CN109796520A - A kind of extracting method of Nosiheptide fine work - Google Patents
A kind of extracting method of Nosiheptide fine work Download PDFInfo
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Abstract
The present invention provides a kind of extracting methods of Nosiheptide fine work, belong to purification technique field.The present invention filters after first mixing Nosiheptide fermentation liquid and filter aid, obtains Nosiheptide filter cake；Then it washed, be granulated and dried, and Nosiheptide dry granular is extracted using methylene chloride, leaching liquor is subjected to de- carbon, concentration and crystallization and obtains nosipeptide crude product, added ethyl alcohol mashing, obtain Nosiheptide fine work after filtration drying.Extracting method provided by the invention can effectively extract Nosiheptide from fermentation liquid, and remove remaining culture medium residual, water and other impurities in fermentation liquid, improve the yield and purity of product, it reduces in product use process since there are issuable toxic reactions for other impurities, improves the safety of medication.Embodiment the result shows that, the Nosiheptide fine work yield obtained using extracting method of the present invention is up to 80.7%, and content is up to 897u/mg.
Description
Technical field
The present invention relates to purification technique field, in particular to a kind of extracting method of Nosiheptide fine work.
Background technique
Nosiheptide (Nosiheptide) contains element sulphur abundant, belongs to polypeptide antibiotics, is mainly produced by S.actuosus
It is raw；Nosiheptide has good inhibiting effect to gram-positive bacteria, by answering with what 23S RNA and ribosomal protein L 11 were formed
Zoarium is combined closely, to inhibit elongation factor activity and the hydrolysis of GTP, is finally inhibited the synthesis of protein, is made bacterium
Growth is suppressed.Nosiheptide is since solubility is low in enteron aisle for it, it is not easy to be absorbed, thus can be protected in enteron aisle by animal
Hold higher drug concentration, it has the characteristics that dosage is low, scope of restraining fungi is wide, does not remain in animal body, can be obviously promoted chicken,
The growth of the animals such as pig, fish, therefore be a kind of ideal novel nonabsorbable animal feed additive.
Nosiheptide is obtained by biological fermentation process, and soya-bean oil, beancake powder, starch and ammonium sulfate, sodium sulphate are specifically utilized
Equal grains chemical raw material, the cometabolism effect by producing bacterium is synthesized into, after the completion of metabolism, since Nosiheptide is insoluble
Yu Shui is primarily present in the mycelia of production bacterium, is mixed, can not be directly obtained with mycelium, remaining culture medium etc.
Pure Nosiheptide product.
Currently, majority is directly used using the treatment fluid after fermenting as Nosiheptide product in industry, but there is training in this product
Base residual, water and other possible impurity are supported, so that Nosiheptide purity is lower.And may to generate toxicity anti-for excessive impurity
It answers, so that the safety of medication substantially reduces.
Summary of the invention
In view of this, it is an object of that present invention to provide a kind of preparation methods of Nosiheptide fine work.Use extraction of the invention
The yield and purity is high for the Nosiheptide fine work that method obtains, medication it is highly-safe.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides a kind of extracting methods of Nosiheptide fine work, comprising the following steps:
(1) it is filtered after mixing Nosiheptide fermentation liquid and filter aid, obtains Nosiheptide filter cake；
(2) the Nosiheptide filter cake is successively washed, is granulated and dried, obtain Nosiheptide dry granular；
(3) the Nosiheptide dry granular is extracted using methylene chloride, obtains leaching liquor；
(4) de- carbon, concentration and crystallization are successively carried out to the leaching liquor, obtain nosipeptide crude product；
(5) it is successively beaten, filtered and dried after mixing the nosipeptide crude product and ethyl alcohol, obtain Nosiheptide essence
Product.
Preferably, the chemical titer of Nosiheptide fermentation liquid is 2000~2500u/mL in the step (1).
Preferably, the filter aid in the step (1) is perlite, and the dosage of the filter aid is the 3 of fermentating liquid volume
~5%.
Preferably, in the step (2) Nosiheptide dry granular partial size≤1.2mm.
Preferably, the volume ratio of methylene chloride and Nosiheptide dry granular is 5~10:1 in the step (3).
