CN105777603B - A method of extracting L- hydroxyproline from L- hydroxyproline fermentation liquid - Google Patents
A method of extracting L- hydroxyproline from L- hydroxyproline fermentation liquid Download PDFInfo
- Publication number
- CN105777603B CN105777603B CN201610205738.0A CN201610205738A CN105777603B CN 105777603 B CN105777603 B CN 105777603B CN 201610205738 A CN201610205738 A CN 201610205738A CN 105777603 B CN105777603 B CN 105777603B
- Authority
- CN
- China
- Prior art keywords
- hydroxyproline
- exchange resin
- liquid
- fermentation liquid
- extracting
- 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.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D207/16—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
Abstract
The method that the invention discloses a kind of to extract L- hydroxyproline from L- hydroxyproline fermentation liquid, this method is by L- hydroxyproline fermentation liquid micro-filtration, it is neutral that micro-filtrate adjusts pH, again through sun, resin anion (R.A.) absorption, nanofiltration obtains nanofiltration clear liquid, a condensing crystallizing is after decoloration up to product, the present invention also sends a mother liquor back to Zhiyang, resin anion (R.A.) absorption process circulation, the present invention changes the mode that conventional ion absorption exchange process directly adsorbs amino acid, it is changed to adsorbing contaminant, then it is cleaned by nanofiltration, primary crystallization is up to product, the purity of L- hydroxyproline can be greatlyd improve, content, yield and separative efficiency, reduce wastewater flow rate, and generated waste is prepared into feed, avoid environmental pollution.
Description
Technical field
The present invention relates to a kind of extraction process technology field of amino acid, more particularly to it is a kind of from L- hydroxyproline fermentation liquid
The middle method for extracting L- hydroxyproline.
Background technique
Hydroxyproline (Hydroxyproline, Hyp) is imino acid, is the product after L-PROLINE hydroxylating, point
Minor is C5H9NO3.The white flaky crystal of hydroxyproline or crystalline powder, slightly sweet, fusing point is 274 DEG C, soluble easily in water, micro-
It is dissolved in ethyl alcohol.Hydroxyproline has 4 kinds of stereoisomers, and trans- -4- hydroxy-proline is relatively conventional in nature, trans- -4- hydroxyl
Proline is found in animal collagen earliest.Trans- -4-Hydroxyproline is widely used in medicine, chemical industry, animal feed, battalion
Feeding and beauty culture etc..
Currently, the separation and Extraction hydroxyproline from fermentation liquid reported both at home and abroad, generallys use ion-exchange process, will remove
The fermentation liquid of thallus upper prop under the conditions of specific pH is removed, allows hydroxyproline to be adsorbed on ion exchange column, then uses deionization
Water sufficiently washs ion exchange column, removes impurity, then elutes hydroxyproline from ion exchange column with eluent, obtain
Hydroxyproline solution obtains hydroxyproline product using crystallization and drying.This method has the following deficiencies: 1. hydroxyproline
Oxidizable, yield is unstable；2. thick hydroxyproline impurity is more, color is deep, influence to refine yield and product quality.3. water consumption
Greatly, soda acid dosage is big, can generate a large amount of waste residue and waste water in process of production, causes pollution very serious to environment.Energy
Enough overcome these defects, realizes the slag and effluent zero-emission for extracting production process, thoroughly eradicate hydroxyproline production process middle ring
The purification production technology of border pollution problem is both at home and abroad there is not yet any report.
Summary of the invention
Present invention seek to address that the deficiencies in the prior art, and one kind is provided and extracts L- hydroxyl dried meat from L- hydroxyproline fermentation liquid
The method of propylhomoserin, this method can efficiently isolate finished product and improve purity and recovery rate.
