CN109929896A - A kind of production technology of ursodesoxycholic acid - Google Patents
A kind of production technology of ursodesoxycholic acid Download PDFInfo
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Abstract
The invention discloses a kind of production technologies of ursodesoxycholic acid,, using chenodeoxycholic acid as substrate, the thallus by 7 α-steroid dehydrogenase and/or containing 7 α-steroid dehydrogenase carries out catalysis reaction for it, and resulting product passes through ceramic membrane filter, zymoprotein or thallus are removed, 7- Ketolithocholsaeure is obtained；Again using obtained 7- Ketolithocholsaeure as substrate, thallus by 7 β-steroid dehydrogenase and/or containing 7 β-steroid dehydrogenase carries out catalysis reaction, resulting product passes through ceramic membrane filter, remove zymoprotein or thallus, obtain ursodeoxycholic acid crude, crude product is removed into small molecular protein impurity by ultrafiltration membrane again, ursodesoxycholic acid ultrafiltration membrane clear liquid is obtained, using nanofiltration membrane concentration, resin decolorization, concentration, drying.Compared with prior art, the present invention can improve the quality and yield of ursodesoxycholic acid, reduce production cost.
Description
Technical field
The invention belongs to chemical fields, and in particular to a kind of production technology of ursodesoxycholic acid.
Background technique
Ursodesoxycholic acid, main component are 3a, and 7-5 β of beta-dihydroxy-cholestane-24- acid, are organic compound, odorless,
Bitter, it is readily soluble in ethyl alcohol, it is insoluble in chloroform；It is readily soluble in glacial acetic acid, it is dissolved in sodium hydroxide test solution.Medically for increasing
Add bile acid secretion, and change bile component, reduce bile cholesterol and cholesterol ester, the gallbladder be conducive in gall stone is solid
Alcohol gradually dissolves.Ursodesoxycholic acid is the principle active component of rare Chinese medicine bear gall, and " Chinese Pharmacopoeia " version two is classified as gallbladder
Stone dissolves medicine, and " National essential drugs list (Basic medical and health institutions be equipped with use part) " 2009 editions are classified as disease in the liver and gallbladder
Medication, convas tablet.
Ursodesoxycholic acid is one kind of animal bile acids, other common Cholic acids compounds there are also cholic acid lithocholic acid,
Deoxycholic aicd, chenodeoxycholic acid and hyodesoxycholic acid etc..Wherein, ursodesoxycholic acid and chenodeoxycholic acid epimer each other,
It is exactly that 7- hydroxyl structure configurations are different that the two, which is uniquely distinguished,.7- hydroxyls of ursodesoxycholic acid are beta comfiguration, and chenodeoxycholic acid
7- hydroxyls are a configuration.7- hydroxyls of chenodeoxycholic acid can be oxidized to ketone group, i.e. 7- Ketolithocholsaeure, and 7- ketone groups lead to again
It crosses reduction reaction and is converted to β hydroxyl, i.e. ursodesoxycholic acid.Ursodesoxycholic acid, chenodeoxycholic acid and 7- Ketolithocholsaeure three's molecule
Structural formula is as shown in Figure 1.
Currently, clinically ursodesoxycholic acid is mainly used in the various liver and gallbladder for the treatment of, disease of digestive tract.With molecular biosciences
It learns and ursodesoxycholic acid basis and clinical research is constantly progressive, it has been found that ursodesoxycholic acid is promoting immunological regulation, controlling
Treating coronary heart disease etc. also has positive effect.Therefore, go deep into research, the utility value of ursodesoxycholic acid is increasingly recognized
Know and be valued by people, the demand of ursodesoxycholic acid is also being increased year by year.
But the source of natural bear gall is increasingly reduced, people, which have been working hard, to be sought to obtain bear deoxidation divided by natural bear gall
Alternative except cholic acid.In early days, ursodesoxycholic acid is former for starting by the cholic acid of the separation and Extraction from ox, sheep bile
Material, then by complicated chemical method production, wherein need to generate ursodeoxycholic through redox after chenodeoxycholic acid is made
Acid.But this method and step is more, the production time is long, at high cost.After the eighties in last century, zymetology, microbial fermentation processes hair
Exhibition is got up.With zymetology, microbial fermentation processes by chenodeoxycholic acid isomerization, or bear is produced by lithocholic acid β-hydroxylation and is deoxygenated
Cholic acid, still, this method still has that step is more, the production time is long, defect at high cost.
