CN107915749A - A kind of method of purification of cephalosporin C zymotic fluids - Google Patents
A kind of method of purification of cephalosporin C zymotic fluids Download PDFInfo
- Publication number
- CN107915749A CN107915749A CN201610888216.5A CN201610888216A CN107915749A CN 107915749 A CN107915749 A CN 107915749A CN 201610888216 A CN201610888216 A CN 201610888216A CN 107915749 A CN107915749 A CN 107915749A
- Authority
- CN
- China
- Prior art keywords
- tank
- ceramic membrane
- filtrate
- nanofiltration
- ultrafiltration
- 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.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
- C07D501/14—Compounds having a nitrogen atom directly attached in position 7
- C07D501/16—Compounds having a nitrogen atom directly attached in position 7 with a double bond between positions 2 and 3
- C07D501/20—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids
- C07D501/24—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids with hydrocarbon radicals, substituted by hetero atoms or hetero rings, attached in position 3
- C07D501/26—Methylene radicals, substituted by oxygen atoms; Lactones thereof with the 2-carboxyl group
- C07D501/28—Methylene radicals, substituted by oxygen atoms; Lactones thereof with the 2-carboxyl group with the 7-amino radical acylated by an aliphatic carboxylic acid, which is substituted by hetero atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
- C07D501/02—Preparation
- C07D501/12—Separation; Purification
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
Abstract
The invention discloses a kind of method of purification of cephalosporin C zymotic fluids, comprise the following steps：(1) Cephalosporin C fermentation liquid is taken, keeps 5~25 DEG C of broth temperature, adjusts pH=2.2~3.2, adds 1 ‰~4 ‰ v/v formaldehyde, 1 ‰~5 ‰ w/w oxalic acid, are uniformly mixed；(2) ceramic membrane is crossed；(3) ultrafiltration；(4) nanofiltration, takes filtrate.The method of the present invention is easy, efficient, easy to operate, solve thes problems, such as that existing method exists, suitable for promoting and applying.
Description
Technical field
The present invention relates to a kind of method of purification of cephalosporin C zymotic fluids.
Background technology
In the method for existing purification of cephalosporin C zymotic fluids, enterprise is mostly using plate-type hyperfiltration, sheet frame, interval ceramic membrane
Deng filter type, the degraded of feed liquid process feed liquid potency is fast, and the distribution fluctuation of feed liquid potency is larger, to follow-up resin separation purification mistake
Journey produces certain influence, and then may cause large effect to cephalosporin final product quality.Also, preceding method is in life
It is also big there are labor intensity during production, the drawbacks such as work situation is severe.
Therefore, it is necessary to explore a kind of easy, method of efficiently purifying Cephalosporin C fermentation liquid.
The content of the invention
To solve the above problems, the present invention provides a kind of method of purification of cephalosporin C zymotic fluids, including following step
Suddenly：
(1) take Cephalosporin C fermentation liquid, keep 5~25 DEG C of broth temperature, adjust pH=2.2~3.2, add 1 ‰~
4 ‰ formaldehyde, 1 ‰~5 ‰ oxalic acid, are uniformly mixed；
(2) ceramic membrane is crossed；
(3) ultrafiltration；
(4) nanofiltration, takes filtrate.
Further, in step (1), the temperature of the zymotic fluid is 10~15 DEG C.
Further, in step (1), pH=2.5~2.8 are adjusted.
Further, in step (1), the reagent of the tune pH is sulfuric acid.
Further, the sulfuric acid is the dilute sulfuric acid of concentration 20%.
Further, in step (1), 3 ‰ oxalic acid are added.
Further, the step of further including ultrafiltration after the nanofiltration.
Cephalosporin C fermentation liquid of the present invention is the zymotic fluid that this area conventional method is prepared, such as under
State method：
After inclined-plane seed is inoculated into first class seed pot, temperature control cultivates 80~100h, by sterile pipes 25~35
Secondary seed tank is moved into, temperature control cultivates 40~80h 25~35, passes through sterile pipes and moves into three-level seeding tank, three-level kind
Sub- tank temperature control is in 25~35,40~80h of culture, then moves into the big tank that ferments, big tank (volume 500 of fermenting by sterile pipes
Cubic meter) control temperature 25~35, zymotic fluid is put into extraction after cultivating 100~130h.Resisting except low concentration in zymotic fluid
Raw element is outer, also has substantial amounts of undecomposed and utilizes complete culture medium, a large amount of other impurities produced during bacterial metabolism.Training
The main component for supporting base is agricultural and sideline product starch, corn pulp, beancake powder etc..
