CN104419740A - Method for fermenting enramycin - Google Patents
Method for fermenting enramycin Download PDFInfo
- Publication number
- CN104419740A CN104419740A CN201310395254.3A CN201310395254A CN104419740A CN 104419740 A CN104419740 A CN 104419740A CN 201310395254 A CN201310395254 A CN 201310395254A CN 104419740 A CN104419740 A CN 104419740A
- Authority
- CN
- China
- Prior art keywords
- salt
- enramycin
- amino acid
- hour
- fermentation
- 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
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method for fermenting enramycin. According to the method, one or more amino acids, such as arginine, phenylalanine, aspartic acid, threonine and lysine, and salts of the amino acid(s) are added into an enramycin fermentation liquid. The method can be used for greatly increasing the yield of enramycin, and is applicable to industrial production.
Description
Technical field
Technical field of microbial fermentation of the present invention, particularly relates to a kind of fermentation process of enramycin.
Background technology
Microbiotic is from adding in animal-feed for the growth promotion history of existing nearly 60 years so far with lower concentration first.Due to drug residue and drug resistance problems, many conventional antibiotic are limited to use.Enramycin, as a kind of novel peptide type microbiotic, has stable, wide spectrum, noresidue with it, does not produce the advantages such as resistance; With its security in public, as special antibiotics fodder additives, its application receives publicity just day by day.
Enramycin (Enramycin) is fermented by fungicidal streptomycete (Streptomyces fungicidicus) and obtains, have another name called enramycin, enramycin etc., by comprising 13 different types of 17 amino acid moleculars and fatty acid molecule formed.For substrate with different amino acid, peptide synthetase (non-ribosomal peptide synthetase is gathered by non-ribosomal, NRPS) the poly-peptide antibiotics catalyzed and synthesized, its alicyclic ring skeleton comprises the multiple amino acids such as aspartic acid, Threonine, ornithine, citrulline, glycine.Enramycin is different according to its terminal aliphatic acid kind, be divided into enramycin A (C107H138C12N26O31) and enramycin B (C108H140C12N26O31) two components, enramycin is then the mixture be grouped into by this two kinds of one-tenth.
Enramycin was researched and developed in 1966 by Japanese Takede Chemical Industries Ltd, was registered and widely used thereafter in many countries.1993, the Ministry of Agriculture of China ratified this medicine and registers in China.2005, domestic production enterprise and offshore company started joint production enramycin premix.Along with other feeding antibiotics are progressively eliminated, the use of enramycin will certainly be more and more extensive, but the industrial production of enramycin faces the low problem of capacity and output always.
Summary of the invention
In order to solve the technical problem that in prior art, the fermentation yield of enramycin is low, the present invention proposes a kind of method improving enramycin fermentation yield.
The present invention relates to a kind of method improving enramycin fermentation yield, be included in the enramycin fermented liquid that fermentation mid-early stage cultivates to ferment at constant temperature and add one or more amino acid and/or its salt.
Wherein, described amino acid and/or its salt are selected from arginine, phenylalanine, aspartic acid, Threonine, Methionin and/or its salt.
Wherein, addition is respectively by percentage to the quality: arginine/or its salt 0.01-0.1%, phenylalanine/or its salt 0.01-0.1%, aspartic acid/or its salt 0.005-0.05%, Threonine/or its salt 0.005-0.05%, Methionin/or its salt 0.01-0.3%.
Wherein, the culture temperature that described ferment at constant temperature is cultivated is 25-30 DEG C; Medium pH is 6.5-7.5.
Wherein, the described time of adding amino acid and/or amino acid salts in substratum, for starting to ferment in 0-120 hour, in being preferably 0-64 hour, is more preferably in 0-24 hour.
Wherein, the substratum that described enramycin fermentation uses is Semen Maydis powder 7-12%, maltodextrin 8-16%, seitan powder 3-6%, corn steep liquor 0.5-2.0%, soya-bean oil 1-3%, calcium carbonate 0.5-1.0% by percentage to the quality.