Its transformation routes of document report mainly have 3 kinds.1. using chenodeoxycholic acid as raw material, through esterification, selective protection
Hydroxyl, Jones reagent oxidation, metallic sodium and Nickel Chloride restore to obtain ursodesoxycholic acid, total recovery 42%.2. with chenodeoxycholic acid
For raw material, in 800 DEG C, the high compound Ni- α-Al2O3 of specificity of selection, ursodeoxycholic is made in catalyst, high pressure, hydrogenating reduction
Acid, total recovery 65%.3. being obtained bear with the reduction of hydrogen-Raney nickel high pressure or electrochemical reduction using 7- Ketolithocholsaeure as raw material and being gone
Oxycholic acid, yield are greater than 90%.Wherein the synthesis of 7- Ketolithocholsaeure can be using chenodeoxycholic acid as raw material, and acetone and water are medium,
It is made through N- bromo-succinimide (NBS) oxidation, yield is up to 89%.1. reaction step is more for route, and route is long, yield compared with
It is low；Route 2. and 3. severe reaction conditions, need high pressure, the high requirements on the equipment, and route 3. in product purity be only 92.5%.
Currently used chemical method produces ursodesoxycholic acid, mainly there is following defect:
(1), cost of investment is high, high production cost；
(2), chemical synthesis needs the conditions such as high temperature, high pressure, and uses a large amount of organic solvents, and production process is dangerous；
(3), reaction step is more, and route is long, and the yield of product is not high；
(4), seriously polluted, environmental issue is severe.
Ye You related scientific research mechanism proposition at present synthesizes ursodesoxycholic acid with whole-cell catalytic, but in catalyzing and synthesizing,
The separation of purpose product and the full cell of catalyst is the obstacle for being difficult to go beyond, uncomfortable because the viscosity of catalysate is very high
Traditional plate compression is preferably used, and more sewage can be brought, the degree of automation is also low；And ultrafiltration membrane is not used to remove removing protein,
A large amount of impurity can be brought to later purification, product quality purity is not high, and needs to consume a large amount of organic solvent.
It is process safety, reliable in view of this, still up for proposing that a kind of production cost is low, invest it is small, product quality and
The new process of high income.
Summary of the invention
The technical problem to be solved by the present invention is in view of the deficiencies of the prior art, provide one kind to extract with membrane separation technique
The technique of ursodesoxycholic acid, to provide a kind of production technology of ursodesoxycholic acid with high purity, yield is high.
In order to reach above-mentioned goal of the invention, the technical solution adopted by the present invention is as follows:
A kind of production technology of ursodesoxycholic acid, it includes the following steps:
(1) chenodeoxycholic acid is urged by 7 α-steroid dehydrogenase or the thallus containing 7 α-steroid dehydrogenase
Change, obtains catalysate A；
(2) the catalysate A for obtaining step (1) obtains the permeate of the Ketolithocholsaeure containing 7- through micro-filtrate membrane filtration；
(3) in the permeate of the Ketolithocholsaeure containing 7- obtained to step (2), 7 β-steroid dehydrogenase is added or contains 7 β-
The thallus of steroid dehydrogenase carries out catalysis reaction, obtains catalysate B；
(4) the catalysate B for obtaining step (3) obtains the permeate containing ursodesoxycholic acid through micro-filtrate membrane filtration；
(5) permeate containing ursodesoxycholic acid for obtaining step (4) passes through ultrafiltration membrance filter, obtains containing ursodesoxycholic acid
Ultrafiltration membrane permeate；
(6) the ultrafiltration membrane permeate containing ursodesoxycholic acid for obtaining step (5) obtains deoxygenating containing bear through nanofiltration membrane
The nanofiltration membrane concentrate of cholic acid；
(7) the nanofiltration membrane concentrate containing ursodesoxycholic acid for obtaining step (6) is contained through ion-exchange resin decolorization
The resin eluent of ursodesoxycholic acid；
(8) the resin eluent containing ursodesoxycholic acid for obtaining step (7) is evaporated, is crystallized to get ursodesoxycholic acid.
Specifically, in step (1), the use of the 7 α-steroid dehydrogenase or the thallus containing 7 α-steroid dehydrogenase
Amount, the volume ratio with chenodeoxycholic acid is that volume ratio is 0.001%~2%, and being catalyzed temperature used is 10-60 DEG C, and the time is
0.5~5h.The effect of 7 α-steroid dehydrogenase is that catalysis chenodeoxycholic acid is converted into 7- Ketolithocholsaeure.
In step (2), the microfiltration membranes are ceramic membrane, and filtering accuracy 500nm, filtration temperature is 80 DEG C, filtration pressure
Power is 0.1MPa.The effect of micro-filtrate membrane filtration is to remove 7 α-steroid dehydrogenase zymoprotein or containing 7 α-steroid dehydrogenase
Thallus.In microfiltration process, when micro-filtrate membrane filtration precision is 5nm, flux is only 500nm filtering accuracy microfiltration membranes
40%, and the driving force for needing 0.8MPa pressure to run as film device；When micro-filtrate membrane filtration precision is 500nm, flux is originated
It is bigger than 50nm by 25%, it is bigger by 20% than the micro-filtration membrane flux of 200nm filtering accuracy, but flux decline ratio is very fast, and has 3% left side
Right high molecular weight protein and pigment penetrates microfiltration membranes, reduces filtrate quality.Filtration temperature is 10~80 DEG C, temperature is preferably 50~
70 DEG C, filter pressure is 0.1~0.8MPa, preferably 0.25~0.4MPa.It further, is 60 DEG C in temperature, pressure is
0.35MPa, when micro-filtrate membrane filtration precision is 50~200nm, filtration flux is larger, and flux decline slowly, and can be concentrated nearly 5 times,
Zymoprotein or 99.9% or more thallus removing rate, filtrate quality is fine.
In step (3), the dosage of the 7 β-steroid dehydrogenase or the thallus containing 7 β-steroid dehydrogenase and contains
The volume ratio of the permeate of 7- Ketolithocholsaeure is 0.001%~2%, and being catalyzed temperature used is 10-60 DEG C, the time is 0.5~
5h.The effect of 7 β-steroid dehydrogenase is that catalysis 7- Ketolithocholsaeure is converted into ursodesoxycholic acid.
In step (4), the microfiltration membranes are ceramic membrane, and filtering accuracy is 5~500nm, and filtration temperature is 10~80 DEG C,
Filter pressure is 0.1~0.8MPa.The effect of micro-filtrate membrane filtration is removal 7 β-steroid dehydrogenase zymoprotein or containing 7 β-class
The thallus of sterol dehydrogenase.
In step (5), the ultrafiltration membrane is rolling ultrafiltration membrane, and molecular cut off is 1000~40000Da, temperature 10
~60 DEG C, pressure is 0.5~2.5MPa.The effect of ultrafiltration membrane is the impurity such as high molecular weight protein, pigment in removal catalysate,
Improve the purity of ursodesoxycholic acid.In ultra-filtration process, when ultrafiltration molecular cut off is 1000Da, flux is only
68% when 20000Da, and the driving force for needing 1.8MPa pressure to run as film device, and ursodeoxycholic acid product can be retained
30%；When ultrafiltration molecular cut off is 40000Da, the small molecular protein and pigment for having 2.5% or so are through microfiltration membranes, drop
Low filtrate quality.Filtration temperature is 10~60 DEG C, and temperature is preferably 30~50 DEG C, and filter pressure is 0.5~2.5MPa, preferably
0.6~1.0MPa.Further, temperature be 35 DEG C, pressure 0.7MPa, ultrafiltration retaining molecular weight be 8000~
When 10000Da, filtration flux is stablized, and can be concentrated nearly 20 times, 99.9% or more small molecular protein removal rate, filtrate liquid quality is very
Good, product recovery rate is up to 96.7%.
In step (6), the nanofiltration membrane be rolling ultrafiltration membrane, molecular cut off be 100~800Da, temperature be 10~
60 DEG C, pressure is 0.5~2.5MPa.The effect of nanofiltration membrane is improved into the dense of the ursodesoxycholic acid of ion exchange resin
Degree reduces resin feeding amount, improves the working efficiency of resin.In nanofiltration process, when nanofiltration retaining molecular weight is 100Da
When, flux is only the 40% of 800Da molecular weight nanofiltration membrane, and the driving force for needing 2.5MPa pressure to run as film device；
When nanofiltration membrane molecular weight is 800Da, flux ratio 300Da is big by 25%, bigger by 40% than the nanofiltration membrane flux of 150Da molecular weight, still
The product for having 5% or so penetrates nanofiltration membrane, product yield decline.Temperature is 10~50 DEG C, and temperature is preferably 30~50 DEG C, pressure
Power is 0.5~2.5MPa, preferably 1.0~2.0MPa.It further, is 30 DEG C, pressure 1.5MPa in temperature, nanofiltration membrane is cut
Stay molecular weight be 150~300Da when, filtration flux stablize, nearly 10 times can be concentrated, 99.5% or more the rejection of product.