Preferably, the time extracted in the step (3) is 8~12h.
Preferably, it is active carbon that the de- carbon of carbon, which takes off agent, in the step (4), and the dosage that the carbon takes off agent is the extraction
The 1~2% of liquid product.
Preferably, the method being concentrated in the step (4) are as follows:
The de- gained carbon of carbon is taken off into liquid and is concentrated into the 10~20% of original volume under vacuum conditions, obtains Nosiheptide concentrate；
The temperature of the concentration is 55~65 DEG C, vacuum degree >=0.03MPa of the concentration.
Preferably, the method crystallized in the step (4) are as follows:
Water is added into Nosiheptide concentrate, is crystallized, Nosiheptide crystal is obtained；
The temperature of the crystallization is 4 DEG C, and the volume ratio of the water and Nosiheptide concentrate is 0.5~1:1.
Preferably, the volume ratio of ethyl alcohol and nosipeptide crude product is 5~6:1 in the step (5).
The present invention provides a kind of extracting method of Nosiheptide fine work, the present invention first mixes Nosiheptide fermentation liquid and filter aid
It is filtered after conjunction, obtains Nosiheptide filter cake；Then washed, be granulated and dried, and using methylene chloride to Nosiheptide dry granular into
Leaching liquor is carried out de- carbon, concentration and crystallization and obtains nosipeptide crude product by row extraction, adds ethyl alcohol mashing, after filtration drying
To Nosiheptide fine work.Extracting method provided by the invention can effectively extract Nosiheptide from fermentation liquid, and remove
Remaining culture medium residual, water and other impurities, improve the yield and purity of product, reduce in product use process in fermentation liquid
Since there are issuable toxic reactions for other impurities, the safety of medication is improved.Embodiment the result shows that, use this hair
The Nosiheptide fine work yield that bright extracting method obtains is up to 80.7%, and content is up to 897u/mg.
Specific embodiment
The present invention provides a kind of extracting methods of Nosiheptide fine work, comprising the following steps:
(1) it is filtered after mixing Nosiheptide fermentation liquid and filter aid, obtains Nosiheptide filter cake；
(2) the Nosiheptide filter cake is successively washed, is granulated and dried, obtain Nosiheptide dry granular；
(3) the Nosiheptide dry granular is extracted using methylene chloride, obtains leaching liquor；
(4) de- carbon, concentration and crystallization are successively carried out to the leaching liquor, obtain nosipeptide crude product；
(5) it is successively beaten, filtered and dried after mixing the nosipeptide crude product and ethyl alcohol, obtain Nosiheptide essence
Product.
The present invention filters after mixing Nosiheptide fermentation liquid and filter aid, obtains Nosiheptide filter cake.In the present invention, described
The chemical titer of Nosiheptide fermentation liquid is preferably 2000~2500u/mL, more preferably 2200~2400u/mL；The present invention is to institute
The source for stating Nosiheptide fermentation liquid does not have special requirement, the Nosiheptide fermentation using this field conventional commercial or voluntarily prepared
Liquid.In the present invention, the filter aid is preferably perlite, and the dosage of the filter aid is preferably the 3 of fermentating liquid volume
~5%, more preferably 4%.
The present invention preferably will Nosiheptide fermentation liquid and filter aid mix after heat, then again to Nosiheptide fermentation liquid and
The hot mixing liquid of filter aid is filtered.In the present invention, the temperature of the heating is preferably 60~65 DEG C, more preferably 62~
64℃；Time of the heating preferably≤3h, more preferably 1~2h；The present invention reduces Nosiheptide fermentation liquid by heating
Viscosity improves the filter effect of Nosiheptide fermentation liquid.In the present invention, the mode of the filtering is preferably plate compression, described
The pressure of plate compression is preferably 0.4~0.5MPa, more preferably 0.45MPa.Nosiheptide fermentation is removed by filtration in the present invention
Moisture in liquid.
