CN108048381A - A kind of preparation method of high-activity lactic acid bacteria solid formulation - Google Patents
A kind of preparation method of high-activity lactic acid bacteria solid formulation Download PDFInfo
- Publication number
- CN108048381A CN108048381A CN201810165716.5A CN201810165716A CN108048381A CN 108048381 A CN108048381 A CN 108048381A CN 201810165716 A CN201810165716 A CN 201810165716A CN 108048381 A CN108048381 A CN 108048381A
- Authority
- CN
- China
- Prior art keywords
- lactic acid
- acid bacteria
- molasses
- solid formulation
- fermentation
- 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.)
- Pending
Links
Classifications
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
The present invention provides a kind of preparation methods of high-activity lactic acid bacteria solid formulation, using molasses, corn pulp as primary carbon source nitrogen source, reduce culture medium cost, by three sections of pH control techniques, greatly increase fermentation level.By reuse after liquid portion processing of the zymotic fluid after centrifugation, effective metabolite of lactic acid bacteria is remained, while good protective effect is played to lactic acid.The preparation method of the present invention, simple for process, operation easily, production efficiency it is high, and the viable count of lactobacillus prepared is high, and stability is good, easily stored.
Description
Technical field
The present invention relates to the production technical field of lactic acid bacteria, more particularly to a kind of system of high-activity lactic acid bacteria solid formulation
Preparation Method.
Background technology
Lactic acid bacteria plays the ecological balance for maintaining various beneficial floras in terms of body enteron aisle is adjusted, and reduces the courage in serum
Sterol concentration is metabolized the lactic acid of generation, makes the acidity of ambient enviroment reduce, and inhibits the growth and breeding of miscellaneous bacteria in enteron aisle, while
Promote the absorption and utilization of nutriment.Therefore, lactic acid bacteria is widely used in light industry, food industry, medicine and feed work
In the various industries such as industry.
Lactobacter growth breeding needs complicated nutriment, traditional lactobacillus-fermented multiselect MRS culture mediums, price
It is expensive.Simultaneously because fermentation later stage nutriment deficiency and metabolite（Mainly lactic acid）Accumulation inhibits thalli growth, causes
Fermentation level is low, and covert adds cost.
Lactic acid bacteria easily inactivates, the resistance to high temperature, hydrochloric acid in gastric juice, cholate etc. in production, processing, preservation and application process
Force difference, particularly high temperature working processes and when passing through gastric environment can mortality, so as to reduce biological action, cause to make
It is poor with the repeatability and stability of effect.
Therefore, by optimization for fermentation technology, improve fermentation level, reduce production cost and by add protective agent and
Specific preparation technique increases the stability and tolerance of lactic acid bacteria, improves the survival rate and activity of lactic acid bacteria, has great meaning
Justice.
The content of the invention
It is an object of the invention to provide the preparation methods of a kind of high fermentation level, the lactic acid bacteria solid formulation of high activity.
A kind of preparation method of high-activity lactic acid bacteria solid formulation, comprises the following steps：
（1）Extracting lactic acid bacterium strain is inoculated in by 5 ~ 10% inoculum concentration in the seed culture medium after sterilizing, and training is stood at 30 ~ 40 DEG C
15 ~ 20h is supported, up to seed liquor after culture；
（2）Seed liquor is inoculated in by 10 ~ 15% inoculum concentration in fermentation medium, 30 ~ 40 DEG C cultivate 40 ~ 48h to get into
Ripe zymotic fluid, 30~50rpm of speed of agitator in fermentation process, after zymotic fluid pH is reduced to 4.5, by every liter of culture base flow sugaring honey
Ratio addition molasses when 5 ~ 10g/ is small, while pH4.5~5.0 are controlled by flowing hydrogenation calcium oxide, it is controlled after the 30h that ferments
PH4.0~4.5；
（3）4000~6000rpm of maturing fermentation liquid is centrifuged into 10~15min, respectively obtains bacterium mud and fermentation broth liquor part, it will
The liquid portion obtains the concentrate of solid content 30%~40% through plate-frame filtering, ultrafiltration, activated carbon decolorizing and vacuum concentration；
（4）To step（3）In bacterium mud in add protective agent, be uniformly mixed, obtain bacterium mud protection agent composition, protective agent each group
It is respectively oligofructose 0.3 ~ 0.5%, beta-cyclodextrin 2~3%, vitamin C 0.1~0.3%, soybean point to account for bacterium mud mass percent
Protein isolate 1~2%, sodium alginate 2~3%；
（5）By step（4）Obtained bacterium mud protection agent composition and step（3）In concentrate mixing, and add account for concentrate
Middle 20~30% resistant starch of solid quality percentage, 0.5~1% carragheen, 0.2~0.5% xanthans are uniformly mixed, and are made
Softwood；
（6）By step（5）Softwood obtained is put into squeeze out and pelletize in finisher,pill, and granulation conditions are under 600~800 revs/min
Reason 10~15 minutes is dried to moisture content 5~8% at 40~50 DEG C to get high-activity lactic acid bacteria solid formulation after granulation；
