CN108004191A - A kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation method and its Zymolysis Equipment - Google Patents
A kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation method and its Zymolysis Equipment Download PDFInfo
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Abstract
The present invention relates to biological field, the present invention provides a kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation method, it includes：First inoculating lactobacillus acidophilus' bacterium solution is fermented on unpasteurized solid medium, is inoculated clostridium butyricum bacterium solution and is fermented.It can utilize lactobacillus acidophilus and clostridium butyricum effectively to suppress the growth of miscellaneous bacteria to the coordinate repression of miscellaneous bacteria, effectively control the miscellaneous bacteria in production process, and can be under conditions of raw material be unsterilised, lactobacillus acidophilus and clostridium butyricum are fermented, reduce cost, it is highly practical.In addition, the present invention also provides a kind of Zymolysis Equipment, it can be used for anaerobic bacteria or facultative anaerobic bacteria, the solid state fermentation production of such as clostridium butyricum, lactobacillus acidophilus, this equipment is by air outlet pipe, by Volatile Metabolites, such as hydrogen, carbon dioxide are discharged into outside air kind, metabolic inhibition is released, viable bacteria growth and breeding ability can be discharged, improves fermentation viable count.
Description
Technical field
The present invention relates to biological technical field, in particular to a kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation
Method and its equipment.
Background technology
Lactobacillus acidophilus is generally obtained by liquid state fermentation, stringenter to equipment and control requirement, in high temperature drying mistake
It is easily dead in journey, it is general using lyophilized technique in production, it is with high costs.Clostridium butyricum can both be obtained by liquid state fermentation
, it can also be obtained by solid state fermentation, but domestic production at present is not horizontal universal high, and cost is higher.Solid state fermentation produces
The easy pollution microbes of clostridium butyricum, can also reduce the utility value of raw material.
The content of the invention
It is an object of the invention to provide a kind of lactobacillus acidophilus and the production method of clostridium butyricum mixed fermentation, it can
Using lactobacillus acidophilus and clostridium butyricum to the coordinate repression of miscellaneous bacteria, it can effectively suppress the growth of miscellaneous bacteria, effectively control
Miscellaneous bacteria in production process.
Another object of the present invention is to provide the production equipment of a kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation, its
Produced available for the solid state fermentation of anaerobic bacteria or facultative anaerobic bacteria, viable bacteria growth and breeding ability can be improved.
What the present invention was realized in：
A kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation method, it includes：On unpasteurized solid medium first
Inoculating lactobacillus acidophilus' bacterium solution is fermented, and is inoculated clostridium butyricum bacterium solution and is fermented.
A kind of Zymolysis Equipment, it is used for above-mentioned lactobacillus acidophilus and clostridium butyricum mixed fermentation method, it includes closed
Container body, container body are equipped with unidirectional gas outlet passage.
The invention has the advantages that：
The present invention provides a kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation method, it includes：In unpasteurized solid-state
First inoculating lactobacillus acidophilus' bacterium solution is fermented on culture medium, is inoculated clostridium butyricum bacterium solution and is fermented.It can utilize thermophilic
Lactobacillus lactis and clostridium butyricum can effectively suppress the growth of miscellaneous bacteria, effectively control produced to the coordinate repression of miscellaneous bacteria
Miscellaneous bacteria in journey, and lactobacillus acidophilus and clostridium butyricum can be fermented, reduced under conditions of raw material is unsterilised
This, it is highly practical.In addition, the present invention also provides a kind of Zymolysis Equipment, it can be used for the solid-state of anaerobic bacteria or facultative anaerobic bacteria hair
Ferment produces, and this equipment is by air outlet pipe, and by Volatile Metabolites, such as hydrogen, carbon dioxide are discharged into outside air kind,
Metabolic inhibition is released, viable bacteria growth and breeding ability can be discharged, improves fermentation viable count.
Brief description of the drawings
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it will be appreciated that the following drawings illustrate only certain embodiments of the present invention, therefore be not construed as pair
The restriction of scope, for those of ordinary skill in the art, without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is structure diagram of the Zymolysis Equipment of first embodiment of the invention offer under the first visual angle；
Fig. 2 is structure diagram of the Zymolysis Equipment of first embodiment of the invention offer under the second visual angle；
Fig. 3 is structure diagram of the Zymolysis Equipment of first embodiment of the invention offer under the 3rd visual angle；
Fig. 4 is the structure diagram of the head cover for the Zymolysis Equipment that first embodiment of the invention provides.
Icon：10- Zymolysis Equipment；100- container bodies；110- head covers；200- air outlet pipes；201- ventholes.
Embodiment
, below will be in the embodiment of the present invention to make the purpose, technical scheme and advantage of the embodiment of the present invention clearer
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, builds according to normal condition or manufacturer
The condition of view carries out.Reagents or instruments used without specified manufacturer, is the conventional production that can be obtained by commercially available purchase
Product.