After obtaining Nosiheptide filter cake, the present invention is successively washed, is granulated and dried to the Nosiheptide filter cake, obtains that
Western peptide dry granular.In the present invention, the volume of the water scouring water is preferably 5~10 times of the Nosiheptide filter cake volume, more excellent
It is selected as 6~8 times；The water scouring water is preferably deionized water.The present invention does not have special requirement to the mode of the washing, makes
With water-washing method well known to those skilled in the art, in embodiments of the present invention, preferably by the Nosiheptide filter cake and water
In investment pretreatment tank, is washed, after the completion of washing, filtered out water using plate compression under stirring conditions；Institute
The pressure for stating plate compression is preferably 0.4~0.5MPa, more preferably 0.45MPa.The present invention can be removed described by washing
Water-solubility impurity in Nosiheptide filter cake.
After the completion of washing, the present invention is successively granulated and dries to the filter cake after washing, obtains Nosiheptide dry granular.This hair
Bright it is preferable to use pelletizers to be granulated to the Nosiheptide filter cake, and the sieve diameter of the pelletizer is preferably 1.2mm.This hair
Bright to be dried it is preferable to use fluidizing fluid-bed, the fluidizing fluid-bed inlet air temperature is preferably 100~120 DEG C, more preferably
It is 105~115 DEG C；The fluidizing fluid-bed leaving air temp is preferably 35~60 DEG C, and more preferably 40~50 DEG C.In the present invention
In, the partial size of the Nosiheptide dry granular preferably≤1.2mm, more preferably≤1.0mm.
After obtaining Nosiheptide dry granular, the present invention extracts the Nosiheptide dry granular using methylene chloride, is extracted
Liquid.In the present invention, the volume ratio of the methylene chloride and Nosiheptide dry granular is preferably 5~10:1, more preferably 5:1；It is described
The time of extraction is preferably 8~12h, more preferably 10~11h.The present invention is transferred to Nosiheptide in leaching liquor by extraction.
After obtaining leaching liquor, the present invention successively carries out de- carbon, concentration and crystallization to the leaching liquor, and it is thick to obtain Nosiheptide
Product.In the present invention, it is preferably active carbon that the de- carbon of the carbon, which takes off agent, and the dosage that the carbon takes off agent is preferably the leaching liquor
The 1~2% of volume, more preferably 1.4~1.8%.The present invention removed by the de- effect of carbon that carbon takes off agent pigment in leaching liquor,
Remain the impurity such as fermentation material oil.
In the present invention, the method for the concentration is preferred are as follows: the de- gained carbon of carbon is taken off liquid and is concentrated into original under vacuum conditions
The 10~20% of volume obtain Nosiheptide concentrate.
The present invention carbon is more preferably taken off that liquid is concentrated into original volume 15~18%；In the present invention, the temperature of the concentration
Preferably 55~65 DEG C, more preferably 60~62 DEG C；The vacuum degree of the concentration preferably >=0.03MPa, more preferably >=
0.05MPa。
In the present invention, the method for the crystallization is preferred are as follows: water is added into Nosiheptide concentrate, is crystallized, obtains
Nosiheptide crystal.
In the present invention, the temperature of the crystallization is preferably 0~10 DEG C, and more preferably 4 DEG C, the water and Nosiheptide are concentrated
The volume ratio of liquid is preferably 0.5~1:1, more preferably 0.5:1；The present invention preferably crystallizes under stirring conditions.
After the completion of crystallization, the present invention is preferably successively centrifuged and dries to gained crystallized product, obtains nosipeptide crude product；
The present invention does not have particular/special requirement to the mode of the centrifugation, uses centrifugation well known to those skilled in the art；This hair
Bright preferably to be dried under vacuum conditions, the temperature of the drying is preferably 60 DEG C, and the time is preferably 2~2.5h, more preferably
For 2.2~2.4h.
After obtaining nosipeptide crude product, the present invention is successively beaten after mixing the nosipeptide crude product and ethyl alcohol, filters
And drying, obtain Nosiheptide fine work.In the present invention, the volume ratio of the ethyl alcohol and nosipeptide crude product is preferably 5~6:1, more
Preferably 5.5:1；Present invention preferably uses the modes of stirring to be beaten, and the time of the stirring is preferably 6~8h, more preferably
For 6.5~7.5h.The present invention can be such that crude product and ethyl alcohol is sufficiently mixed by mashing, and then remove undesired impurities.
