CN107056861A - A kind of separation from fermentation system obtains the technique and complete set of equipments of Ademetionine - Google Patents
A kind of separation from fermentation system obtains the technique and complete set of equipments of Ademetionine Download PDFInfo
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- CN107056861A CN107056861A CN201710076801.XA CN201710076801A CN107056861A CN 107056861 A CN107056861 A CN 107056861A CN 201710076801 A CN201710076801 A CN 201710076801A CN 107056861 A CN107056861 A CN 107056861A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
- C07H19/167—Purine radicals with ribosyl as the saccharide radical
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
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- G—PHYSICS
- G05—CONTROLLING; REGULATING
- G05D—SYSTEMS FOR CONTROLLING OR REGULATING NON-ELECTRIC VARIABLES
- G05D21/00—Control of chemical or physico-chemical variables, e.g. pH value
- G05D21/02—Control of chemical or physico-chemical variables, e.g. pH value characterised by the use of electric means
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Abstract
The invention discloses the technique that a kind of separation from fermentation system obtains Ademetionine, using two step section pH, the brand-new technique of three UF membranes realizes the separation of Ademetionine.Technique disclosed in this invention, separation obtains the Ademetionine in zymotic fluid, and when input concentration is 50 ~ 140g/L, after separation, SAM concentration can reach 300 g/L, and purity is 99.80%, and 99.80% is reached as high as to SAM recovery rate.
Description
Technical field
The invention belongs to field of biological pharmacy, be related to a kind of separation from fermentation system obtain Ademetionine technique and
Complete set of equipments, and in particular to acquisition adenosine egg ammonia is separated from zymotic fluid using the method for regulation pH technique, and UF membrane
Acid.
Background technology
SAM, also known as S-adenosylmethionine, its English entitled S-Adenosyl-methionine (SAM)
, it is the activity form of methionine.
SAM contains an active methyl, so also known as active methionine, it is widely present in various
In tissue and body fluid, participate in the transmethylation of body, polyamines and synthesize and turn sulfurization.SAM is important in vivo
Methyl donor, it participates in vivo acid, protein, carbohydrate, polysaccharide and phosphatide, nucleic acid and methylated, so as to adjust
Some physiological activities in organism, such as processing of information transmission, protein and modification are saved, if SAM content
It is too low to cause a variety of diseases.
At present, SAM is mainly produced by biological fermentation process.But, for many years, isolated and purified from bio-fermented liquid
SAM is always a technical barrier not solved very well.Due to SAM and the raw material L- methionine (Met) of residual
With close physicochemical property, and in bio-fermented liquid SAM products concentration it is relatively low, use conventional methods and be difficult to separate
Purify SAM.
In the prior art, a kind of cationic ion-exchange resin layer is proposed in Chinese invention patent ZL 200510019206.X
Extraction method is analysed, this method cost height, separation process length, low production efficiency, and SAM easily occur in long-time separation process
Degraded.SAM is separated using trichloroacetic acid solvent precipitation in Chinese invention patent ZL 20081002015.4.But three
Monoxone toxicity is big, excitant strong, and subsequent treatment is complicated, and cost is higher.In the A of Chinese invention patent CN 102617681
Separation concentration is carried out to SAM using liquid film and organic solvent.But organic solvent n-butanol, n-hexyl alcohol, n-octyl alcohol or certain herbaceous plants with big flowers alcohol are to ring
Border pollution is big, while preparing for liquid film is complex.
Research and development new separating and purifying technologies, high efficiency, low cost isolate and purify SAM from bio-fermented liquid, are
Problem urgently to be resolved hurrily in current SAM productions.
The content of the invention
In order to overcome the shortcoming of traditional separation method, the present invention provides a kind of new separating and purifying technology, from biofermentation
High efficiency in liquid, low cost isolate and purify SAM.
One of technical purpose of the present invention obtains the technique of Ademetionine for a kind of separation from fermentation system of offer,
In this technique, two step section pH value, the method for three membrane separation concentrations are employed.