Compared with existing technique, present invention process tool has the advantage that：
1st, suitable process conditions are found, feed liquid is degraded in production most slow；
2nd, the further ultrafiltration of concentrate after nanofiltration, improves feed liquid quality；
3rd, nanofiltration water-dialyzing is used to ceramic membrane moisturizing, is reduced blowdown, is improved yield；
4th, liquid is concentrated by ultrafiltration and continues ceramic membrane filter, improve yield；
5th, ceramic membrane, ultrafiltration, nanofiltration semi-automatic operation, operative employee is easy to operate, and labor intensity is small.
In the present invention, the implication of english abbreviation is as follows：
CPC：Cephalosporin；
D-CPC：Deacetylate cephalosporin；
DO-CPC：Deacetoxycephalosporin C.
Obviously, the above according to the present invention, according to the ordinary technical knowledge and customary means of this area, is not departing from
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The embodiment of form by the following examples, remakes further specifically the above of the present invention
It is bright.But the scope that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to following example.It is all to be based on the above of the present invention
The technology realized belongs to the scope of the present invention.
Brief description of the drawings
Fig. 1 is the process flow chart of the method for the present invention.
Embodiment
In the following embodiments, the specific detection method of indices is as follows：
(1) detection of D-CPC, DO-CPC and CPC
It is as follows using HPLC methods, actual conditions：
A. the preparation of reference substance：
The CPC sodium salt reference substances of accurately weighed (24~26mg), add that pH4.75 sodium-acetate buffers are appropriate, and use is ultrasonic-wave assisted
It is molten, it is transferred to after dissolving in 100mL volumetric flasks, with buffer solution constant volume graduation mark, shakes up.The preparation of reference substance should claim two parts, and two
The relative average debiation of response factor after part reference substance sample introduction should be less than 0.5%.
B. the preparation of sample solution：
Sample stoste is diluted according to required extension rate with sodium acetate buffer, makes the concentration after dilution be
Every milliliter about 200 of potency unit, is filtered with the disposable aspiration needle filter of 0.22um and waits sample introduction into sample introduction bottle.
C. the preparation of zymotic fluid and acidifying solution
After sample is stirred evenly with glass bar, precision weighs 1g into 200mL volumetric flasks, with sodium-acetate buffer constant volume
To graduation mark, shake up, filtered with the disposable aspiration needle filter of 0.22um and wait sample introduction into sample introduction bottle.
D. chromatography, sample introduction 10ul will be injected after reference substance solution and sample solution filtering respectively, instrument is analyzed, analysis
After, record peak area.
E. the measure of density
Zymotic fluid or acidifying solution are put into ultrasonic cleaner ultrasonic 5 minutes to remove bubble, then the capacity with 50mL
Accurate zymotic fluid or the acidifying solution of measuring of bottle is to graduation mark, its accurately weighed quality
F. result is with calculating：
CPC contents (ug/ml)=standard items response factor × AX×DX×1000 (2)
In formula：AX：Sample peak area；
DX：The extension rate of sample；
1000：Reduced unit is ug/mL；
m：The weight (g) of zymotic fluid or acidifying solution；
ρ：The density of zymotic fluid or acidifying solution (g/mL)；
D-CPC%, DO-CPC%, CPC% obtain a result according to peak area % in chromatogram；
(2) Protein Detection
According to the protein content of sample, the sample solution of 1mL or 5mL is drawn, is specially：
Zymotic fluid, acidifying solution：25 times, suction amount 1mL, Coomassie Brillant Blue solution 5mL of extension rate.
Big super dense filtrate, concentrate：10 times, suction amount 1mL, Coomassie Brillant Blue solution 5mL of extension rate.
Big super dilute filtrate, finished product filtrate：5 times, suction amount 1mL, Coomassie Brillant Blue solution 5mL of extension rate
5mL Coomassie Brillant Blue solutions are added, is placed 5 minutes after mixing, absorbance is measured under 595nm, by absorbance
Value is brought into standard curve calculation formula and is calculated as a result, drawing protein concentration.