In the present invention, the mode of the filtering is preferably plate compression, and the pressure of the plate compression is preferably 0.4~
0.5MPa, more preferably 0.45MPa.The present invention is preferably dried under vacuum conditions, and the temperature of the drying is preferably 60
DEG C, the time is preferably 2~2.5h, more preferably 2.2~2.4h.
The extracting method of Nosiheptide fine work provided by the invention is described in detail in conjunction with the embodiments below, still
They cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
A kind of extracting method of Nosiheptide fine work, steps are as follows:
(1) it filters: collecting 10 tons of 2000u/mL Nosiheptide fermentation liquid, the perlite 300kg of fermentating liquid volume 3% is added,
60 DEG C are warming up to, with plate compression, the moisture in fermentation liquid is removed, the filter cake 500kg containing Nosiheptide is obtained；
(2) it washes: in the Nosiheptide filter cake investment pretreatment tank that step (1) is obtained, 5 times of filter cake volume of water is added,
It stirs evenly, again plate compression, obtains Nosiheptide purifying filter cake；
(3) it is granulated: obtaining purifying Nosiheptide filter cake using step (2), be granulated with pelletizer, sieve diameter 1.2mm is obtained
Nosiheptide particle；
(4) dry: the Nosiheptide particle that step (3) is obtained uses fluidizing fluid-bed drying, controls inlet air temperature 100
DEG C, 40 DEG C of leaving air temp, obtain Nosiheptide dry granular；
(5) extract: it is dry to add Nosiheptide for 2.5 tons of methylene chloride of first 5 times of the Nosiheptide dry granular volume into pot for solvent extraction
Grain, extracts 12h, and filtering takes leaching liquor；
(6) carbon is de-: isometric active carbon 25kg being added in leaching liquor, stirs 1h, filters to obtain the de- liquid of carbon；
(7) it is concentrated: the carbon being obtained by filtration being taken off into liquid, is concentrated into original volume under conditions of 60 DEG C, vacuum degree 0.06MPa
10%, stop concentration；
(8) it crystallizes: under 4 DEG C of stirring conditions, in the ratio that purified water/concentrate volume ratio is 0.5:1 into concentrate
125 liters of purified water are added, the crystallization of Nosiheptide is obtained, centrifugation separates Nosiheptide crystallization from liquid, obtains Nosiheptide
Crude product 21.25kg；
(9) dry: the dry 2h under 60 DEG C of vacuum conditions obtains nosipeptide crude product 17.6kg；
(10) it is beaten: being added 88 liters of ethyl alcohol of 5 times of nosipeptide crude product volume, stir 6h；
(11) it filters: plate compression is carried out to the nosipeptide crude product after mashing；
(12) dry: the dry 2h under 60 DEG C of vacuum conditions obtains Nosiheptide fine work 17kg.
Through detecting, the yield 76.0% of gained Nosiheptide fine work, content 895u/mg.