Below to lactobacillus acidophilus provided in an embodiment of the present invention and clostridium butyricum mixed fermentation method and Zymolysis Equipment
It is specifically described.
On the one hand, lactobacillus acidophilus provided by the invention and clostridium butyricum mixed fermentation method, it includes following steps：
First inoculating lactobacillus acidophilus' bacterium solution is fermented on unpasteurized solid medium, inoculates clostridium butyricum bacterium solution
Ferment.
Lactobacillus acidophilus, belongs to lactobacillus, and Gram-positive bacillus, the end of bar is rounded, is primarily present in small intestine,
Lactic acid, acetic acid and some antibiotic to work to harmful bacteria are discharged, are probiotics.Lactobacillus acidophilus is anaerobic bacteria, it is survived
In the whole intestines and stomach of human body, micro-ecological environment is adjusted.
Clostridium butyricum, also known as Miyarisan, clostridium butyricum, butyric, belongs to Bacillaceae, fusobacterium, is
A kind of Gram-positive bacillus of obligate anaerobic, its both ends blunt circle, center section slightly expand, and bacterium is in direct rod shape or slightly
There is bending, single or paired, short chain is accidental to have Filamentous thalline, whole body flagellum.Spore oval, eccentric or secondary end life.It, which has, adjusts
Whole intestinal colony balance, the features such as promoting intestinal beneficial flora propagation, strengthen immune function.
Above-mentioned lactobacillus acidophilus bacterium solution refers to, by 2~4 parts of yeast extract, 1~3 part of brown sugar, 0.5~1.5 part of bean cake powder, 92
~96 parts of water are uniformly mixed, and it is 7~8,110~130 DEG C to adjust initial pH, sterilizes 20~40 minutes, is cooled to 35~39 DEG C, is
Lactobacillus acidophilus liquid culture medium.The Lactobacillus acidophilus species of a glycerol tube preservation are taken, by 0.05~0.15% inoculum concentration,
Strain is connected in lactobacillus acidophilus culture medium, amplification step by step spreads cultivation, and is lactobacillus acidophilus bacterium solution.
Above-mentioned clostridium butyricum bacterium solution refers to, by 1~3 part of glucose, 1~2 part of peptone, 1~2 part of yeast extract, sulfuric acid
0.05~0.15 part of ammonium, 0.05~0.15 part of sodium carbonate, 0.2~0.4 part of corn pulp, 0.03~0.07 part of manganese sulfate, magnesium sulfate
0.03~0.07 part, 93~97 parts of water be uniformly mixed, adjust initial pH7~8,110~130 DEG C, sterilize 20~40 minutes, be cooled to
36~38 DEG C, be clostridium butyricum fluid nutrient medium.The clostridium butyricum strain of a glycerol tube preservation is taken, by 0.05~0.15%
Inoculum concentration, strain is connected in clostridium butyricum culture medium, step by step amplification spread cultivation, be clostridium butyricum bacterium solution.
Above-mentioned solid medium refers to, made of natural solid material, in some embodiments of the invention, Gu
The fermentation raw material of state culture medium can include 23~27 parts of wheatfeed, 30~34 parts of bean cake powder, 9.5~13.5 parts of corn flour,
Preferably, bean cake powder and corn flour cross 40 mesh sieves after broken.Cross 40 mesh sieves and be because wheat bran, dregs of beans, corn etc. all
It is feed particulate material, and particle is larger, if after fermentation, drying, then crushed, then in crushing process, acidophilus breast bar
Bacterium and clostridium butyricum will undergo pulverizer high-speed cruising caused by shearing force and high temperature, influence thalline stability.And if hair
Crushed before ferment, then after fermentation, drying, material is simply bonded together, it is possible to very small with shearing force and heat production
Coarse powder equipment (kibbler roll) is crushed, and avoids shearing force and high temperature, is a kind of protection to thalline.Carried by the present invention
Live products physical arrangement is not seriously damaged made of the method for confession, and viable bacteria can more stablize.
The incubation of lactobacillus acidophilus is all liquid culture mostly, and the environment sheet survived by enterobacteriaceae is as solid-state
Or semisolid, and the lactobacillus acidophilus atrichia after fluid nutrient medium culture, it is impossible to move, thalline is excessively intensive, metabolism production
Thing concentration is high, and the growth of thalline is suppressed.For solid medium compared to liquid culture medium, cost is few again for tool, easy to operate etc.
Advantage.
