CN109929775A - The preparation method of antibacterial microbe leaven - Google Patents
The preparation method of antibacterial microbe leaven Download PDFInfo
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- CN109929775A CN109929775A CN201910115835.4A CN201910115835A CN109929775A CN 109929775 A CN109929775 A CN 109929775A CN 201910115835 A CN201910115835 A CN 201910115835A CN 109929775 A CN109929775 A CN 109929775A
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Abstract
The present invention provides the preparation method of antibacterial microbe leaven, belongs to microorganisms technical field, is included in the environment of protective agent presence, and the microorganism for being used to ferment fixedly is stored in inert carrier；Mentioned microorganism is with total viable count 5 × 107~5 × 1011The concentration of cfu/L is comprised in aqueous suspension；Above-mentioned inert carrier is shellfish material；Above-mentioned protective agent includes sodium glutamate and sodium nitrite.The preparation method of antibacterial microbe leaven provided by the invention can be made can antibacterial anti-corrosion, improve flavor, highly-safe, stability is good, does not generate the leavening of drug resistance, the fermentation agent carrier is not reunited, good dispersion, can decorating site it is more, the fixed site of microorganism is more, and supported quantity is high, immobilization is deep, inactivation rate is low, long shelf-life, can be used for fermentation fertilizer, fermented feed and fermented food field.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to the preparation method of antibacterial microbe leaven.
Background technique
Fermentation, is a kind of means that the mankind dexterously fabricate food, feed etc. using beneficial microbe, hair
Ferment agent is functionally similar to probiotic or probiotics, is mainly used for fermented feed, fermented food, and animal body cannot be inhaled
It receives the substance utilized and is converted to the absorbable higher substance of nutrition utilized, and generate a large amount of protease, lipase, starch
Enzyme, cellulolytic enzyme, B family vitamin and vitamin A, D etc. increase profitable strain quantity, inhibit or kill harmful bacteria, enhancing
Immunity of organisms.
The strain that existing traditional spontaneous fermentation participates in is mainly lactic acid bacteria, micrococcus luteus (mainly staphylococcus and Cook
Bacterium), followed by yeast and mould, wherein lactic acid bacteria and micrococcus luteus dominate microbiota.However because raw material is in spontaneous fermentation
It is preceding not can be carried out pasteurize, thus the growth of putrefactive microorganisms is often detected in spontaneous fermentation, as Escherichia coli, enterobacteria and
Pseudomonad.The presence of putrefactive microorganisms is directly related to safety and the shelf-life of food or feed.Traditional natural fermentation simultaneously
Drawback is a lot of: fermentation period is relatively long, and productivity is low；It is influenced, is fermented volatile by sanitary condition, production season and salt dosage
It loses；Fermented quality is unstable, is unfavorable for batch production, scale and standardized production；Continue to use traditional handicraft, it is difficult to realize extensive
Industrialized production；Strange land production, it is difficult to guarantee the consistency of product quality；Sensory experience is poor, the Toxics such as nitrite
The high Yi Chaobiao of matter content, edible safety are poor.
It is added to biocidal property microbe leaven in fermentation, probiotics such as lactic acid bacteria and micrococcus luteus etc. can be made,
Dominant bacteria is rapidly become after fermentation initial stage rapid growth, same to enterobacteria, enterobacteria and pseudomonad competition, so that these
The growth of corrupt flora is suppressed, and helps to improve microbial quality, shelf life and the safety of product.
Antibiotic creates huge economic benefit to aquaculture, and antibiotic is used as always feed growth-promoting additive wide
General application, however due to the especially exceeded use of extensive use of the antibiotic in animal and fowl fodder, cause various pathogenic microorganisms
It develops drug resistance and the suprainfection of animal, seriously affects livestock birds health situation；Drug in animal products can be caused residual simultaneously
It stays, not only influences the quality and its safety of animal products, it is resistance to antibiotic generation to also result in pathogenic microorganism in mankind's body
Pharmacological property, and then directly threaten human health.Over nearly twenty or thirty year, the ratio of drug resistance strain is continuously increased, and the mankind cure bacterium and pass
The difficulty of infectious diseases is increasing, and effective dosage of required antibiotic climbs up and up, in recent years national to antibiotic
Supervision reinforcement, nonreactive cultivation is the general trend of cultivation industry development from now on, and fermentative feedstuff of microbe is using microorganism
Metabolism and breeding, produce or modulate have green, safety and efficiently many advantages, such as feed, beneficial microbe
In fermentation, numerous metabolites is generated, including the substance with antibiotic effect, therefore, promotes fermented feed
In antibiotic-free, castoff regenerative resource utilization and alleviate good development in terms of people and animals strive.
