilizing pot
19min, Lactobacillus paracasei strain is inoculated in Lactobacillus paracasei seed culture medium according to volume ratio 4% and cultivated.
(2) selenium is added：Each two hour detects zymotic fluid OD values, when reaching 1.2~1.6, and records total fermentation time h1,
And add sodium selenite solution and make it that the Se content of zymotic fluid is 0.6 μ g/g.
(3) centrifuge：After adding selenium, then the h1 durations that ferment, zymotic fluid is centrifuged afterwards.
(4) preserve：By the Lactobacillus paracasei thalline by sodium selenite domestication of precipitation in -18 DEG C of freezen protectives.
(5) potato fluid nutrient medium is prepared：By 210g selenium-rich potatos, peeling is cleaned, is cut into small pieces, adds water
1050ml, boil 32min, after filtering plus water complements to 1050mL, then with 1.7g magnesium sulfate, 1.1g ammonium sulfate and 2.1g phosphoric acid
Hydrogen dipotassium mixes, and the 19min that sterilized under 122 DEG C of high temperature.
(5) it is inoculated with：When Lactobacillus paracasei is produced in enormous quantities, directly with the physiology with zymotic fluid equal volume before centrifugation
The Lactobacillus paracasei thalline of salt solution or phosphate buffer dilution refrigeration, is inoculated into fluid nutrient medium by 8% volume ratio.
(6) centrifuge：Each two hour monitors zymotic fluid OD values, when reaching 1.2~1.6, records total fermentation time h2, and
Addition sodium selenite solution make it that zymotic fluid Se content is 5 μ g/g.After adding selenium, then the h2 durations that ferment, zymotic fluid is centrifuged afterwards, i.e.,
Obtain the selenium-rich Lactobacillus paracasei thalline that Se content is 3024.57 μ g/g.Dry weight yield is 2.03g/L, and selenium high conversion rate reaches
98.13%.
Embodiment 7
(1) Lactobacillus paracasei seed culture medium is prepared：By junket peptone 9.5g, powdered beef 7.5g, dusty yeast 3.5g, glucose
19.5g, magnesium sulfate 0.15g, sodium acetate 4.5g, Triammonium citrate 1.5g, dipotassium hydrogen phosphate 1.5g, manganese sulfate 0.04g, Tween 80
0.9g and distilled water 950mL mixing, regulation pH value is 6.1, and the 23min that sterilized under 119 DEG C of high temperature in high-pressure sterilizing pot,
Lactobacillus paracasei strain is inoculated in into Lactobacillus paracasei seed culture medium according to volume ratio 2% to be cultivated.
(2) selenium is added：Each two hour detects zymotic fluid OD values, when reaching 1.2~1.6, and records total fermentation time h1,
And add sodium selenite solution and make it that the Se content of zymotic fluid is 0.3 μ g/g.
(3) centrifuge：After adding selenium, then the h1 durations that ferment, zymotic fluid is centrifuged afterwards.
(4) preserve：By the Lactobacillus paracasei thalline by sodium selenite domestication of precipitation in -18 DEG C of freezen protectives.
(5) potato fluid nutrient medium is prepared：By 190g selenium-rich potato, peeling is cleaned, is cut into small pieces, adds water 950ml,
28min is boiled, after filtering plus water complements to 950mL, then is mixed with 1.3g magnesium sulfate, 0.9g ammonium sulfate and 1.9g dipotassium hydrogen phosphates
Close, and the 23min that sterilized under 119 DEG C of high temperature.
(5) it is inoculated with：When Lactobacillus paracasei is produced in enormous quantities, directly with the physiology with zymotic fluid equal volume before centrifugation
The Lactobacillus paracasei thalline of salt solution or phosphate buffer dilution refrigeration, is inoculated into fluid nutrient medium by 7% volume ratio.
(6) centrifuge：Each two hour monitors zymotic fluid OD values, when reaching 1.2~1.6, records total fermentation time h2, and
Addition sodium selenite solution make it that zymotic fluid Se content is 9 μ g/g.After adding selenium, then the h2 durations that ferment, zymotic fluid is centrifuged afterwards, i.e.,
The selenium-rich Lactobacillus paracasei thalline that Se content is 6095.61 μ g/g is obtained, dry weight yield is 1.69g/L, and selenium high conversion rate reaches
91.03%.
The Se content and dry weight yield and selenium obtained by above-described embodiment by conventional method measuring and calculation converts
Rate can be seen that the cultural method and inorganic selenium effectively be converted into Organic Selenium, obtain the higher selenium-rich pair cheese of Se content
Lactobacillus thalline, it had both been ensure that containing Se form be Organic Selenium so as to obtain organic selenium-rich Lactobacillus paracasei product, can drop
Low selenium toxicity, the physiologically active and absorptivity of selenium are improved, and can plays the physiological function of probiotics, and enhancing body supports to disease
Imperial ability.