It is because lactobacillus acidophilus fermenting carbohydrate produces substantial amounts of acid, peroxide in unpasteurized solid medium top fermentation
Change a variety of antibacterial substances such as hydrogen, diacetyl, bacteriocin, reduce fermentation substrate environmental pH, suppress the growth of pathogenic bacteria.And butyric acid
Clostridium can promote the growth of beneficial Bifidobacterium, lactic acid bacteria, bacteroid, and effectively suppress harmful bacteria staphylococcus, read
Pearl bacterium, klebsiella, campylobacter, Pseudomonas aeruginosa, Escherichia coli, shigella dysenteriae and salmonella typhi and spoilage organisms it is numerous
Grow.Clostridium butyricum and Bifidobacterium can suppress the growth of clostridium, and compared with individually culture, and when two bacterium co-incubations shows
Go out stronger Ant agonism.
For solid medium not by sterilizing, first inoculating lactobacillus acidophilus, are inoculated with clostridium butyricum afterwards, are because clostridium butyricum
During the fermentation, a large amount of butyric acid and hydrogen are produced, the metabolite of generation causes culture medium to be gradually bonded together.And acidophilus
Lactobacillus during the fermentation, will not produce substantial amounts of emplastic.If first it is inoculated with clostridium butyricum, then in inoculation acidophilus breast
During bacillus, mixing is uneven, is unfavorable for fermenting.If first inoculating lactobacillus acidophilus, inoculate clostridium butyricum, then in inoculation butyric acid
When clostridium, mixing ratio is more uniform, beneficial to fermentation.
Further, above-mentioned fermentation raw material has further included 1~3 part of calcium carbonate, 0.5~1.5 part of sodium carbonate, manganese sulfate 0.01
~0.05 part, 0.02~0.08 part of magnesium sulfate.
Above-mentioned inoculating lactobacillus acidophilus' bacterium solution first on unpasteurized solid medium carries out fermentation and refers to, by solid state rheology
80~96 parts of base, 30~50 parts of water, 4~6 parts of lactobacillus acidophilus bacterium solution, are fitted into Zymolysis Equipment, coefficient after mixing
87%~93%, ferment, then, inoculate 4~6 parts of the bacterium solution of clostridium butyricum, after mixing, be again loaded into fermentation and set
In standby, then ferment.After fermentation, it is dried with low temperature fluidized bed, in drying process, temperature of charge is no more than 50 DEG C, does
Moisture is less than 12% after dry.With 40 mesh rotor-vibrating screens sieve, screenings, packed products, oversize oppositely rolling roller crusher into
After 40 mesh sieves, screenings, packed products after row second-time breakage.It is because it only has an of short duration roller using kibbler roll
Journey is pressed through, heat production is small, and shearing force is small, thalline is destroyed small.And because before fermentation, with regard to raw material is crushed, institute
After fermenting and drying, kibbler roll can be used.
Further, in some embodiments of the invention, the mixing ratio of lactobacillus acidophilus bacterium solution and clostridium butyricum bacterium solution
For (0.8~1.2):(0.8~1.2).In other words, the lactobacillus acidophilus bacterium solution being inoculated with above-mentioned solid medium and butyric acid shuttle
The mixing ratio of bacterium bacterium solution is equal or close to 1：1.
Further, on solid state rheology, when the fermentation time of the bacterium solution of inoculating lactobacillus acidophilus is 6~12 small, inoculation
When fermentation time after the bacterium solution of clostridium butyricum is 24~36 small.
On the other hand, the present invention provides a kind of Zymolysis Equipment, it is used for above-mentioned lactobacillus acidophilus and clostridium butyricum mixing
Fermentation process, it includes container body, and container body is equipped with unidirectional gas outlet passage.
Further, in some embodiments of the invention, unidirectional gas outlet passage is air outlet pipe, one end position of air outlet pipe
In in container body, the other end of air outlet pipe is arranged on outside container body, and one end outside container body of air outlet pipe is equipped with list
Sealed to vent valve.
Specifically, one end that air outlet pipe is arranged in container body can be equipped with multiple ventholes, and the aperture of venthole is 0.5
~2mm.
Head cover is additionally provided with the top of container body, air outlet pipe can be connected with head cover.
The feature and performance of the present invention are described in further detail with reference to embodiments.
First embodiment
Attached drawing 1~2 is refer to, the present invention provides a kind of Zymolysis Equipment 10, it includes container body 100, container body 100
Equipped with unidirectional gas outlet passage.
Further, in the present embodiment, container body 100 is column structure, can use stainless steel, glass material
Matter or porcelain, the diameter of container body 100 can be 300~1000mm, high 300~1000mm, bottom and circumferential sealing.
Container body 100 is used to accommodate fermentation raw material progress fermentation process, and container body 100 is equipped with unidirectional outlet passageway
It is to during the fermentation, ventilate, discharges the hydrogen produced in clostridium butyricum fermentation process, while release hydrogen to fourth
The metabolic inhibition that sour clostridium produces.Clostridium butyricum is strict anaerobes, mainly in material submerged fermentation, if be not vented logical
Road, the hydrogen that bottom produces will jack up material, until outside ejection tank.