Summary of the invention
The purpose of the present invention is to provide a kind of carriers not to reunite, good dispersion, and the fixed site of microorganism is more, supported quantity
It is high, inactivation rate is low, long shelf-life, can antibacterial anti-corrosion, improve flavor, antibacterial microorganism that is highly-safe, not generating drug resistance hair
The preparation method of ferment agent, made leavening can be used for fermentation fertilizer, fermented feed and fermented food field.
The technical solution that the present invention is taken to achieve the above object are as follows:
The preparation method of antibacterial microbe leaven, this method protective agent there are in the environment of, will be used for ferment it is micro-
It is stored on inert carrier to biological fixation；Mentioned microorganism is with total viable count 5 × 107~5 × 1011The concentration of cfu/L by comprising
In aqueous suspension；Above-mentioned inert carrier is shellfish material；Above-mentioned protective agent includes sodium glutamate and sodium nitrite.The fermentation
Agent assigns its positive charge using shellfish material as inert carrier after amino acid modification, mutually inhales with surface for the strain of negative electrical charge
Draw and combine integral, to achieve the purpose that immobilized microorganism, microbe leaven is reduced in long-time saves with this
Caused by inactivation rate, extend the holding time and validity period of leavening.
Preferably, shellfish material is the shellfish material of amino acid modification；Amino acid is L-arginine.Due to microbial bacteria
Kind surface is in elecrtonegativity, and shellfish material with water-wet behavior itself, is difficult under the action of hydrone active force and hydrogen bond
It is in combination, therefore amino acid modification is carried out to shellfish material, shellfish material surface cationic charge density can be increased, thus
The stability to microbial cell immobilization is improved, achievees the purpose that for strain to be fixed on shellfish material.
Further preferably, the shellfish material of amino acid modification is made by following steps: it is 1 that concentration, which is added, in shellfish material
In the amino acid solution of~5mg/L, under 500~800W power after 15~30min of mixing ultrasonic treatment, it is centrifuged simultaneously water washing and precipitating
To neutrality, it is dried in vacuo 12 at 70~90 DEG C~for 24 hours to obtain the final product.
Still further preferably, the weight ratio of shellfish material and amino acid solution, with amino acid basis, shellfish material: amino acid
=1:0.5~1.5.
Still further preferably, change pretreatment by calcining and water before shellfish material modification；Calcination temperature is 400~600 DEG C,
Time is 1~2h, oxygen-free environment；Water change temperature be 130~180 DEG C, the time be 12~for 24 hours, reaction kettle used be polytetrafluoroethyl-ne
Alkene reaction kettle.Discarded shell forms irregular porous structure, specific surface area and hole after the processing such as high-temperature calcination extruding
Gap increases, internal capillary and mesoporous extruding, therefore has stronger thermal stability and adsorption capacity, is a kind of excellent carrier and suction
Attached dose, adsorb and be sustained excellent effect.
Still further preferably, pre-treatment step is as follows: after shell is cleaned up, being placed in 0.1~0.5% hydrochloric acid solution
10~12h of middle immersion is subsequently placed in the tube furnace that temperature is 400~600 DEG C and carries out anaerobic 1~2h of calcining, after taking out cooling,
It crushed 100~120 meshes, 3~6 times of amounts be then added into conch meal, the solution of potassium carbonate that concentration is 1~2%, 400
Then mixed solution is sent into ptfe autoclave, 130~180 by 10~30min of ultrasonic dissolution under~500W power
At DEG C reaction 12~for 24 hours, then solution is centrifuged, precipitating is taken to be washed to neutrality, at 60~90 DEG C dry 4~6h after, gained
Shellfish material is spare.Change by anaerobic calcining, weak caustic solution water, CaO, SiO in shellfish material can be made2Equal ingredients generate soluble
Salt, so that the ash content of charcoal reduces in shellfish material, to improve its absorption property.
Preferably, microorganism includes Lactobacillus saki, staphylococcus xylosus, Lactobacillus casei, bacillus licheniformis, cloth
Lardy saccharomycete, streptococcus thermophilus and lactobacillus acidophilus.Multi-cultur es are used cooperatively, and can not only accelerate course of fermentation, but also can produce
The flavor substance of various alcohols, acids, ketone is given birth to, the bad smells substances such as indoles, pyrimidine are reduced, improves the sense of feed or food
Official's experience, at the same metabolite can antibacterial, anti-corrosion, inhibit the generation of biogenic amine in fermentation process, substitute antibiotics and anti-corrosion
The use of agent improves the edible safety of product.
Preferably, the aqueous suspension containing microorganism is made by following steps: being respectively inoculated with microorganism fungus kind
In independent culture solution after sterilizing, 12 are cultivated under conditions of temperature is 25~40 DEG C, shaking speed is 50~300r/min
It after~72h, obtains the bacterium solution of each strain respectively, each bacterium solution mixing is then taken to compound to obtain the final product；Above-mentioned each bacterium solution is in compositional liquor
Volume accounting are as follows: Lactobacillus saki 10~15%, staphylococcus xylosus 15~25%, Lactobacillus casei 20~25%, lichens bud
Spore bacillus 15~25%, Bu Ladi saccharomycete 15~20%, streptococcus thermophilus 5~15% and lactobacillus acidophilus 5~15%, bacterium solution
Volume accounting adds up to 100%.
