CN108841763A - A kind of preparation method of zinc dense plants lactobacillus additive - Google Patents
A kind of preparation method of zinc dense plants lactobacillus additive Download PDFInfo
- Publication number
- CN108841763A CN108841763A CN201810803962.9A CN201810803962A CN108841763A CN 108841763 A CN108841763 A CN 108841763A CN 201810803962 A CN201810803962 A CN 201810803962A CN 108841763 A CN108841763 A CN 108841763A
- Authority
- CN
- China
- Prior art keywords
- zinc
- preparation
- medium
- culture
- dense plants
- 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
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (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)
Abstract
The present invention relates to field of feed additive technology, and in particular to a kind of preparation method of zinc dense plants lactobacillus additive；Include the steps that for resistance to high zinc lactobacillus plantarum strain being used for the preparation of zinc dense plants lactobacillus additive；Wherein, the preparation of the zinc dense plants lactobacillus additive includes the following steps：Slant strains culture, level liquid seed culture, secondary liquid seed culture, fermentation tank culture, emulsification, freeze-drying；The preparation method of zinc dense plants lactobacillus additive of the invention securely and reliably, contains organic Zinc compounds and probiotics bioactivity maintenance higher level.
Description
Technical field
The present invention relates to field of feed additive technology, and in particular to a kind of preparation side of zinc dense plants lactobacillus additive
Method.
Background technique
Zinc is one of microelement necessary to growth of animal, participates in animal body composition, is participated in the activation factor of enzyme
Substance in vivo metabolism maintains the normal physiological function of body and influences cellular replication and differentiation as regulatory factor.
Currently, the zinc added in internal feed is mainly the inorganic forms such as zinc oxide, zinc sulfate.This addition manner exists
Many problems, if increasing the additive amount of zinc, not only increase the deposition of zinc in excrement, but also meeting if animal absorptivity is low
Certain pollution is caused to environment.In recent years a large number of experiments show that, organic state micro element additive, such as microelement amino
Sour chelate, has the characteristics that be easy by animal absorptions, small toxicity, animal weight gain is obvious, biological value is high, in pig, fowl, instead
Preferable effect is achieved on the animals such as hay animal and fish.However, there is production in additive of micromineral-AA chelate
At high cost, the problems such as manufacturing process is not perfect, it is difficult to which large-scale promotion uses.
Most study is saccharomycete in terms of zinc-rich microorganism at present, however when microbial process being used to carry out zinc-rich,
Its inorganic zinc concentration needs to control in a certain range, otherwise will affect the growth of microorganism or causes biologically active
Saccharomycete mortality, while also will affect the health of host, that is, it needs to reach one between the additive amount and high zinc-rich amount of zinc
A balance.Thus, how to develop a kind of safe and reliable, probiotics bacterial flour additive agent containing organic Zinc compounds, the additive
Have effects that also ensure the bioactivity of probiotics while zinc supplementation, becomes current technical problem urgently to be resolved.
Summary of the invention
In order to solve the above technical problems, the object of the present invention is to provide it is a kind of safe and reliable, contain organic Zinc compounds and benefit
Raw bacterium bioactivity maintains the preparation method of the zinc dense plants lactobacillus additive of higher level.