In the process, the first time pH value regulation technical issues that need to address are that impurity is excessive in solution fermentation system
The problem of, by adjusting pH value for 1.5-2.0 and ultrafiltration membrance filter, can effectively to the cell fragment in feed liquid, foreign protein and
Pigment is removed.
In the process, second pH value regulation technical issues that need to address are, the raw material L- egg ammonia of SAM and residual
Sour (Met) has close physicochemical property, in the prior art, is difficult to realize UF membrane, by adjusting pH value for 5.0-6.0,
The hydration radiuses of the SAM of chirality in the solution can be effectively made to change, so as to realize SAM UF membrane.
Another technical purpose of the present invention there is provided the complete set of equipments that the separation from fermentation system obtains Ademetionine,
By the complete set of equipments, efficiently separating for the zymotic fluid containing SAM can be achieved.Wherein, the complete set of equipments is employed in the present invention and made
Technique, i.e. two step section pH value, the method for three membrane separation concentrations.
Specifically, the technical scheme is that：
A kind of separation from fermentation system obtains the technique of Ademetionine, wherein, its step is followed successively by：
（1）By containing SAM zymotic fluid add acid, regulation pH value be 1.5-2, stirring to have precipitation generation；
（2）Mixed fermentation liquid is subjected to ultrafiltration by the tubular ultra-filtration membrane of wide runner；
（3）Acid or alkali are added in material after ultrafiltration transmission, regulation pH value is 5.0-6.0；
（4）PH value ultrafiltration permeate will be mixed up by one-level nanofiltration membrane, and material wash and further with pure water
Concentration；
（5）The concentrate of one-level NF membrane is connected in series through into two grades of NF membranes to be concentrated.
2. technique according to claim 1, it is characterised in that the membrane material of the milipore filter is PVDF, internal diameter is
12mm, 5 ~ 50 DEG C of operating temperature, operating pressure is≤0.5MPa, and membrane aperture is 0.1 μm；The membrane material of NF membrane is PVDF, is cut
70 ~ 150Da of molecular weight, 5 ~ 50 DEG C of operating temperature are stayed, operating pressure is 4.1 MPa.
Technique of the present invention, wherein, SAM concentration is 50 ~ 140g/L in the zymotic fluid.
Technique of the present invention, wherein, the acid is one kind in sulfuric acid, hydrochloric acid；The alkali is sodium hydroxide, ammoniacal liquor
In one kind.
A kind of separation from fermentation system obtains the complete automation equipment of Ademetionine, wherein, this is set of automatic
Change equipment and comprise at least two soda acid tanks, two head tanks, three groups of film devices, power-equipment, its connecting pipeline of each equipment room
And valve, the system of Automated condtrol.
Complete automation equipment of the present invention, wherein, the equipment containing modular charging bucket device 1., pump 2., ultrafiltration
Film and membrane module 3., soda acid tank arrangement 4., soda acid tank arrangement 5., charging bucket device 6., pump 7., NF membrane and membrane module 8., supercharging
Pump 9., NF membrane and membrane module 10., connecting pipeline, the system of Automated condtrol.
Complete automation equipment of the present invention, wherein, 1. the charging bucket device, at least contains, the charging positioned at top
Through hole, the through hole that adds water, acid adding through hole plus alkali through hole；Go out material through hole positioned at bottom.
Complete automation equipment of the present invention, wherein, 6. the charging bucket device, at least contains, the charging positioned at top
Through hole, the through hole that adds water, acid adding through hole plus alkali through hole；Go out material through hole positioned at bottom.
Complete automation equipment of the present invention, wherein, 1. charging bucket device goes out material through hole and Beng② UNICOMs, and pump is 2. with surpassing
Filter membrane and membrane module 3. charging aperture UNICOM；Charging bucket device 6. with milipore filter and membrane module 3. discharge port end UNICOM；Charging bucket device is 6.
Discharging opening and Beng⑦ UNICOMs；Pump 7. with NF membrane and membrane module 8. charging aperture UNICOM；8. discharging opening and the increasing of NF membrane and membrane module
Press pump 9. UNICOM, booster pump discharge port end and NF membrane and membrane module 10. feed inlet end UNICOM.