Preferably, steps are as follows for immobilization: after carrier is mixed with protective agent, the suspension containing microorganism is added,
PH to 6~8 is adjusted, and maintains the temperature at 25~30 DEG C, then immobilizes aeration 6~12h of culture, aeration quantity is 150~
200L/h after the completion of immobilization, is centrifuged, after taking precipitating normal saline flushing, as under the rate of 3000~5000r/min
Microbe leaven.Microorganism is fixed on a inert carrier in the presence of protective agent, and addition aeration operation is so that micro- life
Object strain can be accelerated domestication, increase supported quantity of the microorganism on carrier, can make work of microorganism during preservation
Bacterium number is as much as possible to be preserved, and ensures greatest benefit when leavening uses.
Preferably, the weight ratio of carrier and the suspension containing microorganism is 6~12:1；Carrier and protectant weight
Than for 5~10:0.05；The weight ratio of sodium glutamate and sodium nitrite that protective agent includes is 0.5~1:1.Protectant addition,
So that between carrier and microorganism immobilization not merely only exist charge effect, cation in protective agent is with shellfish material
Positive charge mutual exclusion so that carboxylic acid and nitric acid group are exposed to the one side closer to shellfish material, and occurs with amino acid thereon
It is coupled, amino acid residue is promoted to be exposed to the surface of molecule, forms surface layer mucous membrane, increase the fixation of carrier and microbial cell
Site promotes microorganism to be adhered tightly to the surface of carrier, then extends by the compatibility between amino acid and microbial cell
Into hole, microorganism is fixed on carrier by protectant cation in side, so that microbial immobilization degree is deepened,
Product stability can greatly promote, while carrier upper amino acid being made to supply microorganism as energy source again, protect most possibly
Microbial activity is held, inactivation rate is reduced, extends storage life.
Preferably, purposes of the leavening in fermentation fertilizer, fermented feed and fermented food field.
The invention has the benefit that
1) the made leavening of the present invention is used cooperatively using multi-cultur es, can accelerate course of fermentation, but can produce alcohols,
The flavor substance of acids, ketone reduces the bad smells substances such as indoles, pyrimidine, improves the sensory experience of feed or food, simultaneously
Metabolite can antibacterial, anti-corrosion, inhibit the generation of biogenic amine in fermentation process, the use of substitute antibiotics and preservative mentions
The safety of high product, does not generate drug resistance；
2) the made leavening of the present invention has apparent antibacterial anti-corrosion effect, and Spawn incubation method used is simple, facilitates investment
Commercial practice acts synergistically between strain, and in addition to Fermentation Function, the made fertilizer of the leavening can promote plant growth, raises
Material and food can promote animal body growth and development, adjust function of intestinal canal, have a good application prospect；
3) after amino-acid modified modification, microorganism is immobilized, fixed bit in the present invention using conch meal as carrier
Point is more, and immobilization is deep, and supported quantity is high, and microbial inactivation rate is low, stability energy consumption, extends storage life；
4) present invention uses shell for support material, which does not reunite and good dispersion, and decorating site is more, not only
Pollution of the waste shell to environment is alleviated, while providing the new way using shell, improves the utility value of shell, it is real
The effective use of resource is showed.
Present invention employs above-mentioned technical proposals to provide the preparation method of antibacterial microbe leaven, compensates for the prior art
Deficiency, reasonable design, easy operation.
Specific embodiment
Technical solution of the present invention is described in further detail below in conjunction with specific embodiment:
Embodiment 1:
The preparation method of antibacterial microbe leaven, this method protective agent there are in the environment of, will be used for ferment it is micro-
It is stored on inert carrier to biological fixation；Mentioned microorganism is with total viable count 5 × 107~5 × 1011The concentration of cfu/L by comprising
In aqueous suspension；Above-mentioned inert carrier is shellfish material；Above-mentioned protective agent includes sodium glutamate and sodium nitrite.The fermentation
Agent assigns its positive charge using shellfish material as inert carrier after amino acid modification, mutually inhales with surface for the strain of negative electrical charge
Draw and combine integral, to achieve the purpose that immobilized microorganism, microbe leaven is reduced in long-time saves with this
Caused by inactivation rate, extend the holding time and validity period of leavening.
Shellfish material is the shellfish material of amino acid modification；Amino acid is L-arginine.Since microorganism fungus kind surface is in
Elecrtonegativity, and itself with water-wet behavior shellfish material, be difficult under the action of hydrone active force and hydrogen bond it is in combination,
Therefore amino acid modification is carried out to shellfish material, shellfish material surface cationic charge density can be increased, to improve to micro-
The stability of biological cell immobilization achievees the purpose that for strain to be fixed on shellfish material.