Embodiment 2
A kind of extracting method of Nosiheptide fine work, steps are as follows:
(1) it filters: collecting 5 tons of 2500u/mL Nosiheptide fermentation liquid, the perlite 150kg of fermentating liquid volume 3% is added, rise
Temperature, with plate-frame filtering, the moisture in fermentation liquid is removed, the filter cake 250kg containing Nosiheptide is obtained to 65 DEG C；
(2) it washes: in the Nosiheptide filter cake investment pretreatment tank that step (1) is obtained, 8 times of filter cake volume of water is added,
It stirs evenly, again plate-frame filtering, obtains Nosiheptide purifying filter cake；
(3) it is granulated: obtaining Nosiheptide purifying filter cake using step (2), be granulated with pelletizer, sieve diameter 1.2mm is obtained
Nosiheptide particle；
(4) dry: the Nosiheptide particle that step (3) is obtained uses fluidizing fluid-bed drying, controls inlet air temperature 110
DEG C, 55 DEG C of leaving air temp, obtain Nosiheptide dry granular；
(5) extract: it is dry to add Nosiheptide for 1.25 tons of methylene chloride of first 10 times of the Nosiheptide dry granular volume into pot for solvent extraction
Grain, extracts 10h, and filtering takes leaching liquor；
(6) carbon is de-: the active carbon 12.5kg of 1.5 times of volumes of leaching liquor is added, stirs 1h, filters to obtain the de- liquid of carbon；
(7) it is concentrated: the carbon being obtained by filtration being taken off into liquid and is concentrated into original volume under the conditions of 55 DEG C, vacuum degree 0.05MPa
15%, stop concentration；
(8) it crystallizes: under 4 DEG C of stirrings, adding in purified water/concentrate volume ratio 0.8:1 ratio into concentrate
Enter purified water 65L, obtain the crystallization of Nosiheptide, centrifugation separates Nosiheptide crystallization from liquid, obtains nosipeptide crude product
10kg；
(9) dry: dry 2h obtains the dry crude product 8.2kg of Nosiheptide under 60 DEG C of vacuum conditions；
(10) it is beaten: 5.5 times of nosipeptide crude product volume of ethyl alcohol 60L is added, stir 8h；
(11) it filters: plate compression is carried out to the nosipeptide crude product after mashing；
(12) dry: the dry 2h under 60 DEG C of vacuum conditions obtains Nosiheptide fine work 8kg.
Through detecting, the yield of gained Nosiheptide fine work is 78.6%, content 892u/mg.
Embodiment 3
A kind of extracting method of Nosiheptide fine work, steps are as follows:
(1) it filters: collecting 20 tons of 2100u/mL Nosiheptide fermentation liquid, the perlite of fermentating liquid volume 5% is added, is warming up to
65 DEG C, with plate-frame filtering, the moisture in fermentation liquid is removed, 1.0 tons of filter cake containing Nosiheptide are obtained；
(2) it washes: in the Nosiheptide filter cake investment pretreatment tank that step (1) is obtained, 10 times of filter cake volume of water is added,
It stirs evenly, again plate-frame filtering, obtains Nosiheptide purifying filter cake；
(3) it is granulated: obtaining Nosiheptide purifying filter cake using step (2), be granulated with pelletizer, sieve diameter 1.2mm is obtained
Nosiheptide particle；
(4) dry: the Nosiheptide particle that step (3) is obtained uses fluidizing fluid-bed drying, controls inlet air temperature 120
DEG C, 60 DEG C of leaving air temp, obtain Nosiheptide dry granular；
(5) extract: 5 tons of methylene chloride of first 8 times of the Nosiheptide dry granular volume into pot for solvent extraction add Nosiheptide dry granular,
12h is extracted, filtering takes leaching liquor；
(6) carbon is de-: the active carbon 50kg of 2 times of volumes of leaching liquor is added, stirs 1h, filters to obtain the de- liquid of carbon；
(7) it is concentrated: the carbon being obtained by filtration being taken off into liquid and is concentrated into original volume under the conditions of 65 DEG C, vacuum degree 0.03MPa
20%, stop concentration；
(8) it crystallizes: under 4 DEG C of stirrings, being added in purified water/concentrate volume ratio 1:1 ratio into concentrate
Purified water 250L obtains the crystallization of Nosiheptide, and centrifugation separates Nosiheptide crystallization from liquid, obtains nosipeptide crude product
45.5kg；
(9) dry: the dry 2h under 60 DEG C of vacuum conditions obtains the dry crude product 37.2kg of Nosiheptide；
(10) it is beaten: 6 times of nosipeptide crude product volume of ethyl alcohol 186L is added, stir 7h；
(11) it filters: plate compression is carried out to the nosipeptide crude product after mashing；
(12) dry: the dry 2h under 60 DEG C of vacuum conditions obtains Nosiheptide fine work 36kg.
Through detecting, the yield of gained Nosiheptide fine work is 80.7%, content 897u/mg.