The method of the present invention of embodiment 1
Zymotic fluid respectively enters 4 160m through plate heat exchanger (cooling to 10-15 DEG C)3Souring tank, treats souring tank liquid level
When reaching 30%, tank body stirring is opened, while opens -10 DEG C of recirculated waters of tank body, keeps feed temperature not rise.Treat souring tank liquid
When position reaches 50%, start to be slowly added to 3 ‰ (w/w) oxalic acid, add 20% dilute sulfuric acid and be adjusted to pH3.0-3.2, add 1.5 ‰
(v/v) formaldehyde, it is to be mixed uniformly after, acidified pump squeezes into shaking screen, into ceramic membrane head tank, treats that tank inner volume reaches 30%
When, -10 DEG C of recirculated waters of tank body are opened, keep feed temperature not rise.Treat liquid level to 50m3When, squeeze into film through ceramic film raw material pump
Preceding right angle strainer, into ceramic membranous system, (ceramic membranous system, using continuous filtered model, 1-14 groups ceramics film cascade mistake
Filter, and realize online benefit sour water (control production process pH will not rise) method, tubular heat exchanger is equipped with film group, keeps production
Process feed temperature will not rise, and 1-5 group outputs penetrating fluid enters the high unit storage tank of ceramic membrane, and 8-14 groups enter the low list of ceramic membrane
Position storage tank.6th, 7 groups of high and low unit storage tanks can switch at any time, and output bacteria residue gets in and squeezes into big bacterium through pump after small bacteria residue tank
Slag ladle, then squeeze into spray drying workshop by bacteria residue pump and handled)；The high unit filtrate of ceramic membrane is squeezed into by ultrafiltration charging pump
Nine bag filters (50 μ) arrive film group cartridge filter (25 μ) afterwards, ultrafiltration membrane system are finally entered, after film group is opened in equipment
Cooling system automatically turns on, and ensures that feed temperature does not rise, and sour water is mended using online, to control material liquid pH and concentrate potency,
So that Pollution of Ultrafiltration Membrane is minimum, potency loss smaller, ultrafiltration output dialyzate enters finished product filtrate tank after tubular heat exchanger,
The detection of finished product filtrate is qualified to be extracted by filtrate delivery pump through dilute sulfuric acid and filtrate mixer to resin, and output concentrate enters
Liquid storage tank is concentrated, next group zymotic fluid is applied mechanically, handles again；The low unit filtrate of ceramic membrane squeezes into nine pocket types by nanofiltration charging pump
Filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally to nanofiltration membrane system, cooling system in equipment after film group is opened
Automatically turn on, ensure that production process feed temperature does not rise, nanofiltration dislysate enters sodium thin liquid storage tank, as ceramic membrane moisturizing
With nanofiltration concentrate, which enters to hand over to resin into finished product filtrate tank after the high unit storage tank of ceramic membrane further cleans, to be extracted.