The shellfish material of amino acid modification is made by following steps: the ammonia that concentration is 1.5mg/L is added in shellfish material
In base acid solution, under 500W power after mixing ultrasonic treatment 30min, it is centrifuged and water washing and precipitating is to neutrality, it is true at 70 DEG C
Sky is dried for 24 hours to obtain the final product.
The weight ratio of shellfish material and amino acid solution, with amino acid basis, shellfish material: amino acid=1:0.8.
Change pretreatment by calcining and water before shellfish material modification；Calcination temperature is 400 DEG C, time 2h, oxygen-free environment；
It is 130 DEG C that water, which changes temperature, and the time is that for 24 hours, reaction kettle used is ptfe autoclave.Discarded shell is swollen through high-temperature calcination
After the processing such as change, irregular porous structure is formed, specific surface area and hole increase, internal capillary and mesoporous extruding, therefore
There are stronger thermal stability and adsorption capacity, is a kind of excellent carrier and adsorbent, absorption and sustained release excellent effect.
Pre-treatment step is as follows: after shell is cleaned up, being placed in 0.2% hydrochloric acid solution and impregnates 12h, be subsequently placed in
Temperature calcines 2h to carry out anaerobic in 400 DEG C of tube furnace, and after taking out cooling, crushing is sieved with 100 mesh sieve, and then adds into conch meal
Enter 3 times of amounts, the solution of potassium carbonate that concentration is 1%, then mixed solution is sent into poly- by the ultrasonic dissolution 30min under 400W power
It in tetrafluoroethene reaction kettle, reacts at 130 DEG C for 24 hours, is then centrifuged solution, precipitating is taken to be washed to neutrality, dried at 70 DEG C
After dry 6h, gained shellfish material is spare.Change by anaerobic calcining, weak caustic solution water, CaO, SiO in shellfish material can be made2Deng at
It is mitogenetic at solubility salt so that in shellfish material charcoal ash content reduce, to improve its absorption property.
Microorganism includes Lactobacillus saki, staphylococcus xylosus, Lactobacillus casei, bacillus licheniformis, Bu Ladi yeast
Bacterium, streptococcus thermophilus and lactobacillus acidophilus.Multi-cultur es are used cooperatively, and can not only accelerate course of fermentation, but also can produce various alcohol
Class, acids, ketone flavor substance, reduce indoles, the bad smells substance such as pyrimidine, improve the sensory experience of feed or food,
Simultaneously metabolite can antibacterial, anti-corrosion, inhibit the generation of biogenic amine in fermentation process, substitute antibiotics and preservative make
With improving the edible safety of product.
Aqueous suspension containing microorganism is made by following steps: after microorganism fungus kind is inoculated in sterilizing respectively
In independent culture solution, after cultivating for 24 hours under conditions of temperature is 30 DEG C, shaking speed is 100r/min, each strain is obtained respectively
Bacterium solution, then take the mixing of each bacterium solution to compound to obtain the final product；Volume accounting of the above-mentioned each bacterium solution in compositional liquor are as follows: Lactobacillus saki
10%, staphylococcus xylosus 15%, Lactobacillus casei 20%, bacillus licheniformis 20%, Bu Ladi saccharomycete 20%, thermophilic chain
Coccus 10% and lactobacillus acidophilus 5%, bacterium solution volume accounting add up to 100%.
Steps are as follows for immobilization: after carrier is mixed with protective agent, the suspension containing microorganism is added, adjusts pH to 6,
And 25 DEG C are maintained the temperature at, and aeration culture 6h, aeration quantity 200L/h are then immobilized, after the completion of immobilization, in
It is centrifuged under the rate of 3000r/min, after taking precipitating normal saline flushing, as microbe leaven.Microorganism is in protective agent
In the presence of, fixed addition aeration operation enables microorganism fungus kind to be accelerated domestication on a inert carrier, increases micro- life
Supported quantity of the object on carrier can make viable count of microorganism during preservation is as much as possible to preserve, and ensure
Greatest benefit when leavening uses.
The weight ratio of carrier and the suspension containing microorganism is 6:1；Carrier and protectant weight ratio are 5:0.05；It protects
The weight ratio of sodium glutamate and sodium nitrite that shield agent includes is 0.5:1.Protectant addition, so that between carrier and microorganism
Ground immobilization not merely only exists charge effect, the positive charge mutual exclusion of cation and shellfish material in protective agent, so that carboxylic acid
It is exposed to the one side closer to shellfish material with nitric acid group, and is coupled with amino acid thereon, amino acid residue is promoted
Be exposed to the surface of molecule, form surface layer mucous membrane, increase the fixation site of carrier and microbial cell, by amino acid with it is micro-
Compatibility between biological cell promotes microorganism to be adhered tightly to the surface of carrier, then extends in hole, protectant sun
Microorganism is fixed on carrier by ion in side, so that microbial immobilization degree is deepened, product stability can mention significantly
It rises, while carrier upper amino acid being made to supply microorganism as energy source again, keep microbial activity most possibly, reduce and lose
Motility rate extends storage life.
