CN106086120A - A kind of streptomycete fermentation culture medium - Google Patents
A kind of streptomycete fermentation culture medium Download PDFInfo
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- CN106086120A CN106086120A CN201610628331.9A CN201610628331A CN106086120A CN 106086120 A CN106086120 A CN 106086120A CN 201610628331 A CN201610628331 A CN 201610628331A CN 106086120 A CN106086120 A CN 106086120A
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- culture medium
- straw
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/46—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin
- C12P19/54—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin the cyclohexyl radical being bound directly to a nitrogen atom of two or more radicals, e.g. streptomycin
Abstract
The invention belongs to field of microbial culture technology, disclosing a kind of streptomycete fermentation culture medium, it is prepared by the raw material of following percentage by weight: straw zymolyte 7.3 9.2%, Rhizoma Solani tuber osi diffusion juice 4.2 6.8%, bean cake hydrolysate 3.3 5.5%, fishbone powder hydrolyzed solution 2.6 4.7%, Testa Tritici 2.3 3.7%, glucose 1.5 2.6%, dipotassium hydrogen phosphate 0.1 0.2%, magnesium sulfate 0.05 0.08%, sodium chloride 0.02 0.03%, remaining is water.Fermentation medium of the present invention is with low cost, improves enterprise profit.
Description
Technical field
The invention belongs to field of microbial culture technology, be specifically related to a kind of streptomycete fermentation culture medium.
Background technology
Streptomyces (streptomyces), its substrate mycelium multi-branched, general tabula is sparse, seldom ruptures, and often produces
Various water solublity or fat-soluble pigment；Aerial hyphae is slightly thicker than substrate mycelium, is wrapped by epitheca, and aerial hyphae partial differentiation becomes
Straight, flexible, hook and loop to loose spacious or the fibrillae of spores of tight spiral, contain the spore of about 50 often, and short person contains 5～10
Spore；Spore is arthrospore, mycelia fracture form, and have sloughs epitheca, and have carries epitheca or remnants, and epitheca determines spore
Surface texture；G+C mole content in DNA is 69～76%.This genus strain number is most, and taxonomic identification is relatively difficult, typically
Think the shape of fibrillae of spores, the surface texture of spore, the color of spore and in organic culture medium, whether produce Melanoidins be
Topmost classification indication.They distributed poles in soil are wide, and riotous growth on synthetic medium mostly, minority is plant cause
Pathogenic bacteria.Because many kinds are the producing strains of antibiotic and produce the most species of antibiotic and famous (such as streptomycin).
Streptomycin is a kind of antibiotics extracted from the culture fluid of ash streptomycete.Belong to Aminoglycoside alkali compounds,
It is combined with tubercule bacillus thalline ribonucleoprotein (RNP) body protein, serves the effect of interference tubercule bacillus protein synthesis,
Thus kill or suppress the effect of growth of bacillus tubercle.Owing to the pain reaction of streptomycin intramuscular injection is smaller, suitably face
Bed uses, as long as application selects proper, dosage is the most proper, and most patients can (general 2 months left with long term injections
Right).So, the application mainstay that recent decades, it was still in antituberculosis therapy.
It is main path that streptomycete fermentation method prepares streptomycin, but the cost of culture medium governs sending out of enterprise
Exhibition, the most relatively multiple enterprises uses PDA culture medium or yeast dextrose culture-medium, and cost of material is high, and the profit of enterprise is substantially reduced.
Developing the fermentation medium of a kind of low cost is our technical issues that need to address.
Summary of the invention
Present invention aim to address the defect that prior art streptomycete fermentation culture medium cost is high, it is provided that a kind of low cost
Streptomycete fermentation culture medium.
The present invention is achieved by the following technical solution:
A kind of streptomycete fermentation culture medium, it is prepared by following raw material:
Straw zymolyte 7.3-9.2%, Rhizoma Solani tuber osi diffusion juice 4.2-6.8%, bean cake hydrolysate 3.3-5.5%, fishbone powder hydrolyzed solution
2.6-4.7%, Testa Tritici 2.3-3.7%, glucose 1.5-2.6%, dipotassium hydrogen phosphate 0.1-0.2%, magnesium sulfate 0.05-0.08%, chlorination
Sodium 0.02-0.03%, remaining is water.
Described straw zymolyte is prepared according to following technique:
Rice straw is put into and pulverizer carries out pulverizing obtains powder of straw, be then paved into the flat bed of 1cm thickness, then ultraviolet
Line irradiates 15min, then puts in container, the water soaking of interpolation double weight 2 hours, adds subsequently and accounts for powder of straw 0.5% weight
The cellulase of part, is warming up to 50 DEG C, keeps 50 DEG C to hydrolyze 12 hours, and then 100 DEG C of enzyme denaturing, are finally concentrated into cream by enzymolysis solution
Shape, to obtain final product.