Wherein, said method comprising the steps of: slant pore is cultivated: by aseptic for fungicidal streptomycete access slant pore substratum, temperature 25-30 DEG C, relative humidity 40-60% constant temperature culture 8-12 days; Seed and fermentation culture: spore suspension is accessed seed culture medium, cultivate 40-60 hour, then aseptic access fermention medium for 25-30 DEG C, inoculum size counts 5-15% with volume percent, cultivates 180-240 hour for 25-30 DEG C; In fermentation period, use ammoniacal liquor to regulate fermentation pH6.5-7.5.
The present invention, by certain fermentation period, adds one or several amino acid (or amino acid salts), makes the biosynthesizing of enramycin have abundanter, more balanced substrate combination, thus significantly improve the fermentation yield of enramycin.The present invention is applicable to suitability for industrialized production, has very high practical value and meaning.
Embodiment
Below in conjunction with embodiment, the invention will be further described.
Following examples miospore is cultivated and seed culture all adopts unified training method, temperature error≤0.5 DEG C, relative humidity deviation≤10%, pH error≤0.5.
Slant pore is cultivated: by fungicidal streptomycete (Streptomyces fungicidicus), (biotechnology company limited is opened in Zhengzhou one hundred, enramycin produces fungus strain Streptomyces fungicidious mutant strain, article number: BKSFJ06) aseptic access slant pore substratum, temperature 28 DEG C, relative humidity 40-60% constant temperature culture 10 days; Slant pore substratum comprises composition: W-Gum 50g/L, maltodextrin 30g/L, peptone 8g/L, fish peptone 6g/L, agar 20g/L.
Seed fermentation is cultivated: will containing the 3x10 that has an appointment
10the spore suspension aseptic access 10L seed culture medium of individual spore, cultivates 48 hours for 28 DEG C; Seed culture medium comprises composition: W-Gum 50g/L, maltodextrin 30g/L, corn steep liquor 15g/L, seitan powder 10g/L, calcium carbonate 6g/L, pH7.2.
Other parameter of each embodiment:
Embodiment 1: by aseptic for cultured seed liquor access 60L fermention medium, inoculum size 6L, cultivate 180 hours for 25 DEG C, fermenting process ammoniacal liquor regulates pH6.5; The composition that fermention medium comprises by percentage to the quality Semen Maydis powder 7%, maltodextrin 12%, seitan powder 4%, corn steep liquor 1.0%, soya-bean oil 1%, calcium carbonate 1.0%, ammoniacal liquor regulates pH7.5.
Embodiment 2: by aseptic for cultured seed liquor access 50L fermention medium, inoculum size 7L, cultivate 180 hours for 25 DEG C, fermenting process ammoniacal liquor regulates pH6.5; The composition that fermention medium comprises by percentage to the quality Semen Maydis powder 10%, maltodextrin 8%, seitan powder 4%, corn steep liquor 1.5%, soya-bean oil 2%, calcium carbonate 1.0%, ammoniacal liquor regulates pH7.2.
Embodiment 3: by aseptic for cultured seed liquor access 50L fermention medium, inoculum size 4L, cultivate 200 hours for 28 DEG C, fermenting process ammoniacal liquor regulates pH6.8; The composition that fermention medium comprises by percentage to the quality Semen Maydis powder 10%, maltodextrin 12%, seitan powder 6%, corn steep liquor 1.2%, soya-bean oil 3%, calcium carbonate 0.8%, ammoniacal liquor regulates pH7.0.
Embodiment 4: by aseptic for cultured seed liquor access 60L fermention medium, inoculum size 7L, cultivate 220 hours for 28 DEG C, fermenting process ammoniacal liquor regulates pH7.0; The composition that fermention medium comprises by percentage to the quality Semen Maydis powder 12%, maltodextrin 12%, seitan powder 4%, corn steep liquor 0.5%, soya-bean oil 2%, calcium carbonate 1.0%, ammoniacal liquor regulates pH6.8.
Embodiment 5: by aseptic for cultured seed liquor access 80L fermention medium, inoculum size 4L, cultivate 220 hours for 28 DEG C, fermenting process ammoniacal liquor regulates pH7.0; The composition that fermention medium comprises by percentage to the quality Semen Maydis powder 10%, maltodextrin 14%, seitan powder 5%, corn steep liquor 2.0%, soya-bean oil 2%, calcium carbonate 1.0%, ammoniacal liquor regulates pH6.5.
Embodiment 6: by aseptic for cultured seed liquor access 60L fermention medium, inoculum size 6L, cultivate 240 hours for 30 DEG C, fermenting process ammoniacal liquor regulates pH7.5; The composition that fermention medium comprises by percentage to the quality Semen Maydis powder 8%, maltodextrin 16%, seitan powder 3%, corn steep liquor 1.5%, soya-bean oil 2%, calcium carbonate 1.0%, ammoniacal liquor regulates pH6.5.
The present invention adopts HPLC external standard method to the detection of the final enramycin content of fermented liquid: get 60g enramycin fermented liquid, add 120ml methyl alcohol, with 1M salt acid for adjusting pH to 3.0-4.5, place jolting 2-4 hour on shaking table, shaking speed 220-230rpm, 2000 leave the heart gets supernatant sample detection in 20 minutes.Adopt HPLC method, use Agilent company hplc device, adopt Gemini C18 post (4.6x150mm, 5mm; Phenomenex), with acetonitrile (chromatographic grade): 50mM phosphate sodium dihydrogen buffer solution=30:70, flow velocity 1mL/min, detect at 267nm place, column temperature 35 DEG C, enramycin content in quantitative analysis sample, sample size 10 μ L.
Embodiment 1
Fermentation within the 0th hour, 24 hours, 64 hours, 120 hours, be added to respectively 0.01%, 0.05%, 0.1% arginine or its salt (account for total fermented liquid mass percent, lower same), control group does not add any amino acid, survey the enramycin content of each group of fermented liquid when having fermented respectively, result is as shown in table 1:
Table 1 different time adds the arginine of different concns or the enramycin content (g/L) of its salt secondary fermentation liquid
Embodiment 2
Phenylalanine or its salt of 0.01%, 0.05%, 0.1% within 0th hour, 24 hours, 64 hours, 120 hours, is added respectively in fermentation, control group does not add any amino acid, survey the enramycin content of each group of fermented liquid when having fermented respectively, result is as shown in table 2:
Table 2 different time adds the phenylalanine of different concns or the enramycin content (g/L) of its salt secondary fermentation liquid
Embodiment 3
Fermentation 0 hour, within 24 hours, 64 hours, 120 hours, be added to 0.005%, 0.01%, 0.05% aspartic acid respectively, control group does not add any amino acid, and survey the enramycin content of each group of fermented liquid when having fermented respectively, result is as shown in table 3:
Table 3 different time adds the enramycin content (g/L) of the aspartic acid secondary fermentation liquid of different concns
Embodiment 4
Fermentation 0 hour, within 24 hours, 64 hours, 120 hours, be added to respectively 0.005%, 0.02%, 0.05% Threonine or its salt, control group does not add any amino acid, survey the enramycin content of each group of fermented liquid when having fermented respectively, result is as shown in table 4:
Table 4 different time adds the Threonine of different concns or the enramycin content (g/L) of its salt or its salt secondary fermentation liquid
Embodiment 5
Fermentation 0 hour, within 24 hours, 64 hours, 120 hours, be added to respectively 0.01%, 0.1%, 0.3% Methionin or its salt, control group does not add any amino acid, survey the enramycin content of each group of fermented liquid when having fermented respectively, result is as shown in table 5:
Table 5 different time adds the Methionin of different concns or the enramycin content (g/L) of its salt secondary fermentation liquid
Embodiment 6
0.03% Methionin, 0.03% phenylalanine, 0.01 noisy aspartic acid, 0.01% Threonine is added in 0 hour-120 hours respectively in fermentation, control group does not add any amino acid, survey the enramycin content of name group fermented liquid when having fermented respectively, result is as shown in table 6:
Table 6 different time adds the enramycin content (g/L) of multiple propylhomoserin secondary fermentation liquid
Above example illustrates, fermentation is after mid-early stage, different time added one or more amino acid, and in fermented liquid, the content of the enramycin of final gained is all significantly tested higher than control group.