Complete automation equipment of the present invention, wherein, the control system of the automation includes the online inspection of pH value
Survey with and control system, by the pH value of on-line checking batch can 1. with batch can 6., and control enter tank 1. with batch can 6. soda acid
The folding of mouth valve, realizes the On-line Control of pH value；Also include pump provided with flow sensor ~, pressure sensor and control
The pressure and data on flows imported and exported in device processed, flow sensor and pressure sensor collection pump, and transmit to controller, control
Data reception module, processing module and communication module are provided with device processed, data reception module receives the number of two sensor transmissions
It is believed that being transferred to processing module after breath, finally transmitted again via communication module to service terminal；Service terminal receives controller hair
The data message come, and it is shown in its display screen.
In a preferred embodiment of the invention, in the Ademetionine feed liquid that concentration is 50 ~ 140g/L, add
Hydrochloric acid solution, mixing, pH value is adjusted to 1.5-2.0, and produces precipitation, feed liquid is crossed into membrane material for PVDF, internal diameter is 12mm,
5 ~ 50 DEG C of operating temperature, operating pressure is≤0.5MPa, and membrane aperture is 0.1 μm of milipore filter, and filtrate is with salt acid for adjusting pH value 5.0-
6.0；It is PVDF, 70 ~ 150Da of molecular cut off by membrane material, 5 ~ 50 DEG C of operating temperature, operating pressure is the one of 4.1 MPa
It is 200g/L that level NF membrane, which carries out nanofiltration to SAM concentration, and purity is 99%；Filtrate is by secondary nanofiltration, and SAM concentration is 300g/L,
Purity is 99.8%.
In the another embodiment of the present invention, concentration passes through certainly for 50 ~ 140g/L Ademetionine feed liquid
1. dynamic valve automatically flows into batch can, then the acid by automated system sour tank 4. it is automatic the pH value of feed liquid is adjusted to 1.5 with acid ~
2.0, milipore filter is 2. pumped into finally by pump 3. the impurity in feed liquid is removed, while adding pure water automatically by automatic valve to thing
3. material enters milipore filter after being cleaned, and 6. the permeate of milipore filter 3. automatically flows into batch can, then passes through automated system
The pH value of feed liquid is adjusted to 5.0 ~ 6.0 automatically by the soda acid of sour tank 4., 5., then 8. 7. NF membrane is pumped into material progress by pump
One-level is cleaned and concentrated, and the SAM of primary concentration concentration can reach 200g/L, and purity can reach 99%, the efflux of NF membrane 8.
Two grades of special NF membranes are 9. pumped into by booster pump again SAM concentration, which is 300 g/L, 10. to be reached to material progress secondary concentration, it is pure
Spend for 99.80%.
The beneficial effects of the present invention are
（1）SAM separation concentration is carried out by the way of UF membrane, the technique belongs to physical process, eliminates chemical reagent pair
The pollution of product, operation is simple and reliable, and usability is strong, considerably reduces production cost, easily realizes industry amplification.
（2）All operations of the present invention have carried out integrated modular, by computer centralized Control, the steady quality of product,
Save human cost simultaneously.Pump is provided with flow sensor, pressure sensor and controller, and flow sensor and pressure are passed
The pressure imported and exported and data on flows in sensor collection pump, and transmit to controller, provided with data reception module, place in controller
Module and communication module are managed, data reception module is transferred to processing module after receiving the data message of two sensor transmissions, most
Transmitted again via communication module to service terminal afterwards；Service terminal receives the data message that controller is sent, and it is aobvious to be shown in it
Display screen.
（3）The film device that the present invention is used, it is strong with contamination resistance compared with conventional roll or narrow flow road milipore filter,
Clean easy characteristic.