Further, attached drawing 3~4 is refer to, unidirectional gas outlet passage is air outlet pipe 200, is specially multiple air outlet pipes
200, one end of air outlet pipe 200 is located in container body 100, and the other end of air outlet pipe 200 is arranged on outside container body 100, outlet
Sealed equipped with one-way ventilating valve one end outside container body 100 of pipe 200.
Specifically, one end that air outlet pipe 200 is arranged in container body 100 can be equipped with multiple ventholes 201, venthole
201 can be such that the caused exhaust gas in solid medium discharges in time, prevent the gas of thalline fermentation discharge from suppressing thalline
Growth.The aperture of venthole 201 is 0.5~2mm.When aperture is less than 0.5mm, venting capability deficiency；When aperture is more than 2mm,
Fermentation materials, through Kong Guanzhong, easily block air outlet pipe 200 from hole.
Further, the top of container body 100 is additionally provided with head cover 110, and air outlet pipe 200 can be connected with head cover 110.Cause
It is up to arrange for gas, air outlet pipe 200 is arranged at top is conducive to the discharge of gas.
10 advantage of Zymolysis Equipment provided by the invention is,, must in fermentation process if in fermenting in liquid fermentation fermentor
Hydrogen must be discharged outside toward tank, otherwise, pressure inside the tank will be constantly improve, and the hydrogen of high concentration suppresses clostridium butyricum growth and breeding.Such as
Fruit is fermented in common solid-state Zymolysis Equipment, if hydrogen can not be excluded, the hydrogen of Zymolysis Equipment bottom will be thing
Outside material ejection tank, pollute, cause fermentation to fail.The equipment of invention design, solves hydrogen in solid state fermentation container and arranges
The problem of putting, relieves growth inhibition of the hydrogen to clostridium butyricum, keeps solid material to be all the time in round
The environment of one enclosed oxygen isolation, the growth of clostridium butyricum is exactly the environment for needing enclosed oxygen isolation.
Second embodiment
The present embodiment provides a kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation method.
1 part, 94 parts 3 parts of yeast extract, 2 parts of brown sugar, bean cake powder water are uniformly mixed, adjust initial pH7.5,121 DEG C, sterilizing 30
Minute, 37 DEG C are cooled to, is lactobacillus acidophilus liquid culture medium.The Lactobacillus acidophilus species of a glycerol tube preservation are taken, are pressed
0.1% inoculum concentration, strain is connected in lactobacillus acidophilus culture medium, and amplification step by step spreads cultivation, and is lactobacillus acidophilus bacterium solution, is examined
The viable count of Lactobacillus lactis is surveyed, it is 2.2 hundred million CFU/g to draw lactobacillus acidophilus bacterium solution viable count.
By 2 parts of glucose, 1.5 parts of peptone, 1.5 parts of yeast extract, 0.1 part of ammonium sulfate, 0.1 part of sodium carbonate, corn pulp
0.3 part, 0.05 part of manganese sulfate, 0.05 part of magnesium sulfate, 95 parts of water be uniformly mixed, adjust initial pH7.5,121 DEG C, sterilize 30 minutes,
37 DEG C are cooled to, is clostridium butyricum fluid nutrient medium.Take the clostridium butyricum strain of a glycerol tube preservation, connect by 0.1%
Kind amount, strain is connected in clostridium butyricum culture medium, and amplification step by step spreads cultivation, and is clostridium butyricum bacterium solution.Detect clostridium butyricum
Viable count, it is 0.5 hundred million CFU/g to draw clostridium butyricum active bacteria number.
Wheat bran, dregs of beans, corn are crushed, all cross 40 mesh sieves.By 25 parts of wheatfeed, 32 parts of bean cake powder, corn flour
11.5 parts, 0.5 part of sodium chloride, 2 parts of calcium carbonate, 1 part of sodium carbonate, 0.01 part of manganese sulfate, 0.05 part of magnesium sulfate be uniformly mixed, mixing
Solid material (solid medium) is unsterilised, and the quantity for detecting miscellaneous bacteria and mould is drawn, before fermentation, the mould of hybrid solid-state material
Sum is less than 50,000 CFU/g, and miscellaneous bacteria sum is less than 100,000 CFU/g.
88 parts of hybrid solid-state material, 40 parts of water, 5 parts of lactobacillus acidophilus bacterium solution are taken, loads first embodiment after mixing
In the Zymolysis Equipment of offer, coefficient 90%, top cover, sealing, when fermentation 6 is small, inoculates 5 parts of clostridium butyricum bacterium solution,
After mixing, be again loaded into the Zymolysis Equipment, top cover, then ferment 26 it is small when.