Embodiment 2:
The preparation method of antibacterial microbe leaven, the specific steps of which are as follows:
1) take respectively Lactobacillus saki, staphylococcus xylosus, Lactobacillus casei, bacillus licheniformis, Bu Ladi saccharomycete,
After activation, microorganism fungus kind is inoculated in the independent culture solution after sterilizing for the strain of streptococcus thermophilus and lactobacillus acidophilus, in
After cultivating 54h under conditions of temperature is 33 DEG C, shaking speed is 80r/min, the bacterium solution of each strain is obtained respectively, then takes each bacterium
Liquid mixing compounds, above-mentioned each bacterium solution volume accounting in compositional liquor spare to get suspension are as follows: Lactobacillus saki 15%, wood
Sugared staphylococcus 15%, Lactobacillus casei 20%, bacillus licheniformis 15%, Bu Ladi saccharomycete 15%, streptococcus thermophilus
10% and lactobacillus acidophilus 10%, bacterium solution volume accounting add up to 100%；
2) it after cleaning up shell, is placed in 0.3% hydrochloric acid solution and impregnates 10h, being subsequently placed in temperature is 600 DEG C
Anaerobic is carried out in tube furnace and calcines 1h, after taking out cooling, crushed 120 meshes, and 5 times of amounts, concentration are then added into conch meal
For 1.5% solution of potassium carbonate, then it is anti-to be sent into polytetrafluoroethylene (PTFE) by the ultrasonic dissolution 20min under 500W power for mixed solution
It answers in kettle, 12h is reacted at 160 DEG C, is then centrifuged solution, precipitating is taken to be washed to neutrality, after drying 4h at 90 DEG C, gained
Conch meal is spare；
3) conch meal is added in the amino acid solution that concentration is 5mg/L, the mixing ultrasonic treatment 15min under 800W power
Afterwards, simultaneously water washing and precipitating is dried in vacuo 12h at 90 DEG C to neutrality to get inert carrier, above-mentioned conch meal and amino acid for centrifugation
The weight ratio of solution, with amino acid basis, shellfish material: amino acid=1:1.5；
4) after mixing carrier with protective agent, the suspension containing microorganism is added, adjusts pH to 7.5, and keep temperature
At 30 DEG C, aeration culture 12h, aeration quantity 150L/h are then immobilized, after the completion of immobilization, in the speed of 5000r/min
It is centrifuged under rate, after taking precipitating normal saline flushing, the weight ratio of as microbe leaven, above-mentioned carrier and suspension is
12:1, carrier and protectant weight ratio 8.5:0.05, the weight ratio 0.8 of sodium glutamate and sodium nitrite that protective agent includes:
1。
Embodiment 3:
The preparation method of antibacterial microbe leaven, the specific steps of which are as follows:
1) take respectively Lactobacillus saki, staphylococcus xylosus, Lactobacillus casei, bacillus licheniformis, Bu Ladi saccharomycete,
After activation, microorganism fungus kind is inoculated in the independent culture solution after sterilizing for the strain of streptococcus thermophilus and lactobacillus acidophilus, in
After cultivating 36h under conditions of temperature is 30 DEG C, shaking speed is 240r/min, the bacterium solution of each strain is obtained respectively, is then taken each
Bacterium solution mixing compounds, above-mentioned each bacterium solution volume accounting in compositional liquor spare to get suspension are as follows: Lactobacillus saki 15%,
Staphylococcus xylosus 15%, Lactobacillus casei 20%, bacillus licheniformis 15%, Bu Ladi saccharomycete 15%, streptococcus thermophilus
10% and lactobacillus acidophilus 10%, bacterium solution volume accounting add up to 100%；
2) it after cleaning up shell, is placed in 0.2% hydrochloric acid solution and impregnates 10h, being subsequently placed in temperature is 500 DEG C
Anaerobic is carried out in tube furnace and calcines 1h, after taking out cooling, crushed 120 meshes, and 4 times of amounts, concentration are then added into conch meal
For 1.5% solution of potassium carbonate, then it is anti-to be sent into polytetrafluoroethylene (PTFE) by the ultrasonic dissolution 15min under 500W power for mixed solution
It answers in kettle, 18h is reacted at 150 DEG C, is then centrifuged solution, precipitating is taken to be washed to neutrality, after drying 4h at 80 DEG C, gained
Conch meal is spare；
3) conch meal is added in the amino acid solution that concentration is 3mg/L, the mixing ultrasonic treatment 20min under 600W power
Afterwards, simultaneously water washing and precipitating is dried in vacuo 18h at 80 DEG C to neutrality to get inert carrier, above-mentioned conch meal and amino acid for centrifugation
The weight ratio of solution, with amino acid basis, shellfish material: amino acid=1:1；
4) after mixing carrier with protective agent, the suspension containing microorganism is added, adjusts pH to 7, and maintain the temperature at
25 DEG C, aeration training 10h, aeration quantity 180L/h are then immobilized, after the completion of immobilization, under the rate of 5000r/min
Centrifugation, after taking precipitating normal saline flushing, the weight ratio of as microbe leaven, above-mentioned carrier and suspension is 8.5:1,
Carrier and protectant weight ratio 9.5:0.05, the weight ratio 1:1 of sodium glutamate and sodium nitrite that protective agent includes.