Brief description of the drawings
Fig. 1：Separation obtains the complete set of equipments of Ademetionine from fermentation system
Wherein, it is 1. charging bucket device；2. it is pump；3. it is milipore filter and membrane module；4. it is soda acid tank arrangement；5. it is canned for soda acid
Put；6. it is charging bucket device；7. it is pump；8. it is NF membrane and membrane module；9. it is booster pump；10. it is NF membrane and membrane module；PG is
Pressure gauge；AIR is pneumatic operated valve；~ it is flowmeter, it is control cabinet, is control main frame, is display screen.
The chemical constitution of Fig. 2 SAMs.
Embodiment
Embodiment 1 this example demonstrates that from fermentation system separation obtain Ademetionine complete set of equipments composition and
Annexation
The schematic diagram of the complete set of equipments is as shown in Figure 1.The specific equipment containing modular charging bucket device 1., pump 2., ultrafiltration
Film and membrane module 3., soda acid tank arrangement 4., soda acid tank arrangement 5., charging bucket device 6., pump 7., NF membrane and membrane module 8., supercharging
Pump 9., NF membrane and membrane module 10., the system of connecting pipeline and Automated condtrol.
1. charging bucket device contains, feed-throughs, the through hole that adds water, acid adding through hole plus alkali through hole positioned at top；Positioned at bottom
Go out material through hole.6. charging bucket device contains, feed-throughs, the through hole that adds water, acid adding through hole plus alkali through hole positioned at top；Positioned at bottom
Go out material through hole.
Wherein, 1. charging bucket device goes out material through hole and Beng② UNICOMs, pump 2. with milipore filter and membrane module 3. charging aperture UNICOM；Material
Tank arrangement 6. with milipore filter and membrane module 3. discharge port end UNICOM；Charging bucket device 6. discharging opening and Beng⑦ UNICOMs；Pump 7. with nanofiltration
Film and membrane module 8. charging aperture UNICOM；NF membrane and membrane module 8. discharging opening and booster pump 9. UNICOM, booster pump discharge port end with
NF membrane and membrane module 10. feed inlet end UNICOM.
The control device of automation, including pH value on-line checking with and control system, by on-line checking batch can 1. with
The pH value of batch can 6., and control enter folding of the tank 1. with batch can 6. soda acid mouthful valve, realize the On-line Control of pH value；Also wrap
Include pump provided with flow sensor ~, pressure sensor and controller, enter in flow sensor and pressure sensor collection pump
The pressure and data on flows of outlet, and transmit to controller, data reception module, processing module and communication are provided with controller
Module, data reception module is transferred to processing module after receiving the data message of two sensor transmissions, finally again via communication
Module transfer is to service terminal；Service terminal receives the data message that controller is sent, and is shown in its display screen.
Embodiment 2 from fermentation system this example demonstrates that separate the technique of Ademetionine
In concentration in 50 ~ 140g/L Ademetionine feed liquid, to add hydrochloric acid solution, mixing adjusts pH value to 1.5-
2.0, and produce precipitation, feed liquid is crossed into membrane material for PVDF, internal diameter is 12mm, 5 ~ 50 DEG C of operating temperature, operating pressure for≤
0.5MPa, membrane aperture is 0.1 μm of milipore filter, and filtrate is with salt acid for adjusting pH value 5.0-6.0；It is PVDF, retention by membrane material
70 ~ 150Da of molecular weight, 5 ~ 50 DEG C of operating temperature, operating pressure carries out nanofiltration to SAM concentration for 4.1 MPa one-level NF membrane
For 200g/L, purity is 99%；Filtrate is 300g/L by secondary nanofiltration, SAM concentration, and purity is 99.8%.
Embodiment 3 in the equipment of embodiment this example demonstrates that implement the situation of present invention process
Take 20m3SAM fermentation treatment fluids, the concentration of Ademetionine is 80g/L, automatically flows into batch can 1. by automatic valve, so
The pH value of feed liquid is adjusted to 1.5 ~ 2.0 with acid automatically by the acid of sour tank 4. by automated system afterwards, is 2. pumped into finally by pump
3. special milipore filter removes to the cell fragment in feed liquid, foreign protein and pigment, while adding pure water automatically by automatic valve
Material is cleaned.6. the permeate of special milipore filter 3. automatically flows into batch can, then by automated system acid
The pH value of feed liquid is automatically adjusted to 5.0 ~ 6.0 with acid, then be 7. pumped into special NF membrane 8. by pump by the soda acid of tank 4., 5., is received
10. 9. the efflux of filter membrane 8. be pumped into two grades of special NF membranes by booster pump again, obtains the g/L of SAM concentration 280 in material,
Purity 99.50%, yield is 89%.