To achieve the above object, using following technical scheme: one kind extracting L- from L- hydroxyproline fermentation liquid to the present invention
The method of hydroxyproline, which is characterized in that including following operating procedure:
Step 1: L- hydroxyproline fermentation liquid micro-filtration is obtained micro-filtration clear liquid and micro-filtration trapped fluid, micro-filtration clear liquid pH is adjusted
Value is 6~8, is then adsorbed with storng-acid cation exchange resin, then with weak-base anion-exchange resin adsorb to obtain from
Son exchange liquid；
Step 2: ion exchange liquid is crossed, the nanofiltration membrane that molecular weight is 800~1000Da obtains nanofiltration clear liquid and nanofiltration is cut
Stay liquid；
Step 3: adjusting nanofiltration retentate fluid pH value is 6~8, is then adsorbed with storng-acid cation exchange resin, then used
Weak-base anion-exchange resin adsorbs to obtain ion exchange liquid, and ion exchange liquid repeats step 2；
Step 4: it is 1/8~1/6 before concentration that 50~70 DEG C, which are concentrated into volume, by after the active carbon decoloring of nanofiltration clear liquid,
20~30 DEG C of centrifugations of cooling obtain the finished product and a mother liquor of L- hydroxyproline；
Step 5: it is 6~8 that a mother liquor, which adjusts pH value, then adsorbed with storng-acid cation exchange resin, then with weak
Alkalescence anion-exchange resin adsorbs to obtain ion exchange liquid, and ion exchange liquid repeats step 2.
The step 1 middle strong acidity cation exchange resin is in styrene-divinylbenzene copolymer with sulfonic
Cation exchange resin.
It is preferably C100 type strong acid that sulfonic cation exchange resin is had in the styrene-divinylbenzene copolymer
One of property cation exchange resin, 732 type storng-acid cation exchange resins.
In the step 1 adjust micro-filtration clear liquid pH value be 7, make fermentation liquid partial neutral, under this condition highly acid sun from
Sub-exchange resin and weak-base anion-exchange resin are weaker to the absorption of L- hydroxyproline, but to other inorganic salt impurities and less
Measure that miscellaneous amino acid adsorbed effect is obvious, can go out most of impurity in addition to L-PROLINE.
Weak-base anion-exchange resin is polyacrylic acid series anion exchange resin in the step 3.
The polyacrylic acid series anion exchange resin is preferably A830 weak-base anion-exchange resin.
Upper flow velocity when the step 3 middle strong acidity cationic exchange resin adsorption is 0.6~0.8 times strong per hour
The volume ratio of acid cation exchange resin volume, storng-acid cation exchange resin and weak-base anion-exchange resin is 3:
4~3:5.
Thickening temperature is preferably 55 DEG C in the step 4, and concentration volume is preferably 1/7 be concentrated into before concentration, described de-
The method of color is to be added to account for the injection type active carbon of nanofiltration clear liquid 0.3%~0.6% and decolourize at 20~40 DEG C of temperature 1~2h.
The a collection of micro-filtration trapped fluid obtained in the step 1 and the 1/3 batch of mother liquor obtained in the step 4 mixing are done
It is dry, L- hydroxyproline feed can be made.
The beneficial effects of the present invention are: the present invention changes traditional handicraft ion exchange resin absorption L- hydroxyproline to inhale
Attached impurity then extracts L- hydroxyproline by the method for condensing crystallizing, and the purity for not only effectively increasing product, yield are also
Effect is used for generated waste to produce feed, avoids environmental pollution.Specifically have the beneficial effect that:
(1), the problem low for traditional handicraft recovery rate, the present invention by by nanofiltration retentate fluid and a mother liquor send back to from
Sub-exchange resin absorption process, is effectively reduced the loss of raw material, and the recovery rate of L- hydroxyproline is greater than 80%.
(2), the present invention is compared with traditional handicraft, and spent ion exchange resin absorption is a small amount of impurity rather than traditional suction
Attached major part L- hydroxyproline, the dosage of resin greatly reduce, and are greatly saved for eluting and regenerated deionized water, reduction
The generation of waste water.
(3), micro-filtration trapped fluid COD high, traditional handicraft is often untreated to discharge, and pollutes environment, the present invention provides
Micro-filtration trapped fluid and partial mother liquid are mixed, is dry, L- hydroxyproline feed is made, while solving micro-filtration trapped fluid and mother liquor
Pollution problem, realize the purpose of zero-emission.
Detailed description of the invention
Fig. 1 is process flow chart of the invention；
It is described in detail below with reference to the embodiment of the present invention referring to attached drawing.
Specific embodiment
Below with reference to embodiment, the invention will be further described:
The method that the invention discloses a kind of to extract L- hydroxyproline from L- hydroxyproline fermentation liquid, those skilled in the art
Member can use for reference present disclosure, be suitably modified realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications
It will be apparent from for a person skilled in the art, be considered as being included in the present invention.Method of the invention has been led to
It crosses preferable case study on implementation to be described, those skilled in the art can obviously not depart from the content of present invention, spirit and scope to this
The method of invention and with being modified or changes and combinations appropriate, carrys out implementation and application the technology of the present invention.