After fermentation, it is dried with low temperature fluidized bed, in drying process, temperature of charge is no more than 50 DEG C, after dry
Moisture is less than 12%.With the sieving of 40 mesh rotor-vibrating screens, screenings, packed products, oversize two are carried out with oppositely rolling roller crusher
It is secondary it is broken after after 40 mesh sieves, screenings, packed products.It is repeated, until all passing through 40 mesh sieves.
After fermentation, it is 4,200,000,000 CFU/g to detect live lactobacillus acidophilus number in finished product, and clostridium butyricum active bacteria number is 1,100,000,000 CFU/
G, clostridium butyricum gemma number are 1,000,000,000 CFU/g.
3rd embodiment
A kind of lactobacillus acidophilus provided in this embodiment and clostridium butyricum mixed fermentation method, principle are carried with second embodiment
What is supplied is identical, different difference lies in parameter, and difference is as follows：
0.5 part, 92 parts 2 parts of yeast extract, 1 part of brown sugar, bean cake powder water are uniformly mixed, adjust initial pH7,120 DEG C, sterilizing 35
Minute, 36 DEG C are cooled to, is lactobacillus acidophilus liquid culture medium.The Lactobacillus acidophilus species of a glycerol tube preservation are taken, are pressed
0.08% inoculum concentration, strain is connected in lactobacillus acidophilus culture medium, and amplification step by step spreads cultivation, and is lactobacillus acidophilus bacterium solution.
By 1 part of glucose, 1 part of peptone, 1 part of yeast extract, 0.05 part of ammonium sulfate, 0.05 part of sodium carbonate, corn pulp 0.2
Part, 0.03 part of manganese sulfate, 0.03 part of magnesium sulfate, 93 parts of water are uniformly mixed, and are adjusted initial pH7.5,121 DEG C, are sterilized 30 minutes, cooling
It is clostridium butyricum fluid nutrient medium to 36 DEG C.The clostridium butyricum strain of a glycerol tube preservation is taken, by 0.1% inoculum concentration,
Strain is connected in clostridium butyricum culture medium, amplification step by step spreads cultivation, and is clostridium butyricum bacterium solution.
Wheat bran, dregs of beans, corn are crushed, all cross 40 mesh sieves.By 25 parts of wheatfeed, 32 parts of bean cake powder, corn flour
11.5 parts, 0.5 part of sodium chloride, 2 parts of calcium carbonate, 1 part of sodium carbonate, 0.01 part of manganese sulfate, 0.05 part of magnesium sulfate be uniformly mixed, mixing
Solid material (solid medium) is unsterilised.
88 parts of hybrid solid-state material, 40 parts of water, 5 parts of lactobacillus acidophilus bacterium solution are taken, loads first embodiment after mixing
In the Zymolysis Equipment of offer, coefficient 87%, top cover, sealing, when fermentation 8 is small, inoculates 4 parts of clostridium butyricum bacterium solution,
After mixing, be again loaded into the Zymolysis Equipment, top cover, then ferment 24 it is small when.
Fourth embodiment
A kind of lactobacillus acidophilus provided in this embodiment and clostridium butyricum mixed fermentation method, principle are implemented with the second~tri-
Example provides identical, different difference lies in parameter, and difference is as follows：
1.5 parts, 96 parts 3 parts of yeast extract, 2 parts of brown sugar, bean cake powder water are uniformly mixed, adjust initial pH7,123 DEG C, sterilizing 40
Minute, 38 DEG C are cooled to, is lactobacillus acidophilus liquid culture medium.The Lactobacillus acidophilus species of a glycerol tube preservation are taken, are pressed
0.12% inoculum concentration, strain is connected in lactobacillus acidophilus culture medium, and amplification step by step spreads cultivation, and is lactobacillus acidophilus bacterium solution,
The viable count of Lactobacillus lactis is detected, it is 2.2 hundred million CFU/g to draw lactobacillus acidophilus bacterium solution viable count.
By 2 parts of glucose, 1.5 parts of peptone, 2 parts of yeast extract, 0.12 part of ammonium sulfate, 0.15 part of sodium carbonate, corn pulp
0.4 part, 0.07 part of manganese sulfate, 0.07 part of magnesium sulfate, 97 parts of water be uniformly mixed, adjust initial pH7.5,121 DEG C, sterilize 40 minutes,
38 DEG C are cooled to, is clostridium butyricum fluid nutrient medium.Take the clostridium butyricum strain of a glycerol tube preservation, connect by 0.11%
Kind amount, strain is connected in clostridium butyricum culture medium, and amplification step by step spreads cultivation, and is clostridium butyricum bacterium solution.Detect clostridium butyricum
Viable count, it is 0.5 hundred million CFU/g to draw clostridium butyricum active bacteria number.