I.e. when acidifying solution pH is transferred between 3.0-3.2, zymotic fluid average index is as shown in table 1 below：
Table 1
The method of the present invention of embodiment 2
Zymotic fluid respectively enters 4 160m through plate heat exchanger (cooling to 10-15 DEG C)3Souring tank, treats souring tank liquid level
When reaching 30%, tank body stirring is opened, while opens -10 DEG C of recirculated waters of tank body, keeps feed temperature not rise.Treat souring tank liquid
When position reaches 50%, start to be slowly added to 3 ‰ oxalic acid, add 20% dilute sulfuric acid and be adjusted to pH2.8-3.0, add ‰ formaldehyde of 1-4, treat
After stirring evenly, acidified pump squeezes into shaking screen, into ceramic membrane head tank, when tank inner volume reaches 30%, open tank body-
10 DEG C of recirculated waters, keep feed temperature not rise.Treat liquid level to 50m3When, squeeze into the preceding right angle of film through ceramic film raw material pump and filter
Device, into ceramic membranous system, (ceramic membranous system, my company use continuous filtered model, 1-14 group ceramic membrane cascade filtrations, and
Realize online benefit sour water (control production process pH will not rise) method, be equipped with tubular heat exchanger in film group, keep production process
Feed temperature will not rise, and 1-5 group outputs penetrating fluid enters the high unit storage tank of ceramic membrane, and 8-14 groups enter the low unit storage of ceramic membrane
Tank, 6,7 groups of high and low unit storage tanks can switch at any time, and output bacteria residue gets in and squeezes into big bacteria residue tank through pump after small bacteria residue tank,
Squeeze into spray drying workshop by bacteria residue pump again to be handled)；The high unit filtrate of ceramic membrane squeezes into nine bags by ultrafiltration charging pump
Formula filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally enters ultrafiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that feed temperature does not rise, my company uses online benefit sour water, to control material liquid pH and concentrate to imitate
Valency, so that Pollution of Ultrafiltration Membrane is minimum, potency loss smaller, ultrafiltration output dialyzate enters finished product filtrate after tubular heat exchanger
Tank, the detection of finished product filtrate is qualified to be extracted by filtrate delivery pump through dilute sulfuric acid and filtrate mixer to resin, and output concentrate
Into concentration liquid storage tank, next group zymotic fluid is applied mechanically, is handled again；The low unit filtrate of ceramic membrane squeezes into nine by nanofiltration charging pump
Bag filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally to nanofiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that production process feed temperature does not rise, and nanofiltration dislysate enters sodium thin liquid storage tank, is mended as ceramic membrane
Water is used, and nanofiltration concentrate, which enters to hand over to resin into finished product filtrate tank after the high unit storage tank of ceramic membrane further cleans, to be extracted.
I.e. when acidifying solution pH is transferred between 2.8-3.0, zymotic fluid average index is as shown in table 2 below：
Table 2
The method of the present invention of embodiment 3
Zymotic fluid respectively enters 4 160m through plate heat exchanger (cooling to 10-15 DEG C)3Souring tank, treats souring tank liquid level
When reaching 30%, tank body stirring is opened, while opens -10 DEG C of recirculated waters of tank body, keeps feed temperature not rise.Treat souring tank liquid
When position reaches 50%, start to be slowly added to 3 ‰ oxalic acid, add 20% dilute sulfuric acid and be adjusted to pH2.5-2.8, add ‰ formaldehyde of 1-4, treat
After stirring evenly, acidified pump squeezes into shaking screen, into ceramic membrane head tank, when tank inner volume reaches 30%, open tank body-
10 DEG C of recirculated waters, keep feed temperature not rise.Treat liquid level to 50m3When, squeeze into the preceding right angle of film through ceramic film raw material pump and filter
Device, into ceramic membranous system, (ceramic membranous system, my company use continuous filtered model, 1-14 group ceramic membrane cascade filtrations, and
Realize online benefit sour water (control production process pH will not rise) method, be equipped with tubular heat exchanger in film group, keep production process
Feed temperature will not rise, and 1-5 group outputs penetrating fluid enters the high unit storage tank of ceramic membrane, and 8-14 groups enter the low unit storage of ceramic membrane
Tank, 6,7 groups of high and low unit storage tanks can switch at any time, and output bacteria residue gets in and squeezes into big bacteria residue tank through pump after small bacteria residue tank,
Squeeze into spray drying workshop by bacteria residue pump again to be handled)；The high unit filtrate of ceramic membrane squeezes into nine bags by ultrafiltration charging pump
Formula filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally enters ultrafiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that feed temperature does not rise, my company uses online benefit sour water, to control material liquid pH and concentrate to imitate
Valency, so that Pollution of Ultrafiltration Membrane is minimum, potency loss smaller, ultrafiltration output dialyzate enters finished product filtrate after tubular heat exchanger
Tank, the detection of finished product filtrate is qualified to be extracted by filtrate delivery pump through dilute sulfuric acid and filtrate mixer to resin, and output concentrate
Into concentration liquid storage tank, next group zymotic fluid is applied mechanically, is handled again；The low unit filtrate of ceramic membrane squeezes into nine by nanofiltration charging pump
Bag filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally to nanofiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that production process feed temperature does not rise, and nanofiltration dislysate enters sodium thin liquid storage tank, is mended as ceramic membrane
Water is used, and nanofiltration concentrate, which enters to hand over to resin into finished product filtrate tank after the high unit storage tank of ceramic membrane further cleans, to be extracted.
