CN101062934A - Method for extracting natamycin from fermentation technique culture - Google Patents
Method for extracting natamycin from fermentation technique culture Download PDFInfo
- Publication number
- CN101062934A CN101062934A CN 200710015831 CN200710015831A CN101062934A CN 101062934 A CN101062934 A CN 101062934A CN 200710015831 CN200710015831 CN 200710015831 CN 200710015831 A CN200710015831 A CN 200710015831A CN 101062934 A CN101062934 A CN 101062934A
- Authority
- CN
- China
- Prior art keywords
- tennecetin
- extracting
- fermentation technique
- technique culture
- solid
- 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.)
- Granted
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method to extract myprozine from ferment technique culture, which comprises the following steps: adjusting the pH value of the culture to 10. 0-14. 0 with basic substance under the condition of 0-35 deg. c; stirring evenly; separating solid and liquid; collecting filter liquor; getting aqueous phase extract of myprozine; acidifying at normal temperature; adjusting the pH value of the filter liquor to 5. 0-9. 0; depositing myprozine; getting crystalline liquid of myprozine; proceeding solid-liquid separation; getting the product. This method possesses the advantages of low cost, environmental protection and low demand for zymotic fluid and device.
Description
Technical field:
The present invention relates to a kind of method of from fermentation technique culture, directly extracting tennecetin.
Background technology:
Tennecetin [Natamycin claims pimaricin (Pimaricin), tennecetin (Tenecetin) again] is a kind of polyene macrolide antibiotics; Molecular formula C
33H
47NO
13Molecular weight 665.75 dalton; Tennecetin is a kind of amphoteric substance, and iso-electric point is pH6.5, and two kinds of configurations of the general existence of its structure are respectively enol form and ketone form structure.The structural formula of tennecetin is:
Because tennecetin energy efficient, narrow spectrum inhibition yeast and mould also has good physical and chemical stability and extremely low toxicity, thereby tennecetin is used among medical treatment (external application), food, feed, the grain storage, shown good prospect in the application aspect particularly fresh-keeping at food raw material, the food antiseptic.But at present because its high production cost is greatly limited tennecetin product commercial applications, only as high-end aseptic applications.Because water insoluble organic solvent that is dissolved under the certain condition under the tennecetin usual conditions,, therefore be necessary to develop a kind of economically viable tennecetin extraction process so its extracting method can not economical operation.
Tennecetin is by being produced through biological fermentation by streptomycete, and streptomycete commonly used comprises the disclosed Natal of GB-A844289 streptomycete (Streptomyces natalensis), the disclosed brown yellow spore streptomycete of WO93/03171 (Streptomyces gilvosporeus) and exactly exerts and adds streptomycete (Streptomyceschattanoganis) etc.The fermentation unit of tennecetin generally can reach 2g-15g/L.Just can from natamycin fermentation liquor, separation and purification go out tennecetin by suitable microbiotic extractive technique.United States Patent (USP) 3,892,850, WO92/07998, WO92/1058 etc. have announced the extracting method of tennecetin, more than the method announced of these patents all need earlier from fermented liquid enrichment or isolate mycelium, and then utilize organic solvent such as methyl alcohol, Virahol etc. from enrichment or isolated mycelium, to extract tennecetin.The United States Patent (USP) 3,892,850 of the discoverer Struky of tennecetin has been announced the method for extracting tennecetin by distillation behind the acidifying methanol extraction mycelium, precipitation.The method that patent WO92/07998, WO92/1058 announce is: in isolated mycelium, add methyl alcohol, regulate then the pH value to 1.0-4.5 so that tennecetin be extracted in the organic phase; Remove by filter mycelium, to 6.0-9.0 the tennecetin crystallization is separated out by regulating extraction liquid pH value.
Because tennecetin is water insoluble, in fermented liquid, adhere to and mycelium surface with solid form, therefore the extracting method in the above patent reduces the organic solvent consumption for the concentration that improves organic solvent in the extracting solution, they before utilizing organic solvent extraction tennecetin all earlier by a kind of moisture that reduces in the fermented liquid in the solid-liquid separating methods such as centrifugal, concentrated, filtration, mycelium in the enrichment fermented liquid is so that the usefulness of extraction, so just caused the waste of power, manpower expense.Simultaneously all these extracting method all need to use inflammable, volatile organic solvents such as methyl alcohol, Virahol, and this has just caused, and higher solvent loss, recovery cost and higher safety features require, high expenses of environmental protection.More than these two product costs that all caused the organic solvent extracting method to produce higher.
WO97/29207 has announced a kind of method that does not need organic solvent to extract tennecetin.This method uses the method for physics or chemistry to make mycelial cell fragmentation, degraded in the fermented liquid earlier, tennecetin solid suspension in the fermented liquid makes the tennecetin solid separate with cell debris by application of weight gradient centrifugation technology then and obtains product at aqueous phase.This method has the short advantage of operational path, but this technology reaches the requirement that strictness was all once all being arranged to the tennecetin crystallographic dimension in the fermented liquid simultaneously to the content requirement of tennecetin in the fermented liquid higher (greater than 7 grams per liters).In addition, extract purifying specialty angle from microbiotic, the particle that utilizes the gravity gradient centrifugation technique thoroughly to separate the different grain size size can propose high requirement to separating device, also needs the equipment maintenance cost of great number simultaneously.From above 2 application that can find out this extracting method certain limitation is arranged.
Known research to the tennecetin dissolution characteristics at present is confined to the scope of organic solvent, and the extracting method of most tennecetins is also set up on this basis.
Summary of the invention:
The purpose of this invention is to provide a kind of method of from fermentation technique culture, extracting tennecetin, extracting tennecetin with non-organic solvent method from fermented liquid is feature, solved the organic solvent method and extracted the tennecetin that tennecetin causes and extract the production cost problem of higher that organic solvent waste height, operation length cause in the production process, a kind of economy, environmental protection be provided and be suitable for the extracting method that industrialization is produced.
The objective of the invention is to realize as follows: we studied tennecetin under different condition in water solubleness and change of stability trend, find that tennecetin solubleness in water under neutrallty condition is extremely low, but under acidic conditions, reduce along with pH and alkaline condition under along with pH its solubleness that raises raises gradually, according to this characteristic, we work out the method that does not need organic solvent directly to extract tennecetin from the tennecetin fermentation technique culture, and this method comprises: rising fermented liquid pH value makes tennecetin water-soluble; Remove mycelium, collect the tennecetin aqueous extract; Reducing extracting liquid pH value separates out the tennecetin precipitation; Recovery, the drying precipitated tennecetin solid that obtains.The concrete steps of this method are:
A. under 0-35 ℃ of condition, fermentation technique culture is stirred adjusting pH value to 10.0-14.0 with alkaline matter;
B. solid-liquid separation is collected filtrate and is obtained the tennecetin aqueous extract;
C. acidifying is regulated filtrate pH value to 5.0-9.0 under the normal temperature, and the tennecetin precipitation is separated out the crystal solution that obtains tennecetin;
D. crystal solution obtains the tennecetin product through solid-liquid separation.
The tennecetin fermentation technique culture is a natamycin fermentation liquor, or natamycin fermentation liquor is starched or bacterium slag or dry mycelium by the dense bacterium of partial dehydration that solid-liquid separation technique obtains.Preferred natamycin fermentation liquor.Make suspension before bacterium slag or the alkalization of dry mycelium earlier with water or diluted alkaline.
The alkaline matter of a step is sodium hydroxide or potassium hydroxide or ammoniacal liquor, is the industrial extremely alkaline material of regulator solution commonly used.
In a step tennecetin fermentation technique culture is regulated the preferred 11.5-12.5 of pH value with alkaline matter.
When in a step tennecetin fermentation technique culture being alkalized with alkaline matter the preferred 5-25 of temperature ℃.
The solid-liquid separating method of b step adopts general microbiotic to extract the solid-liquid separating method that the purifying professional and technical personnel can utilize,, centrifuging centrifugal, cross flow filter as Plate Filtration, gravity, remove mycelium, collect filtrate and obtain the tennecetin aqueous extract.
The gac that the tennecetin aqueous extract that b step solid-liquid separation obtains adds 0.1-2.5% (W/V) stirs the processing of decolouring more than 30 minutes.
Filtrate pH value process is regulated in the acidifying of c step, the temperature of filtrate can remain on 5-25 ℃, with industrial acid commonly used, example hydrochloric acid, sulfuric acid, phosphoric acid etc., or its acidic solution for preparing is regulated filtrate pH value, reduce the solubleness of tennecetin, make tennecetin form precipitation and separate out the crystal solution that obtains tennecetin at aqueous phase.
The solid-liquid separation of d step crystallization liquid is selected any conventional solid-liquid separating method for use, as centrifuging, suction filtration, Plate Filtration etc., crystal solution is carried out solid-liquid separation obtain the tennecetin crystal.
The tennecetin product that the d step obtains through solid-liquid separation, after salt-free water washing 2～3 times, adopt conventional drying means under 50-80 ℃ of condition, to be dried to moisture between 6.0-9.0%, can obtain off-white color to faint yellow, content greater than 80% tennecetin solid.Conventional drying means is as heating forced air drying, vacuum-drying etc.
The present invention is with by industrial microorganism culturing means commonly used, cultivation can be produced the streptomycete of tennecetin, as brown yellow spore streptomycete, Natal streptomycete, exactly exert tennecetins such as adding streptomycete and produce bacterium, the fermented liquid or the fermented liquid that obtain tennecetin are object through pretreated product, produce the tennecetin finished product.This extracting method has and does not use that organic solvent, cost are low, environmental protection, the characteristic of fermented liquid and equipment is required characteristics such as low.
Embodiment:
Embodiment 1
Get natamycin fermentation liquor 3.0L, content 4.6g/l.Earlier fermented liquid is cooled to 15 ℃; Regulate pH to 12.0 back with the KOH solution of 4mol/L again and stirred 30 minutes, keep 15 ℃ of temperature in this process; The fermented liquid of alkalization after under 3000 rotating speeds centrifugal 15 minutes, is collected centrifuged supernatant, regulate supernatant liquor pH to 6.0-7.0 with the HCl solution of 1.0mol/L then and tennecetin is precipitated separate out.The suction filtration crystal solution with twice of an amount of salt-free water washing after, 60 ℃ of forced air dryings 3 hours tennecetin 11.90 grams, content 84.5%.
Embodiment 2
With tennecetin mycelium 200 grams, wherein tennecetin content is 8.9%, adds to stir in 2 liters of no salt solution mycelium is fully suspended, and is cooled to 5 ℃.Regulate pH to 11.5 back with the ammoniacal liquor of 2mol/L and stirred 30 minutes, keep 15 ℃ of temperature in this process, suction filtration isolate slag is collected filtrate.Use the H of 0.5mol/L then
2SO
4Solution is regulated filtrate pH to 6.0-7.0 separates out the tennecetin precipitation.The suction filtration crystal solution with twice of an amount of salt-free water washing after, 50 ℃ ,-the 0.09MPa condition under vacuum-drying 3 hours tennecetin 15.0 grams, content 85.3%.
Embodiment 3
Get natamycin fermentation liquor 1000L, content 3.8g/l.Earlier fermented liquid is cooled to 20 ℃; Regulate pH to 12.0 back with the NaOH solution of 4mol/L again and stirred 30 minutes, keep 15 ℃ of temperature; After adding 2Kg flocculating aids stirs in the fermented liquid of alkalization, filter with flame filter press.Collect filtrate, regulate filtrate pH to 6.0-7.0 with the HCl solution of 1.0mol/L then, the tennecetin precipitation is separated out.Use the link-suspended basket centrifuge precipitation separation, again with after twice of an amount of salt-free water washing, 60 ℃ of forced air dryings 3 hours, tennecetin 3136 grams, content 82.4%.
Claims (8)
1, a kind of method of extracting tennecetin from fermentation technique culture is characterized in that this method may further comprise the steps:
A. under 0-35 ℃ of condition, fermentation technique culture is stirred adjusting pH value to 10.0-14.0 with alkaline matter;
B. solid-liquid separation is collected filtrate and is obtained the tennecetin aqueous extract;
C. acidifying is regulated filtrate pH value to 5.0-9.0 under the normal temperature, and the tennecetin precipitation is separated out the crystal solution that obtains tennecetin;
D. crystal solution obtains the tennecetin product through solid-liquid separation.
2, the method for from fermentation technique culture, extracting tennecetin according to claim 1, it is characterized in that: the tennecetin fermentation technique culture is a natamycin fermentation liquor, or natamycin fermentation liquor is starched or bacterium slag or dry mycelium by the dense bacterium of partial dehydration that solid-liquid separation technique obtains.
3, the method for extracting tennecetin from fermentation technique culture according to claim 2, it is characterized in that: the tennecetin fermentation technique culture is a natamycin fermentation liquor.
4, the method for extracting tennecetin from fermentation technique culture according to claim 1, it is characterized in that: the alkaline matter of a step is sodium hydroxide or potassium hydroxide or ammoniacal liquor.
5, the method for extracting tennecetin from fermentation technique culture according to claim 1 is characterized in that: in a step tennecetin fermentation technique culture is regulated the pH value to 11.5-12.5 with alkaline matter.
6, the method for extracting tennecetin from fermentation technique culture according to claim 1 is characterized in that: the temperature when in a step tennecetin fermentation technique culture being alkalized with alkaline matter remains on 5-25 ℃.
7, the method for extracting tennecetin from fermentation technique culture according to claim 1 is characterized in that: the gac that the tennecetin aqueous extract that b step solid-liquid separation obtains adds 0.1-2.5% (W/V) stirs the processing of decolouring more than 30 minutes.
8, the method for from fermentation technique culture, extracting tennecetin according to claim 1, it is characterized in that: the tennecetin product that the d step obtains through solid-liquid separation, after salt-free water washing 2～3 times, adopt conventional drying means under 50-80 ℃ of condition, to be dried to moisture between 6.0-9.0%, obtain off-white color to faint yellow, content greater than 80% tennecetin solid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710015831A CN101062934B (en) | 2007-05-31 | 2007-05-31 | Method for extracting natamycin from fermentation technique culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710015831A CN101062934B (en) | 2007-05-31 | 2007-05-31 | Method for extracting natamycin from fermentation technique culture |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101062934A true CN101062934A (en) | 2007-10-31 |
| CN101062934B CN101062934B (en) | 2010-05-19 |
Family
ID=38964241
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200710015831A Active CN101062934B (en) | 2007-05-31 | 2007-05-31 | Method for extracting natamycin from fermentation technique culture |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101062934B (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101307299B (en) * | 2008-03-11 | 2010-06-02 | 湖北大学 | Method for breeding high yield strain of natamycin and high-efficiency fermenting, abstracting and purifying method of natamycin |
| CN102040638A (en) * | 2009-10-20 | 2011-05-04 | 华北制药集团新药研究开发有限责任公司 | Method for preparing nonsolvent of high-purity natamycin |
| CN102242170A (en) * | 2011-04-22 | 2011-11-16 | 南通奥凯生物技术开发有限公司 | Production technology of natamycin |
| CN103665074A (en) * | 2014-01-07 | 2014-03-26 | 厦门大学 | Extraction and purification method for natamycin in fermentation broth |
| CN106676150A (en) * | 2017-03-31 | 2017-05-17 | 山东鲁抗医药股份有限公司 | Method for producing natamycin by using streptomyces gilvosporeus through fermentation |
| CN107880086A (en) * | 2017-11-24 | 2018-04-06 | 浙江工业大学 | A kind of preparation method of water-soluble natamycin sodium salt |
| CN108659075A (en) * | 2018-06-19 | 2018-10-16 | 苏州汉德瑞生物工程有限公司 | A kind of preparation method of novel polymolecularity Natamycin |
| CN112585150A (en) * | 2018-08-16 | 2021-03-30 | 帝斯曼知识产权资产管理有限公司 | Novel epolyonic amphomacrolides and process for purifying natamycin |
| CN117587087A (en) * | 2023-11-22 | 2024-02-23 | 浙江新银象生物工程有限公司 | Preparation method of liquid natamycin |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992010580A1 (en) * | 1990-12-07 | 1992-06-25 | E.I. Du Pont De Nemours And Company | Natamycin recovery |
| US5591438A (en) * | 1990-12-07 | 1997-01-07 | Bio-Technical Resources L.P. | Natamycin recovery |
| ES2085031T3 (en) * | 1991-08-05 | 1996-05-16 | Bio Tech Resources | FERMENTATION PROCEDURE TO PRODUCE NATAMYCIN. |
| US5942611A (en) * | 1995-01-19 | 1999-08-24 | Cultor Ltd. | Process for natamycin recovery |
| CN1515678A (en) * | 2003-08-25 | 2004-07-28 | 天津科技大学 | Preparation method of natamycin |
| CN100393738C (en) * | 2005-07-05 | 2008-06-11 | 广州市微生物研究所 | Natamycin extracting and purifying method |
-
2007
- 2007-05-31 CN CN200710015831A patent/CN101062934B/en active Active
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101307299B (en) * | 2008-03-11 | 2010-06-02 | 湖北大学 | Method for breeding high yield strain of natamycin and high-efficiency fermenting, abstracting and purifying method of natamycin |
| CN102040638A (en) * | 2009-10-20 | 2011-05-04 | 华北制药集团新药研究开发有限责任公司 | Method for preparing nonsolvent of high-purity natamycin |
| CN102040638B (en) * | 2009-10-20 | 2014-06-25 | 华北制药集团新药研究开发有限责任公司 | Method for preparing nonsolvent of high-purity natamycin |
| CN102242170A (en) * | 2011-04-22 | 2011-11-16 | 南通奥凯生物技术开发有限公司 | Production technology of natamycin |
| CN103665074A (en) * | 2014-01-07 | 2014-03-26 | 厦门大学 | Extraction and purification method for natamycin in fermentation broth |
| CN103665074B (en) * | 2014-01-07 | 2016-05-18 | 厦门大学 | The method for extraction and purification of natamycin in a kind of zymotic fluid |
| CN106676150A (en) * | 2017-03-31 | 2017-05-17 | 山东鲁抗医药股份有限公司 | Method for producing natamycin by using streptomyces gilvosporeus through fermentation |
| CN107880086A (en) * | 2017-11-24 | 2018-04-06 | 浙江工业大学 | A kind of preparation method of water-soluble natamycin sodium salt |
| CN107880086B (en) * | 2017-11-24 | 2021-05-07 | 浙江工业大学 | Preparation method of water-soluble natamycin sodium salt |
| CN108659075A (en) * | 2018-06-19 | 2018-10-16 | 苏州汉德瑞生物工程有限公司 | A kind of preparation method of novel polymolecularity Natamycin |
| CN112585150A (en) * | 2018-08-16 | 2021-03-30 | 帝斯曼知识产权资产管理有限公司 | Novel epolyonic amphomacrolides and process for purifying natamycin |
| CN112585150B (en) * | 2018-08-16 | 2024-03-29 | 帝斯曼知识产权资产管理有限公司 | New epoxypolyene ampholytic macrolides and process for purifying natamycin |
| CN117587087A (en) * | 2023-11-22 | 2024-02-23 | 浙江新银象生物工程有限公司 | Preparation method of liquid natamycin |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101062934B (en) | 2010-05-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101062934B (en) | Method for extracting natamycin from fermentation technique culture | |
| CN102408462B (en) | Preparation method of erythromycin thiocyanate | |
| CN107474088B (en) | Extraction process for industrial mass production of spinosad | |
| CN102382158B (en) | A kind of preparation method of high purity amphotericin B | |
| CN102040638B (en) | Method for preparing nonsolvent of high-purity natamycin | |
| CN102659882B (en) | Method for extracting tartaric acid acetylisovaleryl tylosion | |
| CN112515032A (en) | Extraction method of selenoprotein in cardamine violifolia, selenoprotein obtained by extraction method and application of selenoprotein | |
| CN107513042B (en) | Method for extracting shenqinmycin by non-chemical solvent | |
| CN101974075A (en) | Method for extracting polymyxin B and E from fermentation technique culture | |
| CN106554273B (en) | Method for purifying long-chain dicarboxylic acid in fermentation liquor | |
| CN102286044A (en) | Preparation method of natamycin laminar crystal | |
| CN106008624B (en) | A kind of method for crystallising improving avilamycin active principle A, B content | |
| CN101709072B (en) | Method for efficiently extracting and purifying natamycin | |
| CN1257982C (en) | Method for separating and refining polyhydroxy fatty acid ester in bacteria cell from bacteria fermentation liquor | |
| CN204162637U (en) | A kind of Matachrom extraction element | |
| CN102816751A (en) | High-activity chitosanase and preparation method thereof | |
| CN112760240B (en) | Method for extracting natural green pigment from penicillium fungus | |
| CN101514155A (en) | Method for separating and extracting pyruvic acid from fermentation broth by an ion exchange method | |
| CN101942491A (en) | Method for efficiently separating and extracting welan gum from welan gum fermentation liquor | |
| CN112778792B (en) | Method for extracting natural green pigment from trichoderma fungus | |
| CN103641893A (en) | Method for recovering colistin sulfate from colistin sulfate slag | |
| CN115772198A (en) | Novel method for desugaring abamectin | |
| CN102358716B (en) | Method of extracting succinic acid from fermentation broth | |
| CN101321771B (en) | Process for the preparation of a potassium salt of penicillin | |
| CN103304396B (en) | Method for separating and purifying 1403C from fermentation liquid of 1403C |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |