CN106074622A - A kind of preparation method of fixing lactic acid bacteria crude granule - Google Patents
A kind of preparation method of fixing lactic acid bacteria crude granule Download PDFInfo
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- CN106074622A CN106074622A CN201610492239.4A CN201610492239A CN106074622A CN 106074622 A CN106074622 A CN 106074622A CN 201610492239 A CN201610492239 A CN 201610492239A CN 106074622 A CN106074622 A CN 106074622A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
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- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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Abstract
The invention discloses the preparation method of a kind of fixing lactic acid bacteria crude granule, belong to lactein preparing technical field.The present invention takes Dens Draconis and pulverizes and sieves; after calcining stands with lactic acid ethanol solution, glycine betaine; distillating filtering obtains filtrate and is dried; add dithyl sulfate, griseofulvin and distilled water mixing; filtrate is filtered to obtain in cooling; mix sterilizing with MRS culture medium, sterilized water etc. and obtain culture medium; inoculating lactobacillus acidophilus cultivates; modification screening is irradiated with ultraviolet; by it through ferment in second time; filter to obtain filtrate, carry out covering protection by carrageenan solutions, refilter to obtain the air-dried prepared fixing lactic acid bacteria crude granule of filtrate.The invention has the beneficial effects as follows: preparation process of the present invention is simple, products obtained therefrom bacterium timeliness length, antimetabolic performance spectrum width good, antibacterial；Using consumption to decrease more than 26.5%, occur without precipitation in storage process, activity is higher than other products more than 23.6%.
Description
Technical field
The present invention relates to the preparation method of a kind of fixing lactic acid bacteria crude granule, belong to lactein preparing technical field.
Background technology
Lactein is a kind of active polypeptide or protein produced by lactic acid bacteria.Major part bacteriocin is to nearly edge
The antibacterial of relation has inhibitory action, and minority bacteriocin antimicrobial spectrum is the widest.Can be divided mainly into heat-staple small-molecular peptides, heat sensitive
High molecular weight protein and lantibiotics.Lactein is the preparation that a kind of pure lactic acid bacteria produces, and is through sterilization, true by skimmed milk
Empty concentrate, be dried after inoculating lactic acid bacteria fermentation and the lactic acid thalline made and the mixing powder of metabolite thereof, can cure
Medicinal or food additive, function of dominant type milk product.In addition to lactic acid bacteria has the health care such as stomach invigorating, whole intestinal to human body except itself,
Can produce many benefit materials in its metabolic process, the specific function of lactein causes the very big pass of scientific and technological industry circle among these
Note.Due to lactein stable in properties, do not limited by processing conditions, so range of application is the most extensive, can be applicable to include sending out
The fields such as ferment milk, health food, cosmetics and food fresh keeping.
Along with socioeconomic development and the raising of living standards of the people, people are to healthy attention degree also day increasingly
By force, no matter from the perspective of food preservative, antistaling agent, ablastins or economic animal fertilizer synergist develop, natural food
Anticorrosion, antistaling agent replace or part substituted chemistry preservative, do not produce drug resistance and can be by genetic engineering modified antibacterial
Element substitutes original antibiotic, and substituting original feed additive without toxin residual feed additive is trend of the times.But lactein
The shortcoming that there is self, the weak point such as big in consumption, titer timeliness low, antibacterial is short, antimetabolic poor performance, narrow antimicrobial spectrum, with
And dominant strain selection-breeding difficulty, strain improvement research need to strengthen；Antimicrobial spectrum is the narrowest, and dissolubility is bad in aqueous,
Storage process is understood due to precipitation loss of activity.
Summary of the invention
The technical problem to be solved:, antimetabolic performance short for current lactein consumption timeliness big, antibacterial
Difference, narrow antimicrobial spectrum, understand due to precipitation the drawback of loss of activity, it is provided that one takes Dens Draconis and pulverizes and sieves in storage process,
After calcining stands with lactic acid ethanol solution, glycine betaine, distillating filtering obtains filtrate and is dried, and addition dithyl sulfate, sallow are mould
Element and distilled water mix, and filtrate is filtered to obtain in cooling, mix sterilizing with MRS culture medium, sterilized water etc. and obtain culture medium, and inoculation is addicted to acid
Lactobacillus is cultivated, and irradiates modification screening through ultraviolet, by it through ferment in second time, filters to obtain filtrate, carry out by carrageenan solutions
Covering protection, refilters to obtain the filtrate method that air-dries prepared fixing lactic acid bacteria crude granule.Preparation process of the present invention is simple, has
Effect solves that antibacterial timeliness is short, antimetabolic poor performance, narrow antimicrobial spectrum problem, uses consumption little, occurs without precipitation in storage process,
Activity is high.