I.e. when acidifying solution pH is transferred between 2.5-2.8, zymotic fluid average index is as shown in table 3 below：
Table 3
The method of the present invention of embodiment 4
Zymotic fluid respectively enters 4 160m through plate heat exchanger (cooling to 10-15 DEG C)3Souring tank, treats souring tank liquid level
When reaching 30%, tank body stirring is opened, while opens -10 DEG C of recirculated waters of tank body, keeps feed temperature not rise.Treat souring tank liquid
When position reaches 50%, start to be slowly added to 3 ‰ oxalic acid, add 20% dilute sulfuric acid and be adjusted to pH2.2-2.5, add ‰ formaldehyde of 1-4, treat
After stirring evenly, acidified pump squeezes into shaking screen, into ceramic membrane head tank, when tank inner volume reaches 30%, open tank body-
10 DEG C of recirculated waters, keep feed temperature not rise.Treat liquid level to 50m3When, squeeze into the preceding right angle of film through ceramic film raw material pump and filter
Device, into ceramic membranous system, (ceramic membranous system, my company use continuous filtered model, 1-14 group ceramic membrane cascade filtrations, and
Realize online benefit sour water (control production process pH will not rise) method, be equipped with tubular heat exchanger in film group, keep production process
Feed temperature will not rise, and 1-5 group outputs penetrating fluid enters the high unit storage tank of ceramic membrane, and 8-14 groups enter the low unit storage of ceramic membrane
Tank, 6,7 groups of high and low unit storage tanks can switch at any time, and output bacteria residue gets in and squeezes into big bacteria residue tank through pump after small bacteria residue tank,
Squeeze into spray drying workshop by bacteria residue pump again to be handled)；The high unit filtrate of ceramic membrane squeezes into nine bags by ultrafiltration charging pump
Formula filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally enters ultrafiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that feed temperature does not rise, my company uses online benefit sour water, to control material liquid pH and concentrate to imitate
Valency, so that Pollution of Ultrafiltration Membrane is minimum, potency loss smaller, ultrafiltration output dialyzate enters finished product filtrate after tubular heat exchanger
Tank, the detection of finished product filtrate is qualified to be extracted by filtrate delivery pump through dilute sulfuric acid and filtrate mixer to resin, and output concentrate
Into concentration liquid storage tank, next group zymotic fluid is applied mechanically, is handled again；The low unit filtrate of ceramic membrane squeezes into nine by nanofiltration charging pump
Bag filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally to nanofiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that production process feed temperature does not rise, and nanofiltration dislysate enters sodium thin liquid storage tank, is mended as ceramic membrane
Water is used, and nanofiltration concentrate, which enters to hand over to resin into finished product filtrate tank after the high unit storage tank of ceramic membrane further cleans, to be extracted.
I.e. between acidifying solution pH is transferred to 2.2-2.5, zymotic fluid index is as shown in table 4 below：
Table 4
The screening of 5 oxalic acid addition of embodiment
According to the method for embodiment 1, be separately added into 1 ‰, 2 ‰, 3 ‰, 4 ‰, 5 ‰ oxalic acid, other operations are constant, pair plus
The feed liquid for entering different oxalic acid is detected, as a result as shown in table 5 below：
Table 5
Consider each factor, be preferably added to 3 ‰ oxalic acid.
The screening of 6 broth temperature of embodiment
According to the method for embodiment 1, broth temperature is controlled in 20-25 DEG C, 15-25 DEG C, 10-15 DEG C, 5-10 respectively
DEG C, other operations are constant, and feed liquid is detected, as a result as shown in table 6 below：
Table 6
Consider each factor, preferable temperature is 10-15 DEG C.
The method of the present invention of embodiment 7
Zymotic fluid respectively enters 4 160m through plate heat exchanger (cooling to 10-15 DEG C)3Souring tank, treats souring tank liquid level
When reaching 30%, tank body stirring is opened, while opens -10 DEG C of recirculated waters of tank body, keeps feed temperature not rise.Treat souring tank liquid
When position reaches 50%, start to be slowly added to 3 ‰ oxalic acid, add 20% dilute sulfuric acid and be adjusted to pH2.2-2.5, add ‰ formaldehyde of 1-4, treat
After stirring evenly, acidified pump squeezes into shaking screen, into ceramic membrane head tank, when tank inner volume reaches 30%, open tank body-
10 DEG C of recirculated waters, keep feed temperature not rise.Treat liquid level to 50m3When, squeeze into the preceding right angle of film through ceramic film raw material pump and filter
Device, into ceramic membranous system, (ceramic membranous system, my company use continuous filtered model, 1-14 group ceramic membrane cascade filtrations, and
Realize online benefit sour water (control production process pH will not rise) method, be equipped with tubular heat exchanger in film group, keep production process
Feed temperature will not rise, and 1-5 group outputs penetrating fluid enters the high unit storage tank of ceramic membrane, and 8-14 groups enter the low unit storage of ceramic membrane
Tank, 6,7 groups of high and low unit storage tanks can switch at any time, and output bacteria residue gets in and squeezes into big bacteria residue tank through pump after small bacteria residue tank,
Squeeze into spray drying workshop by bacteria residue pump again to be handled)；The high unit filtrate of ceramic membrane squeezes into nine bags by ultrafiltration charging pump
Formula filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally enters ultrafiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that feed temperature does not rise, my company uses online benefit sour water, to control material liquid pH and concentrate to imitate
Valency, so that Pollution of Ultrafiltration Membrane is minimum, potency loss smaller, ultrafiltration output dialyzate enters finished product filtrate after tubular heat exchanger
Tank, the detection of finished product filtrate is qualified to be extracted by filtrate delivery pump through dilute sulfuric acid and filtrate mixer to resin, and output concentrate
Into concentration liquid storage tank, next group zymotic fluid is applied mechanically, is handled again；The low unit filtrate of ceramic membrane squeezes into nine by nanofiltration charging pump
Bag filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally to nanofiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that production process feed temperature does not rise, and nanofiltration dislysate enters sodium thin liquid storage tank, is mended as ceramic membrane
Water is used, and nanofiltration concentrate is handed over to resin into finished product filtrate tank and extracted.
As a result it is as shown in table 7 below：
Table 7
The method of the present invention of embodiment 8
Zymotic fluid respectively enters 4 160m through plate heat exchanger (cooling to 10-15 DEG C)3Souring tank, treats souring tank liquid level
When reaching 30%, tank body stirring is opened, while opens -10 DEG C of recirculated waters of tank body, keeps feed temperature not rise.Treat souring tank liquid
When position reaches 50%, start to be slowly added to 3 ‰ oxalic acid, add 20% dilute sulfuric acid and be adjusted to pH2.2-2.5, add ‰ formaldehyde of 1-4, treat
After stirring evenly, acidified pump squeezes into shaking screen, into ceramic membrane head tank, when tank inner volume reaches 30%, open tank body-
10 DEG C of recirculated waters, keep feed temperature not rise.4. treat liquid level to 50m3When, squeeze into the preceding right angle of film through ceramic film raw material pump and filter
Device, into ceramic membranous system, (ceramic membranous system, my company use continuous filtered model, 1-14 group ceramic membrane cascade filtrations, and
Realize online benefit sour water (control production process pH will not rise) method, be equipped with tubular heat exchanger in film group, keep production process
Feed temperature will not rise, and 1-5 group outputs penetrating fluid enters the high unit storage tank of ceramic membrane, and 8-14 groups enter the low unit storage of ceramic membrane
Tank, 6,7 groups of high and low unit storage tanks can switch at any time, and output bacteria residue gets in and squeezes into big bacteria residue tank through pump after small bacteria residue tank,
Squeeze into spray drying workshop by bacteria residue pump again to be handled)；The high unit filtrate of ceramic membrane squeezes into nine bags by ultrafiltration charging pump
Formula filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally enters ultrafiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that feed temperature does not rise, my company uses online benefit sour water, to control material liquid pH and concentrate to imitate
Valency, so that Pollution of Ultrafiltration Membrane is minimum, potency loss smaller, ultrafiltration output dialyzate enters finished product filtrate after tubular heat exchanger
Tank, the detection of finished product filtrate is qualified to be extracted by filtrate delivery pump through dilute sulfuric acid and filtrate mixer to resin, and output concentrate
Into concentration liquid storage tank, next group zymotic fluid is applied mechanically, is handled again；The low unit filtrate of ceramic membrane squeezes into nine by nanofiltration charging pump
Bag filter (50 μ) arrives film group cartridge filter (25 μ) afterwards, finally to nanofiltration membrane system, cools down after film group is opened in equipment
System automatically turns on, and ensures that production process feed temperature does not rise, and nanofiltration dislysate enters sodium thin liquid storage tank, is mended as ceramic membrane
Water is used, and nanofiltration concentrate, which enters to hand over to resin into finished product filtrate tank after the high unit storage tank of ceramic membrane further cleans, to be extracted.