Wherein, step（1）Described in seed culture based component be：60 ~ 80g/L of molasses, 10 ~ 15g/L of yeast extract, corn pulp 10
~ 15g/L, 1 ~ 2g/L of potassium dihydrogen phosphate, 0.5 ~ 1g/L of magnesium sulfate, 0.2 ~ 0.5g/L of manganese sulfate, Tween-80 0.5~1%, carbonic acid
8 ~ 10g/L of calcium, step（2）Described in fermentation medium components be：150 ~ 200g/L of molasses, 20 ~ 30g/L of yeast extract, corn
Starch 30 ~ 50g/L, 2 ~ 3g/L of potassium dihydrogen phosphate, 0.5 ~ 1g/L of magnesium sulfate, 0.2 ~ 0.5g/L of manganese sulfate, Tween-80 0.5~1%,
20 ~ 30g/L of calcium carbonate；
In one of the embodiments, step（1）Described in lactic acid bacteria culturers for lactobacillus acidophilus, lactobacillus plantarum, Bao Jiali
Any one in sub- lactobacillus, Bifidobacterium；
In one of the embodiments, step（1）Described in seed culture based component be：Molasses 70g/L, yeast extract 12g/L,
Corn pulp 15g/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 10g/L；
In one of the embodiments, step（2）Described in the ingredient of fermentation medium be：Molasses 200g/L, yeast extract
25g/L, corn pulp 50g/L, potassium dihydrogen phosphate 3g/L, magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate
30g/L；
In one of the embodiments, step（4）It is respectively oligofructose that middle protective agent each component, which accounts for bacterium mud mass percent,
0.5%th, beta-cyclodextrin 3%, vitamin C 0.3%, soybean protein isolate 2%, sodium alginate 3%；
The preparation method of above-mentioned high-activity lactic acid bacteria solid formulation, first by using molasses, corn pulp in the fermentation medium
Culture medium cost is reduced Deng inexpensive carbon source nitrogen source, secondly by addition calcium carbonate and Discrete control pH, is reduced to lactic acid bacteria
Inhibit, so as to improve fermentation level.By adding protective agent and resistant starch, the stabilization during lactic acid bacteria preserves is added
Property, and substantially increase its survival rate after gastric environment.
The preparation method of above-mentioned high-activity lactic acid bacteria solid formulation, simple for process, operation easily, production efficiency it is high, and make
Standby lactic acid bacteria solid formulation activity is high, and stability is good.
Specific embodiment
In order to make the foregoing objectives, features and advantages of the present invention clearer and more comprehensible, below to the specific reality of the present invention
The mode of applying is described in detail.Many details are elaborated in the following description in order to fully understand the present invention.But
The invention can be embodied in many other ways as described herein, and those skilled in the art can be without prejudice to this hair
Similar improvement is done in the case of bright intension, therefore the present invention is not limited to the specific embodiments disclosed below.
The preparation method of the high-activity lactic acid bacteria solid formulation of one embodiment, comprises the following steps：
Step S100, extracting lactic acid bacterium strain is inoculated in by 5 ~ 10% inoculum concentration in the seed culture medium after sterilizing, at 30 ~ 40 DEG C
15 ~ 20h of quiescent culture, up to seed liquor after culture；
Wherein, seed culture based component is：60 ~ 80g/L of molasses, 10 ~ 15g/L of yeast extract, 10 ~ 15g/L of corn pulp, di(2-ethylhexyl)phosphate
1 ~ 2g/L of hydrogen potassium, 0.5 ~ 1g/L of magnesium sulfate, 0.2 ~ 0.5g/L of manganese sulfate, Tween-80 0.5~1%, 8 ~ 10g/L of calcium carbonate；
Preferably, lactic acid bacteria culturers are arbitrary in lactobacillus acidophilus, lactobacillus plantarum, lactobacillus bulgaricus, Bifidobacterium
It is a kind of；
Preferably, seed culture based component is：Molasses 70g/L, yeast extract 12g/L, corn pulp 15g/L, potassium dihydrogen phosphate 2g/
L, magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 10g/L；
Step S200, seed liquor is inoculated in by 10 ~ 15% inoculum concentration in fermentation medium, 40 ~ 48h is cultivated at 30 ~ 40 DEG C,
Up to maturing fermentation liquid, 30~50rpm of speed of agitator in fermentation process, after zymotic fluid pH is reduced to 4.5, by every liter of culture base flow
Ratio addition molasses when 5 ~ 10g/ of sugaring honey is small, while pH4.5~5.0 are controlled by flowing hydrogenation calcium oxide, it is controlled after the 30h that ferments
PH4.0~4.5 processed；
Wherein, fermentation medium components are：150 ~ 200g/L of molasses, 20 ~ 30g/L of yeast extract, 30 ~ 50g/L of corn pulp, phosphoric acid
2 ~ 3g/L of potassium dihydrogen, 0.5 ~ 1g/L of magnesium sulfate, 0.2 ~ 0.5g/L of manganese sulfate, Tween-80 0.5~1%, 20 ~ 30g/L of calcium carbonate；
Preferably, the ingredient of fermentation medium is：Molasses 200g/L, yeast extract 25g/L, corn pulp 50g/L, potassium dihydrogen phosphate
3g/L, magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 30g/L；
Step S300,4000~6000rpm of maturing fermentation liquid is centrifuged into 10~15min, respectively obtains bacterium mud and fermentation broth liquor
Part by the liquid portion through plate-frame filtering, ultrafiltration, activated carbon decolorizing and vacuum concentration, obtains the dense of solid content 30%~40%
Contracting liquid；
Step S400, protective agent is added in the bacterium mud into step S300, be uniformly mixed, obtained bacterium mud protection agent composition, protect
Shield agent each component account for bacterium mud mass percent be respectively oligofructose 0.3 ~ 0.5%, beta-cyclodextrin 2~3%, vitamin C 0.1~