Wheat bran, dregs of beans, corn are crushed, all cross 40 mesh sieves.By 25 parts of wheatfeed, 32 parts of bean cake powder, corn flour
11.5 parts, 0.5 part of sodium chloride, 2 parts of calcium carbonate, 1 part of sodium carbonate, 0.01 part of manganese sulfate, 0.05 part of magnesium sulfate be uniformly mixed, mixing
Solid material (solid medium) is unsterilised.
88 parts of hybrid solid-state material, 40 parts of water, 5 parts of lactobacillus acidophilus bacterium solution are taken, loads common fermentation after mixing and sets
In standby, coefficient 90%, top cover, sealing, when fermentation 8 is small, inoculates 6 parts of clostridium butyricum bacterium solution, after mixing, then
It is secondary to be fitted into the Zymolysis Equipment, top cover, then ferment 26 it is small when.
After fermentation, it is dried with low temperature fluidized bed, in drying process, temperature of charge is no more than 50 DEG C, after dry
Moisture is less than 12%.With the sieving of 40 mesh rotor-vibrating screens, screenings, packed products, oversize two are carried out with oppositely rolling roller crusher
It is secondary it is broken after after 40 mesh sieves, screenings, packed products.It is repeated, until all passing through 40 mesh sieves.
After fermentation, it is 2,000,000,000 CFU/g to detect live lactobacillus acidophilus number in finished product, and clostridium butyricum active bacteria number is 500,000,000 CFU/
G, clostridium butyricum gemma number are 500,000,000 CFU/g.
5th embodiment
Verify the lactobacillus acidophilus in the second~tetra- embodiment and the fermentation time of clostridium butyricum mixed fermentation.
The fermentation process provided according to first embodiment, 1 part, 94 parts 3 parts of yeast extract, 2 parts of brown sugar, bean cake powder water are mixed
Uniformly, initial pH7.5 is adjusted, 121 DEG C, sterilizes 30 minutes, is cooled to 37 DEG C, is lactobacillus acidophilus liquid culture medium.Take one
The Lactobacillus acidophilus species of glycerol tube preservation, by 0.1% inoculum concentration, strain are connected in lactobacillus acidophilus culture medium, step by step
Amplification spreads cultivation, and is lactobacillus acidophilus bacterium solution, detects the viable count of lactobacillus acidophilus bacterium solution.
By 2 parts of glucose, 1.5 parts of peptone, 1.5 parts of yeast extract, 0.1 part of ammonium sulfate, 0.1 part of sodium carbonate, corn pulp
0.3 part, 0.05 part of manganese sulfate, 0.05 part of magnesium sulfate, 95 parts of water be uniformly mixed, adjust initial pH7.5,121 DEG C, sterilize 30 minutes,
37 DEG C are cooled to, is clostridium butyricum fluid nutrient medium.Take the clostridium butyricum strain of a glycerol tube preservation, connect by 0.1%
Kind amount, strain is connected in clostridium butyricum culture medium, and amplification step by step spreads cultivation, and is clostridium butyricum bacterium solution, detects clostridium butyricum
Viable count.
Wheat bran, dregs of beans, corn are crushed, all cross 40 mesh sieves.By 25 parts of wheatfeed, 32 parts of bean cake powder, corn flour
11.5 parts, 0.5 part of sodium chloride, 2 parts of calcium carbonate, 1 part of sodium carbonate, 0.01 part of manganese sulfate, 0.05 part of magnesium sulfate be uniformly mixed, detection
Total number of molds and miscellaneous bacteria sum.
Hybrid solid-state material (solid medium) is unsterilised.Take 88 parts of the hybrid solid-state material, 40 parts of water, lactobacillus acidophilus
5 parts of bacterium solution, is fitted into stainless steel barrel, fills after mixing, top cover, is sealed by fermentation.Every 1 hour, sampling detection
Live lactobacillus acidophilus number.The detection is tested in triplicate.
5 parts of the bacterium solution of clostridium butyricum is inoculated, is fitted into after mixing in stainless steel barrel, does not fill, apart from lid mouth 10cm,
Top cover, continues to ferment.A sampling clostridium butyricum active bacteria number of detection and gemma number when 3 is small.Detection experiment repeats
Three times.
2. interpretation of result
Before fermentation, about 100,000,000 CFU/g of viable count of lactobacillus acidophilus bacterium solution, about 0.5 hundred million CFU/g of viable count of clostridium butyricum,
The total number of molds of hybrid solid-state material is less than 50,000 CFU/g, and miscellaneous bacteria sum is less than 100,000 CFU/g.
