CN106278952B - One kind arginic techniques of separation and Extraction L from zymotic fluid - Google Patents
One kind arginic techniques of separation and Extraction L from zymotic fluid Download PDFInfo
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- CN106278952B CN106278952B CN201610655175.5A CN201610655175A CN106278952B CN 106278952 B CN106278952 B CN 106278952B CN 201610655175 A CN201610655175 A CN 201610655175A CN 106278952 B CN106278952 B CN 106278952B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C277/00—Preparation of guanidine or its derivatives, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups
- C07C277/08—Preparation of guanidine or its derivatives, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups of substituted guanidines
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Abstract
The invention belongs to technical field of amino acid production, discloses one kind arginic techniques of separation and Extraction L from zymotic fluid, and the technique includes micro-filtration, the flocculation step such as miscellaneous, ultrafiltration and isoelectric point crystallization.Present invention process step is simple, and it is high to obtain L arginine purity.
Description
Technical field
The invention belongs to technical field of amino acid production, is related to a kind of work of the separation and Extraction L-arginine from zymotic fluid
Skill.
Background technology
Arginine（Arginine, Arg）Chemical scientific name：(S)-2-Amino-5-guanidinopentanoic acid, it is a kind of aliphatic alkaline
Polarity alpha amino acid containing guanidine radicals, it is positively charged in physiological conditions；L-arginine is the coding amino in protein synthesis
Acid.For mammal, arginine is classified as the half necessary or necessary amino acid of conditionity, depend on biology stage of development and
Depending on health status, natural product are largely present in nucleoprotamine, are also the basic composition of various protein.
L-arginine is described as " magical molecule " by scientist, and it is one of basic amino acid, for the adult of health,
For nonessential amino acid, but arginine is the amino acid for maintaining infant growth essential；It is in ornithine circulation
Between metabolin, ammonia can be promoted to be transformed into urea, so as to reduce Blood Ammonia Concentration, when body development is not yet ripe or body stress bar
Under part, such as lack arginine, body just can not complete physiological function；Patient, which lacks arginine, can cause blood ammonia too high or even confused
Fan, baby lack arginine and are then unable to normal development and growth.The doctor of L-arginine has been accepted from Nobel Prize in medicine in 1998
After learning effect, L-arginine causes the mad interest in medical drug field, and Columbia Univ USA is once likened to the heart
" the magical bullet " of vascular system；The discovery of L-arginine makes heart disease, immunologic function, obesity, gene growth shortage, high blood
The treatment of the symptoms such as pressure, sexuality decline and mankind's aging obtains very big improvement.
In terms of food, L-arginine is indispensable flavor enhancement in the food industry, and configuration nutritional support is used
With the important source material of special support herbal cuisine；L-arginine-Pidolidone salt can be as the flavouring for avoiding sodium patient.With L- essence ammonia
Acid has the characteristics that antibiotic property is strong, nontoxic for cationic surfactant prepared by raw material, is the excellent anti-corrosion of food, cosmetics
Agent.Equally, as half necessary amino acid of mammal, animal feed additive is can act also as, promotes growing animal into long hair
The physiological mechanism educated and kept fit.
The production of L-arginine is initially to extract to obtain from protein hydrolyzate, and this method hydrolysis time is long, yield is low, carries
Pure difficult, cost height, pollution are larger, are not suitable for large-scale production.As amino acid industry progressively develops, microbe fermentation method turns into
The most promising production technology of amino acids production, in terms of L-arginine production, microbe fermentation method also shows potential excellent
Gesture, but in microbial fermentation solution extraction process, it is not high that L-arginine shows as yield.Patented technology before the present inventor
" a kind of Clean L-arginine production technology ", extract the arginine of different purity rank, product three times using power technologies such as three-levels
Rank is different, and enterprise income also can be accordingly different, and raising arginine purity is our technical issues that need to address.
The content of the invention
In order to overcome the defects of prior art arginine extraction process purity is low, technique is cumbersome, the invention provides one kind
The technique of separation and Extraction L-arginine from zymotic fluid.