For solving above-mentioned technical problem, the present invention uses the technical scheme as described below to be:
(1) taking Dens Draconis to put in pulverizer and pulverize, cross 20 mesh sieves, the granule after sieving is put in Muffle furnace, in 200～
Calcine 5～8min at 250 DEG C, cool to room temperature with the furnace, by solid-to-liquid ratio 1:2～3, the granule after calcining is soaked in mass fraction
Be in the lactic acid ethanol solution of 10%, then be placed in ultrasonator, under 23～24KHz vibrate 10～15min, with backward its
Middle addition calcined particle quality 3～the glycine betaine of 6%, stir and be placed on 40～50 DEG C, stands 3～4h；
(2) after above-mentioned standing terminates, being put into by mixture in distilling apparatus, at 105～110 DEG C, distillation 1～3h, the coldest
But to room temperature, then filter, use distilled water flushing filtrate until flushing liquor pH is neutral, filtrate is put into vacuum and does
In dry case, at 70～80 DEG C, it is vacuum dried 2～3h；
(3) 15:2:1 in mass ratio, takes above-mentioned dried filtrate, dithyl sulfate and griseofulvin and puts in container, to
Container adds volume of mixture 4～the distilled water of 6 times in container, container is heated to 50～60 DEG C, with 200r/min
Stop heating after stirring 3～5h, be passed through 5 DEG C of hydrogen from container bottom until the mixture temperature in container is down to 10～15 DEG C,
Filter, collect filtrate, obtain fixing lactic acid bacteria element carrier；
(4) count by weight, take the fixing lactic acid bacteria element carrier of 45～50 parts of above-mentioned gained, 30～35 parts of MRS culture medium,
12～15 parts of sterilized water, 4～6 parts of peptones and 8～12 parts of yeast extracts, stir, moist heat sterilization 12 at 110～120 DEG C
～15min, obtain culture medium, by inoculum concentration 13～16%, bacillus acidophilus is seeded in culture medium, and culture medium is moved to perseverance
In temperature incubator, at 30～37 DEG C, cultivate 18～22h；
(5) after above-mentioned cultivation terminates, culture medium is poured in glass fermentation tank, use ultraviolet to irradiate glass fermentation tank, set
Temperature is 30～37 DEG C, with 140～160r/min stirring fermentations 2～3h, stops stirring subsequently, after continuing fermentation 3～4h, will send out
Mixture in ferment tank filters, and uses the distilled water flushing filtrate 8～10min of 10mL/min, filtrate after rinsing
Put into and the carrageenan solutions that mass fraction is 10% stands 20～30min, then filter, collect filtrate and put into air-dry machine
In air-dry, i.e. can get fixing lactic acid bacteria crude granule.
The application process of the present invention: take the 1.0～2.0g fixing lactic acid bacteria crude granules present invention prepared, use warm water
Take after mixing it with water, be administered orally after taking after mixing it with water, 3 times on the one, can effectively treat enteral abnormal fermentation, dyspepsia, enteritis etc..This immobilization breast
Acid rhzomorph granule, antibacterial timeliness length, antimetabolic performance spectrum width good, antibacterial, it is worthy to be popularized and uses.
The present invention is compared with additive method, and Advantageous Effects is:
(1) preparation process of the present invention is simple, products obtained therefrom antibacterial timeliness length, antimetabolic performance spectrum width good, antibacterial；
(2) use consumption decrease more than 26.5%, in storage process without precipitation occur, activity higher than other products 23.6% with
On.