In step (7), the ion exchange resin be acrylic type strong-base anion-exchange resin, flow velocity be 2~
6BV/h, preferable flow rate are 3~5BV/h, and temperature is 20-80 DEG C, and preferable temperature is 40~50 DEG C.It is 4BV/h, temperature in flow velocity
When being 50 DEG C, the viscosity of feed liquid is smaller, and the effect of decoloration is best, not only can guarantee decolorizing effect, but also can guarantee higher production effect
Rate, while energy consumption is relatively low, and the yield of ursodesoxycholic acid is more than 99.6%.
The utility model has the advantages that
1, the present invention uses ceramic membrane filter in the production technology of ursodesoxycholic acid, can effectively remove high molecular weight protein or bacterium
Body cell improves product quality.Ceramic membrane can be resistant to high temperature, high pressure, chemical attack, and service life is longer；On the other hand
Also avoid pollution of the solid waste to environment.
2, the present invention uses ultrafiltration membrance filter in the production technology of ursodesoxycholic acid, can effectively remove high molecular weight protein,
The purity of product is greatly improved, the feed loading of ion exchange resin below is reduced, while reducing the dosage of organic solvent.
3, the present invention uses nanofiltration membrane pre-concentration in the production technology of ursodesoxycholic acid, and the dosage of resin is greatly decreased,
The evaporated water for reducing 80% or more simultaneously, reduces production energy consumption, while also reducing production cost.Nanofiltration membrane precision is high,
The yield of ursodesoxycholic acid can be improved.
4, present invention process uses membrane separation plant and ion-exchange unit, reduces the occupied area of equipment, reduces
Capital construction cost has done a large amount of Optimization Work to the parameter of new equipment and traditional handicraft, obtains optimal processing parameter, protects
The energy-efficient operation of production is demonstrate,proved, while the quality of product is higher.The production technology relative energy-saving, compare traditional mode of production
Technique, high degree of automation can save 50% labour cost, remarkable in economical benefits.
Detailed description of the invention
The present invention is done with reference to the accompanying drawings and detailed description and is further illustrated, of the invention is above-mentioned
And/or otherwise advantage will become apparent.
Fig. 1 is ursodesoxycholic acid, chenodeoxycholic acid and 7- Ketolithocholsaeure molecular structural formula.
Fig. 2 is the production technological process of ursodesoxycholic acid of the present invention.
Specific embodiment
According to following embodiments, the present invention may be better understood.
In following embodiment, the enzyme activity determination of 7 α-steroid dehydrogenase is using chenodeoxycholic acid as substrate, 3mL's
Include in reaction mixture: the 0.2mMNAD+ of 2.89mL (50mM kaliumphosphate buffer, pH8.0 in prepare), the 150mM of 10 μ L
Chenodeoxycholic acid, the 100 diluted enzyme solutions of μ L measure light absorption value at 340nm and increase.Unit of enzyme activity's (unit/mL) calculates public
Formula are as follows: [△ A340/ minutes × 3 (mL) × thick enzyme extension rate]/[6.22 × 0.1 (mL)].The enzyme activity of measurement is 205u/mg.
The enzyme activity determination of 7 β-steroid dehydrogenase wraps in the reaction mixture of a 3mL using ursodesoxycholic acid as substrate
Contain: the 0.2mMNADP+ of 2.89mL (50mM kaliumphosphate buffer, pH8.0 in prepare), the 150mM ursodesoxycholic acid of 10 μ L, 100
The diluted enzyme solution of μ L measures light absorption value at 340nm and increases.Unit of enzyme activity (unit/mL) calculation formula are as follows: [△ A340/ points
Clock × 3 (mL) × thick enzyme extension rate]/[6.22 × 0.1 (mL)].The enzyme activity of measurement is 205u/mg.
Embodiment 1
Ursodesoxycholic acid is prepared according to flow chart as shown in Figure 1:
(1) chenodeoxycholic acid is catalyzed by 7 α-steroid dehydrogenase, additive amount is chenodeoxycholic acid volume
0.001%, catalytic temperature is 20 DEG C, obtains catalysate A；
(2) by catalysate A that step (1) obtains through micro-filtrate membrane filtration (microfiltration membranes are ceramic membrane, filtering accuracy 5nm,
Filtration temperature is 20 DEG C, filter pressure 0.8MPa), obtain the permeate of the Ketolithocholsaeure containing 7-；
(3) in the permeate of the Ketolithocholsaeure containing 7- obtained to step (2), 7 β-steroid dehydrogenase is added, additive amount is
The 0.001% of the permeate volume of the Ketolithocholsaeure containing 7-, catalytic temperature are 20 DEG C, carry out catalysis reaction, obtain catalysate B；
(4) by catalysate B that step (3) obtains through micro-filtrate membrane filtration (microfiltration membranes are ceramic membrane, filtering accuracy 5nm,
Filtration temperature is 20 DEG C, filter pressure 0.8MPa), obtain the permeate containing ursodesoxycholic acid；
(5) by ultrafiltration membrance filter, (ultrafiltration membrane is roll-to-roll ultrafiltration to the permeate containing ursodesoxycholic acid for obtaining step (4)
Film, molecular cut off 40000Da, temperature are 10~60 DEG C, pressure 0.5MPa), obtain the ultrafiltration membrane containing ursodesoxycholic acid
Permeate；
(6) through nanofiltration membrane, (nanofiltration membrane is rolling to the ultrafiltration membrane permeate containing ursodesoxycholic acid for obtaining step (5)
Ultrafiltration membrane, molecular cut off 800Da, temperature are 10 DEG C, pressure 2.5MPa), it is dense to obtain the nanofiltration membrane containing ursodesoxycholic acid
Contracting liquid；
(7) the nanofiltration membrane concentrate containing ursodesoxycholic acid for obtaining step (6) is through ROHM AND HAAS AMBERLITE
FPA98Cl ion-exchange resin decolorization (acrylic type strong-base anion-exchange resin, flow velocity 2BV/h, temperature are 20 DEG C),
Obtain the resin eluent containing ursodesoxycholic acid；
(8) the resin eluent containing ursodesoxycholic acid for obtaining step (7) is evaporated, is crystallized to get ursodesoxycholic acid.
The yield for the ursodesoxycholic acid that the present embodiment obtains is 98.6%, and the purity of ursodesoxycholic acid is 96.6%, product
The content of middle albumen is 3.1%, pigment content 0.3%, and ceramic membrane is because of filtering accuracy height, and temperature is low, and flux is smaller, filtering
Time is longer.Because protein content is higher, cause the flux of nanofiltration membrane also smaller.
Embodiment 2
Ursodesoxycholic acid is prepared according to flow chart as shown in Figure 1:
(1) chenodeoxycholic acid is catalyzed by the thallus containing 7 α-steroid dehydrogenase, additive amount is chenodeoxycholic
The 2% of sour volume, catalytic temperature are 60 DEG C, obtain catalysate A；
(2) the catalysate A for obtaining step (1) is through micro-filtrate membrane filtration (filtering accuracy 500nm, filtration temperature 80
DEG C, filter pressure 0.1MPa), obtain the permeate of the Ketolithocholsaeure containing 7-；
(3) in the permeate of the Ketolithocholsaeure containing 7- obtained to step (2), the bacterium containing 7 β-steroid dehydrogenase is added
Body, additive amount are the 2% of the permeate volume of the Ketolithocholsaeure containing 7-, and catalytic temperature is 60 DEG C, carry out catalysis reaction, are catalyzed
Product B；
(4) through micro-filtrate membrane filtration, (microfiltration membranes are ceramic membrane to the catalysate B for obtaining step (3), and filtering accuracy is
50nm, filtration temperature are 80 DEG C, filter pressure 0.1MPa), obtain the permeate containing ursodesoxycholic acid；
(5) by ultrafiltration membrance filter, (ultrafiltration membrane is roll-to-roll ultrafiltration to the permeate containing ursodesoxycholic acid for obtaining step (4)
Film, molecular cut off 1000Da, temperature are 60 DEG C, pressure 2.5MPa), it obtains the ultrafiltration membrane containing ursodesoxycholic acid and penetrates
Liquid；
(6) through nanofiltration membrane, (nanofiltration membrane is rolling to the ultrafiltration membrane permeate containing ursodesoxycholic acid for obtaining step (5)
Ultrafiltration membrane, molecular cut off 100Da, temperature are 10 DEG C, pressure 2.5MPa), it is dense to obtain the nanofiltration membrane containing ursodesoxycholic acid
Contracting liquid；
(7) the nanofiltration membrane concentrate containing ursodesoxycholic acid for obtaining step (6) is through ROHM AND HAAS AMBERLITE
FPA98Cl ion-exchange resin decolorization (acrylic type strong-base anion-exchange resin, flow velocity 6BV/h, temperature are 60 DEG C),
Obtain the resin eluent containing ursodesoxycholic acid；
(8) the resin eluent containing ursodesoxycholic acid for obtaining step (7) is evaporated, is crystallized to get ursodesoxycholic acid.