The present invention is achieved by the following technical solution：
A kind of technique of the separation and Extraction L-arginine from zymotic fluid, it comprises the following steps：
Step 1）：L-arginine zymotic fluid collects filtered solution through filtration sterilization of microfiltration membranes；Then addition accounts for filtered fluid
1%（w/w）Flocculant, 200 turns/min stirring 15min, then stand 120min, microfiltration membranes secondary filter, remove precipitation, receive
Collect clear liquid A；Clear liquid A is condensed into a quarter of original volume, then slow cooling is 10.5 with ammoniacal liquor regulation pH to 20 DEG C
Etc. electric solution；
Step 2）：Step 1）The electric solution such as gained settles 3 hours, centrifugation, collects sediment A, pure by being put into sediment A
Change in water, then add flocculant, 200 turns/min stirs 15min, and then microfiltration membranes filter three times, removes precipitation, collects clear liquid
B, clear liquid B is continued into ultrafiltration membrance filter, filtered fluid is collected, filtered fluid is condensed into 1/3rd of original volume, then use ammonia
The electric solution of grade that water regulation pH is 10.5；Temperature control settles 8 hours at 20 DEG C；The L-arginine of crystallization is obtained, by crystallization
L-arginine is dried, and is produced.
The microfiltration membranes once filter, secondary filter and the condition that filters three times are：Microfiltration membranes are inorganic ceramic membrane,
Molecular cut off is 10000Da, and micro-filtration temperature is 50 DEG C；The condition of the ultrafiltration membrance filter is：Ultrafiltration retaining molecular weight is
300Da, ultrafiltrate temperature are 40 DEG C.
The flocculant is prepared according to following technique：
Step（1）：Add the purified water for accounting for twice of mass of starch into starch, be uniformly mixing to obtain suspension, then add
With the sodium carboxymethylcellulose of the quality such as starch, 200 turns/min stirring 30min, it is subsequently placed in 80 DEG C and is dried to moisture 5%
（w/w）Hereinafter, modified starch is obtained；
Step（2）：According to 5:1:0.2 mass ratio takes chitosan, sodium humate and pectin, is mixed evenly, so
2M sodium hydrate aqueous solution is added afterwards, and 30KHz ultrasonic 10min, last 500 turns/min stir 3min, obtain being modified shell and gather
Sugar；
Step（3）：Corn stalk powder, sepiolite are crushed respectively to obtain corn stalk powder and sepiolite powder, by maize straw
Powder, sepiolite powder and diatomite are according to 3:1:1 mass ratio is mixed evenly, and is added to and accounts for the dense of 5 times of weight of diatomite
Spend in the sodium hydroxide solution for 1M, 200 turns/min stirring 120min, then filter, dry, it is 10-20 mesh to be ground into particle diameter
Powder；
Step（4）：By modified starch, modification of chitosan and step（3）The powder of gained is according to 5:3:2 mass ratio mixes
Conjunction stirs, and is subsequently placed in closed reactor, rises to 150 DEG C with 2 DEG C/s heating rate, is incubated 30min, take out, throws
Enter into agitator, 500 turns/min stirring 3min, obtain compound, be placed in baking oven, 80 DEG C of drying 90min, take out, crush
Produce.
The beneficial effect that the present invention obtains mainly includes：
The present invention provides a kind of novel flocculant, is adapted to amino acid zymotic fluid purifying, avoids chemical substance, flocculation purification effect
Fruit is good, and cost is cheap, environment friendly and pollution-free；Flocculant of the present invention is this using agricultural wastes and natural mineral matter as primary raw material
Raw material sources are extensive and cost is low, and prepare porous material, and flocculation purification effect is good, realize agricultural wastes recovery profit
With；The present invention is modified to starch and chitosan so that flocculant aperture, adsorption capacity and specific surface area carry significantly
It is high；Using flocculant process twice in amino acid-extraction, operating procedure is simplified, saves cost, and cause purifying
Better, the flocculant after use is also used as fertilizer preparation, turns waste into wealth.
Embodiment
The present invention will be further explained using specific embodiment, but be should not be construed as to this hair below
The limitation of bright initiative spirit.