After fermentation, the exponential phase of lactobacillus acidophilus is 6~12h, so being managed when selection 6~12 is small for lactobacillus acidophilus
Think fermentation time, the exponential phase of clostridium butyricum is 18~24h, after the 18h that ferments, that is, initially forms a small amount of gemma, is sending out
After ferment 24h, gemma rate is more than 70%, and when fermenting 36h, gemma rate is more than 90%, and afterwards, gemma rate is not further added by.So choosing
It is clostridium butyricum fermentation time to select 24h~36h.
To sum up, the present invention provides a kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation method, it includes：Unpasteurized
First inoculating lactobacillus acidophilus' bacterium solution is fermented on solid medium, is inoculated clostridium butyricum bacterium solution and is fermented.It being capable of profit
With lactobacillus acidophilus and clostridium butyricum to the coordinate repression of miscellaneous bacteria, it can effectively suppress the growth of miscellaneous bacteria, effectively control life
Miscellaneous bacteria during production, and lactobacillus acidophilus and clostridium butyricum can be fermented under conditions of raw material is unsterilised, reduce
Cost, it is highly practical.In addition, the present invention also provides a kind of Zymolysis Equipment, it can be used for anaerobic bacteria or facultative anaerobic bacteria, such as butyric acid
The solid state fermentation production of clostridium, lactobacillus acidophilus etc., this equipment is by air outlet pipe, by Volatile Metabolites, such as hydrogen, two
Carbonoxide etc. is discharged into outside air kind, releases metabolic inhibition, can discharge viable bacteria growth and breeding ability, improves fermentation viable bacteria
Number.
The foregoing is only a preferred embodiment of the present invention, is not intended to limit the invention, for the skill of this area
For art personnel, the invention may be variously modified and varied.Within the spirit and principles of the invention, that is made any repaiies
Change, equivalent substitution, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of lactobacillus acidophilus and clostridium butyricum mixed fermentation method, it is characterised in that it includes：Trained in unpasteurized solid-state
Support first inoculating lactobacillus acidophilus' bacterium solution on base to ferment, inoculate clostridium butyricum bacterium solution and ferment.
2. according to the method described in claim 1, it is characterized in that, the fermentation time for being inoculated with the lactobacillus acidophilus bacterium solution is 6
~12 it is small when, be inoculated with the fermentation time after the clostridium butyricum bacterium solution for 24~36 it is small when.
3. according to the method described in claim 1, it is characterized in that, the lactobacillus acidophilus bacterium solution and the clostridium butyricum bacterium
The inoculum concentration of liquid is 0.05~0.15%.
4. according to the method described in claim 1, it is characterized in that, the lactobacillus acidophilus bacterium solution and the clostridium butyricum bacterium solution
Mixing ratio be (0.8~1.2):(0.8~1.2).
5. according to the method described in claim 1, it is characterized in that, the fermentation raw material of the solid medium includes wheatfeed
23~27 parts, 30~34 parts of bean cake powder, 9.5~13.5 parts of corn flour, it is preferable that the wheatfeed, the bean cake powder and institute
Corn flour is stated after broken, crosses 40 mesh sieves.
6. according to the method described in claim 5, it is characterized in that, the fermentation raw material has further included 1~3 part of calcium carbonate, carbon
Sour 0.5~1.5 part of sodium, 0.01~0.05 part of manganese sulfate, 0.02~0.08 part of magnesium sulfate.
7. a kind of Zymolysis Equipment, it is used for such as claim 1~6 any one of them lactobacillus acidophilus and clostridium butyricum mixing hair
Fermenting process, it is characterised in that it includes closed container body, and the container body is equipped with unidirectional gas outlet passage.
8. Zymolysis Equipment according to claim 7, it is characterised in that the unidirectional gas outlet passage is air outlet pipe, described
One end of air outlet pipe is located in the container body, and the other end of the air outlet pipe is arranged on outside the container body, the outlet
Sealed equipped with one-way ventilating valve one end outside the container body of pipe.
9. Zymolysis Equipment according to claim 8, it is characterised in that the air outlet pipe is arranged on one in the container body
End is equipped with multiple ventholes, and the aperture of the venthole is 0.5~2mm.