The yield for the ursodesoxycholic acid that the present embodiment obtains is 90.1%, and the purity of ursodesoxycholic acid is 99.6%, product
The content of middle albumen is 0.2%, pigment content 0.2%；The flux of ceramic membrane is larger, but because ultrafiltration retaining molecular weight compared with
It is small, a part of ursodesoxycholic acid is retained, causes yield lower.
Embodiment 3
Ursodesoxycholic acid is prepared according to flow chart as shown in Figure 1:
(1) chenodeoxycholic acid is catalyzed by 7 α-steroid dehydrogenase, additive amount is chenodeoxycholic acid volume
0.05%, catalytic temperature is 20 DEG C, obtains catalysate A；
(2) through micro-filtrate membrane filtration, (microfiltration membranes are ceramic membrane to the catalysate A for obtaining step (1), and filtering accuracy is
200nm, filtration temperature are 20 DEG C, filter pressure 0.2MPa), obtain the permeate of the Ketolithocholsaeure containing 7-；
(3) in the permeate of the Ketolithocholsaeure containing 7- obtained to step (2), 7 β-steroid dehydrogenase is added, additive amount is
The 0.05% of the permeate volume of the Ketolithocholsaeure containing 7-, catalytic temperature are 20 DEG C, carry out catalysis reaction, obtain catalysate B；
(4) through micro-filtrate membrane filtration, (microfiltration membranes are ceramic membrane to the catalysate B for obtaining step (3), and filtering accuracy is
200nm, filtration temperature are 20 DEG C, filter pressure 0.2MPa), obtain the permeate containing ursodesoxycholic acid；
(5) by ultrafiltration membrance filter, (ultrafiltration membrane is roll-to-roll ultrafiltration to the permeate containing ursodesoxycholic acid for obtaining step (4)
Film, molecular cut off 20000Da, temperature are 40 DEG C, pressure 0.8MPa), it obtains the ultrafiltration membrane containing ursodesoxycholic acid and penetrates
Liquid；
(6) through nanofiltration membrane, (nanofiltration membrane is rolling to the ultrafiltration membrane permeate containing ursodesoxycholic acid for obtaining step (5)
Ultrafiltration membrane, molecular cut off 300Da, temperature are 40 DEG C, pressure 2.0MPa), it is dense to obtain the nanofiltration membrane containing ursodesoxycholic acid
Contracting liquid；
(7) the nanofiltration membrane concentrate containing ursodesoxycholic acid for obtaining step (6) is through ROHM AND HAAS AMBERLITE
FPA98Cl ion-exchange resin decolorization (acrylic type strong-base anion-exchange resin, flow velocity 4BV/h, temperature are 40 DEG C),
Obtain the resin eluent containing ursodesoxycholic acid；
(8) the resin eluent containing ursodesoxycholic acid for obtaining step (7) is evaporated, is crystallized to get ursodesoxycholic acid.
The yield for the ursodesoxycholic acid that the present embodiment obtains is 99.3%, and the purity of ursodesoxycholic acid is 98.3%, product
The content 1.5% of middle albumen, pigment content 0.2%；The filtration temperature and pressure of ceramic membrane are lower, cause filtration flux inclined
Low, ultrafiltration retaining molecular weight is larger, has part small molecular protein transmission, causes rear end nanofiltration membrane flux lower, final products
Protein content it is higher.
Embodiment 4
Ursodesoxycholic acid is prepared according to flow chart as shown in Figure 1:
(1) chenodeoxycholic acid is catalyzed through the thallus containing 7 α-steroid dehydrogenase, additive amount is chenodeoxycholic acid
The 0.5% of volume, catalytic temperature are 50 DEG C, obtain catalysate A；
(2) through micro-filtrate membrane filtration, (microfiltration membranes are ceramic membrane to the catalysate A for obtaining step (1), and filtering accuracy is
50nm, filtration temperature are 50 DEG C, filter pressure 0.6MPa), obtain the permeate of the Ketolithocholsaeure containing 7-；
(3) in the permeate of the Ketolithocholsaeure containing 7- obtained to step (2), the bacterium containing 7 β-steroid dehydrogenase is added
Body, additive amount are the 0.5% of the permeate volume of the Ketolithocholsaeure containing 7-, and catalytic temperature is 50 DEG C, carry out catalysis reaction, are urged
Change product B；
(4) through micro-filtrate membrane filtration, (microfiltration membranes are ceramic membrane to the catalysate B for obtaining step (3), and filtering accuracy is
50nm, filtration temperature are 50 DEG C, filter pressure 0.6MPa), obtain the permeate containing ursodesoxycholic acid；
(5) by ultrafiltration membrance filter, (ultrafiltration membrane is roll-to-roll ultrafiltration to the permeate containing ursodesoxycholic acid for obtaining step (4)
Film, molecular cut off 10000Da, temperature are 30 DEG C, pressure 1.0MPa), it obtains the ultrafiltration membrane containing ursodesoxycholic acid and penetrates
Liquid；
(6) through nanofiltration membrane, (nanofiltration membrane is rolling to the ultrafiltration membrane permeate containing ursodesoxycholic acid for obtaining step (5)
Ultrafiltration membrane, molecular cut off 300Da, temperature are 30 DEG C, pressure 2.5MPa), it is dense to obtain the nanofiltration membrane containing ursodesoxycholic acid
Contracting liquid；
(7) the nanofiltration membrane concentrate containing ursodesoxycholic acid for obtaining step (6) is through ROHM AND HAAS AMBERLITE
FPA98Cl ion-exchange resin decolorization (acrylic type strong-base anion-exchange resin, flow velocity 2BV/h, temperature are 50 DEG C),
Obtain the resin eluent containing ursodesoxycholic acid；
(8) the resin eluent containing ursodesoxycholic acid for obtaining step (7) is evaporated, is crystallized to get ursodesoxycholic acid.