Embodiments described above is part of the embodiment of the present invention, rather than whole embodiments.The reality of the present invention
The detailed description for applying example is not intended to limit the scope of claimed invention, but is merely representative of the selected implementation of the present invention
Example.Based on the embodiment in the present invention, what those of ordinary skill in the art were obtained under the premise of creative work is not made
Every other embodiment, belongs to the scope of protection of the invention.
Claims (10)
1. a kind of strain domestication method of Lactobacillus paracasei, it is characterised in that it comprises the following steps：
Lactobacillus paracasei strain is inoculated on Lactobacillus paracasei seed culture medium and cultivated；
When the OD values of nutrient solution reach 1.2~1.6, incubation time h1 is recorded, and add sodium selenite solution and cause nutrient solution
In Se content be 0.2~1 μ g/g, then carry out domestication culture h1 durations.
2. the strain domestication method of Lactobacillus paracasei according to claim 1, it is characterised in that the secondary cheese breast bar
Bacterium seed culture medium count in parts by weight including：9~11 parts of junket peptone, 7~9 parts of powdered beef, 3~5 parts of dusty yeast, glucose 19~
21 parts, 0.1~0.3 part of magnesium sulfate, 4~6 parts of sodium acetate, 1~3 part of Triammonium citrate, 1~3 part of dipotassium hydrogen phosphate, manganese sulfate
0.03~0.06 part, 900~1100 parts of 0.8~1.2 part of Tween 80 and distilled water, and the Lactobacillus paracasei seed is trained
The pH value for supporting base is 6~6.4.
3. the strain domestication method of Lactobacillus paracasei according to claim 2, it is characterised in that before inoculation by described in
Lactobacillus paracasei seed culture is based on sterilizing in high-pressure sterilizing pot, it is preferable that sterilising temp is 115~125 DEG C, sterilization time
For 18~25min.
4. the strain domestication method of Lactobacillus paracasei according to claim 1, it is characterised in that the secondary cheese breast bar
The inoculum concentration of bacterium strain is the 1~5% of the volume of the Lactobacillus paracasei seed culture medium.
5. the strain domestication method of Lactobacillus paracasei according to claim 1, it is characterised in that after also including to culture
Obtained nutrient solution is centrifuged, and the Lactobacillus paracasei thalline freezen protective that will be obtained after centrifugation, it is preferable that storage temperature
For -19~-17 DEG C.
6. a kind of cultural method of Lactobacillus paracasei, it is characterised in that it comprises the following steps：
It will be obtained after the strain domestication method of the Lactobacillus paracasei described in Claims 1 to 5 any one is tamed
Lactobacillus paracasei thalline be inoculated into the potato liquid culture medium containing selenium-rich potato and cultivated；
When the OD values of the nutrient solution in potato liquid culture medium reach 1.2~1.6, incubation time h2 is recorded, and add Asia
Selenic acid sodium solution make it that the Se content in nutrient solution is 0.6~10 μ g/g, then carries out cultivating h2 durations.
7. the cultural method of Lactobacillus paracasei according to claim 6, it is characterised in that the potato liquid culture
Base be count in parts by weight 190~220 parts of selenium-rich potatos are mixed with 900~1100 parts after boil 25~35min, after filtering
Add water mend to 900~1100 parts, then with 1~2 part of magnesium sulfate, 0.8~1.2 part of ammonium sulfate and 1.8~2.2 parts of dipotassium hydrogen phosphates
It is mixed to get, it is preferable that carry out high-temperature sterilization before inoculation to the potato liquid culture medium, sterilising temp is 115~125
DEG C, sterilization time is 18~25min.
8. the cultural method of Lactobacillus paracasei according to claim 6, it is characterised in that to the Lactobacillus paracasei
Thalline carries out inoculated and cultured in potato liquid culture medium again after being first diluted with physiological saline or phosphate buffer, preferably
Ground, the physiological saline of dilution or the volume of the phosphate buffer are equal to the body of the nutrient solution after carrying out domestication culture
Product.
9. the cultural method of Lactobacillus paracasei according to claim 6, it is characterised in that the Lactobacillus paracasei bacterium
The inoculum concentration of body is the 5~10% of the volume of the potato liquid culture medium.