It is found that it can be effectively by Nosiheptide from fermentation liquid using extracting method provided by the invention according to above embodiments
In extract, and the yield of gained Nosiheptide fine work and purity are higher.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of extracting method of Nosiheptide fine work, which comprises the following steps:
(1) it is filtered after mixing Nosiheptide fermentation liquid and filter aid, obtains Nosiheptide filter cake；
(2) the Nosiheptide filter cake is successively washed, is granulated and dried, obtain Nosiheptide dry granular；
(3) the Nosiheptide dry granular is extracted using methylene chloride, obtains leaching liquor；
(4) de- carbon, concentration and crystallization are successively carried out to the leaching liquor, obtain nosipeptide crude product；
(5) it is successively beaten, filtered and dried after mixing the nosipeptide crude product and ethyl alcohol, obtain Nosiheptide fine work.
2. extracting method according to claim 1, which is characterized in that the chemistry of Nosiheptide fermentation liquid in the step (1)
Potency is 2000~2500u/mL.
3. extracting method according to claim 1, which is characterized in that the filter aid in the step (1) is perlite, institute
The dosage for stating filter aid is the 3~5% of fermentating liquid volume.
4. extracting method according to claim 1, which is characterized in that the partial size of Nosiheptide dry granular in the step (2)≤
1.2mm。
5. extracting method according to claim 1, which is characterized in that methylene chloride and Nosiheptide are dry in the step (3)
The volume ratio of grain is 5~10:1.
6. extracting method according to claim 1, which is characterized in that the time extracted in the step (3) is 8~12h.
7. extracting method according to claim 1, which is characterized in that it is living that the de- carbon of carbon, which takes off agent, in the step (4)
Property charcoal, the carbon take off agent dosage be the leaching liquor volume 1~2%.
8. extracting method according to claim 1, which is characterized in that the method being concentrated in the step (4) are as follows:
The de- gained carbon of carbon is taken off into liquid and is concentrated into the 10~20% of original volume under vacuum conditions, obtains Nosiheptide concentrate；
The temperature of the concentration is 55~65 DEG C, vacuum degree >=0.03MPa of the concentration.
9. extracting method according to claim 1 or 8, which is characterized in that the method crystallized in the step (4) are as follows:
Water is added into Nosiheptide concentrate, is crystallized, Nosiheptide crystal is obtained；
The temperature of the crystallization is 4 DEG C, and the volume ratio of the water and Nosiheptide concentrate is 0.5~1:1.
10. extracting method according to claim 1, which is characterized in that ethyl alcohol and nosipeptide crude product in the step (5)
Volume ratio is 5~6:1.
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|---|---|---|---|---|
| CN1840684A (en) * | 2006-01-26 | 2006-10-04 | 浙江工商大学 | Process for fermentation preparation of nosiheptide by using streptomycete |
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| CN101519434A (en) * | 2009-04-03 | 2009-09-02 | 安徽省皖北药业股份有限公司 | Extraction process of nosiheptide raw powder |
| CN105198958A (en) * | 2015-09-29 | 2015-12-30 | 浙江汇能动物药品有限公司 | Nosiheptide finemeal extracting method |
| CN105420319A (en) * | 2015-12-24 | 2016-03-23 | 山东胜利生物工程有限公司 | Preparation method of pure nosiheptide |
| CN107619429A (en) * | 2016-07-15 | 2018-01-23 | 牡丹江佰佳信生物科技有限公司 | A kind of Nosiheptide method of purification |
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2019
- 2019-02-20 CN CN201910126539.4A patent/CN109796520A/en active Pending
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| CN1840684A (en) * | 2006-01-26 | 2006-10-04 | 浙江工商大学 | Process for fermentation preparation of nosiheptide by using streptomycete |
| CN101299010A (en) * | 2008-06-06 | 2008-11-05 | 浙江汇能动物药品有限公司 | Method for testing content of nosiheptide preparation |
| CN101519434A (en) * | 2009-04-03 | 2009-09-02 | 安徽省皖北药业股份有限公司 | Extraction process of nosiheptide raw powder |
| CN105198958A (en) * | 2015-09-29 | 2015-12-30 | 浙江汇能动物药品有限公司 | Nosiheptide finemeal extracting method |
| CN105420319A (en) * | 2015-12-24 | 2016-03-23 | 山东胜利生物工程有限公司 | Preparation method of pure nosiheptide |
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