The preparation method of zinc dense plants lactobacillus additive of the invention, including resistance to high zinc lactobacillus plantarum strain is used for
The step of prepared by zinc dense plants lactobacillus additive；Wherein, the preparation of the zinc dense plants lactobacillus additive includes following step
Suddenly：
S2.1, slant strains culture：By resistance to high zinc zinc dense plants lactobacillus strain streak inoculation in sugared content 2-5%, zinc
On content 20-500ug/ml, the modified MRS solid medium that pH value is 5.5-7.0,35-40 DEG C of cultivation temperature, Anaerobic culturel
36-48h；
S2.2, level liquid seed culture：The bacterium colony on modified MRS solid medium is washed by the physiological saline of sterilizing
Lower mixing, draw 2.5ml bacterium solution be inoculated in sugared content 2-5%, Zn content 20-500ug/ml, pH value 5.5-7.0 250mL change
In good MRS fluid nutrient medium, 35-40 DEG C of cultivation temperature, Anaerobic culturel 15-24h obtains level liquid seed culture fluid；
S2.3, secondary liquid seed culture：By 200ml level liquid seed culture fluid is inoculated in sugared content 2-5%, zinc contains
In the 15-30L MRS liquid culture medium for measuring 20-500ug/ml, pH value 5.5-7.0,35-40 DEG C of cultivation temperature, revolving speed 10-
50rpm cultivates 5-10h, obtains secondary liquid seed culture fluid；
S2.4, fermentation tank culture：20L secondary liquid seed culture fluid is inoculated in sugared content 2-5%, Zn content 20-
1000ug/ml, initial pH value 5.5-7.0 1000-2000L MRS liquid culture medium in, 35-40 DEG C of cultivation temperature, revolving speed
10-50rpm controls pH5.0-6.5 with alkali in fermentation process, cultivates 8-16h, obtains zinc dense plants lactobacillus culture solution；
S2.5, zinc dense plants lactobacillus medium centrifugal to be collected, obtained bacterium mud is emulsified with protective agent, is lyophilized,
It is grinding to obtain zinc dense plants lactobacillus additive.
The experiment proved that it is grinding to obtain zinc dense plants lactobacillus additive, every kilogram 1000-4000 containing zinc milligrams.
Further, the resistance to high zinc lactobacillus plantarum strain is obtained through following steps：
S1.1, lactobacillus plantarum is inoculated into the modified MRS solid culture containing different zinc concentrations using the method for applying plate
On in base, to determine the tolerance of its zinc ion；
S1.2, using determining zinc ion tolerable concentration as initial concentration, lactobacillus plantarum is inoculated into changing containing zinc ion
It is cultivated on good MRS fluid nutrient medium, it is flat in the modified MRS solid containing identical zinc ion concentration then to choose bacterium solution with oese
Scribing line culture is carried out on plate culture medium, this is a generation；Biggish bacterium colony accesses the modified MRS containing zinc ion on picking solid plate
It is cultivated on fluid nutrient medium, then chooses bacterium solution with oese and trained in the modified MRS solid plate containing identical zinc ion concentration
It supports and carries out scribing line culture on base, this was two generations.Such passage and attenuation culture 40-60 generation；
S1.3, the bacterial strain after domestication is subjected to separation screening, is inoculated into MRS liquid culture medium and cultivates, filters out
The strong strain of resistance to zinc ability and preservation, as resistance to high zinc lactobacillus plantarum strain after domestication；
Further, in step S1.1 and step S1.2, modified MRS solid medium and MRS liquid culture medium
Sugared content is 2-5%, Zn content 20-1000ug/ml, pH value 5.5-7.0, cultivation temperature are 35-40 DEG C；Modified MRS liquid
Body culture medium Anaerobic culturel 12-24h, modified MRS solid medium Anaerobic culturel 36-48h.
Further, the condition of determining lactobacillus plantarum zinc ion tolerance is in step S1.1：It is single after cultivating 36-48h
Colony diameter is less than the corresponding zinc ion concentration of 0.5mm.
Further, zinc contains in the MRS liquid culture medium of the same generation and modified MRS solid medium in step S1.2
Measure identical, the Zn content of MRS liquid culture medium and modified MRS solid medium that wherein 1-15 generation uses is 20-
The Zn content of MRS liquid culture medium and modified MRS solid medium that 300ug/ml, 15-35 generation use is 300-
The Zn content of MRS liquid culture medium and modified MRS solid medium that 600ug/ml, 35-50 generation use is 600-
1000ug/ml,
Further, when lactobacillus plantarum is after modified MRS cultured on solid medium 36-48h, when colony diameter is in 1-
When 2mm, start to improve Zn content in the medium；
Further, the alkali used in step S2.4 is the sodium carbonate of 20-40% or sodium hydroxide.
Further, the protective agent in step S2.5 includes one of trehalose, sucrose, glycerol or a variety of, wherein institute
The mass fraction for stating trehalose is 10-20%, and the mass fraction of the sucrose is 2-7%, and the mass fraction of the glycerol is 2-
5%.