Described Rhizoma Solani tuber osi diffusion juice is prepared according to following technique:
By peeling potatoes, it is cut into bulk, is then added in distilled water, boil 45min, natural cooling, then use gauze mistake
Filter, collects filtrate, to obtain final product；Wherein, block Rhizoma Solani tuber osi is 1:3 with the mass ratio of distilled water.
Described bean cake hydrolysate is prepared according to following technique:
Being put into by bean cake in reactor, then add the hydrochloric acid that concentration is 5M of double weight, 300rpm stirring hydrolysis 2 is little
Time, then adding ammonia, the pH of regulation solution is 7.1, to obtain final product.
Described fishbone powder hydrolyzed solution is prepared according to following technique:
Being placed in retort by fishbone powder, add the hydrochloric acid of 5M, stirring hydrolysis 12 hours at a temperature of 50 DEG C, mixing speed is
200 turns/min, using in ammonia and remaining hydrochloric acid after reaction terminating, the pH controlling solution is 7.1, to obtain final product.
The beneficial effect that the present invention obtains mainly includes several aspect:
The present invention has carried out pulverizing and enzymolysis processing to agricultural wastes straw so that nitrogen, phosphorus, potassium, calcium, magnesium and fibrination
Sugar etc. is utilized effectively；Bean cake belongs to agricultural wastes, and it contains substantial amounts of protein, fat, sugar and vitamin etc., but
Being that bacterial strain utilization rate is relatively low, after different biochemical treatments, improve the leaching rate of each nutrient, bacterial strain utilization rate carries significantly
High；Containing carbon source, nitrogen source and the trace element needed for thalli growth in Rhizoma Solani tuber osi diffusion juice；Fishbone powder has been carried out at hydrolysis
Reason, improves the nitrogen source of abundance；Extraction process uses acid-base neutralization mode, while balance pH degree, adds ammonium chloride
The supply in nitrogen source, kills two birds with one stone；Nutrient media components of the present invention uses garbage material, with low cost, it is achieved that to turn waste into wealth,
Improve the added value of industry, enterprise profit is greatly improved；The each material combination of culture medium of the present invention is reasonable, with low cost, can be complete
Replacing market and commonly use culture medium, streptomycete fermentation efficiency improves the most accordingly.
Detailed description of the invention
Hereinafter the present invention is further explained by employing specific embodiment, but should not be construed as the present invention is created
The restriction of new spirit.
Embodiment 1
A kind of streptomycete fermentation culture medium, it is prepared by the raw material of following percentage by weight:
Straw zymolyte 7.3%, Rhizoma Solani tuber osi diffusion juice 6.8%, bean cake hydrolysate 5.5%, fishbone powder hydrolyzed solution 2.6%, Testa Tritici 2.3%,
Glucose 2.6%, dipotassium hydrogen phosphate 0.2%, magnesium sulfate 0.05%, sodium chloride 0.03%, remaining is water；
Its preparation technology is: add in water by each raw material, adjusts pH7.1-7.5,121 DEG C of insulations sterilizing in 20 minutes, is cooled to 32
DEG C, to obtain final product.
Described straw zymolyte is prepared according to following technique:
Rice straw is put into and pulverizer carries out pulverizing obtains powder of straw, be then paved into the flat bed of 1cm thickness, then ultraviolet
Line irradiates 15min, and uitraviolet intensity is 3000uW/cm2, then put in container, the water soaking of interpolation double weight 2 hours,
Add the cellulase (enzyme activity is 3000U/g) accounting for powder of straw 0.5% weight portion subsequently, be warming up to 50 DEG C, keep 50 DEG C of hydrolysis
12 hours, then 100 DEG C of enzyme denaturing, finally enzymolysis solution is concentrated into paste (water content 20% weight portion), to obtain final product；
Described Rhizoma Solani tuber osi diffusion juice is prepared according to following technique:
By peeling potatoes, it is cut into bulk, is then added in distilled water, boil 45min, natural cooling, then use gauze mistake
Filter, collects filtrate, to obtain final product；Wherein, block Rhizoma Solani tuber osi is 1:3 with the mass ratio of distilled water；
Described bean cake hydrolysate is prepared according to following technique:
Being put into by bean cake in reactor, then add the hydrochloric acid that concentration is 5M of double weight, 300rpm stirring hydrolysis 2 is little
Time, then adding ammonia, the pH of regulation solution is 7.1, to obtain final product；
Described fishbone powder hydrolyzed solution is prepared according to following technique:
Being placed in retort by fishbone powder, add the hydrochloric acid of 5M, stirring hydrolysis 12 hours at a temperature of 50 DEG C, mixing speed is
200 turns/min, using in ammonia and remaining hydrochloric acid after reaction terminating, the pH controlling solution is 7.1, to obtain final product.