It is as shown in table 8 below that nanofiltration concentrate enters output filtrate index after the high unit storage tank of ceramic membrane crosses ultrafiltration again：
Table 8
In conclusion the method for the present invention is easy, efficient, easy to operate, solve the problems, such as that existing method exists, fit
In popularization and application.
Claims (7)
- A kind of 1. method of purification of cephalosporin C zymotic fluids, it is characterised in that：Comprise the following steps：（1）Cephalosporin C fermentation liquid is taken, keeps 5~25 DEG C of broth temperature, adjusts pH=2.2~3.2, adds 1 ‰~4 ‰ v/v Formaldehyde, 1 ‰~5 ‰ w/w oxalic acid, are uniformly mixed；（2）Cross ceramic membrane；（3）Ultrafiltration；（4）Nanofiltration, takes filtrate.
- 2. according to the method described in claim 1, it is characterized in that：Step（1）In, the temperature of the zymotic fluid is 10~15 ℃。
- 3. according to the method described in claim 1, it is characterized in that：Step（1）In, adjust pH=2.5~2.8.
- 4. according to claim 1-3 any one of them methods, it is characterised in that：Step（1）In, the reagent of the tune pH is sulphur Acid.
- 5. according to the method described in claim 4, it is characterized in that：The sulfuric acid is the dilute sulfuric acid of concentration 20%.
- 6. according to claim 1-5 any one of them methods, it is characterised in that：Step（1）In, add 3 ‰ oxalic acid.
- 7. according to right 1-6 any one of them methods, it is characterised in that：The step of ultrafiltration being further included after the nanofiltration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610888216.5A CN107915749A (en) | 2016-10-11 | 2016-10-11 | A kind of method of purification of cephalosporin C zymotic fluids |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610888216.5A CN107915749A (en) | 2016-10-11 | 2016-10-11 | A kind of method of purification of cephalosporin C zymotic fluids |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107915749A true CN107915749A (en) | 2018-04-17 |
Family
ID=61891959
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610888216.5A Pending CN107915749A (en) | 2016-10-11 | 2016-10-11 | A kind of method of purification of cephalosporin C zymotic fluids |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107915749A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111185117A (en) * | 2020-01-03 | 2020-05-22 | 山东润德生物科技有限公司 | Material separation system based on cyclic separation in ceramic membrane separation technology |
| CN111850078A (en) * | 2020-07-21 | 2020-10-30 | 伊犁川宁生物技术有限公司 | Fermentation method of cephalosporin C |
| CN111876462A (en) * | 2020-07-21 | 2020-11-03 | 伊犁川宁生物技术有限公司 | Fermentation method of cephalosporin C |
| CN113234093A (en) * | 2021-04-13 | 2021-08-10 | 伊犁川宁生物技术股份有限公司 | 3-deacetoxy cephalosporin C sodium salt and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0044736A2 (en) * | 1980-07-21 | 1982-01-27 | Fujisawa Pharmaceutical Co., Ltd. | Process for the preparation of deacetylcephalosporin C |
| CN104278071A (en) * | 2013-07-03 | 2015-01-14 | 上海凯鑫分离技术有限公司 | Extraction method of cephalosporin C |
| CN105622559A (en) * | 2016-02-03 | 2016-06-01 | 昆山东大智汇技术咨询有限公司 | Gibberellin fermentation liquid membrane extraction water crystallization process |
| CN105669807A (en) * | 2016-03-08 | 2016-06-15 | 厦门世达膜科技有限公司 | Vitamin B12 fermentation liquid separation, purification and concentration technique |
| CN105732746A (en) * | 2016-03-15 | 2016-07-06 | 昆山东大智汇技术咨询有限公司 | Membrane-method extracting and purifying process of nystatin fermentation liquor |
-
2016
- 2016-10-11 CN CN201610888216.5A patent/CN107915749A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0044736A2 (en) * | 1980-07-21 | 1982-01-27 | Fujisawa Pharmaceutical Co., Ltd. | Process for the preparation of deacetylcephalosporin C |