Further, lyophilisation condition includes the following steps in step S2.5：
1) -45 DEG C~-50 DEG C 3~4h of pre-freeze；
2) after 240~360min is warming up to -25~-15 DEG C, 840-960min is maintained；
3) after 4-6h is warming up to 0-5 DEG C, 3-5h is maintained；
4) after 120~240min is warming up to 10~15 DEG C, 120~240min is maintained；
5) after 60-90min is warming up to 25~28 DEG C, 240~360min is maintained.
Further, in MRS liquid culture medium and modified MRS solid medium, the zinc of addition is inorganic zinc, described
Inorganic zinc includes zinc oxide, sulfate, carbonate or chloride salt, and nitrogen source is yeast extract, yeast extract, peptone, beef
One or more of powder is soaked, carbon source is one or more of glucose, lactose, sucrose, maltose.
According to the above aspect of the present invention, the present invention has at least the following advantages：Zinc dense plants lactobacillus additive of the invention, through trying
The growth for showing that store pig can be effectively facilitated is tested, feedstuff-meat ratio is reduced, improves food conversion ratio, and can be significantly reduced
Zn-ef ficiency excretion in grow-finish swine excrement, while the effect of can also play probiotics, reduce the large intestine in fattening swine excrement
Bacillus quantity, pig healthy growth can be promoted by adjusting intestinal flora balance to prove zinc dense plants lactobacillus additive, be promoted
Into domestic animal and environmental protection harmonious development, the economic value of cultivation is improved.
The above description is only an overview of the technical scheme of the present invention, in order to better understand the technical means of the present invention,
And can be implemented in accordance with the contents of the specification, with presently preferred embodiments of the present invention, detailed description is as follows below.
Specific embodiment
With reference to embodiment, the embodiment of the present invention is furthur described in detail.Following embodiment is used for
Illustrate the present invention, but is not intended to limit the scope of the invention.
Embodiment 1
A preferred embodiment of the present invention provides a kind of preparation method of resistance to high zinc lactobacillus plantarum strain, including following step
Suddenly：
1) lactobacillus plantarum is inoculated into the modified MRS solid medium containing different zinc concentrations using the method for applying plate
On, to determine the tolerance of its zinc ion；
2) using determining zinc ion tolerable concentration as initial concentration, lactobacillus plantarum is inoculated into the improvement containing zinc ion
It is cultivated on MRS fluid nutrient medium, then chooses bacterium solution in the modified MRS solid plate containing identical zinc ion concentration with oese
Scribing line culture is carried out on culture medium, this is a generation；Biggish bacterium colony accesses the modified MRS liquid containing zinc ion on picking solid plate
It is cultivated on body culture medium, then chooses bacterium solution in the modified MRS solid plate culture containing identical zinc ion concentration with oese
Scribing line culture is carried out on base, this was two generations.Such passage and attenuation culture 40-60 generation；
3) by after domestication bacterial strain carry out separation screening, be inoculated into MRS liquid culture medium and cultivate, filter out by
The strong strain of resistance to zinc ability and preservation, as resistance to high zinc lactobacillus plantarum strain after domestication；
The sugared content of aforementioned modified MRS solid medium and MRS liquid culture medium is 2-5%, Zn content 20-
1000ug/ml, pH value 5.5-7.0, cultivation temperature are 35-40 DEG C；MRS liquid culture medium Anaerobic culturel 12-24h, changes
Good MRS solid medium Anaerobic culturel 36-48h.