Streptomycin is prepared in fermentation: by streptomyces griseus (Chinese industrial Microbiological Culture Collection administrative center, preserving number
CICC11002) (concentration is 0.5 × 10 to seed culture fluid9Cfu/ml) 2L accesses the 30L fermentation tank equipped with 15L fermentation medium
In, 400rpm, ventilation 2000 L/h, 32 DEG C ferment 96 hours；Fermentation processes: at 48-96 hour, add glucose,
Control the concentration of glucose in fermentation liquid and be not less than 1% mass fraction；At 48-96 hour, control pH was 7.1-7.2；Within 96 hours, take
Sample carries out HPLC mensuration, and the titer of streptavidin is 867U/ml.
Control medium: yeast extract 7.0-9.0%, glucose 6.0-8.0%, corn starch 3.0-5.0%, potassium dihydrogen phosphate
0.8-1.2%, magnesium sulfate 0.05-0.08%, sodium chloride 0.02-0.03%, remaining is water；Control medium is used to ferment,
Ibid, sampling in 96 hours carries out HPLC mensuration to technique, and the titer of streptavidin is 706U/ml.
Simultaneously by fermentation medium is carried out cost of material accounting, the present invention is only about the 60% of matched group cost, joint
Yue Liao enterprise puts into, and improves enterprise's net income.
Embodiment 2
A kind of streptomycete fermentation culture medium, it is prepared by the raw material of following percentage by weight:
Straw zymolyte 9.2%, Rhizoma Solani tuber osi diffusion juice 4.2%, bean cake hydrolysate 3.3%, fishbone powder hydrolyzed solution 4.7%, Testa Tritici 2.3%,
Glucose 1.5%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.05%, sodium chloride 0.03%, remaining is water；
Its preparation technology is: add in water by each raw material, adjusts pH7.1-7.5,121 DEG C of insulations sterilizing in 20 minutes, is cooled to 32
DEG C, to obtain final product.
Described straw zymolyte is prepared according to following technique:
Rice straw is put into and pulverizer carries out pulverizing obtains powder of straw, be then paved into the flat bed of 1cm thickness, then ultraviolet
Line irradiates 15min, and uitraviolet intensity is 3000uW/cm2, then put in container, the water soaking of interpolation double weight 2 hours,
Add the cellulase (enzyme activity is 3000U/g) accounting for powder of straw 0.5% weight portion subsequently, be warming up to 50 DEG C, keep 50 DEG C of hydrolysis
12 hours, then 100 DEG C of enzyme denaturing, finally enzymolysis solution is concentrated into paste (water content 20% weight portion), to obtain final product；
Described Rhizoma Solani tuber osi diffusion juice is prepared according to following technique:
By peeling potatoes, it is cut into bulk, is then added in distilled water, boil 45min, natural cooling, then use gauze mistake
Filter, collects filtrate, to obtain final product；Wherein, block Rhizoma Solani tuber osi is 1:3 with the mass ratio of distilled water；
Described bean cake hydrolysate is prepared according to following technique:
Being put into by bean cake in reactor, then add the hydrochloric acid that concentration is 5M of double weight, 300rpm stirring hydrolysis 2 is little
Time, then adding ammonia, the pH of regulation solution is 7.1, to obtain final product；
Described fishbone powder hydrolyzed solution is prepared according to following technique:
Being placed in retort by fishbone powder, add the hydrochloric acid of 5M, stirring hydrolysis 12 hours at a temperature of 50 DEG C, mixing speed is
200 turns/min, using in ammonia and remaining hydrochloric acid after reaction terminating, the pH controlling solution is 7.1, to obtain final product.
Streptomycin is prepared in fermentation: by streptomyces griseus (CGMCC No.1692) seed culture fluid, (concentration is 1 × 109
Cfu/ml) 2L accesses equipped with in the 30L fermentation tank of 15L fermentation medium, 400rpm, ventilation 2000 L/h, 32 DEG C of fermentations 96
Hour；Fermentation processes: at 48-96 hour, adds glucose, controls the concentration of glucose in fermentation liquid and is not less than 1% mass
Mark；At 48-96 hour, control pH was 7.1-7.2；Sampling in 96 hours carries out HPLC mensuration, and the titer of streptavidin is 981U/
ml。
Control medium: soybean cake powder 8.0%, glucose 5.0%, corn starch 3.5%, ammonium sulfate 3.5%, potassium dihydrogen phosphate
0.8%, magnesium sulfate 0.05%, sodium chloride 0.02%, remaining is water；Using control medium to ferment, ibid, 96 is little for technique
Time sampling carry out HPLC mensuration, the titer of streptavidin is 739U/ml.