The yield for the ursodesoxycholic acid that the present embodiment obtains is 99.3%, and the purity of ursodesoxycholic acid is 99.7%, product
The content of middle albumen is 0.15%, pigment content 0.05%；The flux of the ceramic membrane of the product is larger, the albumen and color of product
Cellulose content is very low, and quality is higher, but the filter pressure of ceramic membrane is higher, and the molecular cut off of nanofiltration membrane is lower, causes filtration pressure
Power is also high, and the flow velocity of resin is low, and the resin consumptions of unit production capacity are big, and production energy consumption is higher.
Embodiment 5
Ursodesoxycholic acid is prepared according to flow chart as shown in Figure 1:
(1) chenodeoxycholic acid is catalyzed by the thallus containing 7 α-steroid dehydrogenase, additive amount is chenodeoxycholic
The 1% of sour volume, catalytic temperature are 40 DEG C, obtain catalysate A；
(2) through micro-filtrate membrane filtration, (microfiltration membranes are ceramic membrane to the catalysate A for obtaining step (1), and filtering accuracy is
50nm, filtration temperature are 60 DEG C, filter pressure 0.35MPa), obtain the permeate of the Ketolithocholsaeure containing 7-；
(3) in the permeate of the Ketolithocholsaeure containing 7- obtained to step (2), the bacterium containing 7 β-steroid dehydrogenase is added
Body, additive amount are the 1% of the permeate volume of the Ketolithocholsaeure containing 7-, and catalytic temperature is 40 DEG C, carry out catalysis reaction, are catalyzed
Product B；
(4) through micro-filtrate membrane filtration, (microfiltration membranes are ceramic membrane to the catalysate B for obtaining step (3), and filtering accuracy is
50nm, filtration temperature are 60 DEG C, filter pressure 0.35MPa), obtain the permeate containing ursodesoxycholic acid；
(5) by ultrafiltration membrance filter, (ultrafiltration membrane is roll-to-roll ultrafiltration to the permeate containing ursodesoxycholic acid for obtaining step (4)
Film, molecular cut off 10000Da, temperature are 35 DEG C, pressure 0.7MPa), it obtains the ultrafiltration membrane containing ursodesoxycholic acid and penetrates
Liquid；
(6) through nanofiltration membrane, (nanofiltration membrane is rolling to the ultrafiltration membrane permeate containing ursodesoxycholic acid for obtaining step (5)
Ultrafiltration membrane, molecular cut off 150Da, temperature are 30 DEG C, pressure 1.5MPa), it is dense to obtain the nanofiltration membrane containing ursodesoxycholic acid
Contracting liquid；
(7) the nanofiltration membrane concentrate containing ursodesoxycholic acid for obtaining step (6) is through ion-exchange resin decolorization (acrylic acid
Type strong-base anion-exchange resin, flow velocity 4BV/h, temperature are 50 DEG C), obtain the resin eluent containing ursodesoxycholic acid；
(8) the resin eluent containing ursodesoxycholic acid for obtaining step (7) is evaporated, is crystallized to get ursodesoxycholic acid.
The yield for the ursodesoxycholic acid that the present embodiment obtains is 99.7%, and the purity of ursodesoxycholic acid is 99.8%, product
The content of middle albumen is 0.08%, pigment content 0.07%；The flux of the ceramic membrane of the product is larger, the albumen and color of product
Cellulose content is very low, and quality is higher.