Embodiment 4:
The preparation method of antibacterial microbe leaven, wherein optimizing to step 3), specific Optimized Measures are as follows: by shellfish
Shell powder is added in the amino acid solution that concentration is 3mg/L, under 600W power after mixing ultrasonic treatment 20min, is centrifuged and washes
It is precipitated to neutrality, is dried in vacuo 18h at 80 DEG C to get inert carrier, the weight ratio of above-mentioned conch meal and amino acid solution,
With amino acid basis, shellfish material: amino acid=1:1, the cycloalkanes of the tannic acid containing 0.04mM and 0.06mM in amino acid solution
Sour sodium, the addition of the two can be using the combination for carrying out chemical bond in molecule between carboxyl and calcium carbonate, in calcium carbonate molecular surface
One layer of monomolecular film is formed, and then calcium carbonate molecular surface energy is reduced, so that attractive interaction dies down between calcium carbonate molecule, is changed
The agglomeration being apt between conch meal enhances the dispersibility of conch meal, while the two gos deep into shell material under ultrasonic wave auxiliary
In the hole of material, and swelling fracture occurs, hole dilatation is increased, increases the decorating site on shellfish material, promote amino
The formation of chemical bond between acid and shellfish material, so that reaction efficiency and reaction product performance get a promotion.
The present embodiment is to optimize test on the basis of embodiment 3, consistent in other steps and embodiment 3, is made
Antibacterial microbe leaven.
Comparative example 1:
This comparative example is to compare test on the basis of embodiment 3, in embodiment 3 the difference is that: step
4) carried out in microorganism it is fixed when, be not added with include sodium glutamate and sodium nitrite protective agent.Other steps and embodiment 3
In it is consistent, antibacterial microbe leaven is made.
Comparative example 2:
This comparative example is to compare test on the basis of embodiment 3, in embodiment 3 the difference is that: step
4) when progress microorganism is fixed in, Air Exposure is not carried out.It is consistent in other steps and embodiment 3, antibacterial microorganism hair is made
Ferment agent.
Embodiment 5:
The bacteriostatic test of antibacterial microbe leaven
Embodiment 3 and 4, the made leavening of comparative example 1 and 2 for taking identical quantity respectively, after being saved 30 days at 4 DEG C,
It activates under the same conditions using MS culture medium respectively, cultivate 12h, be centrifuged, take supernatant respectively to Escherichia coli, golden yellow Portugal
Grape coccus, salmonella, Aspergillus flavus carry out bacteriostatic test, as a result as follows to be not added with the culture medium of leavening as blank group
Table 1.
The bacteriostatic test result of 1 microbe leaven of table
| Antibacterial circle diameter mm | Escherichia coli | Staphylococcus aureus | Salmonella | Aspergillus flavus |
| Embodiment 3 | 27 | 28 | 25 | 23 |
| Embodiment 4 | 31 | 29 | 29 | 24 |
| Comparative example 1 | 23 | 26 | 21 | 19 |
| Comparative example 2 | 19 | 27 | 19 | 17 |
| Blank group | - | - | - | - |
As seen from the above table, except to staphylococcus aureus fungistatic effect difference it is unobvious in addition to, other strains it is antibacterial
In effect, embodiment 3 and 4 is overall, and preferably then effect is poor for comparative example, is due to being using protective agent, so that producing in comparative example 1
Product stability is bad, and immobilized microorganism is easy inactivation in preservation；Since aeration operation is not used to microorganism in comparative example 2
It is tamed, so that the micro organism quantity being fixed on carrier is less, so fungistatic effect is poor.
Embodiment 6:
Application of the antibacterial microbe leaven in fermented feed
Embodiment 3 and 4, the made leavening of comparative example 1 and 2 of identical quantity are taken respectively, and ferment high temperature under the same conditions
Then the maize cob meal and dregs of beans to sterilize is grouped feeding dorking, test period 45d.As a result such as the following table 2.