It should be noted that above embodiment only in order to the present invention to be described and unrestricted, the present invention is also not limited in above-mentioned citing, and all do not depart from technical scheme and the improvement thereof of the spirit and scope of the present invention, and it all should be encompassed in right of the present invention.
Claims (9)
1. an enramycin fermentation process, is characterized in that, in the enramycin substratum that ferment at constant temperature is cultivated, add one or more amino acid and/or its salt.
2. the method for claim 1, is characterized in that, described amino acid and/or its salt are: arginine, phenylalanine, aspartic acid, Threonine, Methionin and/or its salt.
3. method as claimed in claim 2, it is characterized in that, by percentage to the quality described, the amount of adding is respectively arginine/or its salt 0.01-0.1%, phenylalanine/or its salt 0.01-0.1%, aspartic acid/or its salt 0.005-0.05%, Threonine/or its salt 0.005-0.05%, Methionin/or its salt 0.01-0.3%.
4. the method for claim 1, is characterized in that, the culture temperature of described ferment at constant temperature is 25-30 DEG C, and medium pH is 6.5-7.5.
5. the method as described in any one of claim 1-4, is characterized in that, the described time of adding amino acid and/or its salt in substratum ferments in 0-120 hour for starting.
6. method as claimed in claim 5, it is characterized in that, the described time of adding amino acid and/or its salt in substratum ferments in 0-64 hour for starting.
7. method as claimed in claim 6, it is characterized in that, the described time of adding amino acid and/or its salt in substratum ferments in 0-24 hour for starting.
8. the method as described in any one of claim 1-4, it is characterized in that, the substratum that described enramycin fermentation uses is Semen Maydis powder 7-12%, maltodextrin 8-16%, seitan powder 3-6%, corn steep liquor 0.5-2.0%, soya-bean oil 1-3%, calcium carbonate 0.5-1.0% by percentage to the quality.
9. method as claimed in claim 8, is characterized in that comprising the following steps: slant pore is cultivated: by aseptic for fungicidal streptomycete access slant pore substratum, temperature 25-30 DEG C, relative humidity 40-60% constant temperature culture 8-12 days;
Seed and fermentation culture: spore suspension is accessed seed culture medium, cultivate 40-60 hour, then aseptic access fermention medium for 25-30 DEG C, inoculum size counts 5-15% with volume percent, cultivates 180-240 hour for 25-30 DEG C;
In fermentation period 0-120 hour, fill into amino acid and/or its salt, use ammoniacal liquor to regulate fermentation pH6.5-7.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310395254.3A CN104419740A (en) | 2013-09-03 | 2013-09-03 | Method for fermenting enramycin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310395254.3A CN104419740A (en) | 2013-09-03 | 2013-09-03 | Method for fermenting enramycin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104419740A true CN104419740A (en) | 2015-03-18 |
Family
ID=52969864
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310395254.3A Pending CN104419740A (en) | 2013-09-03 | 2013-09-03 | Method for fermenting enramycin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104419740A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105506040A (en) * | 2015-12-23 | 2016-04-20 | 安徽丰原发酵技术工程研究有限公司 | Method for fermentation production of enramycin |
| CN105779535A (en) * | 2016-05-16 | 2016-07-20 | 新疆天富阳光生物科技有限公司 | Culture medium for fermenting and producing enramycin and fermentation method |
| CN110157757A (en) * | 2019-02-15 | 2019-08-23 | 河北圣雪大成制药有限责任公司 | Output increased method, culture medium and the separation method of enramycin one pack system |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101899490A (en) * | 2010-07-14 | 2010-12-01 | 山东胜利股份有限公司 | Method for producing enramycin by using microbial fermentation |
| CN102250993A (en) * | 2011-07-18 | 2011-11-23 | 南京工业大学 | Optimized process for producing enramycin through fermenting streptomyces |
-
2013
- 2013-09-03 CN CN201310395254.3A patent/CN104419740A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101899490A (en) * | 2010-07-14 | 2010-12-01 | 山东胜利股份有限公司 | Method for producing enramycin by using microbial fermentation |
| CN102250993A (en) * | 2011-07-18 | 2011-11-23 | 南京工业大学 | Optimized process for producing enramycin through fermenting streptomyces |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105506040A (en) * | 2015-12-23 | 2016-04-20 | 安徽丰原发酵技术工程研究有限公司 | Method for fermentation production of enramycin |
| CN105506040B (en) * | 2015-12-23 | 2019-02-12 | 安徽丰原发酵技术工程研究有限公司 | A kind of method of producing enramycin by fermentation |
| CN105779535A (en) * | 2016-05-16 | 2016-07-20 | 新疆天富阳光生物科技有限公司 | Culture medium for fermenting and producing enramycin and fermentation method |
| CN110157757A (en) * | 2019-02-15 | 2019-08-23 | 河北圣雪大成制药有限责任公司 | Output increased method, culture medium and the separation method of enramycin one pack system |
| CN110157757B (en) * | 2019-02-15 | 2022-11-25 | 河北圣雪大成制药有限责任公司 | Yield improving method of enramycin monocomponent, culture medium and separation method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103146606B (en) | Microbial agent for high-efficiency straw fermentation | |
| CN101597578B (en) | Enramycin producing strain and extraction method by using macroporous resin | |
| CN104342390B (en) | A kind of Sinorhizobium meliloti strain and combinations thereof and application | |
| CN105670976B (en) | Fermentation medium of bacillus amyloliquefaciens and application thereof | |
| CN103030441B (en) | Novel pest-resistant liquid compound microbial fertilizer | |
| CN103194410B (en) | Paenibacillus mucilaginosus and method for producing compound microorganism bacterium agent by utilizing same | |
| CN103614323B (en) | A kind of substratum of bacillus amyloliquefaciens and application | |
| CN102936608B (en) | Method for producing avilamycin by fermenting | |
| CN104419740A (en) | Method for fermenting enramycin | |
| WO2015001575A4 (en) | Plant growth promoting formulation of piriformospora indica and azotobacter chroococcum with talcum powder | |
| CN104131054B (en) | Fermentation culture medium and fermentation method for improving enramycin yield | |
| CN103131652B (en) | Rhizobium japonicum culture medium and method for preparing liquid rhizobium japonicum agent by adopting rhizobium japonicum culture medium | |
| CN103205479A (en) | Culture medium used for producing echinocandin B | |
| Li et al. | Establishment of beet molasses as the fermentation substrate for industrial vitamin B12 production by Pseudomonas denitrificans | |
| CN102154419A (en) | Fermentation medium for improving enramycin yield | |
| CN101921250A (en) | Bisabolane sesquiterpene derivative and preparation method and application thereof | |
| CN103540534B (en) | The industrial method for culturing of high-protein desert algae | |
| CN102618604B (en) | Preparation method of cyclic lipopeptide compound | |
| CN101880703A (en) | Method for fermenting daptomycin by adding caprate | |
| CN102559522B (en) | Bread yeast with high nucleic acid and preparation method thereof | |
| Ming et al. | The solid fermentation state’s optimization of Trichoderma Harzianum M1 | |
| CN104404112A (en) | Method for producing avilamycin by material supplementation in microbial fermentation process | |
| CN109161507A (en) | A kind of Corynebacterium glutamicum of high yield L-Orn and its application | |
| CN103103240A (en) | Strain culture method capable of improving yield of nosiheptide | |
| CN103642739A (en) | Streptomces aureofaciens for producing keratinase and application method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150318 |
|
| RJ01 | Rejection of invention patent application after publication |