Embodiment 4 in the equipment of embodiment this example demonstrates that implement the situation of present invention process
Take 20m3SAM fermentation treatment fluids, the concentration of Ademetionine is 120g/L, and batch can is automatically flowed into 1. by automatic valve,
Then the sour tank automatic pH value by feed liquid of acid 4. is adjusted to 1.5 ~ 2.0 with acid by automated system, finally by pump 2. pump
Enter special milipore filter 3. to remove the cell fragment in feed liquid, foreign protein and pigment, while adding automatically by automatic valve pure
Water is cleaned to material.6. the permeate of special milipore filter 3. automatically flows into batch can, then passes through automated system handle
The pH value of feed liquid is automatically adjusted to 5.0 ~ 6.0 with acid, then be 7. pumped into special NF membrane 8. by pump by the soda acid of sour tank 4., 5.,
10. 9. the efflux of NF membrane 8. be pumped into two grades of special NF membranes by booster pump again, obtains the g/ of SAM concentration 298 in material
L, purity 99.60%, yield is 89.50%.
Embodiment 5 in the equipment of embodiment this example demonstrates that implement the situation of present invention process
Take 20m3SAM fermentation treatment fluids, the concentration of Ademetionine is 140g/L, and batch can is automatically flowed into 1. by automatic valve,
Then the sour tank automatic pH value by feed liquid of acid 4. is adjusted to 1.5 ~ 2.0 with acid by automated system, finally by pump 2. pump
Enter special milipore filter 3. to remove the cell fragment in feed liquid, foreign protein and pigment, while adding automatically by automatic valve pure
Water is cleaned to material.6. the permeate of special milipore filter 3. automatically flows into batch can, then passes through automated system handle
The pH value of feed liquid is automatically adjusted to 5.0 ~ 6.0 with acid, then be 7. pumped into special NF membrane 8. by pump by the soda acid of sour tank 4., 5.,
10. 9. the efflux of NF membrane 8. be pumped into two grades of special NF membranes by booster pump again, obtains the g/ of SAM concentration 300 in material
L, purity 99.60%, yield is 90%.
Claims (10)
1. a kind of technique that separation obtains Ademetionine from fermentation system, it is characterised in that its step is followed successively by：
（1）By containing SAM zymotic fluid add acid, regulation pH value be 1.5-2, stirring to have precipitation generation；
（2）Mixed fermentation liquid is subjected to ultrafiltration by the tubular ultra-filtration membrane of wide runner；
（3）Acid or alkali are added in material after ultrafiltration transmission, regulation pH value is 5.0-6.0；
（4）PH value ultrafiltration permeate will be mixed up by one-level nanofiltration membrane, and material wash and further with pure water
Concentration；
（5）The concentrate of one-level NF membrane is connected in series through into two grades of NF membranes to be concentrated.
2. technique according to claim 1, it is characterised in that the membrane material of the milipore filter is PVDF, and internal diameter is 12mm,
5 ~ 50 DEG C of operating temperature, operating pressure is≤0.5MPa, and membrane aperture is 0.1 μm；The membrane material of the NF membrane is PVDF, retention
70 ~ 150Da of molecular weight, 5 ~ 50 DEG C of operating temperature, operating pressure is 4.1 MPa.
3. technique according to claim 1, it is characterised in that SAM concentration is 50 ~ 140 g/L in the zymotic fluid.
4. technique according to claim 1, it is characterised in that the acid is one kind in sulfuric acid, hydrochloric acid；The alkali is hydrogen
One kind in sodium oxide molybdena, ammoniacal liquor.