Embodiment 2
The present embodiment provides a kind of preparation methods of zinc dense plants lactobacillus additive, include the following steps：
1) slant strains culture：By resistance to high zinc zinc dense plants lactobacillus strain streak inoculation in sugared content 2-5%, Zn content
On 20-500ug/ml, the modified MRS culture medium slant that pH value is 5.5-7.0,35-40 DEG C of cultivation temperature, Anaerobic culturel
36-48h；
2) level liquid seed culture：The physiological saline of 5ml sterilizing is added into inclined-plane lactobacillus plantarum test tube, by inclined-plane
On bacterium colony wash lower mixing, draw 2.5ml bacterium solution and be inoculated in sugared content 2-5%, Zn content 20-500ug/ml, pH value 5.5-7.0
250mL MRS liquid culture medium triangular flask in, 35-40 DEG C of cultivation temperature, Anaerobic culturel 15-24h obtains level liquid
Seed culture fluid；
3) secondary liquid seed culture：200ml level liquid seed culture fluid is inoculated in sugared content 2-5%, Zn content
20-500ug/ml, pH value 5.5-7.0 15-30L MRS liquid culture medium fermentor in, 35-40 DEG C of cultivation temperature, revolving speed
10-50rpm cultivates 5-10h, obtains secondary liquid seed culture fluid；
4) fermentation tank culture：20L secondary liquid seed culture fluid is inoculated in sugared content 2-5%, Zn content 20-
1000ug/ml, initial pH value 5.5-7.0 1000-2000L MRS liquid culture medium fermentor in, cultivation temperature 35-40
DEG C, revolving speed 10-50rpm controls pH5.0-6.5 with alkali in fermentation process, and zinc is added in batches, cultivates 8-16h, obtains zinc-rich plant
Object lactobacillus culture solution；
5) zinc dense plants lactobacillus medium centrifugal is collected, obtained bacterium mud is emulsified with protective agent, is lyophilized, powder
It is broken.It is grinding to obtain zinc dense plants lactobacillus additive, every kilogram 1000-4000 containing zinc milligrams.
Embodiment 3
The present embodiment provides a kind of detection sides that zinc dense plants lactobacillus additive influences grow-finish pig growth performance
Method and result.
1, experimental animal
Experimental animal：" Du × length × big " tri-crossbreeding 120 for choosing weight about 18kg, according to genetic background phase
With, weight, close, the not identical principle of nest is randomly divided into 3 groups, every group 4 repetition, each repetition 10.
2, test material
1) zinc dense plants lactobacillus bacterium powder product
2 method of embodiment is selected to prepare zinc dense plants lactobacillus bacterium powder product, Zn content 3000mg/kg.
2) daily ration is tested
Diet Formula includes based on mass fraction：Corn 55.24%, barley 7%, expanded soybean 6.23%, decortication dregs of beans
20.68%, wheat-middlings 6%, cotton dregs 4%, soya-bean oil 0.5%, salt 0.35% and vitamin and microelement.
In control group daily ration, the additive capacity of Zn-ef ficiency is in every kilogram of daily ration：100-200mg/kg, the addition of Zn-ef ficiency
Form is zinc sulfate；In test group daily ration the addition form of Zn-ef ficiency be zinc dense plants lactobacillus bacterium powder, additive capacity with compare
Group is identical.
3, test method
1) Animal experiment
" Du × length × greatly " tri-crossbreeding 120 for choosing weight about 18kg, is randomly divided into 3 groups, every group of 4 repetitions,
Each repetition 10.Control group, test group prepare Diet by this formula.Test is divided into early period according to the growth phase of pig
(20-40kg), mid-term (40-80kg) and 3 stages of later period (80-120kg).
Start in experiment, the 26th day (20-40kg), the 72nd day (40-80kg) and experiment terminate first 1 day (80-120kg)
20：00 fracture supplies water, next day 08：00 weigh on an empty stomach to all test pigs as unit of repetition, settles accounts corresponding experimental period
Feed consumption situation, calculate average weight gain, daily ingestion amount and feed-weight ratio.
Suitable temperature humidity is provided in experimental period, floor husbandry is divulged information, and is freely eaten, drinking-water.Immune disinfectant program is pressed
It is carried out according to pig farm conventional method.
2) Zn-ef ficiency utilization rate
From on-test, every 10 days, as unit of repetition, uniform collection excrement was baked to (65 DEG C), resurgence, system
At air-drying sample.Feed is sampled simultaneously, feed and excrement collected all need to crush, cross 40 mesh of aperture (0.45mm),
It is sealed.Mineral Concentrations are measured with Agilient ICP-MS 7500a inductively-coupled plasma spectrometer.
3) sero-immunity index determining
Vena cave blood sampling is carried out in the different phase of test, is collected whole blood (anticoagulant heparin), is separated serum, protected at -20 DEG C
It deposits.Serum IgA, interleukin-1, interleukin-6, tumor necrosis factor-alpha are detected, kit is purchased from Beijing China English biological study institute,
Detecting instrument is 7160 full automatic biochemical apparatus of Hitachi, and r-911 Full automatic exempts from calculating instrument.