Detailed description of the invention
First taking Dens Draconis to put in pulverizer and pulverize, cross 20 mesh sieves, the granule after sieving is put in Muffle furnace, in
Calcine 5～8min at 200～250 DEG C, cool to room temperature with the furnace, by solid-to-liquid ratio 1:2～3, the granule after calcining is soaked in matter
Amount mark is in the lactic acid ethanol solution of 10%, then is placed in ultrasonator, vibration 10～15min under 23～24KHz, with
After be added thereto to calcined particle quality 3～the glycine betaine of 6%, stir and be placed on 40～50 DEG C, stand 3～4h；So
After after above-mentioned standing terminates, mixture is put in distilling apparatus, at 105～110 DEG C distill 1～3h, naturally cool to
Room temperature, then filter, use distilled water flushing filtrate until flushing liquor pH is neutral, filtrate is put into vacuum drying oven
In, at 70～80 DEG C, it is vacuum dried 2～3h；15:2:1 in mass ratio, take above-mentioned dried filtrate, dithyl sulfate and
Griseofulvin is put in container, adds volume of mixture 4～the distilled water of 6 times in container, heat container in container
To 50～60 DEG C, to stop heating after 200r/min stirring 3～5h, it is passed through 5 DEG C of hydrogen from container bottom until mixed in container
Compound temperature is down to 10～15 DEG C, filters, and collects filtrate, obtains fixing lactic acid bacteria element carrier；Count the most by weight,
Take the fixing lactic acid bacteria element carrier of 45～50 parts of above-mentioned gained, 30～35 parts of MRS culture medium, 12～15 parts of sterilized water, 4～6
Part peptone and 8～12 parts of yeast extracts, stir, and at 110～120 DEG C, moist heat sterilization 12～15min, obtain culture medium, press
Inoculum concentration 13～16%, is seeded to bacillus acidophilus in culture medium, and culture medium is moved in constant incubator, 30～37
18～22h are cultivated at DEG C；Finally after above-mentioned cultivation terminates, culture medium is poured in glass fermentation tank, use ultraviolet to irradiate glass
Glass fermentation tank, design temperature is 30～37 DEG C, with 140～160r/min stirring fermentations 2～3h, stops stirring subsequently, continues to send out
After ferment 3～4h, the mixture in fermentation tank is filtered, uses the distilled water flushing filtrate 8～10min of 10mL/min,
After rinsing, filtrate is put into and is stood 20～30min in the carrageenan solutions that mass fraction is 10%, then filters, and collects
Screening is put in air-dry machine and is air-dried, and i.e. can get fixing lactic acid bacteria crude granule.
Example 1
First taking Dens Draconis to put in pulverizer and pulverize, cross 20 mesh sieves, the granule after sieving is put in Muffle furnace, in 225
Calcining 6min at DEG C, cool to room temperature with the furnace, by solid-to-liquid ratio 1:2, the granule after calcining is soaked in mass fraction is 10%
In lactic acid ethanol solution, then being placed in ultrasonator, vibrate under 23KHz 10min, is added thereto to calcined particle subsequently
The glycine betaine of quality 3%, stirs and is placed on 40 DEG C, stands 3h；Then, after above-mentioned standing terminates, mixture is put into steaming
In distillation unit, at 105 DEG C, distilling 1h, naturally cool to room temperature, then filter, using distilled water flushing filtrate until rushing
Washing liquid pH is neutral, filtrate is put in vacuum drying oven, is vacuum dried 2h at 70 DEG C；15:2:1 in mass ratio, takes
State dried filtrate, dithyl sulfate and griseofulvin and put in container, in container, add volume of mixture 4 in container
Distilled water again, is heated to 50 DEG C to container, to stop heating after 200r/min stirring 3h, is passed through 5 DEG C from container bottom
Hydrogen, until the mixture temperature in container is down to 10 DEG C, filters, and collects filtrate, obtains fixing lactic acid bacteria element carrier；
Count the most by weight, take the fixing lactic acid bacteria element carrier of 45 parts of above-mentioned gained, 30 parts of MRS culture medium, 12 parts of sterilized water, 4
Part peptone and 8 parts of yeast extracts, stir, and at 110 DEG C, moist heat sterilization 12min, obtains culture medium, by inoculum concentration 13%, and will
Bacillus acidophilus is seeded in culture medium, and culture medium is moved in constant incubator, cultivates 18h at 30 DEG C；Last upper
Stating pours in glass fermentation tank by culture medium after cultivation terminates, and uses ultraviolet to irradiate glass fermentation tank, and design temperature is 30 DEG C,
With 140r/min stirring fermentation 2h, stop stirring subsequently, after continuing fermentation 3h, the mixture in fermentation tank is filtered, makes
By distilled water flushing filtrate 8min of 10mL/min, after rinsing, filtrate puts into the carrageenan solutions that mass fraction is 10%
Middle standing 20min, then filter, collect filtrate and put in air-dry machine air-dried, i.e. can get fixing lactic acid bacteria crude granule.
Take the fixing lactic acid bacteria crude granule that the present invention is prepared by 1.0g, with warm boiled water, be administered orally after taking after mixing it with water, one
Day three times, can effectively treat enteral abnormal fermentation, dyspepsia, enteritis etc..This fixing lactic acid bacteria crude granule, antibacterial timeliness
Long, antimetabolic performance spectrum width good, antibacterial, is worthy to be popularized and uses.