0.3%th, soybean protein isolate 1~2%, sodium alginate 2~3%；
Preferably, it is respectively oligofructose 0.5%, beta-cyclodextrin 3%, vitamin C that protective agent each component, which accounts for bacterium mud mass percent,
0.3%th, soybean protein isolate 2%, sodium alginate 3%；
Step S500, the bacterium mud protection agent composition and step obtained step S400（3）In concentrate mixing, and add account for
20~30% resistant starch of solid quality percentage, 0.5~1% carragheen, 0.2~0.5% xanthans in concentrate, mixing are equal
It is even, softwood is made；
Step S600, by softwood made from step S500, be put into squeeze out and pelletize in finisher,pill, granulation conditions for 600~800 turns/
Processing 10~15 minutes under minute, are dried to moisture content 5~8% at 40~50 DEG C to get high-activity lactic acid bacteria solid-state system after granulation
Agent；
Main advantages of the present invention are：
1st, using molasses as primary carbon source, corn pulp is as nitrogen source, compared to common MRS culture mediums, reduce culture medium into
This, while in molasses, compared to sugared part of single kind, lactic acid bacteria life is more advantageous to comprising sugars such as sucrose, glucose, fructose
It is long；
2nd, it is short for the seed culture time, it is necessary to nutrition it is less, seed culture medium compare fermentation medium, nutritional ingredient concentration
It is lower, it on the one hand prevents from growing due to hyperosmosis lactic acid bacteria inhibiting caused by nutritional ingredient excessive concentration, on the other hand prevent
The speed of growth of lactic acid bacteria is too fast, and product such as lactic acid of generation etc. is excessive, so as to which lactic acid bacteria inhibiting is grown；
3rd, by flowing sugaring honey, control carbon source supply, so as to the speed of growth of lactic acid bacteria is controlled in suitable scope, neither
Lactic acid bacteria can will not be made to overgrow, and then be subject to metabolite because of overnutrition because subalimentation reduces fermentation efficiency
Inhibit, reduce the ability and activity of continued propagation breeding；
4th, a large amount of lactic acid can be generated during lactobacter growth, pH is caused to reduce, too low pH lactic acid bacteria inhibiting can give birth in turn
It is long.One aspect of the present invention adds calcium carbonate in basal medium, with the lactic acid of generation in earlier fermentation, ensures lactic acid early period
Bacterium adds calcium hydroxide during the fermentation there are one good growing environment, controls pH, while generates calcium lactate；
5th, sucrose is easier to be hydrolyzed into glucose and fructose in acid condition in molasses, and the growth for molasses and lactic acid bacteria is special
Property, present invention employs three sections of unique pH control technologies, earlier fermentation nature pH growths, but pass through the calcium carbonate in culture medium
A part of lactic acid is neutralized, slows down pH decrease speeds, after pH drops to 4.5, calcium hydroxide control pH4.5~5.0 is added, ensures
The growth of lactic acid bacteria will not be suppressed because of too low pH, after the 30h that ferments, control pH4.0~4.5, allow sucrose in molasses more
Glucose and fructose are easily converted to, so as to be utilized by lactic acid bacteria.Method is regulated and controled by this pH, optimal control lactic acid bacteria
The speed of growth substantially increases fermentation level；
6th, in common lactic acid bacteria production technology, merely with the lactic acid thalline in bacterium mud after the completion of fermentation, and the liquid after centrifuging
Body portion is not utilized well, is dealt with improperly or even can be caused environmental pollution.Containing actually in liquid portion includes
Bulk fermentation product including calcium lactate.Most of effect that lactic acid bacteria plays in human or animal's body is exactly based on these products
Realize, thus the present invention by liquid portion by plate-frame filtering, ultrafiltration, activated carbon decolorizing and vacuum concentration, remove therein
It after the substances such as pigment, colloid, is back to use in subsequent technique, on the one hand remains useful component therein, lactic acid bacteria can be increased and consolidated
The application effect of state preparation, the on the other hand presence of these ingredients also play a very good protection to lactic acid bacteria；
7th, the stability of lactic acid bacteria can be increased by the synergistic effect of Multiple components in protective agent, reduce the work during preserving
Property loss, particularly xylo-oligosaccharide with lactic acid bacteria together into enteron aisle lactic acid bacteria can be promoted to improve the effect of intestinal environment.
8th, the characteristic for being not easy to be degraded in gastric environment using resistant starch by the viscous glutinous effect of carragheen, xanthans, is used
Resistant starch can dramatically increase lactic acid bacteria and cross the survival rate after stomach in lactic acid bacteria is wrapped in.
The preparation method of the high-activity lactic acid bacteria solid formulation of the present invention, simple for process, operation easily, production efficiency it is high,
And the lactic acid bacteria solid formulation activity prepared is high, stability is good.
With reference to specific embodiment, the invention will be further elaborated.
Embodiment 1
A kind of preparation method of high-activity lactic acid bacteria solid formulation, comprises the following steps：
（1）Lactobacillus acidophilus species is taken to be inoculated in by 5% inoculum concentration in the seed culture medium after sterilizing, in 40 DEG C of quiescent cultures
15h, up to seed liquor after culture；
Wherein, seed culture based component is：Molasses 80g/L, yeast extract 15g/L, corn pulp 15g/L, potassium dihydrogen phosphate 2g/L,
Magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 10g/L；
（2）Seed liquor is inoculated in by 15% inoculum concentration in fermentation medium, 48h are cultivated at 40 DEG C to get maturing fermentation liquid,
Speed of agitator 50rpm in fermentation process, after zymotic fluid pH is reduced to 4.5, by ratios of every liter of culture base flow sugaring honey 10g/ when small
Example addition molasses, while pH4.5 is controlled by flowing hydrogenation calcium oxide, control pH4.0 after the 30h that ferments；
Wherein, fermentation medium components are：Molasses 200g/L, yeast extract 20g/L, corn pulp 30g/L, potassium dihydrogen phosphate 3g/
L, magnesium sulfate 0.5g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 20g/L；
（3）Maturing fermentation liquid 4000rpm is centrifuged into 10min, bacterium mud and fermentation broth liquor part are respectively obtained, by the liquid portion
Through plate-frame filtering, ultrafiltration, activated carbon decolorizing and vacuum concentration, the concentrate of solid content 30% is obtained；
（4）To step（3）In bacterium mud in add protective agent, be uniformly mixed, obtain bacterium mud protection agent composition, protective agent each group
Point account for bacterium mud mass percent be respectively oligofructose 0.5%, beta-cyclodextrin 3%, vitamin C 0.1%, soybean protein isolate 1%,
Sodium alginate 3%；
（5）By step（4）Obtained bacterium mud protection agent composition and step（3）In concentrate mixing, and add account for concentrate
Middle 30% resistant starch of solid quality percentage, 0.5% carragheen, 0.2% xanthans are uniformly mixed, and softwood is made；
（6）By step（5）Softwood obtained is put into squeeze out and pelletize in finisher,pill, and granulation conditions are to handle 10 under 800 revs/min
Minute, it is dried after granulation at 50 DEG C to moisture content 8% to get high-activity lactic acid bacteria solid formulation；
Embodiment 2
A kind of preparation method of high-activity lactic acid bacteria solid formulation, comprises the following steps：
（1）Lactobacillus plantarum strain is taken to be inoculated in by 10% inoculum concentration in the seed culture medium after sterilizing, in 30 DEG C of quiescent cultures
20h, up to seed liquor after culture；
Wherein, seed culture based component is：Molasses 80g/L, yeast extract 15g/L, corn pulp 10g/L, potassium dihydrogen phosphate 2g/L,
Magnesium sulfate 0.5g/L, manganese sulfate 0.2g/L, Tween-80 1%, calcium carbonate 8g/L；
（2）Seed liquor is inoculated in by 10% inoculum concentration in fermentation medium, 40h are cultivated at 30 DEG C to get maturing fermentation liquid,
Speed of agitator 30rpm in fermentation process, after zymotic fluid pH is reduced to 4.5, by ratios of every liter of culture base flow sugaring honey 10g/ when small
Example addition molasses, while pH5.0 is controlled by flowing hydrogenation calcium oxide, control pH4.5 after the 30h that ferments；
Wherein, fermentation medium components are：Molasses 150g/L, yeast extract 20g/L, corn pulp 50g/L, potassium dihydrogen phosphate 2g/
L, magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 0.5%, calcium carbonate 30g/L；
（3）Maturing fermentation liquid 4000rpm is centrifuged into 15min, bacterium mud and fermentation broth liquor part are respectively obtained, by the liquid portion
Through plate-frame filtering, ultrafiltration, activated carbon decolorizing and vacuum concentration, the concentrate of solid content 40% is obtained；
（4）To step（3）In bacterium mud in add protective agent, be uniformly mixed, obtain bacterium mud protection agent composition, protective agent each group
Point account for bacterium mud mass percent be respectively oligofructose 0.3%, beta-cyclodextrin 2%, vitamin C 0.3%, soybean protein isolate 2%,
Sodium alginate 2%；
（5）By step（4）Obtained bacterium mud protection agent composition and step（3）In concentrate mixing, and add account for concentrate
Middle 20% resistant starch of solid quality percentage, 1% carragheen, 0.2% xanthans are uniformly mixed, and softwood is made；
（6）By step（5）Softwood obtained is put into squeeze out and pelletize in finisher,pill, and granulation conditions are to handle 15 under 600 revs/min
Minute, it is dried after granulation at 40 DEG C to moisture content 5% to get high-activity lactic acid bacteria solid formulation；
Embodiment 3
A kind of preparation method of high-activity lactic acid bacteria solid formulation, comprises the following steps：
（1）Bulgarian lactobacillus strain is taken to be inoculated in by 10% inoculum concentration in the seed culture medium after sterilizing, in 30 DEG C of standings
20h is cultivated, up to seed liquor after culture；
Wherein, seed culture based component is：Molasses 80g/L, yeast extract 10g/L, corn pulp 10g/L, potassium dihydrogen phosphate 1g/L,
Magnesium sulfate 1g/L, manganese sulfate 0.2g/L, Tween-80 0.5%, calcium carbonate 10g/L；