4) in excrement Escherichia coli flora measurement
In the different phase of test, the fresh excrement sample of every group of 10 growing and fattening pigs of searching for food at random is encapsulated with aseptic plastic bag, is adopted
Count of bacteria is carried out with spread plate.
5) data statistic analysis
Test data handles data using 20.0 statistical analysis software of SPSS, using one-way analysis of variance Oneway
ANOVA carries out significance test, and carries out Multiple range test using DuncanShi method, with p<0.05 as significant difference judgement mark
Standard, test result are indicated with " average ± standard deviation ".
4, result and analysis
1) influence of the zinc dense plants lactobacillus additive to grow-finish pig growth performance
The results are shown in Table 1, and daily gain, end weight conspicuousness are higher than control group (P between testing each group<0.05), feed-weight ratio is aobvious
Work property is lower than control group (P<0.05).Illustrate that zinc dense plants lactobacillus bacterium powder product has certain promotion to make growing-finishing pigs performance
With reduction feed-weight ratio is improved food conversion ratio.
Influence (n=4) of the 1 zinc dense plants lactobacillus bacterium powder product of table to grow-finish pig growth performance
| First starting weight/kg | End weight/kg | Average daily gain/kg | Average daily gain/kg | Feed-weight ratio | |
| Control group | 18.33±0.64 | 99.1±1.49b | 0.62±0.09b | 1.12±0.07 | 1.81±0.03a |
| Test group | 18.43±0.63 | 113.71±2.11a | 0.74±0.09a | 1.20±0.08 | 1.68±0.13b |
2) influence that zinc dense plants lactobacillus additive drains Zn-ef ficiency in grow-finish swine excrement
The results are shown in Table 2, the significant (P of each stage Zn-ef ficiency excretion group difference<0.05), test group Zn-ef ficiency is drained
Amount is substantially less than control group.By taking the excretion of 20-40kg stage zinc as an example, test group and the significant (P of control group group difference<
0.05), test group reduces 28.38% or so than control group.This illustrates that zinc dense plants lactobacillus additive of the present invention can be significant
The excretion for reducing Zn-ef ficiency in the excrement of store pig improves the utilization rate of zinc, mitigates the pollution to environment.
The influence (n=4) that 2 zinc dense plants lactobacillus bacterium powder product of table drains store pig Zn-ef ficiency
3) influence of the zinc dense plants lactobacillus bacterium powder product to Escherichia coli flora in grow-finish swine excrement
The results are shown in Table 4.Pass through three different growing stages (20-40kg, 40-80kg, the 80- tested it
Weight 120kg) carries out the quantity of Escherichia coli in excrement in detection fattening pig, shows that the Escherichia coli quantity in test group is equal
Lower than control group (p<0.05), illustrate to can significantly reduce in fattening pig enteron aisle using zinc dense plants lactobacillus additive of the invention
Escherichia coli quantity.
Influence (lg (CFU/g)) of the 4 zinc dense plants lactobacillus additive of table to microbial flora in grow-finish swine excrement
The above is only a preferred embodiment of the present invention, it is not intended to restrict the invention, it is noted that for this skill
For the those of ordinary skill in art field, without departing from the technical principles of the invention, can also make it is several improvement and
Modification, these improvements and modifications also should be regarded as protection scope of the present invention.