10. a kind of Lactobacillus paracasei, it is characterised in that it is as the Lactobacillus paracasei described in claim 6~9 any one
Cultural method culture obtain.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020063515A1 (en) * | 2018-09-30 | 2020-04-02 | 内蒙古伊利实业集团股份有限公司 | Novel application of lactobacillus paracasei k56 capable of regulating balance of gastrointestinal flora |
| CN111004753A (en) * | 2019-12-30 | 2020-04-14 | 苏州硒泰克生物科技有限公司 | Lactobacillus paracasei capable of highly producing selenocysteine and application thereof |
| CN111286471A (en) * | 2019-12-12 | 2020-06-16 | 湖北华扬科技发展有限公司 | Method for high-density culture of lactobacillus paracasei by using soybean meal hydrolysate |
| CN112458029A (en) * | 2020-12-21 | 2021-03-09 | 苏州微克生活科技有限公司 | Preparation method and application of high-activity lactobacillus paracasei freeze-dried powder |
| CN113100129A (en) * | 2021-04-14 | 2021-07-13 | 中国水产科学研究院黄海水产研究所 | Culture medium and culture method for long-term and continuous culture of back mouth insects |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100728585B1 (en) * | 2005-05-16 | 2007-06-15 | 주식회사 메디오젠 | lactic ferments with selenium and manufacturing method thereof |
| CN103805537A (en) * | 2013-12-30 | 2014-05-21 | 青岛碧水蓝天生物技术有限公司 | Method for producing organic selenium-enriched lactic acid bacteria |
| CN105154368A (en) * | 2015-09-28 | 2015-12-16 | 内蒙古蒙牛乳业（集团）股份有限公司 | Lactobacillus plantarum multiplying culture medium and lactobacillus plantarum multiplying method |
| CN105713867A (en) * | 2016-04-28 | 2016-06-29 | 陈鹤 | Lactobacillus brevis selenium-rich fermentation method and application |
| CN106978365A (en) * | 2017-02-27 | 2017-07-25 | 陕西理工学院 | The preparation method and a kind of fermentation preparation of a kind of selenium-rich cheese production zymophyte powder |
-
2017
- 2017-11-22 CN CN201711176088.2A patent/CN107699528A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100728585B1 (en) * | 2005-05-16 | 2007-06-15 | 주식회사 메디오젠 | lactic ferments with selenium and manufacturing method thereof |
| CN103805537A (en) * | 2013-12-30 | 2014-05-21 | 青岛碧水蓝天生物技术有限公司 | Method for producing organic selenium-enriched lactic acid bacteria |
| CN105154368A (en) * | 2015-09-28 | 2015-12-16 | 内蒙古蒙牛乳业（集团）股份有限公司 | Lactobacillus plantarum multiplying culture medium and lactobacillus plantarum multiplying method |
| CN105713867A (en) * | 2016-04-28 | 2016-06-29 | 陈鹤 | Lactobacillus brevis selenium-rich fermentation method and application |
| CN106978365A (en) * | 2017-02-27 | 2017-07-25 | 陕西理工学院 | The preparation method and a kind of fermentation preparation of a kind of selenium-rich cheese production zymophyte powder |
Non-Patent Citations (1)
| Title |
|---|
| 高玉荣等: "《新型功能性大豆发酵食品》", 31 December 2015, 中国纺织出版社 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020063515A1 (en) * | 2018-09-30 | 2020-04-02 | 内蒙古伊利实业集团股份有限公司 | Novel application of lactobacillus paracasei k56 capable of regulating balance of gastrointestinal flora |
| US11666613B2 (en) | 2018-09-30 | 2023-06-06 | Inner Mongolia Yili Industrial Group Co., Ltd. | Use of Lactobacillus paracasei subsp. paracasei K56 capable of regulating gastrointestinal flora balance |
| CN111286471A (en) * | 2019-12-12 | 2020-06-16 | 湖北华扬科技发展有限公司 | Method for high-density culture of lactobacillus paracasei by using soybean meal hydrolysate |
| CN111004753A (en) * | 2019-12-30 | 2020-04-14 | 苏州硒泰克生物科技有限公司 | Lactobacillus paracasei capable of highly producing selenocysteine and application thereof |
| CN111004753B (en) * | 2019-12-30 | 2022-09-06 | 苏州硒泰克生物科技有限公司 | Lactobacillus paracasei capable of highly producing selenocysteine and application thereof |
| CN112458029A (en) * | 2020-12-21 | 2021-03-09 | 苏州微克生活科技有限公司 | Preparation method and application of high-activity lactobacillus paracasei freeze-dried powder |
| CN113100129A (en) * | 2021-04-14 | 2021-07-13 | 中国水产科学研究院黄海水产研究所 | Culture medium and culture method for long-term and continuous culture of back mouth insects |
| CN113100129B (en) * | 2021-04-14 | 2021-12-03 | 中国水产科学研究院黄海水产研究所 | Culture medium and culture method for long-term and continuous culture of back mouth insects |
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