Simultaneously by fermentation medium is carried out cost of material accounting, the present invention is only less than the 50% of matched group cost, joint
Yue Liao enterprise puts into, and improves enterprise's net income.
Finally, in addition it is also necessary to be only several specific embodiments of the present invention it is noted that listed above.Obviously, the present invention is not
It is limited to above example, it is also possible to have many deformation.Those of ordinary skill in the art can be direct from present disclosure
The all deformation derived or associate, are all considered as protection scope of the present invention.
Claims (5)
1. a streptomycete fermentation culture medium, it is prepared by the raw material of following percentage by weight:
Straw zymolyte 7.3-9.2%, Rhizoma Solani tuber osi diffusion juice 4.2-6.8%, bean cake hydrolysate 3.3-5.5%, fishbone powder hydrolyzed solution
2.6-4.7%, Testa Tritici 2.3-3.7%, glucose 1.5-2.6%, dipotassium hydrogen phosphate 0.1-0.2%, magnesium sulfate 0.05-0.08%, chlorine
Changing sodium 0.02-0.03%, remaining is water.
Culture medium the most according to claim 1, it is characterised in that described straw zymolyte is according to the preparation of following technique
:
Rice straw is put into and pulverizer carries out pulverizing obtains powder of straw, be then paved into the flat bed of 1cm thickness, then ultraviolet
Line irradiates 15min, then puts in container, the water soaking of interpolation double weight 2 hours, adds subsequently and accounts for powder of straw 0.5% weight
The cellulase of part, is warming up to 50 DEG C, keeps 50 DEG C to hydrolyze 12 hours, and then 100 DEG C of enzyme denaturing, are finally concentrated into cream by enzymolysis solution
Shape, to obtain final product.
Culture medium the most according to claim 1, it is characterised in that described Rhizoma Solani tuber osi diffusion juice is according to the preparation of following technique
: by peeling potatoes, it is cut into bulk, is then added in distilled water, boil 45min, natural cooling, then use gauze mistake
Filter, collects filtrate, to obtain final product；Wherein, block Rhizoma Solani tuber osi is 1:3 with the mass ratio of distilled water.
Culture medium the most according to claim 1, it is characterised in that described bean cake hydrolysate is according to the preparation of following technique
:
Being put into by bean cake in reactor, then add the hydrochloric acid that concentration is 5M of double weight, 300rpm stirring hydrolysis 2 is little
Time, then adding ammonia, the pH of regulation solution is 7.1, to obtain final product.
Culture medium the most according to claim 1, it is characterised in that described fishbone powder hydrolyzed solution is according to the preparation of following technique
: being placed in retort by fishbone powder, add the hydrochloric acid of 5M, stirring hydrolysis 12 hours at a temperature of 50 DEG C, mixing speed is
200 turns/min, using in ammonia and remaining hydrochloric acid after reaction terminating, the pH controlling solution is 7.1, to obtain final product.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111100798A (en) * | 2020-01-16 | 2020-05-05 | 中化农业（临沂）研发中心有限公司 | Aspergillus awamori, microbial inoculum and application thereof |
| CN111154661A (en) * | 2020-01-16 | 2020-05-15 | 中化农业（临沂）研发中心有限公司 | Complex microbial inoculant and application thereof |
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| CN1344699A (en) * | 2000-09-26 | 2002-04-17 | 邓伟 | Microbial streptomycete fertilizer and production process |
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| CN102286572A (en) * | 2011-06-23 | 2011-12-21 | 浙江大学 | Method for preparing fermentable sugar solution from straws |
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| CN1344699A (en) * | 2000-09-26 | 2002-04-17 | 邓伟 | Microbial streptomycete fertilizer and production process |
| CN1618772A (en) * | 2003-11-20 | 2005-05-25 | 郭伟光 | High efficiency biological organic fertilizer and its production technology |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111100798A (en) * | 2020-01-16 | 2020-05-05 | 中化农业（临沂）研发中心有限公司 | Aspergillus awamori, microbial inoculum and application thereof |
| CN111154661A (en) * | 2020-01-16 | 2020-05-15 | 中化农业（临沂）研发中心有限公司 | Complex microbial inoculant and application thereof |
| CN111154661B (en) * | 2020-01-16 | 2021-09-21 | 中化农业（临沂）研发中心有限公司 | Complex microbial inoculant and application thereof |
| CN111100798B (en) * | 2020-01-16 | 2021-09-21 | 中化农业（临沂）研发中心有限公司 | Aspergillus awamori, microbial inoculum and application thereof |
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