Claims (10)
1. a kind of preparation method of zinc dense plants lactobacillus additive, it is characterised in that：Including by resistance to high zinc lactobacillus plantarum bacterium
The step of kind is for the preparation of zinc dense plants lactobacillus additive；Wherein, the preparation of the zinc dense plants lactobacillus additive includes
Following steps：
S2.1, slant strains culture：By resistance to high zinc zinc dense plants lactobacillus strain streak inoculation in sugared content 2-5%, Zn content
On 20-500ug/ml, the modified MRS solid medium that pH value is 5.5-7.0,35-40 DEG C of cultivation temperature, Anaerobic culturel 36-
48h；
S2.2, level liquid seed culture：The bacterium colony on modified MRS solid medium is washed into lower mix by the physiological saline of sterilizing
It is even, draw the 250mL modified MRS that 2.5ml bacterium solution is inoculated in sugared content 2-5%, Zn content 20-500ug/ml, pH value 5.5-7.0
In fluid nutrient medium, 35-40 DEG C of cultivation temperature, Anaerobic culturel 15-24h obtains level liquid seed culture fluid；
S2.3, secondary liquid seed culture：200ml level liquid seed culture fluid is inoculated in sugared content 2-5%, Zn content
20-500ug/ml, pH value 5.5-7.0 15-30L MRS liquid culture medium in, 35-40 DEG C of cultivation temperature, revolving speed 10-
50rpm cultivates 5-10h, obtains secondary liquid seed culture fluid；
S2.4, fermentation tank culture：20L secondary liquid seed culture fluid is inoculated in sugared content 2-5%, Zn content 20-1000ug/
Ml, initial pH value 5.5-7.0 1000-2000L MRS liquid culture medium in, 35-40 DEG C of cultivation temperature, revolving speed 10-
50rpm controls pH5.0-6.5 with alkali in fermentation process, cultivates 8-16h, obtains zinc dense plants lactobacillus culture solution；
S2.5, zinc dense plants lactobacillus medium centrifugal is collected, obtained bacterium mud is emulsified with protective agent, is lyophilized, through powder
It is broken to obtain zinc dense plants lactobacillus additive.
2. the preparation method of zinc dense plants lactobacillus additive according to claim 1, it is characterised in that：The resistance to high zinc
Lactobacillus plantarum strain is obtained through following steps：
S1.1, lactobacillus plantarum is inoculated into the modified MRS solid medium containing different zinc concentrations using the method for applying plate
On, to determine the tolerance of its zinc ion；
S1.2, using determining zinc ion tolerable concentration as initial concentration, lactobacillus plantarum is inoculated into the modified MRS containing zinc ion
It is cultivated on fluid nutrient medium, then chooses bacterium solution with oese and trained in the modified MRS solid plate containing identical zinc ion concentration
It supports and carries out scribing line culture on base, this is a generation；Biggish bacterium colony accesses the modified MRS liquid containing zinc ion on picking solid plate
It is cultivated on culture medium, then chooses bacterium solution in the modified MRS solid plate culture medium containing identical zinc ion concentration with oese
On carry out scribing line culture, this be two generations.Such passage and attenuation culture 40-60 generation；
S1.3, by after domestication bacterial strain carry out separation screening, be inoculated into MRS liquid culture medium and cultivate, filter out by
The strong strain of resistance to zinc ability and preservation, as resistance to high zinc lactobacillus plantarum strain after domestication.
3. the preparation method of zinc dense plants lactobacillus additive according to claim 2, it is characterised in that：Step S1.1 and
In step S1.2, the sugared content of modified MRS solid medium and MRS liquid culture medium is 2-5%, Zn content 20-
1000ug/ml, pH value 5.5-7.0, cultivation temperature are 35-40 DEG C；MRS liquid culture medium Anaerobic culturel 12-24h, changes
Good MRS solid medium Anaerobic culturel 36-48h.
4. the preparation method of zinc dense plants lactobacillus additive according to claim 2, it is characterised in that：In step S1.1
The condition for determining lactobacillus plantarum zinc ion tolerance is：Cultivate 36-48h after, single colonie diameter be less than the corresponding zinc of 0.5mm from
Sub- concentration.
5. the preparation method of zinc dense plants lactobacillus additive according to claim 2, it is characterised in that：In step S1.2
The MRS liquid culture medium of the same generation is identical with Zn content in modified MRS solid medium, the improvement that wherein 1-15 generation uses
The Zn content of MRS fluid nutrient medium and modified MRS solid medium is 20-300ug/ml, the modified MRS liquid that 15-35 generation uses
The Zn content of body culture medium and modified MRS solid medium is 300-600ug/ml, the modified MRS liquid training that 35-50 generation uses
The Zn content for supporting base and modified MRS solid medium is 600-1000ug/ml.
6. the preparation method of zinc dense plants lactobacillus additive according to claim 2, it is characterised in that：When plant cream bar
Bacterium is after modified MRS cultured on solid medium 36-48h, and when colony diameter is in 1-2mm, beginning improves zinc in the medium
Content.
7. the preparation method of zinc dense plants lactobacillus additive according to claim 1, it is characterised in that：In step S2.4
The alkali used is the sodium carbonate of 20-40% or sodium hydroxide.
8. the preparation method of zinc dense plants lactobacillus additive according to claim 1, it is characterised in that：In step S2.5
Protective agent include one of trehalose, sucrose, glycerol or a variety of, wherein the mass fraction of the trehalose be 10-
20%, the mass fraction of the sucrose is 2-7%, and the mass fraction of the glycerol is 2-5%.
9. the preparation method of zinc dense plants lactobacillus additive according to claim 1, it is characterised in that：In step S2.5
Lyophilisation condition includes the following steps：
1) -45 DEG C~-50 DEG C 3~4h of pre-freeze；
2) after 240~360min is warming up to -25~-15 DEG C, 840-960min is maintained；
3) after 4-6h is warming up to 0-5 DEG C, 3-5h is maintained；
4) after 120~240min is warming up to 10~15 DEG C, 120~240min is maintained；
5) after 60-90min is warming up to 25~28 DEG C, 240~360min is maintained.
10. the preparation method of zinc dense plants lactobacillus additive according to claim 1, it is characterised in that：Modified MRS liquid
In body culture medium and modified MRS solid medium, the zinc of addition is inorganic zinc, and the inorganic zinc includes zinc oxide, sulfate, carbon
Hydrochlorate or chloride salt, nitrogen source are one or more of yeast extract, yeast extract, peptone, beef extract powder, and carbon source is Portugal
One or more of grape sugar, lactose, sucrose, maltose.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810803962.9A CN108841763A (en) | 2018-07-20 | 2018-07-20 | A kind of preparation method of zinc dense plants lactobacillus additive |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810803962.9A CN108841763A (en) | 2018-07-20 | 2018-07-20 | A kind of preparation method of zinc dense plants lactobacillus additive |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108841763A true CN108841763A (en) | 2018-11-20 |
Family
ID=64196712
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810803962.9A Pending CN108841763A (en) | 2018-07-20 | 2018-07-20 | A kind of preparation method of zinc dense plants lactobacillus additive |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108841763A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109438039A (en) * | 2018-11-19 | 2019-03-08 | 黑龙江八农垦大学 | Yeast zinc foliar fertilizer, production method and application |
| CN115181682A (en) * | 2022-02-10 | 2022-10-14 | 江南大学 | Lactobacillus fermentum with high organic zinc enrichment |
| CN115806920A (en) * | 2022-12-27 | 2023-03-17 | 苏州微克生活科技有限公司 | Zinc-rich lactobacillus paracasei and method for producing organic zinc |
| WO2023165607A1 (en) * | 2022-03-03 | 2023-09-07 | 江南大学 | Strain of high organic zinc enrichment bifidobacterium animalis for promoting growth and reproductive development |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101700101A (en) * | 2009-11-24 | 2010-05-05 | 黄建壮 | Zinc-containing microbial feed additive and fabrication method thereof |
| CN101971921A (en) * | 2010-11-12 | 2011-02-16 | 胡文锋 | Zinc-enriched inactivated lactobacillus microecological agent and preparation method thereof |
| CN103602707A (en) * | 2013-04-25 | 2014-02-26 | 胡文锋 | Bio-synthesis method of nano zinc oxide by utilizing lactic acid bacteria and nano zinc oxide composite feed addictive |
| CN107119038A (en) * | 2017-05-05 | 2017-09-01 | 浙江大学 | A kind of Zinc-rich saccharomyces cerevisiae product and its production method |
-
2018
- 2018-07-20 CN CN201810803962.9A patent/CN108841763A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101700101A (en) * | 2009-11-24 | 2010-05-05 | 黄建壮 | Zinc-containing microbial feed additive and fabrication method thereof |
| CN101971921A (en) * | 2010-11-12 | 2011-02-16 | 胡文锋 | Zinc-enriched inactivated lactobacillus microecological agent and preparation method thereof |
| CN103602707A (en) * | 2013-04-25 | 2014-02-26 | 胡文锋 | Bio-synthesis method of nano zinc oxide by utilizing lactic acid bacteria and nano zinc oxide composite feed addictive |
| CN107119038A (en) * | 2017-05-05 | 2017-09-01 | 浙江大学 | A kind of Zinc-rich saccharomyces cerevisiae product and its production method |