Embodiment 1
It is clear that fermentation liquid after taking L- hydroxyproline to ferment through 0.05 micron~0.1 micrometer ceramics film process obtains 10L micro-filtration
Liquid and micro-filtration trapped fluid (giving over to stand-by), adjusting pH is 6~7, then first crosses 1L C100 highly acid with the flow velocity of 12.5ml/min
Cation exchange resin, after 1.5L A830 weak-base anion-exchange resin.By ion exchange liquid, cross molecular weight 800~
The nanofiltration membrane of 1000Da obtains nanofiltration clear liquid and nanofiltration retentate fluid.Nanofiltration retentate fluid adjusts pH and sends back to ion exchange for 6~7
Resin circulating process, nanofiltration clear liquid are added 25 DEG C of decoloration 1.5h of injection type active carbon of its volume 0.5%, light transmittance are obtained by filtration
96% clear liquid.
Above-mentioned destainer is concentrated in vacuo to the 1/8 of its volume at 55 DEG C, is cooled to 20~30 DEG C, 1h is kept the temperature, filters
To wet product 352.1g, mother liquor sends ion exchange resin circulating process back to.
L- hydroxyproline wet product is dried in 60 DEG C, obtains dry product 322.3g, HPLC99.3%, yield 80.6%.
The comparative test of product yield and purity
It is extracted using traditional handicraft: fetching source and the identical fermentation liquid of volume, pH is adjusted to hand over for 3 through C100 highly acidic cation
After changing resin adsorption, being eluted with ammonium hydroxide, condensing crystallizing, last total recovery are 59.2% twice after eluent decoloration,
HPLC97.3%.
Embodiment 2
Fermentation liquid after taking L- hydroxyproline to ferment obtains 99.6L micro-filtration through 0.05 micron~0.1 micrometer ceramics film process
Clear liquid and micro-filtration trapped fluid (giving over to stand-by), adjusting pH is 6~7, then first crosses 732 type strong acid of 9L with the flow velocity of 135ml/min
Property cation exchange resin, after 15L A830 weak-base anion-exchange resin.By ion exchange liquid, cross molecular weight 800~
The nanofiltration membrane of 1000Da obtains nanofiltration clear liquid and nanofiltration retentate fluid.Nanofiltration retentate fluid adjusts pH and sends back to ion exchange for 6~7
Resin circulating process, nanofiltration clear liquid are added 25 DEG C of decoloration 1.5h of injection type active carbon of its volume 0.5%, light transmittance are obtained by filtration
95.7% clear liquid.
Above-mentioned destainer is concentrated in vacuo to the 1/8 of its volume at 55 DEG C, is cooled to 20~30 DEG C, 1h is kept the temperature, filters
To wet product 3670.2g, mother liquor sends ion exchange resin circulating process back to.
L- hydroxyproline wet product is dried in 60 DEG C, obtains dry product 3310.8g, HPLC99.4%, yield 83.1%.
Embodiment 3
It is prepared by the feed of L- hydroxyproline
The spare micro-filtration trapped fluid of a batch and 1/3 batch of mother liquor are mixed, L- hydroxyproline feed product, L- are spray-dried to obtain
Hydroxyproline 30% or so.
The present invention is exemplarily described above, it is clear that present invention specific implementation is not subject to the restrictions described above,
As long as using the various improvement that the inventive concept and technical scheme of the present invention carry out, or not improved directly apply to other fields
It closes, it is within the scope of the present invention.
Claims (9)
1. a kind of method for extracting L- hydroxyproline from L- hydroxyproline fermentation liquid, which is characterized in that walked including following operation
It is rapid:
Step 1: L- hydroxyproline fermentation liquid is obtained micro-filtration clear liquid and micro-filtration through 0.05 micron~0.1 micrometer ceramics membrane microfiltration
Trapped fluid, adjust micro-filtration clear liquid pH value be 6~8, then adsorbed with storng-acid cation exchange resin, then with alkalescent yin from
Sub-exchange resin adsorbs to obtain ion exchange liquid；
Step 2: ion exchange liquid, which is crossed the nanofiltration membrane that molecular weight is 800~1000Da, obtains nanofiltration clear liquid and nanofiltration retentate fluid；
Step 3: adjusting nanofiltration retentate fluid pH value is 6~8, is then adsorbed with storng-acid cation exchange resin, then use weak base
Property anion exchange resin adsorb to obtain ion exchange liquid, ion exchange liquid repeats step 2；
Step 4: it is 1/8~1/6 before concentration, cooling that 50~70 DEG C, which are concentrated into volume, by after the active carbon decoloring of nanofiltration clear liquid
20~30 DEG C of centrifugations obtain the finished product and a mother liquor of L- hydroxyproline；
Step 5: it is 6~8 that a mother liquor, which adjusts pH value, is then adsorbed with storng-acid cation exchange resin, then use alkalescent
Anion exchange resin adsorbs to obtain ion exchange liquid, and ion exchange liquid repeats step 2.