10. Zymolysis Equipment according to claim 8, it is characterised in that the top of the container body is equipped with head cover, described
Air outlet pipe is connected with the head cover.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110241053A (en) * | 2019-07-17 | 2019-09-17 | 厦门昶科生物工程有限公司 | A kind of method of mixing fermentation culture clostridium butyricum |
| CN110353099A (en) * | 2019-09-02 | 2019-10-22 | 河南牧业经济学院 | A kind of production method of clostridium butyricum and clostridium butyricum feed addictive |
| CN111647541A (en) * | 2020-07-09 | 2020-09-11 | 广东大泽农生物科技股份有限公司 | Clostridium butyricum viable bacteria preparation, production method thereof and animal feed additive |
| CN111676162A (en) * | 2020-06-18 | 2020-09-18 | 宜宾学院 | Compound inoculant and its application in composting and fermenting leaves of Ginger japonica |
| CN118813452A (en) * | 2024-06-26 | 2024-10-22 | 湖北蓝谷中微生物技术有限公司 | A method for the joint fermentation of Clostridium butyricum and Lactobacillus amylovora |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928073A (en) * | 2006-09-29 | 2007-03-14 | 杜灵广 | Combination fermentation process of clostridiumbutyricum and lactobacillus acidophilus |
| CN103255093A (en) * | 2013-05-20 | 2013-08-21 | 厦门六维生物科技有限公司 | Preparation method of lactobacillus acidophilus |
| CN104031825A (en) * | 2013-03-07 | 2014-09-10 | 西北民族大学 | Low-temperature anaerobic biogas fermentation tank |
| CN104152522A (en) * | 2014-05-05 | 2014-11-19 | 浙江省农业科学院 | Method for preparing wheat active peptide from mixed bacterium solid state fermentation gluten powder |
| CN205556692U (en) * | 2016-04-20 | 2016-09-07 | 江西正合环保工程有限公司 | Constant temperature anaerobic fermentation tank |
| CN106544293A (en) * | 2016-10-12 | 2017-03-29 | 湖北华扬科技发展有限公司 | A kind of method that use Pichia sp. fermentation bacterium mud produces Clostridium butyricum |
| CN107006720A (en) * | 2017-04-12 | 2017-08-04 | 湖北华扬科技发展有限公司 | A kind of probiotics and its application |
-
2018
- 2018-01-22 CN CN201810058724.XA patent/CN108004191A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928073A (en) * | 2006-09-29 | 2007-03-14 | 杜灵广 | Combination fermentation process of clostridiumbutyricum and lactobacillus acidophilus |
| CN104031825A (en) * | 2013-03-07 | 2014-09-10 | 西北民族大学 | Low-temperature anaerobic biogas fermentation tank |
| CN103255093A (en) * | 2013-05-20 | 2013-08-21 | 厦门六维生物科技有限公司 | Preparation method of lactobacillus acidophilus |
| CN104152522A (en) * | 2014-05-05 | 2014-11-19 | 浙江省农业科学院 | Method for preparing wheat active peptide from mixed bacterium solid state fermentation gluten powder |
| CN205556692U (en) * | 2016-04-20 | 2016-09-07 | 江西正合环保工程有限公司 | Constant temperature anaerobic fermentation tank |
| CN106544293A (en) * | 2016-10-12 | 2017-03-29 | 湖北华扬科技发展有限公司 | A kind of method that use Pichia sp. fermentation bacterium mud produces Clostridium butyricum |
| CN107006720A (en) * | 2017-04-12 | 2017-08-04 | 湖北华扬科技发展有限公司 | A kind of probiotics and its application |
Non-Patent Citations (3)
| Title |
|---|
| 王天云等: "嗜酸乳杆菌固态发酵的初步研究", 《生命科学研究》 * |
| 王艺宙等: "嗜酸乳杆菌和酪酸梭菌协同抑制痢疾杆菌的活性研究", 《中国医科大学学报》 * |
| 谢丽静等: "丁酸梭菌的应用及对食品安全的影响", 《中国食物与营养》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110241053A (en) * | 2019-07-17 | 2019-09-17 | 厦门昶科生物工程有限公司 | A kind of method of mixing fermentation culture clostridium butyricum |
| CN110353099A (en) * | 2019-09-02 | 2019-10-22 | 河南牧业经济学院 | A kind of production method of clostridium butyricum and clostridium butyricum feed addictive |
| CN111676162A (en) * | 2020-06-18 | 2020-09-18 | 宜宾学院 | Compound inoculant and its application in composting and fermenting leaves of Ginger japonica |
| CN111676162B (en) * | 2020-06-18 | 2022-09-13 | 宜宾学院 | Complex microbial inoculant and application thereof in alpinia zerumbet branch and leaf composting fermentation |
| CN111647541A (en) * | 2020-07-09 | 2020-09-11 | 广东大泽农生物科技股份有限公司 | Clostridium butyricum viable bacteria preparation, production method thereof and animal feed additive |
| CN111647541B (en) * | 2020-07-09 | 2021-04-09 | 广东大泽农生物科技股份有限公司 | Clostridium butyricum viable bacteria preparation, production method thereof and animal feed additive |
| CN118813452A (en) * | 2024-06-26 | 2024-10-22 | 湖北蓝谷中微生物技术有限公司 | A method for the joint fermentation of Clostridium butyricum and Lactobacillus amylovora |
| CN118813452B (en) * | 2024-06-26 | 2025-05-27 | 湖北蓝谷中微生物技术有限公司 | A method for the joint fermentation of Clostridium butyricum and Lactobacillus amylovora |
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