Non-Patent Citations (4)
| Title |
|---|
| 梁猛: "《生物工程技术实验指导》", 28 February 2018 * |
| 武冬萌: "富集锌、铁的酵母菌和乳酸菌及其在面包中的应用", 《中国优秀硕士学位论文全文数据库》 * |
| 聂树人: "《难选冶金矿石浸金》", 31 August 1997 * |
| 赵治平: "富硒锌益生菌制剂研制及其在犬上的初步应用", 《中国优秀硕士学位论文全文数据库》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109438039A (en) * | 2018-11-19 | 2019-03-08 | 黑龙江八农垦大学 | Yeast zinc foliar fertilizer, production method and application |
| CN115181682A (en) * | 2022-02-10 | 2022-10-14 | 江南大学 | Lactobacillus fermentum with high organic zinc enrichment |
| CN115181682B (en) * | 2022-02-10 | 2023-10-03 | 江南大学 | Lactobacillus fermentum highly enriched with organic zinc |
| WO2023165607A1 (en) * | 2022-03-03 | 2023-09-07 | 江南大学 | Strain of high organic zinc enrichment bifidobacterium animalis for promoting growth and reproductive development |
| CN115806920A (en) * | 2022-12-27 | 2023-03-17 | 苏州微克生活科技有限公司 | Zinc-rich lactobacillus paracasei and method for producing organic zinc |
| CN115806920B (en) * | 2022-12-27 | 2024-02-09 | 苏州微克生活科技有限公司 | Zinc-rich lactobacillus paracasei and method for producing organic zinc |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106479924B (en) | The preparation and application of a kind of clostridium butyricum and clostridium butyricum active bacteria preparation | |
| CN108841763A (en) | A kind of preparation method of zinc dense plants lactobacillus additive | |
| CN102660478B (en) | Lactobacillus salivarius and freeze-dried preparation thereof and application of freeze-dried preparation | |
| CN103275907B (en) | Bacillus amyloliquefacien and preparation method and application thereof | |
| CN104770575B (en) | A kind of Radix Astragali probiotics and its preparation method and application | |
| CN107164269A (en) | A kind of lactobacillus paracasei, preparation and its application in pig feed | |
| CN105368755A (en) | Acid-yielding Enterococcus faecium, bacteriostatic microecological preparation and application thereof | |
| CN101971920B (en) | Porcine lactobacillus reuteri lyophilized preparation and preparation method thereof | |
| CN104106726A (en) | New probiotic, and probiotic fermentation feed prepared by using it | |
| CN104673726A (en) | Porcine lactobacillus acidophilus freeze-drying preparation and application thereof | |
| CN102391962B (en) | Enterococcus faecium preparation agent and manufacturing technology thereof | |
| CN100584209C (en) | Feed level microbe additive and its preparation and application | |
| CN109022333A (en) | A kind of preparation method and applications of composite microbial fermentation bacteria agent | |
| CN102321552B (en) | Clostridium butyricum used for feeding, and application thereof | |
| CN108085261A (en) | One Accharomyces cerevisiae and its culture and the application in feed | |
| CN105533140A (en) | Making method of lactic acid bacteria and compound enzyme solid state fermented corn germ meal | |
| CN107319161A (en) | A kind of fermented type selenium-enriched plant feed addictive | |
| CN105524855B (en) | A kind of bacillus coagulans and its application with aquatic pathogenic bacterium antagonistic properties | |
| CN113215018A (en) | Lactobacillus plantarum subspecies and application thereof in mixed fermentation of soybean meal and artemisia vulgaris | |
| CN111304119B (en) | Feeding bacillus subtilis for degrading fumonisins and application thereof | |
| PL214583B1 (en) | A new strain of Lactobacillus plantarum S, the use of the strain of Lactobacillus plantarum S and preparation for ensiling bulky feed | |
| CN112790287A (en) | Preparation technology and application of environment-friendly comprehensive deodorization biological feed for fattening pigs | |
| CN106479944B (en) | A kind of bacillus amyloliquefaciens and its application | |
| CN105341425A (en) | Pig feed containing bacteriostatic microbial ecological agent and application thereof | |
| CN112574924B (en) | Bacillus subtilis strain, microecological preparation and application thereof |
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: 20181120 |
|
| RJ01 | Rejection of invention patent application after publication |