（2）Seed liquor is inoculated in by 10% inoculum concentration in fermentation medium, 40h are cultivated at 40 DEG C to get maturing fermentation liquid,
Speed of agitator 30rpm in fermentation process, after zymotic fluid pH is reduced to 4.5, by ratios of every liter of culture base flow sugaring honey 10g/ when small
Example addition molasses, while pH4.8 is controlled by flowing hydrogenation calcium oxide, control pH4.2 after the 30h that ferments；
Wherein, fermentation medium components are：Molasses 200g/L, yeast extract 30g/L, corn pulp 50g/L, potassium dihydrogen phosphate 3g/
L, magnesium sulfate 0.5g/L, manganese sulfate 0.2g/L, Tween-80 0.5%, calcium carbonate 20g/L；
（3）Maturing fermentation liquid 6000rpm is centrifuged into 15min, bacterium mud and fermentation broth liquor part are respectively obtained, by the liquid portion
Through plate-frame filtering, ultrafiltration, activated carbon decolorizing and vacuum concentration, the concentrate of solid content 30% is obtained；
（4）To step（3）In bacterium mud in add protective agent, be uniformly mixed, obtain bacterium mud protection agent composition, protective agent each group
Point account for bacterium mud mass percent be respectively oligofructose 0.3%, beta-cyclodextrin 2%, vitamin C 0.1%, soybean protein isolate 1%,
Sodium alginate 3%；
（5）By step（4）Obtained bacterium mud protection agent composition and step（3）In concentrate mixing, and add account for concentrate
Middle 20% resistant starch of solid quality percentage, 1% carragheen, 0.5% xanthans are uniformly mixed, and softwood is made；
（6）By step（5）Softwood obtained is put into squeeze out and pelletize in finisher,pill, and granulation conditions are to handle 10 under 800 revs/min
Minute, it is dried after granulation at 50 DEG C to moisture content 5% to get high-activity lactic acid bacteria solid formulation；
Embodiment 4
A kind of preparation method of high-activity lactic acid bacteria solid formulation, comprises the following steps：
（1）Bifidobacterium species is taken to be inoculated in by 10% inoculum concentration in the seed culture medium after sterilizing, in 37 DEG C of quiescent cultures
20h, up to seed liquor after culture；
Wherein, seed culture based component is：Molasses 70g/L, yeast extract 12g/L, corn pulp 15g/L, potassium dihydrogen phosphate 2g/L,
Magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 10g/L；
（2）Seed liquor is inoculated in by 15% inoculum concentration in fermentation medium, 48h are cultivated at 37 DEG C to get maturing fermentation liquid,
Speed of agitator 50rpm in fermentation process, after zymotic fluid pH is reduced to 4.5, by ratios of every liter of culture base flow sugaring honey 10g/ when small
Example addition molasses, while pH4.5 is controlled by flowing hydrogenation calcium oxide, control pH4.0 after the 30h that ferments；
Wherein, fermentation medium components are：Molasses 200g/L, yeast extract 25g/L, corn pulp 50g/L, potassium dihydrogen phosphate 3g/
L, magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 30g/L；
（3）Maturing fermentation liquid 6000rpm is centrifuged into 10min, bacterium mud and fermentation broth liquor part are respectively obtained, by the liquid portion
Through plate-frame filtering, ultrafiltration, activated carbon decolorizing and vacuum concentration, the concentrate of solid content 40% is obtained；
（4）To step（3）In bacterium mud in add protective agent, be uniformly mixed, obtain bacterium mud protection agent composition, protective agent each group
It is respectively oligofructose 0.5%, beta-cyclodextrin 3%, vitamin C 0.3%, soybean protein isolate 2%, sea point to account for bacterium mud mass percent
Mosanom 3%；
（5）By step（4）Obtained bacterium mud protection agent composition and step（3）In concentrate mixing, and add account for concentrate
Middle 30% resistant starch of solid quality percentage, 0.5% carragheen, 0.2% xanthans are uniformly mixed, and softwood is made；
（6）By step（5）Softwood obtained is put into squeeze out and pelletize in finisher,pill, and granulation conditions are to handle 10 under 800 revs/min
Minute, it is dried after granulation at 50 DEG C to moisture content 8% to get high-activity lactic acid bacteria solid formulation；
Comparative example 1
A kind of preparation method of high-activity lactic acid bacteria solid formulation, comprises the following steps：
（1）Bifidobacterium species is taken to be inoculated in by 10% inoculum concentration in the seed culture medium after sterilizing, in 37 DEG C of quiescent cultures
20h, up to seed liquor after culture；
Wherein, seed culture based component is：Molasses 70g/L, yeast extract 12g/L, corn pulp 15g/L, potassium dihydrogen phosphate 2g/L,
Magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 10g/L；
（2）Seed liquor is inoculated in by 15% inoculum concentration in fermentation medium, 48h are cultivated at 37 DEG C to get maturing fermentation liquid,
Speed of agitator 50rpm in fermentation process, after zymotic fluid pH is reduced to 4.5, by ratios of every liter of culture base flow sugaring honey 10g/ when small
Example addition molasses, while control pH4.5 by flowing hydrogenation calcium oxide；
Wherein, fermentation medium components are：Molasses 200g/L, yeast extract 25g/L, corn pulp 50g/L, potassium dihydrogen phosphate 3g/
L, magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 30g/L；
（3）Maturing fermentation liquid 6000rpm is centrifuged into 10min, respectively obtains bacterium mud and fermentation broth liquor part；
（4）To step（3）In bacterium mud in add protective agent, be uniformly mixed, obtain bacterium mud protection agent composition, protective agent each group
It is respectively oligofructose 0.5%, beta-cyclodextrin 3%, vitamin C 0.3%, soybean protein isolate 2%, sea point to account for bacterium mud mass percent
Mosanom 3%；
（5）To step（4）It is anti-to account for bacterium mud protective agent mixture quality percentage 30% for addition in obtained bacterium mud protection agent composition
Property starch, 0.5% carragheen, 0.2% xanthans, be uniformly mixed, be made softwood；
（6）By step（5）Softwood obtained is put into squeeze out and pelletize in finisher,pill, and granulation conditions are to handle 10 under 800 revs/min
Minute, it is dried after granulation at 50 DEG C to moisture content 8% to get high-activity lactic acid bacteria solid formulation；
Comparative example 2
A kind of preparation method of high-activity lactic acid bacteria solid formulation, comprises the following steps：
（1）Bifidobacterium species is taken to be inoculated in by 10% inoculum concentration in the seed culture medium after sterilizing, in 37 DEG C of quiescent cultures
20h, up to seed liquor after culture；
Wherein, seed culture based component is：Molasses 70g/L, yeast extract 12g/L, corn pulp 15g/L, potassium dihydrogen phosphate 2g/L,
Magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 10g/L；
（2）Seed liquor is inoculated in by 15% inoculum concentration in fermentation medium, 48h are cultivated at 37 DEG C to get maturing fermentation liquid,
Speed of agitator 50rpm in fermentation process, after zymotic fluid pH is reduced to 4.0, by ratios of every liter of culture base flow sugaring honey 10g/ when small
Example addition molasses, while control pH4.0 by flowing hydrogenation calcium oxide；
Wherein, fermentation medium components are：Molasses 200g/L, yeast extract 25g/L, corn pulp 50g/L, potassium dihydrogen phosphate 3g/
L, magnesium sulfate 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 30g/L；
（3）Maturing fermentation liquid 6000rpm is centrifuged into 10min, respectively obtains bacterium mud and fermentation broth liquor part；
（4）To step（3）In bacterium mud in add protective agent, be uniformly mixed, obtain bacterium mud protection agent composition, protective agent each group
It is respectively oligofructose 0.5%, beta-cyclodextrin 3%, soybean protein isolate 2%, sodium alginate 3% point to account for bacterium mud mass percent；
（5）To step（4）It is anti-to account for bacterium mud protective agent mixture quality percentage 5% for addition in obtained bacterium mud protection agent composition
Property starch, 0.5% carragheen, 0.2% xanthans, be uniformly mixed, be made softwood；
（6）By step（5）Softwood obtained is put into squeeze out and pelletize in finisher,pill, and granulation conditions are to handle 10 under 800 revs/min
Minute, it is dried after granulation at 50 DEG C to moisture content 8% to get high-activity lactic acid bacteria solid formulation；
Above-described embodiment and comparative example and the comparison situation such as following table of the lactic acid bacteria of prior art production：
1st, maturing fermentation liquid lactic acid bacterium number average out to 2.8 × 10 in Examples 1 to 410Cfu/ml is ripe in comparative example 1~2
Zymotic fluid lactic acid bacterium number average out to 6.2 × 109Cfu/ml, comparative example fermentation level is suitable with the prior art, compared with reality
For applying example 1~4, regulate and control using two sections of pH, three sections of exclusive pH modulation process of the present invention effectively provide for lactic acid bacteria
The growing environment stablized, promotes the utilizing status of molasses, and final fermentation level than the prior art improves 4 times or so.
2nd, viable bacteria rate is suitable with the prior art after lactic acid bacteria prepared by comparative example 1~2 preserves 6 months, and Examples 1 to 4 is put down
Equal viable bacteria rate 55.4%, is greatly improved compared with prior art, and a large amount of protectant in the research process of the present invention
Effect is compared, it is found that reuse can be obviously improved the stability of lactic acid bacteria after processing by fermentation broth liquor part, and
It can also mutually be cooperateed with other protective agents, effectively solve the problems, such as that lactic acid bacteria is not easy to maintain.
3rd, comparative example 1~2 is handled by simulate the gastric juice（PH2.0,50u/mL, 2h）Afterwards, viable bacteria rate is suitable with the prior art,
The resistant starch that wherein comparative example 1 is added is more than comparative example 2, this demonstrates the protective effect that resistant starch crosses lactic acid bacteria in stomach.
The mean viable rate 37.4% of Examples 1 to 4 improves about 1.5 times than the prior art, illustrates protective agent in the present invention, reuse
Fermentation broth liquor in ingredient, resistant starch etc. effectively played and protect lactic acid bacteria, can allow more viable lactic acid bacterias into
Enter enteron aisle to play a role.