2. the method according to claim 1 for extracting L- hydroxyproline from L- hydroxyproline fermentation liquid, which is characterized in that
The step 1 middle strong acidity cation exchange resin is to hand in styrene-divinylbenzene copolymer with sulfonic cation
Change resin.
3. the method according to claim 2 for extracting L- hydroxyproline from L- hydroxyproline fermentation liquid, which is characterized in that
It is the exchange of C100 type highly acidic cation that sulfonic cation exchange resin is had in the styrene-divinylbenzene copolymer
One of resin, 732 type storng-acid cation exchange resins.
4. the method according to claim 1 for extracting L- hydroxyproline from L- hydroxyproline fermentation liquid, which is characterized in that
It is 7 that micro-filtration clear liquid pH value is adjusted in the step 1.
5. the method according to claim 1 for extracting L- hydroxyproline from L- hydroxyproline fermentation liquid, which is characterized in that
Weak-base anion-exchange resin is polyacrylic acid series anion exchange resin in the step 3.
6. the method according to claim 5 for extracting L- hydroxyproline from L- hydroxyproline fermentation liquid, which is characterized in that
The polyacrylic acid series anion exchange resin is A830 weak-base anion-exchange resin.
7. the method according to claim 1 for extracting L- hydroxyproline from L- hydroxyproline fermentation liquid, which is characterized in that
Upper flow velocity when the step 3 middle strong acidity cationic exchange resin adsorption be per hour 0.6~0.8 times of highly acid sun from
The volume ratio of sub-exchange resin volume, storng-acid cation exchange resin and weak-base anion-exchange resin is 3:4~3:5.
8. the method according to claim 1 for extracting L- hydroxyproline from L- hydroxyproline fermentation liquid, which is characterized in that
Thickening temperature is 55 DEG C in the step 4, and concentration volume is 1/7 be concentrated into before concentration, and the method for the decoloration is to be added to account for
The injection type active carbon of nanofiltration clear liquid 0.3%~0.6% decolourizes 1~2h at 20~40 DEG C of temperature.
9. L- hydroxyproline is extracted into slave L- hydroxyproline fermentation liquid described in any claim in 8 according to claim 1
Method, which is characterized in that a collection of micro-filtration trapped fluid that the step 1 obtains and the 1/3 batch of mother liquor obtained in the step 4
Mixing, drying, can be made L- hydroxyproline feed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610205738.0A CN105777603B (en) | 2016-04-01 | 2016-04-01 | A method of extracting L- hydroxyproline from L- hydroxyproline fermentation liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610205738.0A CN105777603B (en) | 2016-04-01 | 2016-04-01 | A method of extracting L- hydroxyproline from L- hydroxyproline fermentation liquid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105777603A CN105777603A (en) | 2016-07-20 |
| CN105777603B true CN105777603B (en) | 2019-02-15 |
Family
ID=56394782
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610205738.0A Active CN105777603B (en) | 2016-04-01 | 2016-04-01 | A method of extracting L- hydroxyproline from L- hydroxyproline fermentation liquid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105777603B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106977442A (en) * | 2017-04-28 | 2017-07-25 | 天津科技大学 | A kind of extraction process of hydroxyproline |
| CN106995398A (en) * | 2017-05-10 | 2017-08-01 | 通辽梅花生物科技有限公司 | L proline novel technology for extracting |
| CN107435057B (en) * | 2017-08-12 | 2020-12-04 | 河南巨龙生物医药有限公司 | Method for producing hydroxyproline by adopting escherichia coli JL-HYP fermentation |
| CN107513030B (en) * | 2017-10-19 | 2020-06-16 | 福建师范大学 | Method for separating and purifying L-hydroxyproline from L-hydroxyproline fermentation liquor |