Example 2
First taking Dens Draconis to put in pulverizer and pulverize, cross 20 mesh sieves, the granule after sieving is put in Muffle furnace, in 225
Calcining 6min at DEG C, cool to room temperature with the furnace, by solid-to-liquid ratio 1:3, the granule after calcining is soaked in mass fraction is 10%
In lactic acid ethanol solution, then being placed in ultrasonator, vibrate under 24KHz 13min, is added thereto to calcined particle subsequently
The glycine betaine of quality 5%, stirs and is placed on 45 DEG C, stands 4h；Then, after above-mentioned standing terminates, mixture is put into steaming
In distillation unit, at 108 DEG C, distilling 2h, naturally cool to room temperature, then filter, using distilled water flushing filtrate until rushing
Washing liquid pH is neutral, filtrate is put in vacuum drying oven, is vacuum dried 3h at 75 DEG C；15:2:1 in mass ratio, takes
State dried filtrate, dithyl sulfate and griseofulvin and put in container, in container, add volume of mixture 5 in container
Distilled water again, is heated to 55 DEG C to container, to stop heating after 200r/min stirring 4h, is passed through 5 DEG C from container bottom
Hydrogen, until the mixture temperature in container is down to 13 DEG C, filters, and collects filtrate, obtains fixing lactic acid bacteria element carrier；
Count the most by weight, take the fixing lactic acid bacteria element carrier of 48 parts of above-mentioned gained, 33 parts of MRS culture medium, 14 parts of sterilized water, 5
Part peptone and 10 parts of yeast extracts, stir, and at 115 DEG C, moist heat sterilization 14min, obtains culture medium, by inoculum concentration 15%, and will
Bacillus acidophilus is seeded in culture medium, and culture medium is moved in constant incubator, cultivates 20h at 34 DEG C；Last upper
Stating pours in glass fermentation tank by culture medium after cultivation terminates, and uses ultraviolet to irradiate glass fermentation tank, and design temperature is 35 DEG C,
With 150r/min stirring fermentation 3h, stop stirring subsequently, after continuing fermentation 4h, the mixture in fermentation tank is filtered, makes
By distilled water flushing filtrate 9min of 10mL/min, after rinsing, filtrate puts into the carrageenan solutions that mass fraction is 10%
Middle standing 25min, then filter, collect filtrate and put in air-dry machine air-dried, i.e. can get fixing lactic acid bacteria crude granule.
Take the fixing lactic acid bacteria crude granule that the present invention is prepared by 1.5g, with warm boiled water, be administered orally after taking after mixing it with water, one
Day 3 times, can effectively treat enteral abnormal fermentation, dyspepsia, enteritis etc..This fixing lactic acid bacteria crude granule, antibacterial timeliness length,
Antimetabolic performance spectrum width good, antibacterial, is worthy to be popularized and uses.
Example 3
First taking Dens Draconis to put in pulverizer and pulverize, cross 20 mesh sieves, the granule after sieving is put in Muffle furnace, in 250
Calcining 8min at DEG C, cool to room temperature with the furnace, by solid-to-liquid ratio 1:3, the granule after calcining is soaked in mass fraction is 10%
In lactic acid ethanol solution, then being placed in ultrasonator, vibrate under 24KHz 15min, is added thereto to calcined particle subsequently
The glycine betaine of quality 6%, stirs and is placed on 50 DEG C, stands 4h；Then, after above-mentioned standing terminates, mixture is put into steaming
In distillation unit, at 110 DEG C, distilling 3h, naturally cool to room temperature, then filter, using distilled water flushing filtrate until rushing
Washing liquid pH is neutral, filtrate is put in vacuum drying oven, is vacuum dried 3h at 80 DEG C；15:2:1 in mass ratio, takes
State dried filtrate, dithyl sulfate and griseofulvin and put in container, in container, add volume of mixture 6 in container
Distilled water again, is heated to 60 DEG C to container, to stop heating after 200r/min stirring 5h, is passed through 5 DEG C from container bottom
Hydrogen, until the mixture temperature in container is down to 15 DEG C, filters, and collects filtrate, obtains fixing lactic acid bacteria element carrier；
Count the most by weight, take the fixing lactic acid bacteria element carrier of 50 parts of above-mentioned gained, 35 parts of MRS culture medium, 15 parts of sterilized water, 6
Part peptone and 12 parts of yeast extracts, stir, and at 120 DEG C, moist heat sterilization 15min, obtains culture medium, by inoculum concentration 16%, and will
Bacillus acidophilus is seeded in culture medium, and culture medium is moved in constant incubator, cultivates 22h at 37 DEG C；Last upper
Stating pours in glass fermentation tank by culture medium after cultivation terminates, and uses ultraviolet to irradiate glass fermentation tank, and design temperature is 37 DEG C,
With 160r/min stirring fermentation 3h, stop stirring subsequently, after continuing fermentation 4h, the mixture in fermentation tank is filtered, makes
By distilled water flushing filtrate 10min of 10mL/min, after rinsing, filtrate puts into the OK a karaoke club peptization that mass fraction is 10%
Liquid stands 30min, then filters, collect filtrate and put in air-dry machine air-dried, i.e. can get fixing lactic acid bacteria element
Grain.
Take the fixing lactic acid bacteria crude granule that the present invention is prepared by 2.0g, with warm boiled water, be administered orally after taking after mixing it with water, one
Day 3 times, can effectively treat enteral abnormal fermentation, dyspepsia, enteritis etc..This fixing lactic acid bacteria crude granule, antibacterial timeliness length,
Antimetabolic performance spectrum width good, antibacterial, is worthy to be popularized and uses.
Claims (1)
1. the preparation method of a fixing lactic acid bacteria crude granule, it is characterised in that concrete preparation process is:
(1) taking Dens Draconis to put in pulverizer and pulverize, cross 20 mesh sieves, the granule after sieving is put in Muffle furnace, in 200～
Calcine 5～8min at 250 DEG C, cool to room temperature with the furnace, by solid-to-liquid ratio 1:2～3, the granule after calcining is soaked in mass fraction
Be in the lactic acid ethanol solution of 10%, then be placed in ultrasonator, under 23～24KHz vibrate 10～15min, with backward its
Middle addition calcined particle quality 3～the glycine betaine of 6%, stir and be placed on 40～50 DEG C, stands 3～4h；
(2) after above-mentioned standing terminates, being put into by mixture in distilling apparatus, at 105～110 DEG C, distillation 1～3h, the coldest
But to room temperature, then filter, use distilled water flushing filtrate until flushing liquor pH is neutral, filtrate is put into vacuum and does
In dry case, at 70～80 DEG C, it is vacuum dried 2～3h；
(3) 15:2:1 in mass ratio, takes above-mentioned dried filtrate, dithyl sulfate and griseofulvin and puts in container, to
Container adds volume of mixture 4～the distilled water of 6 times in container, container is heated to 50～60 DEG C, with 200r/min
Stop heating after stirring 3～5h, be passed through 5 DEG C of hydrogen from container bottom until the mixture temperature in container is down to 10～15 DEG C,
Filter, collect filtrate, obtain fixing lactic acid bacteria element carrier；
(4) count by weight, take the fixing lactic acid bacteria element carrier of 45～50 parts of above-mentioned gained, 30～35 parts of MRS culture medium,
12～15 parts of sterilized water, 4～6 parts of peptones and 8～12 parts of yeast extracts, stir, moist heat sterilization 12 at 110～120 DEG C
～15min, obtain culture medium, by inoculum concentration 13～16%, bacillus acidophilus is seeded in culture medium, and culture medium is moved to perseverance
In temperature incubator, at 30～37 DEG C, cultivate 18～22h；
(5) after above-mentioned cultivation terminates, culture medium is poured in glass fermentation tank, use ultraviolet to irradiate glass fermentation tank, set
Temperature is 30～37 DEG C, with 140～160r/min stirring fermentations 2～3h, stops stirring subsequently, after continuing fermentation 3～4h, will send out
Mixture in ferment tank filters, and uses the distilled water flushing filtrate 8～10min of 10mL/min, filtrate after rinsing
Put into and the carrageenan solutions that mass fraction is 10% stands 20～30min, then filter, collect filtrate and put into air-dry machine
In air-dry, i.e. can get fixing lactic acid bacteria crude granule.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090155328A1 (en) * | 2007-12-14 | 2009-06-18 | E. I. Du Pont De Nemours And Company | Films comprising antimicrobial and fungistatic agents |
| CN105478080A (en) * | 2015-12-25 | 2016-04-13 | 常州大学 | Preparation method of biological charcoal |
-
2016
- 2016-06-29 CN CN201610492239.4A patent/CN106074622A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090155328A1 (en) * | 2007-12-14 | 2009-06-18 | E. I. Du Pont De Nemours And Company | Films comprising antimicrobial and fungistatic agents |
| CN105478080A (en) * | 2015-12-25 | 2016-04-13 | 常州大学 | Preparation method of biological charcoal |
Non-Patent Citations (2)
| Title |
|---|
| 田宇: "乳酸菌素产生菌的筛选及其发酵产物性质的初步研究", 《中国优秀硕士学位论文全文数据库，工程科技I辑》 * |
| 高松: "《辽宁中药志 动物、矿物、海洋类》", 31 July 2015 * |
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