| CN104278071A (en) * | 2013-07-03 | 2015-01-14 | 上海凯鑫分离技术有限公司 | Extraction method of cephalosporin C |
| CN105622559A (en) * | 2016-02-03 | 2016-06-01 | 昆山东大智汇技术咨询有限公司 | Gibberellin fermentation liquid membrane extraction water crystallization process |
| CN105669807A (en) * | 2016-03-08 | 2016-06-15 | 厦门世达膜科技有限公司 | Vitamin B12 fermentation liquid separation, purification and concentration technique |
| CN105732746A (en) * | 2016-03-15 | 2016-07-06 | 昆山东大智汇技术咨询有限公司 | Membrane-method extracting and purifying process of nystatin fermentation liquor |
Non-Patent Citations (1)
| Title |
|---|
| 盖来兵: "头孢菌素C发酵液的膜过滤工艺研究", 《中国学位论文全文数据库》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111185117A (en) * | 2020-01-03 | 2020-05-22 | 山东润德生物科技有限公司 | Material separation system based on cyclic separation in ceramic membrane separation technology |
| CN111185117B (en) * | 2020-01-03 | 2021-12-14 | 山东润德生物科技有限公司 | Material separation system based on cyclic separation in ceramic membrane separation technology |
| CN111850078A (en) * | 2020-07-21 | 2020-10-30 | 伊犁川宁生物技术有限公司 | Fermentation method of cephalosporin C |
| CN111876462A (en) * | 2020-07-21 | 2020-11-03 | 伊犁川宁生物技术有限公司 | Fermentation method of cephalosporin C |
| CN113234093A (en) * | 2021-04-13 | 2021-08-10 | 伊犁川宁生物技术股份有限公司 | 3-deacetoxy cephalosporin C sodium salt and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107915749A (en) | A kind of method of purification of cephalosporin C zymotic fluids | |
| CN109689191B (en) | Alternate tangential flow rapid harvest | |
| Vogel et al. | Fermentation and biochemical engineering handbook: principles, process design and equipment | |
| US20180030398A1 (en) | System, apparatus and method for the production of cells and/or cell products | |
| US10494421B2 (en) | System, apparatus and method for biomolecules production | |
| US20130115588A1 (en) | Integrated bioreactor and separation system and methods of use therof | |
| CN111770986A (en) | System and method for preparing biomolecules (e.g., viral vaccines) | |
| EP2898059A1 (en) | Disposable bottle reactor tank | |
| ES2864721T3 (en) | Arrangement and continuous process of a bioreactor for the production and capture of a biopolymer | |
| US20070231886A1 (en) | Reactor plant and process for culturing phototropic microorganisms | |
| CN106916726B (en) | A kind of fermentation process coupled with separation system for continuously fermenting on a large scale | |
| CN205062055U (en) | Phytoplankton reation kettle and adopt this reation kettle's continuous culture to put permanent disguise | |
| CN105969651A (en) | Microorganism oscillating cleaning and recycling system | |
| Behrendt et al. | Mass spectrometry: A tool for on-line monitoring of animal cell cultures | |
| RU2710967C9 (en) | Method for detachable-topping fermentation with high cell density | |
| CN106434291A (en) | Device and method for continuous cultivation and production of lysine | |
| CN116515611A (en) | Online sterile cell sampling system and sampling method | |
| CN108977358A (en) | A kind of closed bioreactor and its cell culture processes | |
| CN105803042A (en) | Method for detecting high-concentration microbial agent | |
| CN206635330U (en) | A kind of virion antigen purifies concentrator | |
| JPWO2020168315A5 (en) | ||
| CN206543476U (en) | Cleaning system and apply its liquid microbial process units | |
| CN209113917U (en) | A kind of fermentor device to be put | |
| CN219807951U (en) | Filter equipment for cell culture | |
| EP0242984A1 (en) | Culture device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180417 |