Comparative example 1
It by 7- ketone deoxycholic aicd, alcohols solvent and nano Pd particle/C catalyst, stirs evenly, hydrogen will be full of in reaction kettle,
8h is reacted under the conditions of pressure 20MPa, 90 DEG C of temperature to get reaction solution using evaporation, is concentrated to get ursodeoxycholic acid product,
Its yield is 78.3%, and the purity of ursodesoxycholic acid is 95.2%, and the content of albumen is 4.2% in product, and pigment content is
0.6%；.The production technology needs to carry out hydrogenation catalyst reaction at 20MPa, and working condition is harsher, dangerous, and pollutes
Environment is serious.
The present invention provides a kind of thinking of the production technology of ursodesoxycholic acid and methods, implement the technical solution
There are many method and approach, the above is only a preferred embodiment of the present invention, it is noted that for the common of the art
For technical staff, various improvements and modifications may be made without departing from the principle of the present invention, these are improved and profit
Decorations also should be regarded as protection scope of the present invention.The available prior art of each component part being not known in the present embodiment is subject to reality
It is existing.
Claims (8)
1. a kind of production technology of ursodesoxycholic acid, which is characterized in that it includes the following steps:
(1) chenodeoxycholic acid is catalyzed by 7 α-steroid dehydrogenase and/or the thallus containing 7 α-steroid dehydrogenase,
Obtain catalysate A；
(2) the catalysate A for obtaining step (1) obtains the permeate of the Ketolithocholsaeure containing 7- through micro-filtrate membrane filtration；
(3) in the permeate of the Ketolithocholsaeure containing 7- obtained to step (2), 7 β-steroid dehydrogenase is added and/or contains 7 β-
The thallus of steroid dehydrogenase carries out catalysis reaction, obtains catalysate B；
(4) the catalysate B for obtaining step (3) obtains the permeate containing ursodesoxycholic acid through micro-filtrate membrane filtration；
(5) permeate containing ursodesoxycholic acid for obtaining step (4) passes through ultrafiltration membrance filter, obtains super containing ursodesoxycholic acid
Filter membrane permeate；
(6) the ultrafiltration membrane permeate containing ursodesoxycholic acid for obtaining step (5) is obtained through nanofiltration membrane containing ursodesoxycholic acid
Nanofiltration membrane concentrate；
(7) the nanofiltration membrane concentrate containing ursodesoxycholic acid for obtaining step (6) obtains going containing bear through ion-exchange resin decolorization
The resin eluent of oxycholic acid；
(8) the resin eluent containing ursodesoxycholic acid for obtaining step (7) is evaporated, is crystallized to get ursodesoxycholic acid.
2. the production technology of ursodesoxycholic acid according to claim 1, which is characterized in that in step (1), 7 α-
The volume ratio of steroid dehydrogenase enzyme dosage and chenodeoxycholic acid is 0.001%~2%, and being catalyzed temperature used is 10-60 DEG C, when
Between be 0.5~5h.
3. the production technology of ursodesoxycholic acid according to claim 1, which is characterized in that in step (2), the micro-filtration
Film is ceramic membrane, and filtering accuracy is 5~500nm, and filtration temperature is 10~80 DEG C, and filter pressure is 0.1~0.8MPa.
4. the production technology of ursodesoxycholic acid according to claim 1, which is characterized in that in step (3), 7 β-
The volume ratio of the permeate of steroid dehydrogenase enzyme dosage and the Ketolithocholsaeure containing 7- is 0.001%~2%, is catalyzed temperature used and is
10-60 DEG C, the time is 0.5~5h.
5. the production technology of ursodesoxycholic acid according to claim 1, which is characterized in that in step (4), the micro-filtration
Film is ceramic membrane, and filtering accuracy is 5~500nm, and filtration temperature is 10~80 DEG C, and filter pressure is 0.1~0.8MPa.
6. the production technology of ursodesoxycholic acid according to claim 1, which is characterized in that in step (5), the ultrafiltration
Film is rolling ultrafiltration membrane, and molecular cut off is 1000~40000Da, and temperature is 10~60 DEG C, and pressure is 0.5~2.5MPa.
7. the production technology of ursodesoxycholic acid according to claim 1, which is characterized in that in step (6), the nanofiltration
Film is rolling ultrafiltration membrane, and molecular cut off is 100~800Da, and temperature is 10~60 DEG C, and pressure is 0.5~2.5MPa.
8. the production technology of ursodesoxycholic acid according to claim 1, which is characterized in that in step (7), the ion
Exchanger resin is acrylic type strong-base anion-exchange resin, and flow velocity is 2~6BV/h, and temperature is 20-80 DEG C.
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