CN106319005A - Fermentation culture medium capable of enhancing nosiheptide yield and culture method thereof - Google Patents
Fermentation culture medium capable of enhancing nosiheptide yield and culture method thereof Download PDFInfo
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- 239000001963 growth medium Substances 0.000 title claims abstract description 30
- FPTCMHOCGKKRGQ-WYOWUDGCSA-N Multhiomycin Natural products CC=C1NC(=O)[C@@H](NC(=O)c2csc(n2)c3cc(O)c(nc3c4csc(n4)[C@H]5CSC(=O)c6[nH]c7cccc(COC(=O)[C@@H](O)C[C@H](NC(=O)c8csc1n8)c9nc(cs9)C(=O)N5)c7c6C)c%10nc(cs%10)C(=O)N[C@@H](C)C(=O)N)[C@H](C)O FPTCMHOCGKKRGQ-WYOWUDGCSA-N 0.000 title claims abstract description 29
- 101800003864 Nosiheptide Proteins 0.000 title claims abstract description 29
- MQWDKYHFGBWGQZ-JQTJYXGUSA-N nosiheptide Chemical compound N([C@H](C(=O)N\C(C=1SC=C(N=1)C(=O)N[C@@H]1CC(O)C(=O)OCC=2C=CC=C3NC(=C(C3=2)C)C(=O)SC[C@H](NC(=O)C=2N=C1SC=2)C=1SC=C(N=1)C1=N2)=C/C)[C@@H](C)O)C(=O)C(N=3)=CSC=3C1=CC(=O)\C2=C1/NC(C(=O)NC(=C)C(N)=O)=CS1 MQWDKYHFGBWGQZ-JQTJYXGUSA-N 0.000 title claims abstract description 29
- 229950006423 nosiheptide Drugs 0.000 title claims abstract description 29
- 238000000855 fermentation Methods 0.000 title claims abstract description 27
- 230000004151 fermentation Effects 0.000 title claims abstract description 27
- 230000002708 enhancing effect Effects 0.000 title abstract 3
- 238000012136 culture method Methods 0.000 title 1
- 239000000843 powder Substances 0.000 claims abstract description 24
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 20
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000000034 method Methods 0.000 claims abstract description 14
- 239000004382 Amylase Substances 0.000 claims abstract description 13
- 102000013142 Amylases Human genes 0.000 claims abstract description 12
- 108010065511 Amylases Proteins 0.000 claims abstract description 12
- 244000068988 Glycine max Species 0.000 claims abstract description 12
- 235000010469 Glycine max Nutrition 0.000 claims abstract description 12
- 235000019418 amylase Nutrition 0.000 claims abstract description 12
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims abstract description 11
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims abstract description 11
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 10
- 239000008103 glucose Substances 0.000 claims abstract description 10
- 239000011780 sodium chloride Substances 0.000 claims abstract description 10
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 9
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229920002472 Starch Polymers 0.000 claims abstract description 8
- 239000008107 starch Substances 0.000 claims abstract description 8
- 235000019698 starch Nutrition 0.000 claims abstract description 8
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 12
- 241000187762 Streptomyces actuosus Species 0.000 claims description 10
- 229940088417 precipitated calcium carbonate Drugs 0.000 claims description 7
- 239000012043 crude product Substances 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
- 239000002054 inoculum Substances 0.000 claims description 5
- 239000002609 medium Substances 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 4
- 229920002261 Corn starch Polymers 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 3
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 claims description 3
- 239000008120 corn starch Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 150000002333 glycines Chemical class 0.000 claims 1
- 210000000582 semen Anatomy 0.000 claims 1
- 229910000019 calcium carbonate Inorganic materials 0.000 abstract description 2
- 241001655322 Streptomycetales Species 0.000 abstract 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 abstract 1
- 235000011130 ammonium sulphate Nutrition 0.000 abstract 1
- 235000010333 potassium nitrate Nutrition 0.000 abstract 1
- 230000000052 comparative effect Effects 0.000 description 6
- 108090000765 processed proteins & peptides Proteins 0.000 description 6
- 244000005700 microbiome Species 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 2
- 102000004139 alpha-Amylases Human genes 0.000 description 2
- 108090000637 alpha-Amylases Proteins 0.000 description 2
- 229940024171 alpha-amylase Drugs 0.000 description 2
- 239000002027 dichloromethane extract Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 229910052938 sodium sulfate Inorganic materials 0.000 description 2
- 235000011152 sodium sulphate Nutrition 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 235000019730 animal feed additive Nutrition 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- FYHXNYLLNIKZMR-UHFFFAOYSA-N calcium;carbonic acid Chemical compound [Ca].OC(O)=O FYHXNYLLNIKZMR-UHFFFAOYSA-N 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a fermentation culture medium capable of enhancing nosiheptide yield. The fermentation culture medium comprises 0.5-1 g/L KNO3, 0.8-1.5 g/L (NH4)2SO4, 3-5 g/L NaCl, 35-45 g/L soybean powder, 1-2.5 g/L yeast powder, 5-6 g/L glucose, 0.01-0.02 g/L amylase, 40-50 g/L starch, 3.5-5 g/L light calcium carbonate, 0.1-0.5 mg/L serine and the balance of water. The invention also discloses a fermentation method for enhancing nosiheptide yield by inoculating active streptomycete CGMCC No.8732 in the culture medium. The method can obviously enhance the nosiheptide yield, obtains objective economic benefits, and has wide application prospects.