Embodiment 1
A kind of technique of the separation and Extraction L-arginine from zymotic fluid, it comprises the following steps：
L-arginine zymotic fluid collects filtered solution through filtration sterilization of microfiltration membranes；Then filtered fluid 1% is accounted for according to addition
（w/w）Flocculant, 200 turns/min stirring 15min, then stand 120min, microfiltration membranes secondary filter, remove precipitation, collect
Clear liquid A；Clear liquid A is condensed into a quarter of original volume, then slow cooling is to 20 DEG C, with ammoniacal liquor regulation pH be 10.5 etc.
Electric solution；
Settled 3 hours etc. electric solution, centrifugation, collect sediment A, will put into sediment A in purified water, then addition accounts for
Sediment A 2%（w/w）Flocculant, 200 turns/min stirring 15min, then microfiltration membranes filter three times, remove precipitation, collect clear
Liquid B, clear liquid B is continued into supermembrane filtering, filtered fluid is collected, filtered fluid is condensed into 1/3rd of original volume, then use ammonia
The electric solution of grade that water regulation pH is 10.5；Temperature control settles 8 hours at 20 DEG C；The L-arginine of crystallization is obtained, by crystallization
L-arginine is dried, and is produced.
Above-mentioned L-arginine product detects through HPLC, product purity 99.5%；The content of L-arginine is in Preliminary fermentation liquid
2732g, it is 2156g to obtain L-arginine product, is converted into the rate of recovery 78.3%；Product purity is higher, can be used as medicine or food
With arginine, product quality and the added value of industry are improved.
The microfiltration membranes once filter, secondary filter and the condition that filters three times are：Microfiltration membranes are inorganic ceramic membrane,
Molecular cut off is 10000Da, and micro-filtration temperature is 50 DEG C；The condition of the ultrafiltration membrance filter is：Ultrafiltration retaining molecular weight is
300Da, ultrafiltrate temperature are 40 DEG C；
The flocculant is prepared according to following technique：
1）Add the purified water for accounting for twice of mass of starch into starch, be uniformly mixing to obtain suspension, then addition and starch
Etc. the sodium carboxymethylcellulose of quality, 200 turns/min stirring 30min are subsequently placed in 80 DEG C and are dried to moisture 5%（Quality hundred
Divide ratio）Hereinafter, modified starch is obtained；
2）According to 5:1:0.2 mass ratio takes chitosan, sodium humate and pectin, is mixed evenly, and then adds
The 2M of twice of mass of chitosan sodium hydrate aqueous solution, 30KHz ultrasonic 10min are accounted for, last 500 turns/min stirs 3min,
Obtain modification of chitosan；
3）Corn stalk powder, sepiolite are crushed respectively to obtain corn stalk powder and sepiolite powder, by corn stalk powder, Hai Pao
Stone flour and diatomite are according to 3:1:1 mass ratio is mixed evenly, and is added to the concentration for accounting for 5 times of weight of diatomite as 1M's
In sodium hydroxide solution, 200 turns/min stirring 120min, then filter, dry, it is 10-20 mesh powders to be ground into particle diameter；
4）By modified starch, modification of chitosan and step 3）Gained powder is according to 5:3:2 mass ratio mixes equal
It is even, it is subsequently placed in closed reactor, rises to 150 DEG C with 2 DEG C/s heating rate, be incubated 30min, take out, put into stirring
In device, 500 turns/min stirring 3min, compound is obtained, is placed in baking oven, 80 DEG C of drying 90min, taken out, crush and produce.
Finally, it is also necessary to it is noted that listed above is only several specific embodiments of the invention.Obviously, this hair
It is bright to be not limited to above example, there can also be many deformations.One of ordinary skill in the art can be from present disclosure
All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Claims (2)
1. a kind of technique of the separation and Extraction L-arginine from zymotic fluid, it comprises the following steps：
Step 1）：L-arginine zymotic fluid collects filtered solution through filtration sterilization of microfiltration membranes；Then addition accounts for filtered fluid weight
1% flocculant, 200 turns/min stirring 15min, 120min is then stood, then by microfiltration membranes secondary filter, remove precipitation, receive
Collect clear liquid A；Clear liquid A is condensed into a quarter of original volume, then slow cooling is adjusted to wait electricity molten to 20 DEG C with ammoniacal liquor
Liquid；
Step 2）：Step 1）The electric solution such as gained settles 3 hours, centrifugation, collects sediment A, will put into purified water in sediment A
In, flocculant, 200 turns/min stirring 15min are then added, then filtered three times with microfiltration membranes, precipitation is removed, collects clear liquid B, will
Clear liquid B carries out ultrafiltration membrance filter, collects filtered fluid, filtered fluid is condensed into 1/3rd of original volume, then be adjusted to ammoniacal liquor
Etc. electric solution, then temperature control is at 20 DEG C, sedimentation 8 hours, obtains the L-arginine of crystallization, and the L-arginine of crystallization is dried
It is dry, produce；
The flocculant is prepared according to following technique：
Step（1）：Add the purified water for accounting for twice of mass of starch into starch, be uniformly mixing to obtain suspension, then add and form sediment
The sodium carboxymethylcellulose of the quality such as powder, 200 turns/min stirring 30min, is subsequently placed in 80 DEG C and is dried to moisture below 5%,
Obtain modified starch；
Step（2）：According to 5:1:0.2 mass ratio takes chitosan, sodium humate and pectin, is mixed evenly, Ran Houtian
Add 2M sodium hydrate aqueous solution, 30KHz ultrasonic 10min, last 500 turns/min stirs 3min, obtains modification of chitosan；
Step（3）：Corn stalk powder, sepiolite are crushed respectively to obtain corn stalk powder and sepiolite powder, by corn stalk powder, sea
Afrodite powder and diatomite are according to 3:1:1 mass ratio is mixed evenly, and is added to that to account for the concentration of 5 times of weight of diatomite be 1M
Sodium hydroxide solution in, 200 turns/min stirring 120min, then filter, dry, it is 10-20 mesh powders to be ground into particle diameter；
Step（4）：By modified starch, modification of chitosan and step（3）Gained powder is according to 5:3:2 mass ratio mixes
Uniformly, it is subsequently placed in closed reactor, rises to 150 DEG C with 2 DEG C/s heating rate, be incubated 30min, take out, put into and stir
Mix in device, 500 turns/min stirring 3min, obtain compound, be placed in baking oven, 80 DEG C of drying 90min, take out, crush and produce.
2. technique according to claim 1, it is characterised in that
The microfiltration membranes once filter, secondary filter and the condition that filters three times are：Microfiltration membranes are inorganic ceramic membrane, retention
Molecular weight is 10000Da, and micro-filtration temperature is 50 DEG C；The condition of the ultrafiltration membrance filter is：Ultrafiltration retaining molecular weight is
300Da, ultrafiltrate temperature are 40 DEG C.
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| CN201610655175.5A CN106278952B (en) | 2016-08-11 | 2016-08-11 | One kind arginic techniques of separation and Extraction L from zymotic fluid |
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| CN106278952B true CN106278952B (en) | 2017-12-22 |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3617091B2 (en) * | 1994-11-30 | 2005-02-02 | 味の素株式会社 | Purification method of basic amino acids |
| CN103145590B (en) * | 2013-03-06 | 2014-05-28 | 山东阜丰发酵有限公司 | Clean L-arginine production technology |
| CN103695487B (en) * | 2013-12-24 | 2015-09-23 | 滨州市生物技术研究院有限责任公司 | A kind of fermentable produces arginine technique |
| CN105418461B (en) * | 2015-11-23 | 2017-10-31 | 江南大学 | A kind of arginic continuous ion exchage extracting process of L |
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Denomination of invention: A process for separating and extracting L-arginine from fermentation broth Effective date of registration: 20211018 Granted publication date: 20171222 Pledgee: China Construction Bank Corporation Junan sub branch Pledgor: SHANDONG FUFENG FERMENTATION Co.,Ltd. Registration number: Y2021370000120 |
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Date of cancellation: 20230414 Granted publication date: 20171222 Pledgee: China Construction Bank Corporation Junan sub branch Pledgor: SHANDONG FUFENG FERMENTATION CO.,LTD. Registration number: Y2021370000120 |