5. a kind of separation from fermentation system obtains the complete automation equipment of Ademetionine, it is characterised in that this is set of
Automation equipment comprises at least two soda acid tanks, two head tanks, three groups of film devices, power-equipment, its connection of each equipment room
Pipeline and valve, the system of Automated condtrol.
6. complete automation equipment according to claim 5, it is characterised in that the equipment contains modular charging bucket device
1., pump 2., milipore filter and membrane module 3., soda acid tank arrangement 4., soda acid tank arrangement 5., charging bucket device 6., pump 7., NF membrane and film
Component 8., booster pump 9., NF membrane and membrane module 10., connecting pipeline, the system of Automated condtrol.
7. equipment according to claim 5, it is characterised in that 1. the charging bucket device, at least contains, the charging positioned at top
Through hole, the through hole that adds water, acid adding through hole plus alkali through hole；Go out material through hole positioned at bottom.
8. equipment according to claim 5, it is characterised in that 6. the charging bucket device, at least contains, the charging positioned at top
Through hole, the through hole that adds water, acid adding through hole plus alkali through hole；Go out material through hole positioned at bottom.
9. equipment according to claim 5, it is characterised in that 1. charging bucket device goes out material through hole and Beng② UNICOMs, pump 2. with
Milipore filter and membrane module 3. charging aperture UNICOM；Charging bucket device 6. with milipore filter and membrane module 3. discharge port end UNICOM；Charging bucket device
6. discharging opening and Beng⑦ UNICOMs；Pump 7. with NF membrane and membrane module 8. charging aperture UNICOM；NF membrane and membrane module 8. discharging opening with
Booster pump 9. UNICOM, booster pump discharge port end and NF membrane and membrane module 10. feed inlet end UNICOM.
10. equipment according to claim 5, it is characterised in that the control system of the automation, including pH value are online
Detection with and control system, by the pH value of on-line checking batch can 1. with batch can 6., and to enter tank 1. 6. sour with batch can for control
The folding of alkali mouthful valve, realizes the On-line Control of pH value；Also include pump provided with flow sensor ~, pressure sensor and
The pressure and data on flows imported and exported in controller, flow sensor and pressure sensor collection pump, and transmit to controller,
Data reception module, processing module and communication module are provided with controller, data reception module two sensor transmissions of reception
Processing module is transferred to after data message, is finally transmitted again via communication module to service terminal；Service terminal receives controller
The data message sent, and it is shown in its display screen.
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| CN201710076801.XA CN107056861B (en) | 2017-02-13 | 2017-02-13 | Process and complete equipment for separating and obtaining ademetionine from fermentation system |
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| CN201710076801.XA CN107056861B (en) | 2017-02-13 | 2017-02-13 | Process and complete equipment for separating and obtaining ademetionine from fermentation system |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107722040A (en) * | 2017-10-10 | 2018-02-23 | 南京志坤环保科技有限公司 | A kind of membrane separating method and device for recycling ceftazidime mother liquor |
| CN110396120A (en) * | 2019-02-13 | 2019-11-01 | 山东惠仕莱生物科技有限公司 | A method of extracting separation s-adenosylmethionine from s-adenosylmethionine fermentation liquid |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1935825A (en) * | 2006-10-13 | 2007-03-28 | 浙江大学 | Method for preparing S-adenosine-L-methionine sulfate |
| CN102747123B (en) * | 2012-07-31 | 2014-08-06 | 无锡福祈制药有限公司 | Process for preparing ademetionine butanedisulfonate |
| CN104178540B (en) * | 2014-07-23 | 2021-05-07 | 浙江震元制药有限公司 | Method for synthesizing ademetionine by biological catalysis method |
-
2017
- 2017-02-13 CN CN201710076801.XA patent/CN107056861B/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107722040A (en) * | 2017-10-10 | 2018-02-23 | 南京志坤环保科技有限公司 | A kind of membrane separating method and device for recycling ceftazidime mother liquor |
| CN110396120A (en) * | 2019-02-13 | 2019-11-01 | 山东惠仕莱生物科技有限公司 | A method of extracting separation s-adenosylmethionine from s-adenosylmethionine fermentation liquid |
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