Description
Technical field
The present invention relates to field of microbial fermentation, be specifically related to a kind of nosiheptide yield of improving
Fermentation medium and cultural method.
Background technology
Nahsi peptide is a kind of novel nonabsorbable animal feed additive, adds in animal feed
Little amount just can be greatly improved the dressing percentage of birds.Because it does not remains,
So will not be negatively affected to the mankind.Its mechanism of action is: the synthesis of suppression protein,
The growth making antibacterial is suppressed.At present, the level of nosiheptide fermentation is relatively low, and cost is high, application
Restricted.
Summary of the invention
The first object of the present invention is that the culture medium providing a kind of optimization is to improve nosiheptide
Yield, for achieving the above object, the present invention uses following technological means:
A kind of fermentation medium improving nosiheptide yield, including: KNO3 0.5-1g/L、
(NH4)2S040.8-1.5g/L, NaCl 3-5g/L, analysis for soybean powder 35-45g/L, yeast powder 1-2.5g/L,
Glucose 5-6g/L, amylase 0.01-0.02g/L, starch 40-50g/L, precipitated calcium carbonate 3.5-5
G/L, serine 0.1-0.5mg/L；Prepare with water.
Wherein, the most described fermentation medium includes: KNO3 1g/L、(NH4)2S04 1.5g/L、
NaCl 4g/L, analysis for soybean powder 40g/L, yeast powder 2g/L, glucose 5g/L, amylase 0.01g/L,
Corn starch 50g/L, precipitated calcium carbonate 5g/L, serine 0.2-0.4mg/L, prepare with water.
Wherein, the particle size range of analysis for soybean powder is 60 ± 5 mesh, can be purchased from 5 constant virtues Kang Shengwang grain and oil food
Product processing factory.
Wherein, yeast powder is purchased from Hubei middle peasant Hua Wei biological engineering company limited.
Wherein, amylase can be selected for α-amylase, and this amylase is commercially available in Shandong Long Kete enzyme
Preparation company limited.
Culture medium of the present invention, control ph scope is advisable with 6.5-7, concrete tune
Control is grasped by those skilled in the art, and this is not particularly limited by the present invention.
Tradition can be used for following (the one of most common of which of culture medium composition of fermenting and producing Nahsi peptide
Kind): glucose 4%, analysis for soybean powder 3%, starch 4%, sodium sulfate 0.1%, sodium sulfate 0.1%,
Sodium chloride 0.25%, calcium carbonate 0.5%, element liquid 5ml/L, but this culture medium should in reality
Yield with middle discovery Nahsi peptide is not ideal.The present invention by be conventionally used to fermentation culture that
The culture medium of western peptide carries out formulation optimization (the especially addition of serine), has obtained a kind of energy
Enough significantly improve the new culture medium of nosiheptide yield.
Invention also provides a kind of method utilizing above-mentioned culture medium fermentation to prepare nosiheptide,
Described method comprises the steps:
(1) S.actuosus CGMCC NO.8732 is seeded to described culture medium,；
(2) postvaccinal culture medium is cultivated at 220-250rpm/min, 28-30 DEG C 67-75
Hour, separation and fermentation product, obtain nosiheptide.