| CN107827803A (en) * | 2017-11-28 | 2018-03-23 | 绍兴厚普生物科技有限责任公司 | A kind of method that L hydroxyprolines are extracted from zymotic fluid |
| CN109971800A (en) * | 2019-05-17 | 2019-07-05 | 南通普悦生物医药有限公司 | The method for digesting legal system L- hydroxy-proline |
| CN114105849A (en) * | 2021-12-02 | 2022-03-01 | 保定九孚生化有限公司 | Method for extracting L-hydroxyproline |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101633634A (en) * | 2009-08-16 | 2010-01-27 | 山东阜丰生物科技开发有限公司 | Method for extracting L-proline from fermentation liquor by membrane separation technology |
| CN102675180A (en) * | 2012-05-31 | 2012-09-19 | 精晶药业有限公司 | Method for extracting proline |
| CN104276990A (en) * | 2013-07-04 | 2015-01-14 | 江南大学 | Method for decoloring L-hydroxyproline fermentation broth by utilizing active carbon |
-
2016
- 2016-04-01 CN CN201610205738.0A patent/CN105777603B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101633634A (en) * | 2009-08-16 | 2010-01-27 | 山东阜丰生物科技开发有限公司 | Method for extracting L-proline from fermentation liquor by membrane separation technology |
| CN102675180A (en) * | 2012-05-31 | 2012-09-19 | 精晶药业有限公司 | Method for extracting proline |
| CN104276990A (en) * | 2013-07-04 | 2015-01-14 | 江南大学 | Method for decoloring L-hydroxyproline fermentation broth by utilizing active carbon |
Non-Patent Citations (2)
| Title |
|---|
| 从骨胶中制备L-羟脯氨酸的研究;吕红线;《山东轻工业学院学报》;19971231;第11卷(第4期);第53-56页 |
| 由骨胶水解液中分离提取L-羟脯氨酸和L-脯氨酸的实验研究;杜英华等;《内蒙古石油化工》;20021231;第27卷;第16-18页 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105777603A (en) | 2016-07-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105777603B (en) | A method of extracting L- hydroxyproline from L- hydroxyproline fermentation liquid | |
| Boonmee et al. | Improved lactic acid production by in situ removal of lactic acid during fermentation and a proposed scheme for its recovery | |
| CN113003898B (en) | Corn soaking water treatment method | |
| CN106397506B (en) | A kind of purification process of high-quality acarbose | |
| CN102432479B (en) | Method for extracting L-valine from L-valine fermentation liquid | |
| CN115160108B (en) | Process for preparing inositol and phosphoric acid | |
| CN103232353A (en) | Method for separating and extracting L-valine from broth with high efficiency | |
| CN103508933B (en) | Separating and purifying method for L-tryptophan | |
| CN106631852A (en) | Method for extracting L-ornithine hydrochloride from L-ornithine fermentation broth | |
| CN106831894A (en) | A kind of method that deacetylation Coupling Adsorption separates D aminoglucose hydrochlorides | |
| CN109678183B (en) | Method for preparing lithium hydroxide from industrial soluble lithium salt | |
| CN105837488B (en) | A kind of hydroxyproline fermentation manufacturing technique | |
| CN106083952A (en) | A kind of method extracting gentamycin sulfate from gentamicin fermentation broth | |
| CN103420826A (en) | Method for extracting succinic acid from fermentation broth | |
| CN106928288B (en) | A kind of preparation method of dihydrostreptomycin sulfate | |
| CN203866227U (en) | System capable of separating and purifying amino acid continuously | |
| CN105669511B (en) | A kind of hydroxyproline refining methd | |
| CN106588641B (en) | A kind of recovery method of 2-KLG primary crystallization mother liquor | |
| CN204933271U (en) | A kind of succinic acid fermentation liquor separation and purification device | |
| CN100339357C (en) | Hind extraction process for producing L-phenylalanine using phenyl-pyruvic acid enzyme method | |
| CN107827803A (en) | A kind of method that L hydroxyprolines are extracted from zymotic fluid | |
| CN102633848A (en) | Method for removing impurities from gentamicin | |
| CN107586310B (en) | Extraction process of flavomycin | |
| CN102578369B (en) | Method for preparing feed by using waste liquor generated in the process of producing L-glutamine | |
| CN101818216B (en) | Method for refining corncob acid hydrolysis solution |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |