CN106701852A - Method and device for preparing GABA (gamma-aminobutyric acid) by use of bio-enzymolysis method - Google Patents
Method and device for preparing GABA (gamma-aminobutyric acid) by use of bio-enzymolysis method Download PDFInfo
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Abstract
The invention relates to a method and a device for preparing GABA (gamma-aminobutyric acid) by use of a bio-enzymolysis method. The method for preparing the GABA by use of the bio-enzymolysis method comprises the following steps of S100, performing decarboxylic reaction and bio-enzymolysis so as to produce the GABA; S200, performing membrane separation, namely enabling the produced GABA to pass through a filter membrane to enter into filtrate, and glutamate decarboxylase and unreacted L-glutamic acid are intercepted and flow back into a reactor; S300, performing collection, namely collecting the filtrate. According to the method, monosodium glutamate residues are used as raw materials for preparing the GABA through enzymolysis, so that full utilization of resources is realized; the glutamate decarboxylase is capable of continuously achieving enzymolysis of the L-glutamic acid and sodium glutamate in the monosodium glutamate residues so as to produce the GABA, continuously filtering out the GABA produced by bio-enzymolysis by virtue of a membrane separation system, enabling the glutamate decarboxylase and the unreacted L-glutamic acid to flow back into the reactor for further decarboxylic reaction. Since the GABA is filtered out, inhibition on the enzyme activity of the glutamate decarboxylase is reduced; by virtue of the recyclable continuous process, the enzyme utilization efficiency and the enzymolysis efficiency are improved, and a relatively high-purity GABA product can be prepared with low cost and high efficiency.
Description
Technical field
The present invention relates to biological technical field, in particular, be related to it is a kind of can serialization operating biology
Enzymatic isolation method prepares the method and device of GABA.
Background technology
GABA (abbreviation GABA) is a kind of naturally occurring nonprotein amino acid, is lactation
Inhibiting nerve important in animal central nervous systems passes on material, about 30% nervous centralis cynapse portion
Position is with GABA as mediator.When GABA lacks in human body, anxiety, uneasiness, tired, sorrow can be produced
The moods such as worry.
GABA as a kind of bioactivator with important physiological function, human brain cortex,
Played an important role in hippocampus, thalamus, basal ganglion and cerebellum, and have to various functions of body and adjust
Section is acted on.Newest research shows that GABA is removed to be had regulation blood pressure, promote ataraxy, promote brain
Portion's blood flow, enhancement brain vigor, trophic nerve cell, increase growth hormone secretion, strong liver profit kidney, prevention
The multiple efficacies such as fat, promotion alcohol metabolism, improvement climacteric metancholia, thus increasingly cause people
Attention.GABA can both be developed into a kind of medicine with notable pharmacological action, can develop again
As a kind of food with health, prospect is very optimistic.Because the naturally occurring amounts of GABA are low, very
Hardly possible a large amount of extraction from some natural animal-plant tissues separates, and the method for currently acquired GABA is chemical conjunction
Into method and the major class of bioanalysis two, wherein bioanalysis includes plants enriched method, microbe fermentation method and biology enzyme
Three kinds of solution etc..
Chemical synthesis is mainly by pyrrolidones that hydrolysis are obtained under strongly alkaline conditions.This synthesis
Method has chemical residue, and the product for obtaining is not belonging to natural products, is mainly used in chemical industry and medicine
Field, is not suitable for being applied in food.
Plants enriched method in bioanalysis is prepared using the endogenous enzymatic conversion in plant tissue metabolic process
GABA.For example, CN 103181523A patents disclose a kind of paddy endogenous enzyme activation method production
The method of GABA food materials high, in clear and definite rice Endogenous glutamate decarboxylase characteristic and activity
On the basis of regulation mechanism, using glutamate decarboxylase activation technique, the rice of GABA content high is produced
Healthy food；It is raw material with the sprouted unpolished rice rich in GABA, the transaminase of GABA is decomposed using passivation
Embryo GABA highed milled rice is stayed in pretreatment, production；Using containing glutamic acid decarboxylase zymoexciter and special buffering
The natural nutrient solution of system is germinateed；It is raw after being fixed using rice bran, rice plumule as GAD sources
Produce enrichment GABA food ingredient, and apply it to GABA nutrition coordinate ground rice production in.This hair
Bright method is easy, can accelerate food material of the exploitation rich in GABA function factors, can as or use
In the functional food of anti-hypertension, enhancement brain function and liver function.This kind of method is to obtain high level
Certain food of GABA is main purpose, rather than for the purpose of obtaining GABA sterlings.
Microbe fermentation method in bioanalysis is production can produce the microorganism of glutamate decarboxylase (GAD)
Bacterium is fermented, before adding the GABA such as Pidolidone (L-Glu) or L-sodium in the medium
Body thing, during fermented and cultured, the decarboxylation of the GAD produced with microorganism, by Pidolidone/L-
Sodium glutamate is converted into GABA, then isolates and purifies and obtain GABA sterlings；If utilize be saccharomycete,
The food security such as lactic acid bacteria and Aspergillus level microbe can be used for food row by GABA products obtained in ferment
Industry.The acquisition of producing microbial high, the control of fermentation production process, isolating and purifying for GABA all constrain
The large-scale promotion application of the method.
Biologic enzymolysis method is with Pidolidone as substrate, in suitable pH and the reaction system of temperature conditionss
Under, Pidolidone decarboxylation is degraded to GABA products using glutamate decarboxylase.Prepared using the method
Efficiency high, product is more single, it is easy to isolate and purify, and is worth with industrial production higher.But due to paddy
Costly, use cost is higher for propylhomoserin decarboxylase (GAD) price.
The content of the invention
The technical problems to be solved by the invention are, there is provided a kind of biologic enzymolysis method prepares the side of GABA
Method and device, solve utilization ratio and enzymolysis effect that existing biologic enzymolysis method prepares enzyme in the technique of GABA
The problems such as rate is not high, production cost is high.
The present invention in order to solve the above problems, the technical scheme for using for：A kind of biological enzymolysis legal system is provided
The method of standby GABA, comprises the following steps：
S100, decarboxylic reaction：Residue of monosodium glutamate and glutamate decarboxylase are added and is equipped with the reactor of buffer solution
Reaction solution is formed, decarboxylic reaction is carried out at a temperature of 30 DEG C~45 DEG C, biological enzymolysis produce GABA；
S200, UF membrane：A part in reaction solution is pumped into film separation system, the GABA of generation wears
Filter membrane enters in filter liquor, and glutamate decarboxylase and unreacted Pidolidone are trapped and flow back into
In reactor；
S300, collection：The filter liquor is collected, GABA powder is dried to obtain, or after chromatography
It is dried to obtain GABA fine powders.
In the method that the biologic enzymolysis method that the present invention is provided prepares GABA, in the step S100,
The pH of reaction solution is 3.0~5.0, and the mass fraction of residue of monosodium glutamate is 5%~30% in reaction solution, in reaction solution
The mass fraction of glutamate decarboxylase is 0.5%~4%.
In the method that the biologic enzymolysis method that the present invention is provided prepares GABA, in the step S200, will
Reaction solution is continuously pumped into film separation system, or, reaction solution is pumped into UF membrane at interval of 10~30min
System is once.
In the method that the biologic enzymolysis method that the present invention is provided prepares GABA, the step S200 also includes：
S201, one-level UF membrane：Reaction solution is separated using the filter membrane that molecular cut off is 50~500KDa,
The first permeate and the first trapped fluid are obtained, first trapped fluid is flowed back into reactor；
S202, secondary membrane are separated：First permeate is used into the filter membrane that molecular cut off is 100~500Da
Separate, obtain the second permeate and the second trapped fluid, second trapped fluid is flowed back into reactor；
Second permeate is the filter liquor.
In the method that the biologic enzymolysis method that the present invention is provided prepares GABA, in the step S201, enter
Mouth pressure is 0.1MPa~0.35MPa；In the step S202, inlet pressure is 1.0MPa~2.5MPa.
In the method that the biologic enzymolysis method that the present invention is provided prepares GABA, the step S200 and S300
Between also include step：
S205, feed supplement：To adding the buffer solution containing residue of monosodium glutamate in the reactor, in maintenance reaction liquid
The mass fraction of residue of monosodium glutamate is 5%~30%.
The present invention also provides a kind of biologic enzymolysis method and prepares the device of GABA, including be sequentially connected：
Reactor, wherein accommodating reaction solution, there is decarboxylation in the residue of monosodium glutamate and glutamate decarboxylase in reaction solution
Reaction, biological enzymolysis produce GABA；
Film separation system, UF membrane is carried out by the part in reaction solution so that glutamate decarboxylase, L-
The GABA that glutamic acid is produced with biological enzymolysis is separated, and the GABA that biological enzymolysis are produced enters through filter membrane
In filter liquor, it is trapped and flows back into reactor；
Collector, collects the filter liquor, for being dried to obtain GABA powder, or divides for chromatographing
GABA fine powders are dried to obtain after.
In the device that the biologic enzymolysis method that the present invention is provided prepares GABA, the film separation system includes
One-level membrane separation device, secondary membrane separator；
The one-level membrane separation device is separated described anti-using the filter membrane that molecular cut off is 50~500KDa
Answer liquid, obtain the first permeate and the first trapped fluid, the reactor and the one-level membrane separation device it
Between be provided with the first return duct, first trapped fluid flows back into reactor by first return duct
In, first permeate enters temporary storage tank；
The secondary membrane separator separates described the using filter membrane that molecular cut off is 100~500Da
One permeate, obtains the second permeate and the second trapped fluid, and the reactor is filled with second UF membrane
The second return duct is provided between putting, second trapped fluid is flowed back into instead by second return duct
In answering device；Second permeate is the filter liquor, flows into the collector.
In the device that the biologic enzymolysis method that the present invention is provided prepares GABA, the reactor and institute are connected
First circulation pump is provided with the pipeline for stating one-level membrane separation device；The temporary storage tank is connected with described two grades
Second circulation pump is provided with the pipeline of membrane separation device.
It is described also including feeder in the device that the biologic enzymolysis method that the present invention is provided prepares GABA
Feeder accommodates the buffer solution containing residue of monosodium glutamate, is set on the connection feeder and the pipeline of the reactor
It is equipped with feed supplement pump.
Implement the present invention, have the advantages that：Residue of monosodium glutamate is prepared GABA by the present invention
Raw material, realize making full use of for resource；Glutamate decarboxylase can continue by the L- in residue of monosodium glutamate
Glutamic acid and sodium glutamate digest into GABA, and produce biological enzymolysis by the way that film separation system is lasting
GABA leach, glutamate decarboxylase and unreacted Pidolidone then flow back into reactor and continue de-
Carboxylic reacts, and leaches GABA and reduces the enzyme activity of glutamate decarboxylase is suppressed, recyclable serialization
Journey improves the utilization ratio and enzymolysis efficiency of enzyme, inexpensive, efficient to prepare purity GABA higher
Product.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to reality
The accompanying drawing to be used needed for example or description of the prior art is applied to be briefly described, it should be apparent that, below
Accompanying drawing in description is only some embodiments of the present invention, for those of ordinary skill in the art,
On the premise of not paying creative work, other accompanying drawings can also be obtained according to these accompanying drawings.
Fig. 1 is the structural representation of the device preferred embodiment that biologic enzymolysis method of the present invention prepares GABA；
Fig. 2 is the flow chart of the method preferred embodiment that biologic enzymolysis method of the present invention prepares GABA.
Specific embodiment
Below in conjunction with embodiment, the technical scheme in the embodiment of the present invention is clearly and completely retouched
State.
Residue of monosodium glutamate is the accessory substance during glutamate production, will extract the supernatant drying after Pidolidone and be
It is available.Glutamate production process is typically to be inoculated with corynebacterium glutamicum etc. in the substrates such as corn starch sugar
After producing the microorganism of glutamic acid, fermented generation Pidolidone, zymotic fluid is adjusted after thalline is removed with acid
PH separates out most of Pidolidone to the isoelectric point of glutamic acid, then in the Pidolidone alkali for separating out
With obtain sodium glutamate.Separated out the content of Pidolidone in the supernatant of Pidolidone up to 1.2%~
1.5%, even as high as 2%, it is that can obtain above-mentioned residue of monosodium glutamate that will extract the supernatant after Pidolidone and dry.
Pidolidone content is up to more than 20% in residue of monosodium glutamate, sodium glutamate content also up to 1%~5%, but
Residue of monosodium glutamate is generally regarded as cheap raw material as feed or fertilizer, causes a large amount of wastes of resource.
Main innovation point of the invention is to prepare the raw material of GABA, taste as enzymolysis using residue of monosodium glutamate
Smart slag is inexpensive and wide material sources, Pidolidone, sodium glutamate in residue of monosodium glutamate under suitable enzymatic hydrolysis condition
GABA can be generated by decarboxylation, in addition, reaction solution recycle stream between reactor and film separation system
It is dynamic, the GABA of generation is persistently leached to avoid its feedback inhibition to glutamate decarboxylase, realize decarboxylation
Reaction, UF membrane, collect continuous prodution process, and can be lasting by adding raw material continuous metaplasia
Produce, substantially reduce cost, improve efficiency.
Fig. 1 shows that biologic enzymolysis method of the present invention prepares the structural representation of the device preferred embodiment of GABA
Figure, as shown in figure 1, including the reactor 10, film separation system 20, the collection that are sequentially connected by pipeline
Device 30, wherein：
Reactor 10, wherein accommodating reaction solution, the residue of monosodium glutamate and glutamate decarboxylase in reaction solution occur de-
Carboxylic reacts, and biological enzymolysis produce GABA；
Film separation system 20, UF membrane is carried out by the part in reaction solution so that glutamate decarboxylase,
The GABA that Pidolidone is produced with biological enzymolysis is separated, and the GABA that biological enzymolysis are produced enters through filter membrane
Enter in filter liquor, be trapped and flow back into reactor 10；
Collector 30, collects filter liquor, for being dried to obtain GABA powder, or for chromatography
After be dried to obtain GABA fine powders.
In another preferred embodiment of the invention, film separation system 20 include one-level membrane separation device 21,
Secondary membrane separator 22；
One-level membrane separation device 21 separates reaction solution using the filter membrane that molecular cut off is 50~500KDa,
The first permeate and the first trapped fluid are obtained, is provided between reactor 10 and one-level membrane separation device 21
First return duct 11, the first trapped fluid is flowed back into reactor 10 by the first return duct 11, and first
Permeate enters temporary storage tank 23；It is provided with connection reactor 10 and the pipeline of one-level membrane separation device 21
First circulation pump 41；Connect and be provided with second on temporary storage tank 23 and the pipeline of secondary membrane separator 22 and follow
Ring pump 42.
Secondary membrane separator 22 separates first and passes through using the filter membrane that molecular cut off is 100~500Da
Liquid, obtains the second permeate and the second trapped fluid, is set between reactor 10 and secondary membrane separator 22
The second return duct 12 is equipped with, the second trapped fluid is flowed back into reactor 10 by the second return duct 12；
Second permeate is filter liquor, flows into collector 30.
In another preferred embodiment of the invention, the device that biologic enzymolysis method prepares GABA also includes mending
Glassware 50, feeder 50 accommodates the buffer solution containing residue of monosodium glutamate, connection feeder 50 and reactor 10
Feed supplement pump 43 is provided with pipeline.
Fig. 2 shows that biologic enzymolysis method of the present invention prepares the flow chart of the method preferred embodiment of GABA,
As shown in Fig. 2 comprising the following steps：
S100, decarboxylic reaction：Residue of monosodium glutamate and glutamate decarboxylase are added into the reactor 10 equipped with buffer solution
Middle formation reaction solution, carries out decarboxylic reaction at a temperature of 30 DEG C~45 DEG C, and biological enzymolysis produce GABA；
S200, UF membrane：A part in reaction solution is pumped into film separation system 20, the GABA of generation
Enter in filter liquor through filter membrane, glutamate decarboxylase and unreacted Pidolidone are trapped and flow back into
In entering reactor 10；
S300, collection：Filter liquor is collected, GABA powder is dried to obtain, or dried after chromatography
Obtain GABA fine powders.
In the step of the present embodiment S100, Pidolidone, sodium glutamate in residue of monosodium glutamate can be with paddy ammonia
There is decarboxylic reaction generation GABA in acid decarboxylase, but because Pidolidone is sl. sol., be dissolved in anti-
Answer the Pidolidone amount in liquid smaller, limit the enzymolysis efficiency of decarboxylic reaction, the paddy ammonia in residue of monosodium glutamate
Sour sodium serves good supplementary function, therefore more in the incipient stage glutamate decarboxylase of this step
Being combined with sodium glutamate carries out decarboxylic reaction, rapid enzymolysis generation GABA.
In the step of the present embodiment S200, using filter membrane crown_interception by the unreacted substrate in reaction solution,
During glutamate decarboxylase is retained and is flowed back into reactor 10, it is to avoid price glutamic acid higher takes off
The loss of carboxylic acid, improves the utilization ratio of enzyme, and therefore the present invention can also add the glutamic acid of higher concentration
Decarboxylase accelerates the speed of decarboxylic reaction, while ensure that the Glutamic Acid decarboxylase of reactor 10 is maintained
In the range of higher concentration so that maintain enzymolysis efficiency higher in reactor 10 all the time；In addition, lasting filter
Going out GABA can avoid its feedback inhibition to glutamate decarboxylase, further increase glutamic acid decarboxylase
The enzymolysis efficiency of enzyme, fast and efficiently obtains GABA.
In the step of the present embodiment S300, filter liquor can be dried to obtain GABA powder, or pass through
Further chromatography goes the removal of impurity, is dried to obtain purity GABA fine powders higher.
In another preferred embodiment of the invention, in addition to foregoing features, the pH of reaction solution is
3.0~5.0, although the pH for carrying out reaction solution with reaction can change within the specific limits, but buffering
The reaction solution that the presence of liquid ensures is under preferably pH, and in the range of the pH, sodium glutamate can also
Well as the substrate of paddy ammonia decarboxylase, supplement to improve the generation speed of GABA well as one
Degree.The mass fraction of residue of monosodium glutamate is 5%~30%, the quality of reaction solution Glutamic Acid decarboxylase in reaction solution
Fraction is 0.5%~4%, is limited to the solubility of Pidolidone in residue of monosodium glutamate, residue of monosodium glutamate in increase reaction solution
Mass fraction can not improve the meltage of Pidolidone in reaction solution, and this is also usual in common process
The reason for adding the glutamate decarboxylase of low concentration, its is expensive for one side, is lost after first use
The characteristics of abandoning so that larger amount of addition cause waste, on the other hand add larger amount of glutamate decarboxylase
Reaction speed can not be significantly improved；In the present embodiment, film separation system 20 can retain glutamate decarboxylase
And the purpose that reactor 10 is realized recycling is flowed back into, adding larger amount of glutamate decarboxylase will not
Improved according into obvious cost, additionally, due to the presence of residue of monosodium glutamate Glutamic Acid sodium, increase glutamic acid decarboxylase
Enzyme can improve the formation speed of GABA, improve the enzymolysis efficiency of substrate.
In another preferred embodiment of the invention, in addition to foregoing features, will in step S200
The process that reaction solution pumps into film separation system 20 can be continuously pumped into, or intermittence is pumped into.Even
Continuous pumping into refers to the continuous firing of film separation system 20, and reaction solution also persistently pumps into film separation system 20, according to
Film separation system 20 goes out liquid speed rate, is mended in film separation system 20 with the speed suitable with liquid speed rate is gone out
Fill reaction solution, trapped fluid is also lasting to flow back into reactor 10.It refers to every one section that intermittence is pumped into
Time, such as 10~30min, reaction solution is pumped into film separation system 20 once, film separation system 20
Complete to pump into the separation of reaction solution in interval time, and trapped fluid flowed back into reactor 10.Nothing
By being continuously pumped into or discontinuity is pumped into, all to separate GABA, and the glutamate decarboxylase and not anti-of flowing back
For the purpose of the Pidolidone answered.
In another preferred embodiment of the invention, in addition to foregoing features, step S200 is also wrapped
Include：
S201, one-level UF membrane：Reaction solution is separated using the filter membrane that molecular cut off is 50~500KDa,
The first permeate and the first trapped fluid are obtained, the first trapped fluid is flowed back into reactor 10；One-level film point
Retained from by the larger macromolecular substances of insoluble matter, molecular weight in reaction solution, cell fragment etc., i.e.
Pidolidone and the larger glutamate decarboxylase of molecular weight without dissolving are trapped, and GABA is small with other
Molecular substance then enters the first permeate.According to the filter membrane type and equipment selected, filter process goes out liquid speed
Rate can be adjusted accordingly, in common 0.5~5m3Reaction solution system in, one-level UF membrane goes out liquid
Speed is 1.0m3/ h~10m3/ h, inlet pressure is 0.1MPa~0.35MPa.
S202, secondary membrane are separated：First permeate is used into the filter membrane that molecular cut off is 100~500Da
Separate, obtain the second permeate and the second trapped fluid, the second trapped fluid is flowed back into reactor 10；The
Two permeate are filter liquor；Glutamate decarboxylase and some other protein matters are trapped, GABA
Very at least part of other small-molecule substances can enter the second permeate.It is similar, common 0.5~
5m3Reaction solution system in, what secondary membrane was separate goes out liquid speed rate for 0.2m3/ h~2m3/ h, inlet pressure is
1.0MPa~2.5MPa.
In another preferred embodiment of the invention, in addition to foregoing features, step S200 and
Also include step between S300：
S205, feed supplement：To adding the buffer solution containing residue of monosodium glutamate, taste in maintenance reaction liquid in reactor 10
The mass fraction of smart slag is 5%~30%.It is similar with the process that reaction solution pumps into film separation system 20, should
Feed supplement process can also be using being continuously pumped into, or discontinuity is pumped into so that has in reaction solution enough
Reaction substrate is present.In addition, material liquid volume during feed supplement should be suitable with the volume of filter liquor, it is to avoid anti-
Answering the volume of liquid has larger fluctuation.
Embodiment 1
1st, to adding 0.5m in reactor 103PH buffer solutions, open heating and stir, treat temperature liter
During to 30 DEG C, 5% residue of monosodium glutamate and 0.5%GAD, enzyme digestion reaction 10min are added thereto to；
2nd, first circulation pump 41 is opened so that enzyme digestion reaction liquid is continuously pumped into one-level membrane separation device 21,
First permeate enters temporary storage tank 23 from through end, and the circulation fluid containing enzyme and substrate is then from circulation end weight
It is new to return to continuation reaction in reactor 10；The retention molecule of the filter membrane wherein in one-level membrane separation device 21
It is 50KDa to measure, and film aquifer yield is 1.0m3/ h, inlet pressure is 0.35MPa.
3rd, after 10min, second circulation pump 42 is opened, by the continuous pump of the first permeate in temporary storage tank 23
Enter secondary membrane separator 22, the second permeate enters in collector 30 from through end, and contain other compared with
Macromolecular and bivalent ions circulation fluid continue to react in then coming back to reactor 10 from circulation end；Wherein
The molecular cut off of the filter membrane in the second membrane separation device 22 is 100Da, and film aquifer yield is 0.2m3/ h,
Inlet pressure is 2.5MPa.
4th, open feed supplement pump 43, with the second permeate identical flow velocity to added in reactor 10 containing
The buffer solution of 10% residue of monosodium glutamate；
5th, persistent loop reaction about 24h, detects without GABA substantially in the second permeate, terminates reaction,
The material in reactor 10 is released, with water backwash membrane separation device；
6th, the GABA solution in collector 30 is taken out, after resin column chromatography for separation, vacuum concentration,
Spray drying obtains GABA powder, is 98.55% through HPLC detection GABA contents.
Embodiment 2
1st, to adding 5m in reactor 103PH buffer solutions, open heating and stir, treat that temperature rises to
At 37 DEG C, 30% residue of monosodium glutamate and 4%GAD, enzyme digestion reaction 30min are added thereto to；
2nd, first circulation pump 41 is opened so that enzyme digestion reaction liquid intermittence pumps into one-level membrane separation device 21,
First permeate enters temporary storage tank 23 from through end, and the circulation fluid containing enzyme and substrate is then from circulation end weight
It is new to return to continuation reaction in reactor 10；The retention molecule of the filter membrane wherein in one-level membrane separation device 21
It is 500KDa to measure, and film aquifer yield is 10m3/ h, inlet pressure is 0.1MPa.
3rd, after 10min, second circulation pump 42 is opened, the first permeate in temporary storage tank 23 is intermittent
Secondary membrane separator 22 is pumped into, the second permeate enters in collector 30 from through end, and contains other
Bigger molecule and bivalent ions circulation fluid continue to react in then coming back to reactor 10 from circulation end；Its
The molecular cut off of the filter membrane in middle secondary membrane separator 22 is 500Da, and the pure water flux of film is 2m3
/ h, inlet pressure is 1.0MPa.
4th, open feed supplement pump 43, with the second permeate identical flow velocity to added in reactor 10 containing
The buffer solution of 15% residue of monosodium glutamate；
5th, persistent loop reaction about 48h, detects without GABA substantially in the second permeate, terminates reaction,
The material in reactor 10 is released, with water backwash membrane separation device；
6th, the GABA solution in collector 30 is taken out, after vacuum concentration, spray drying obtains GABA
Powder, is 90.43% through HPLC detection GABA contents.
Embodiment 3
1st, to adding 2m in reactor 103PH buffer solutions, open heating and stir, treat that temperature rises to
At 40 DEG C, 20% residue of monosodium glutamate and 1.5%GAD, enzyme digestion reaction 15min are added thereto to；
2nd, first circulation pump 41 is opened so that enzyme digestion reaction liquid is continuously pumped into one-level membrane separation device 21,
First permeate enters temporary storage tank 23 from through end, and the circulation fluid containing enzyme and substrate is then from circulation end weight
It is new to return to continuation reaction in reactor 10；The retention molecule of the filter membrane wherein in one-level membrane separation device 21
It is 300KDa to measure, and film aquifer yield is 2.0m3/ h, inlet pressure is 0.15MPa.
3rd, after 10min, second circulation pump 42 is opened, by the continuous pump of the first permeate in temporary storage tank 23
Enter secondary membrane separator 22, the second permeate enters in collector 30 from through end, and contain other compared with
Macromolecular and bivalent ions circulation fluid continue to react in then coming back to reactor 10 from circulation end；Wherein
The molecular cut off of the filter membrane in secondary membrane separator 22 is 220Da, and the pure water flux of film is 0.4m3
/ h, inlet pressure is 1.2MPa.
4th, open feed supplement pump 43, with the second permeate identical flow velocity to added in reactor 10 containing
The buffer solution of 15% residue of monosodium glutamate；
5th, persistent loop reaction about 36h, detects without GABA substantially in the second permeate, terminates reaction,
The material in reactor 10 is released, with water backwash membrane separation device；
6th, the GABA solution in collector 30 is taken out, after vacuum concentration, spray drying obtains GABA
Powder, is 91.58% through HPLC detection GABA contents.
Obviously, embodiment described above is only a part of embodiment of the invention, rather than whole realities
Apply example.Based on the embodiment in the present invention, those of ordinary skill in the art are not making creative work
On the premise of the every other embodiment that is obtained, belong to the scope of protection of the invention.
Claims (10)
1. a kind of method that biologic enzymolysis method prepares GABA, it is characterised in that comprise the following steps：
S100, decarboxylic reaction：Residue of monosodium glutamate and glutamate decarboxylase are added and is equipped with the reactor of buffer solution
Reaction solution is formed, decarboxylic reaction is carried out at a temperature of 30 DEG C~45 DEG C, biological enzymolysis produce GABA；
S200, UF membrane：A part in reaction solution is pumped into film separation system, the GABA of generation wears
Filter membrane enters in filter liquor, and glutamate decarboxylase and unreacted Pidolidone are trapped and flow back into
In reactor；
S300, collection：The filter liquor is collected, GABA powder is dried to obtain, or after chromatography
It is dried to obtain GABA fine powders.
2. the method that biologic enzymolysis method according to claim 1 prepares GABA, it is characterised in that
In the step S100, the pH of reaction solution is 3.0~5.0, and the mass fraction of residue of monosodium glutamate is in reaction solution
5%~30%, the mass fraction of reaction solution Glutamic Acid decarboxylase is 0.5%~4%.
3. the method that biologic enzymolysis method according to claim 1 prepares GABA, it is characterised in that
In the step S200, reaction solution is continuously pumped into film separation system, or, at interval of 10~30min
Reaction solution is pumped into film separation system once.
4. the method that biologic enzymolysis method according to claim 1 prepares GABA, it is characterised in that
The step S200 also includes：
S201, one-level UF membrane：Reaction solution is separated using the filter membrane that molecular cut off is 50~500KDa,
The first permeate and the first trapped fluid are obtained, first trapped fluid is flowed back into reactor；
S202, secondary membrane are separated：First permeate is used into the filter membrane that molecular cut off is 100~500Da
Separate, obtain the second permeate and the second trapped fluid, second trapped fluid is flowed back into reactor；
Second permeate is the filter liquor.
5. the method that biologic enzymolysis method according to claim 4 prepares GABA, it is characterised in that
In the step S201, inlet pressure is 0.1MPa~0.35MPa；In the step S202, import
Pressure is 1.0MPa~2.5MPa.
6. the method that biologic enzymolysis method according to claim 1 prepares GABA, it is characterised in that
Also include step between the step S200 and S300：
S205, feed supplement：To adding the buffer solution containing residue of monosodium glutamate in the reactor, in maintenance reaction liquid
The mass fraction of residue of monosodium glutamate is 5%~30%.
7. a kind of biologic enzymolysis method prepares the device of GABA, it is characterised in that including what is be sequentially connected：
Reactor (10), wherein accommodating reaction solution, the residue of monosodium glutamate and glutamate decarboxylase in reaction solution occur
Decarboxylic reaction, biological enzymolysis produce GABA；
Film separation system (20), UF membrane is carried out by the part in reaction solution so that glutamate decarboxylase,
The GABA that Pidolidone is produced with biological enzymolysis is separated, and the GABA that biological enzymolysis are produced enters through filter membrane
Enter in filter liquor, glutamate decarboxylase and unreacted substrate are trapped and flow back into reactor (10)
In；
Collector (30), collects the filter liquor, for being dried to obtain GABA powder, or for layer
Analysis is dried to obtain GABA fine powders after separating.
8. biologic enzymolysis method according to claim 7 prepares the device of GABA, it is characterised in that
The film separation system (20) includes one-level membrane separation device (21), secondary membrane separator (22)；
The one-level membrane separation device (21) is separated using the filter membrane that molecular cut off is 50~500KDa
The reaction solution, obtains the first permeate and the first trapped fluid, the reactor (10) and the one-level
The first return duct (11) is provided between separator (21), first trapped fluid passes through described first
Return duct (11) is flowed back into reactor (10), and first permeate enters temporary storage tank (23)；
The secondary membrane separator (22) separates institute using the filter membrane that molecular cut off is 100~500Da
The first permeate is stated, the second permeate and the second trapped fluid, the reactor (10) and described is obtained
The second return duct (12) is provided between two membrane separation devices (22), second trapped fluid is by described
Second return duct (12) is flowed back into reactor (10)；Second permeate is described leaching
Liquid, in the inflow collector (30).
9. biologic enzymolysis method according to claim 8 prepares the device of GABA, it is characterised in that
Connect and be provided with first on the reactor (10) and the pipeline of the one-level membrane separation device (21) and follow
Ring pump (41)；Connect and set on the temporary storage tank (23) and the pipeline of the secondary membrane separator (22)
It is equipped with second circulation pump (42).
10. biologic enzymolysis method according to claim 7 prepares the device of GABA, it is characterised in that
Also include feeder (50), the feeder (50) accommodates the buffer solution containing residue of monosodium glutamate, and connection is described
Feed supplement pump (43) is provided with feeder (50) and the pipeline of the reactor (10).
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| CN107287253A (en) * | 2017-06-28 | 2017-10-24 | 岭南师范学院 | The method that immobilized cell produces gamma aminobutyric acid with D101 resins queen post |
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| CN101974151A (en) * | 2010-10-27 | 2011-02-16 | 华东理工大学 | Method for preparing bio-based nylon polybutyrolactam |
| CN102978250A (en) * | 2012-12-20 | 2013-03-20 | 江苏久吾高科技股份有限公司 | Method for producing Gamma-aminobutyric acid through centrifugal mother liquid of glutamic acid |
| CN204237779U (en) * | 2014-11-12 | 2015-04-01 | 江苏新汉菱生物工程股份有限公司 | A kind of new device producing D-cysteine hydrochloride with enzyme process binding film technology |
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| CN101974151A (en) * | 2010-10-27 | 2011-02-16 | 华东理工大学 | Method for preparing bio-based nylon polybutyrolactam |
| CN102978250A (en) * | 2012-12-20 | 2013-03-20 | 江苏久吾高科技股份有限公司 | Method for producing Gamma-aminobutyric acid through centrifugal mother liquid of glutamic acid |
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