The microorganism that the present invention provides is S.actuosus (streptomyces actuosus)
BJX004, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms
The heart (is called for short CGMCC), Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, address 3, China
Institute of microbiology of academy of science, postcode 100101, deposit number CGMCC NO.8732, protects
Hiding January 17 2014 date, the Classification And Nomenclature of this bacterium is: S.actuosus
(streptomyces actuosus)BJX004。
Wherein, described S.actuosus CGMCC NO.8732 preferably carries with seed liquor form
Confession, inoculum concentration be 4-6% (v/v), preferably inoculum concentration be 5% (v/v).
Preferably, described condition of culture is to cultivate at 230-240rpm/min, 28-30 DEG C
72-75 hour.
Wherein, the separation of described tunning is specially and is extracted by fermentation liquid dichloromethane, so
Rear washing dichloromethane extract, washs crude product, drying at room temperature with normal heptane after being condensed into crude product
Obtain fine work.Concrete operation is grasped by those skilled in the art, and this is not made especially by the present invention
Limit.Use above-mentioned separation means, it is possible to further ensure that purity and the yield of product Nahsi peptide.
Using above-mentioned fermentation condition, after fermentation ends, Nahsi peptide yield can reach
Under 5600ug/mL, relatively same culture conditions, (see summary of the invention under aforementioned ordinary culture medium
8 sections) output increased of fermentation culture gained nosiheptide more than 12%.
Detailed description of the invention
In following example, all raw materials used are known commercially available prod, analysis for soybean powder
It is purchased from 5 constant virtues Kang Shengwang oil and foodstuffs processing factory；Yeast powder is purchased from Hubei middle peasant's Hua Wei biological engineering
Company limited；Amylase is α-amylase, is purchased from Shandong Longke Enzyme Co., Ltd..
Embodiment 1
Present embodiments provide a kind of culture medium that can be used for improving nosiheptide yield, described cultivation
The formula of base is as follows:
KNO3 1g/L、(NH4)2S041.5g/L, NaCl 4g/L, analysis for soybean powder 40g/L, yeast
Powder 2g/L, glucose 5g/L, amylase 0.01g/L, corn starch 50g/L and lightweight carbonic acid
Calcium 5g/L, serine 0.4mg/L；Prepare with water.
Embodiment 2
Present embodiments provide a kind of culture medium that can be used for improving nosiheptide yield, described cultivation
The formula of base is as follows:
KNO3 0.5g/L、(NH4)2S040.8g/L, NaCl 3g/L, analysis for soybean powder 35g/L, ferment
Female powder 1g/L, glucose 5g/L, amylase 0.01g/L, starch 40g/L, precipitated calcium carbonate
3.5g/L, serine 0.1mg/L；Prepare with water.
Embodiment 3
Present embodiments provide a kind of culture medium that can be used for improving nosiheptide yield, described cultivation
The formula of base is as follows:
KNO3 0.5g/L、(NH4)2S040.8g/L, NaCl 5g/L, analysis for soybean powder 45g/L, ferment
Female powder 2.5g/L, glucose 6g/L, amylase 0.02g/L, starch 45g/L, precipitated calcium carbonate
4g/L, serine 0.5mg/L；Prepare with water.
Embodiment 4
Present embodiments provide a kind of culture medium that can be used for improving nosiheptide yield, described cultivation
The formula of base is as follows:
KNO3 1g/L、(NH4)2S041.5g/L, NaCl 5g/L, analysis for soybean powder 45g/L, yeast
Powder 2.5g/L, glucose 6g/L, amylase 0.02g/L, starch 50g/L, precipitated calcium carbonate 5
G/L, serine 0.5mg/L；Prepare with water.
Embodiment 5
A kind of method that embodiment improves fermenting and producing nosiheptide, specifically includes following step
Rapid:
(1) seed liquor of S.actuosus CGMCC NO.8732 is seeded to embodiment 1
Prepared culture medium, inoculum concentration is 5% (v/v)；
(2) by postvaccinal culture medium at 220rpm/min, cultivate 72 hours at 28 DEG C,
Separation and fermentation product (fermentation liquid dichloromethane is extracted, then washes dichloromethane extract,
Washing crude product with normal heptane after being condensed into crude product, drying at room temperature obtains fine work) i.e. obtain nosiheptide.
In the present embodiment, the yield of nosiheptide is 5600ug/mL.
Embodiment 6
The present embodiment is also provided that a kind of method of fermenting and producing nosiheptide, with embodiment 5 phase
Ratio, distinctive points is only that the present embodiment uses the preparation-obtained culture medium of embodiment 2, inoculation
Amount is 4% (v/v), and the condition of fermentation culture is at 220rpm/min, cultivates 75 at 30 DEG C
Hour, the yield of nosiheptide is 5450ug/mL.