Embodiment described above only expresses the several embodiments of the present invention, and description is more specific and detailed, but simultaneously
Cannot the limitation to the scope of the claims of the present invention therefore be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the guarantor of the present invention
Protect scope.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (5)
1. a kind of preparation method of high-activity lactic acid bacteria solid formulation, comprises the following steps：
（1）Extracting lactic acid bacterium strain is inoculated in by 5 ~ 10% inoculum concentration in the seed culture medium after sterilizing, and training is stood at 30 ~ 40 DEG C
15 ~ 20h is supported, up to seed liquor after culture；
（2）Seed liquor is inoculated in by 10 ~ 15% inoculum concentration in fermentation medium, 30 ~ 40 DEG C cultivate 40 ~ 48h to get into
Ripe zymotic fluid, 30~50rpm of speed of agitator in fermentation process, after zymotic fluid pH is reduced to 4.5, by every liter of culture base flow sugaring honey
Ratio addition molasses when 5 ~ 10g/ is small, while pH4.5~5.0 are controlled by flowing hydrogenation calcium oxide, it is controlled after the 30h that ferments
PH4.0~4.5；
（3）4000~6000rpm of maturing fermentation liquid is centrifuged into 10~15min, respectively obtains bacterium mud and fermentation broth liquor part, it will
The liquid portion obtains the concentrate of solid content 30%~40% through plate-frame filtering, ultrafiltration, activated carbon decolorizing and vacuum concentration；
（4）To step（3）In bacterium mud in add protective agent, be uniformly mixed, obtain bacterium mud protection agent composition, protective agent each group
It is respectively oligofructose 0.3 ~ 0.5%, beta-cyclodextrin 2~3%, vitamin C 0.1~0.3%, soybean point to account for bacterium mud mass percent
Protein isolate 1~2%, sodium alginate 2~3%；
（5）By step（4）Obtained bacterium mud protection agent composition and step（3）In concentrate mixing, and add account for concentrate
Middle 20~30% resistant starch of solid quality percentage, 0.5~1% carragheen, 0.2~0.5% xanthans are uniformly mixed, and are made
Softwood；
（6）By step（5）Softwood obtained is put into squeeze out and pelletize in finisher,pill, and granulation conditions are under 600~800 revs/min
Reason 10~15 minutes is dried to moisture content 5~8% at 40~50 DEG C to get high-activity lactic acid bacteria solid formulation after granulation；
Wherein, step（1）Described in seed culture based component be：60 ~ 80g/L of molasses, 10 ~ 15g/L of yeast extract, corn pulp 10
~ 15g/L, 1 ~ 2g/L of potassium dihydrogen phosphate, 0.5 ~ 1g/L of magnesium sulfate, 0.2 ~ 0.5g/L of manganese sulfate, Tween-80 0.5~1%, carbonic acid
8 ~ 10g/L of calcium, step（2）Described in fermentation medium components be：150 ~ 200g/L of molasses, 20 ~ 30g/L of yeast extract, corn
Starch 30 ~ 50g/L, 2 ~ 3g/L of potassium dihydrogen phosphate, 0.5 ~ 1g/L of magnesium sulfate, 0.2 ~ 0.5g/L of manganese sulfate, Tween-80 0.5~1%,
20 ~ 30g/L of calcium carbonate.
2. the preparation method of high-activity lactic acid bacteria solid formulation according to claim 1, which is characterized in that step（1）In
The lactic acid bacteria culturers are any one in lactobacillus acidophilus, lactobacillus plantarum, lactobacillus bulgaricus, Bifidobacterium.
3. the preparation method of high-activity lactic acid bacteria solid formulation according to claim 1, which is characterized in that step（1）In
The seed culture based component is：Molasses 70g/L, yeast extract 12g/L, corn pulp 15g/L, potassium dihydrogen phosphate 2g/L, sulfuric acid
Magnesium 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 10g/L.
4. the preparation method of high-activity lactic acid bacteria solid formulation according to claim 1, which is characterized in that step（2）In
The ingredient of the fermentation medium is：Molasses 200g/L, yeast extract 25g/L, corn pulp 50g/L, potassium dihydrogen phosphate 3g/L, sulphur
Sour magnesium 1g/L, manganese sulfate 0.5g/L, Tween-80 1%, calcium carbonate 30g/L.
5. the preparation method of high-activity lactic acid bacteria solid formulation according to claim 1, which is characterized in that step（4）In
It is respectively oligofructose 0.5%, beta-cyclodextrin 3%, vitamin C 0.3%, soybean point that protective agent each component, which accounts for bacterium mud mass percent,
From albumen 2%, sodium alginate 3%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810165716.5A CN108048381A (en) | 2018-02-28 | 2018-02-28 | A kind of preparation method of high-activity lactic acid bacteria solid formulation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810165716.5A CN108048381A (en) | 2018-02-28 | 2018-02-28 | A kind of preparation method of high-activity lactic acid bacteria solid formulation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108048381A true CN108048381A (en) | 2018-05-18 |
Family
ID=62125983
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810165716.5A Pending CN108048381A (en) | 2018-02-28 | 2018-02-28 | A kind of preparation method of high-activity lactic acid bacteria solid formulation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108048381A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108991320A (en) * | 2018-08-14 | 2018-12-14 | 河南金鹿堂药业有限公司 | A kind of biodiasmin and preparation method thereof to ferment automatically |
| CN111349578A (en) * | 2018-12-24 | 2020-06-30 | 中粮生物化学（安徽）股份有限公司 | Preparation method of lactobacillus solid microbial inoculum |
| CN112175858A (en) * | 2019-07-05 | 2021-01-05 | 中粮生物化学（安徽）股份有限公司 | Solid culture medium for lactobacillus plantarum fermentation, lactobacillus plantarum solid microbial inoculum and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101353636A (en) * | 2008-09-11 | 2009-01-28 | 中国水产科学研究院南海水产研究所 | Production method of lactobacillus micro-ecological preparation for cultivation water regulation |
| CN102018217A (en) * | 2009-09-19 | 2011-04-20 | 菲伯纳生物医药有限责任公司 | Composition containing bifidobacterium and application thereof |
| CN102860411A (en) * | 2012-10-16 | 2013-01-09 | 广东海洋大学 | Lactobacillus rhamnosus probiotic and preparation method thereof |
| CN104642746A (en) * | 2015-03-23 | 2015-05-27 | 四川高福记生物科技有限公司 | Preparation method of feeding lactic acid bacteria micro pills |
| CN106282048A (en) * | 2016-08-04 | 2017-01-04 | 华侨大学 | A kind of complex microorganism preparations and its preparation method and application |
| CN107815429A (en) * | 2017-11-14 | 2018-03-20 | 北京好实沃生物技术有限公司 | A kind of resistance to granulation lactobacillus preparation and preparation method thereof |
-
2018
- 2018-02-28 CN CN201810165716.5A patent/CN108048381A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101353636A (en) * | 2008-09-11 | 2009-01-28 | 中国水产科学研究院南海水产研究所 | Production method of lactobacillus micro-ecological preparation for cultivation water regulation |
| CN102018217A (en) * | 2009-09-19 | 2011-04-20 | 菲伯纳生物医药有限责任公司 | Composition containing bifidobacterium and application thereof |
| CN102860411A (en) * | 2012-10-16 | 2013-01-09 | 广东海洋大学 | Lactobacillus rhamnosus probiotic and preparation method thereof |
| CN104642746A (en) * | 2015-03-23 | 2015-05-27 | 四川高福记生物科技有限公司 | Preparation method of feeding lactic acid bacteria micro pills |
| CN106282048A (en) * | 2016-08-04 | 2017-01-04 | 华侨大学 | A kind of complex microorganism preparations and its preparation method and application |
| CN107815429A (en) * | 2017-11-14 | 2018-03-20 | 北京好实沃生物技术有限公司 | A kind of resistance to granulation lactobacillus preparation and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| 程丽娟 袁静: "《发酵食品工艺学》", 31 December 2002 * |
| 臧学丽、胡莉娟: "《实用发酵工程技术》", 31 January 2017 * |
| 袁秀丽等: ""乳酸菌包埋方法的研究"", 《食品与发酵工业》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108991320A (en) * | 2018-08-14 | 2018-12-14 | 河南金鹿堂药业有限公司 | A kind of biodiasmin and preparation method thereof to ferment automatically |
| CN111349578A (en) * | 2018-12-24 | 2020-06-30 | 中粮生物化学（安徽）股份有限公司 | Preparation method of lactobacillus solid microbial inoculum |
| CN111349578B (en) * | 2018-12-24 | 2022-12-23 | 中粮生物科技股份有限公司 | Preparation method of lactobacillus solid microbial inoculum |
| CN112175858A (en) * | 2019-07-05 | 2021-01-05 | 中粮生物化学（安徽）股份有限公司 | Solid culture medium for lactobacillus plantarum fermentation, lactobacillus plantarum solid microbial inoculum and preparation method and application thereof |
| CN112175858B (en) * | 2019-07-05 | 2022-11-18 | 中粮生物科技股份有限公司 | Solid culture medium for lactobacillus plantarum fermentation, lactobacillus plantarum solid microbial inoculum and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Beheshtipour et al. | Supplementation of Spirulina platensis and Chlorella vulgaris algae into probiotic fermented milks | |
| CN101715876B (en) | Preparation method of high-efficiency compound microorganism soybean meal leaven | |
| US11707494B2 (en) | Pre-fermented symbiotic matrix based on a cereal suspension with encapsulated probiotics, manufacture process and corresponding utilization | |
| CN106721932A (en) | A kind of feed biological de- mould dose of livestock and poultry and its application | |
| CN104664154B (en) | Yeast culture and preparation method thereof | |
| CN108048381A (en) | A kind of preparation method of high-activity lactic acid bacteria solid formulation | |
| CN103766486B (en) | A kind of milky-drinks containing bifidus bacillus embedding pearl and preparation method thereof | |
| CN110236046A (en) | A kind of high viable type lactobacillus-fermented blueberry pomace beverage and preparation method thereof | |
| CN110607255A (en) | Preparation method and application of lactobacillus delbrueckii and direct vat set lactobacillus delbrueckii starter | |
| US7326558B2 (en) | Process for treating spirulina | |
| JP2020174667A (en) | Methods for producing lactic acid bacteria with protein-polysaccharide dual coating by using protein hydrolysate | |
| CN108041383B (en) | High-nutrition tremella beverage with typical tremella flavor and preparation method thereof | |
| CN109953322A (en) | The preparation method of dragon fruit ferment | |
| WO2015063282A1 (en) | Use of algae to increase the viable active biomass of lactic acid bacteria | |
| CN102077932A (en) | Method for preparing nutritional jelly rich in highly active intestinal probiotics | |
| CN109182397B (en) | Method for improving yield of reuterin | |
| Fouad et al. | Bio-Labneh fortified with functional microcapsules filled with chickpea flour and probiotics | |
| CN113679035A (en) | Noni enzyme powder and preparation method thereof | |
| Vinderola et al. | The production of lactic acid bacteria starters and probiotic cultures: An industrial perspective | |
| KR101848230B1 (en) | Prebiotics composition for Lactobacillus with improved stability, and method for stabilization using thereof | |
| CN106795539A (en) | The method that oil is produced in microorganism | |
| Sheth et al. | Recent advancements in the production of probiotic fermented beverages | |
| CN107080134A (en) | A kind of preparation method of organic rice fruits and vegetables functional beverage | |
| CN112391317B (en) | Probiotic bacterial strain composition for producing cubilose acid and application | |
| CN104643094A (en) | Diet supplement enriched with probiotics and preparation method of diet supplement |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180518 |
|
| RJ01 | Rejection of invention patent application after publication |