Influence of 2 fermented feed of table to meat chicken growth performance
| Embodiment 3 | Embodiment 4 | Comparative example 1 | Comparative example 2 | |
| Average starting weight/g | 38.95 | 38.91 | 38.86 | 38.94 |
| Average end weight/g | 1347.67 | 1383.17 | 1273.87 | 1287.51 |
| Average daily gain/(g/d/ is only) | 29.1 | 29.9 | 27.5 | 27.7 |
| Average daily gain/(g/d/ is only) | 73.89 | 75.94 | 68.92 | 69.45 |
| Feed-weight ratio/(g:g) | 2.54 | 2.54 | 2.51 | 2.51 |
As seen from the above table, the produced feed of the leavening of embodiment 3 and 4 all has with comparative example aobvious in daily gain, feed intake
Difference is write, is since the microorganism immobilization amount of embodiment 3 and 4 is high, viable bacteria number is more, and ferment effect is good, on food calling, growth promotion
There is apparent gain effect.
Embodiment 7:
Application of the antibacterial microbe leaven in fermentation fertilizer
Embodiment 3 and 4, the made leavening of comparative example 1 and 2 of identical quantity are taken respectively, and ferment high temperature under the same conditions
The corn stover and rice straw to sterilize applies coral usage tree root in seedling stage then under identical planting conditions respectively, examination
Testing the period is 3 months, and observation sapling growth period sweet viburnum height of seedling, trunk perimeter, tree crown degree of enclosing, trunk branch amount are remembered
Record is averaged according to and analyzes, as a result such as the following table 3.
3 sweet viburnum seedling growth status analysis of table
| Sapling height/cm | Trunk perimeter/cm | Tree crown degree of enclosing/cm | Branch amount/ | |
| Embodiment 3 | 102.5 | 46.7 | 106.1 | 66 |
| Embodiment 4 | 104.7 | 45.6 | 108.3 | 69 |
| Comparative example 1 | 98.6 | 41.5 | 101.2 | 53 |
| Comparative example 2 | 98.1 | 40.3 | 98.3 | 58 |
As seen from the above table, sapling height of the produced fermentation fertilizer of embodiment leavening to trees seedling stage, trunk perimeter, tree crown
Degree of enclosing, the growth of trunk branch amount all have preferable promotion growth, illustrate that fermentation fertilizer fertilizer efficiency is stable, full of nutrition, hair
Ferment effect is good.
Embodiment 8:
Application of the antibacterial microbe leaven in fermented food
Embodiment 3 and 4, the made leavening of comparative example 1 and 2 for taking identical quantity respectively, ferment under the same conditions through going out
The carrot and cabbage that bacterium crosses, then the quality to made sauerkraut and sense organ carry out Comprehensive Assessment, and wherein organic acid is mainly surveyed
Determine lactic acid and acetic acid content, concrete outcome such as the following table 4.
4 sauerkraut organoleptic quality Comprehensive Assessment of table
| Embodiment 3 | Embodiment 4 | Comparative example 1 | Comparative example 2 | |
| Raw material availability % | 89.5 | 89.1 | 86.3 | 84.5 |
| Total acidity g/mL | 4.76 | 4.82 | 3.87 | 3.51 |
| Organic acid content mg/mL | 1.94 | 1.96 | 1.75 | 1.77 |
| Mouthfeel | Salubrious sense is obvious | It is mellow, soft | Slightly stimulate | Slightly stimulate |
| Fragrance | Sour is pleasant | Sour is pleasant | Sour is prominent | Sour is prominent |
As seen from the above table, the made fermented food of leavening is in acidity, raw material availability and fragrance, the sensory experience of mouthfeel
On, embodiment performs better, and illustrates that the made fermented food of the made leavening of embodiment has more the market competitiveness.
The prior art of routine techniques dawn known to those skilled in the art in above-described embodiment, therefore herein no longer in detail
It repeats.
The above embodiments are only used to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this field
Member can also make a variety of changes and modification without departing from the spirit and scope of the present invention.Therefore, all equivalent
Technical solution also belong to scope of the invention, scope of patent protection of the invention should be defined by the claims.
Claims (10)
1. the preparation method of antibacterial microbe leaven, it is characterised in that: protective agent there are in the environment of, will be used for ferment
Microorganism is fixedly stored on inert carrier；The microorganism is with total viable count 5 × 107~5 × 1011The concentration of cfu/L is wrapped
It is contained in aqueous suspension；The inert carrier is shellfish material；The protective agent includes sodium glutamate and sodium nitrite.
2. the preparation method of antibacterial microbe leaven according to claim 1, it is characterised in that: the shellfish material is
The shellfish material of amino acid modification；The amino acid is L-arginine.
3. the preparation method of antibacterial microbe leaven according to claim 2, it is characterised in that: the amino acid modification
Shellfish material be made by following steps: shellfish material is added in the amino acid solution that concentration is 1~5mg/L, 500~
Under 800W power after 15~30min of mixing ultrasonic treatment, it is centrifuged and water washing and precipitating is to neutrality, be dried in vacuo at 70~90 DEG C
12~for 24 hours to obtain the final product.