Embodiment 7
The present embodiment is also provided that a kind of method of fermenting and producing nosiheptide, with embodiment 5 phase
Ratio, distinctive points is only that the present embodiment uses the preparation-obtained culture medium of embodiment 3, inoculation
Amount is 6% (v/v), and the condition of fermentation culture is at 250rpm/min, cultivates 67 at 29 DEG C
Hour, the yield of nosiheptide is 5370ug/mL.
Comparative example 1
Compared with Example 5, differing only in of this comparative example, the cultivation used by fermentation culture
In base, do not add serine.
In this comparative example, the yield of nosiheptide is 5000ug/mL.
Comparative example 2
Compared with Example 6, differing only in of this comparative example, the actual conditions of fermentation culture
For at 180rpm/min, cultivate 72 hours at 25 DEG C.
In this comparative example, the yield of nosiheptide is 5230ug/mL.
Obviously, those skilled in the art the present invention can be carried out various change and modification and not
Depart from the spirit and scope of the present invention.So, if these amendments of the present invention and modification belong to
Within the scope of the claims in the present invention and equivalent technologies thereof, then the present invention is also intended to comprise these
Change and including modification.
Claims (10)
1. the fermentation medium that can improve nosiheptide yield, it is characterised in that: including:
KNO30.5-1g/L、(NH4)2S040.8-1.5g/L, NaCl 3-5g/L, analysis for soybean powder 35-45g/L,
Yeast powder 1-2.5g/L, glucose 5-6g/L, amylase 0.01-0.02g/L, starch 40-50g/L,
Precipitated calcium carbonate 3.5-5g/L, serine 0.1-0.5mg/L；Prepare with water.
Culture medium the most according to claim 1, it is characterised in that: including: KNO31g/L、
(NH4)2S041.5g/L, NaCl 4g/L, analysis for soybean powder 40g/L, yeast powder 2g/L, glucose
5g/L, amylase 0.01g/L, corn starch 50g/L, precipitated calcium carbonate 5g/L, serine
0.2-0.4mg/L, prepares with water.
Culture medium the most according to claim 1 and 2, it is characterised in that: described Semen Glycines
The particle size range of powder is 60 ± 5 mesh.
Culture medium the most according to claim 1 and 2, it is characterised in that: described starch
Enzyme is a-amylase.
5. according to the culture medium described in any one of claim 1-4, it is characterised in that: described
The pH value range of culture medium is 6.5-7.
6. one kind utilizes culture medium fermentation described in any one of claim 1-5 to prepare nosiheptide
Method, it is characterised in that: comprise the steps:
(1) S.actuosus CGMCC NO.8732 is seeded to described culture medium,；
(2) postvaccinal culture medium is cultivated at 220-250rpm/min, 28-30 DEG C 67-75
Hour, separation and fermentation product, obtain nosiheptide.
Method the most according to claim 6, it is characterised in that: described S.actuosus
CGMCC NO.8732 provides with seed liquor form, and inoculum concentration is 4-6% (v/v).
Method the most according to claim 7, it is characterised in that: described S.actuosus
CGMCC NO.8732 provides with seed liquor form, and inoculum concentration is 5% (v/v).
Method the most according to claim 6, it is characterised in that: described condition of culture is
Cultivate 72-75 hour at 230-240rpm/min, 28-30 DEG C.
10. according to the method described in any one of claim 6-9, it is characterised in that: described
The separation of tunning is specific as follows: is extracted by fermentation liquid dichloromethane, then washes dichloro
Methane extracting solution, washs crude product with normal heptane after being condensed into crude product, and drying at room temperature obtains fine work.
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Cited By (1)
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| CN106319004A (en) * | 2015-07-09 | 2017-01-11 | 牡丹江佰佳信生物科技有限公司 | Fermentation culture medium capable of enhancing nosiheptide yield and culture method thereof |
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| CN106319004A (en) * | 2015-07-09 | 2017-01-11 | 牡丹江佰佳信生物科技有限公司 | Fermentation culture medium capable of enhancing nosiheptide yield and culture method thereof |
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Denomination of invention: A fermentation medium and cultivation method that can increase the yield of nasilide Granted publication date: 20201027 Pledgee: Mudanjiang New Area Branch of Longjiang Bank Co.,Ltd. Pledgor: MUDANJIANG BIOSEEN BIOLOGICAL CO.,LTD. Registration number: Y2024230000021 |