4. the preparation method of antibacterial microbe leaven according to claim 3, it is characterised in that: the shellfish material with
The weight ratio of amino acid solution, with amino acid basis, shellfish material: amino acid=1:0.5~1.5.
5. the preparation method of antibacterial microbe leaven according to claim 3, it is characterised in that: the shellfish material is repaired
Change pretreatment by calcining and water before decorations；The calcination temperature is 400~600 DEG C, and the time is 1~2h, oxygen-free environment；The water
Change temperature be 130~180 DEG C, the time be 12~for 24 hours, reaction kettle used be ptfe autoclave.
6. the preparation method of antibacterial microbe leaven according to claim 1, it is characterised in that: the microorganism includes
Lactobacillus saki, staphylococcus xylosus, Lactobacillus casei, bacillus licheniformis, Bu Ladi saccharomycete, streptococcus thermophilus and acidophilus
Lactobacillus.
7. the preparation method of antibacterial microbe leaven according to claim 1, it is characterised in that: described to contain microorganism
Aqueous suspension by following steps be made: respectively by microorganism fungus kind be inoculated in sterilizing after independent culture solution in, Yu Wen
After cultivating 12~72h under conditions of degree is 25~40 DEG C, shaking speed is 50~300r/min, the bacterium of each strain is obtained respectively
Then liquid takes each bacterium solution mixing to compound to obtain the final product；Volume accounting of each bacterium solution in compositional liquor are as follows: Lactobacillus saki 10~
15%, staphylococcus xylosus 15~25%, Lactobacillus casei 20~25%, bacillus licheniformis 15~25%, Bu Ladi yeast
Bacterium 15~20%, streptococcus thermophilus 5~15% and lactobacillus acidophilus 5~15%, bacterium solution volume accounting add up to 100%.
8. the preparation method of antibacterial microbe leaven according to claim 1, it is characterised in that: the immobilization step
It is as follows: after carrier is mixed with protective agent, the suspension containing microorganism is added, adjusts pH to 6~8, and maintain the temperature at 25
~30 DEG C, aeration 6~12h of culture is then immobilized, aeration quantity is 150~200L/h, after the completion of immobilization, in 3000~
It is centrifuged under the rate of 5000r/min, after taking precipitating normal saline flushing, as microbe leaven.
9. the preparation method of antibacterial microbe leaven according to claim 1 or 8, it is characterised in that: the carrier with
The weight ratio of suspension containing microorganism is 6~12:1；The carrier and protectant weight ratio are 5~10:0.05；It is described
The weight ratio of sodium glutamate and sodium nitrite that protective agent includes is 0.5~1:1.
10. the preparation method of antibacterial microbe leaven according to claim 1, it is characterised in that: the leavening exists
The purposes of fermentation fertilizer, fermented feed and fermented food field.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136129A (en) * | 2018-08-27 | 2019-01-04 | 南昌大学 | One plant of lactobacillus acidophilus NCU426 |
| CN110467508A (en) * | 2019-08-01 | 2019-11-19 | 山东亿农高科生物科技有限公司 | A kind of facility cultivation acidified soil microorganism renovation agent and preparation method thereof |
| CN113080308A (en) * | 2021-05-17 | 2021-07-09 | 黄石市佳兴生物科技有限公司 | Fermented fluid milk substitute feed and preparation method thereof |
| CN115216432A (en) * | 2022-08-20 | 2022-10-21 | 红河宏斌食品有限公司 | Special production leaven for old jar pickled Chinese cabbage and preparation method thereof |
| CN115612642A (en) * | 2022-09-23 | 2023-01-17 | 东北农业大学 | Lactobacillus sake capable of degrading biogenic amine in broad spectrum and application of lactobacillus sake |
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2019
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136129A (en) * | 2018-08-27 | 2019-01-04 | 南昌大学 | One plant of lactobacillus acidophilus NCU426 |
| CN109136129B (en) * | 2018-08-27 | 2021-08-20 | 南昌大学 | Lactobacillus acidophilus NCU426 |
| CN110467508A (en) * | 2019-08-01 | 2019-11-19 | 山东亿农高科生物科技有限公司 | A kind of facility cultivation acidified soil microorganism renovation agent and preparation method thereof |
| CN113080308A (en) * | 2021-05-17 | 2021-07-09 | 黄石市佳兴生物科技有限公司 | Fermented fluid milk substitute feed and preparation method thereof |
| CN115216432A (en) * | 2022-08-20 | 2022-10-21 | 红河宏斌食品有限公司 | Special production leaven for old jar pickled Chinese cabbage and preparation method thereof |
| CN115612642A (en) * | 2022-09-23 | 2023-01-17 | 东北农业大学 | Lactobacillus sake capable of degrading biogenic amine in broad spectrum and application of lactobacillus sake |
| CN115612642B (en) * | 2022-09-23 | 2023-06-30 | 东北农业大学 | Lactobacillus sake for broad-spectrum degradation of biogenic amine and application thereof |
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