CN103667383B - The preparation method of L-glutaminate - Google Patents

The preparation method of L-glutaminate Download PDF

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CN103667383B
CN103667383B CN201310721139.0A CN201310721139A CN103667383B CN 103667383 B CN103667383 B CN 103667383B CN 201310721139 A CN201310721139 A CN 201310721139A CN 103667383 B CN103667383 B CN 103667383B
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CN103667383A (en
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王威
高树营
李令娣
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SHANDONG MINQIANG BIOTECHNOLOGY Co Ltd
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Abstract

The present invention is the preparation method of L-glutaminate, belongs to extraction and the technical field of refinement of amino acid derivative fermentation liquor, is specifically related to a kind of L-glutaminate and extracts and process for purification.The method comprises following step: the preparation of fermented liquid: slant culture, shaking flask bacterial classification, first order seed are cultivated, secondary seed is cultivated, continue fermentation; Leavened prod Hydrolysis kinetics: fermentation liquor pretreatment, ion-exchange, secondary crystal, decolouring removal of impurities process, nanofiltration membrane decolouring removal of impurities and concentrated, condensing crystal, dry finished product.Beneficial effect of the present invention is, significantly reduces organism and inorganic contaminants content and pigment in glutamine ferment liquid subsequent extracted, refined liquid, improves the yield of glutamine; Improve quality product, reduce production cost, largely alleviate the labour intensity of workman; Greatly reduce the quantity discharged that the solid-liquids such as water, acid, alkali, gac are useless, decrease the pollution to environment.

Description

The preparation method of L-glutaminate
Technical field
The invention belongs to extraction and the technical field of refinement of amino acid derivative fermentation liquor, be specifically related to a kind of L-glutaminate and extract and process for purification.
Background technology
Glutamine (Glutamin) is also referred to as bran amido acid, is the non-essential amino acid that content in human body is the abundantest, and is that unique one can directly by the amino acid of cerebrovascular barrier (BBB).Be stored in human body in skeletal muscle or blood.When injured or ill, glutamine may need to contain the food of Gln to obtain enough amounts by picked-up.Organism generates glutamine by glutamine synthetase catalysis L-glutamic acid and ammonium salt reaction.
L-glutaminate, this product makes accessory substance in food-processing, blending enriching substance.And L glutamine is the important nutritional supplement of body building and bodybuilders.Its (hereinafter referred to as glutamine) is total free aminoacids the abundantest in muscle, accounts for 60% of human body total free aminoacids total amount.Fasting plasma glutamine concentration is 500-750umol/L.Glutamine is not indispensable amino acid, and it can be synthesized by L-glutamic acid, a word used in person's names propylhomoserin, Isoleucine in human body.At disease, nutritional status under the stress situation such as not good or high-intensity exercise, body increases the demand of glutamine, so that self synthesis can not be satisfied the demand.
L-glutaminate is widely used in fields such as medicine, nutritive health-care, food, and demand is large, is referred to as one of most important kind in nonessential amino acid.L-glutamine is the important materials of Development of Novel medicine, clinical treatment and sports nutrition health care, and all body tissues all utilize L-glutamine synthetic cell to starch albumen and nucleoprotein.In physiologically active, outside the Pass having with gastrointestinal tract disease, animal science and animal and veterinary industry, in recent years, again as one of human cytotoxic weapon, strengthening immune system and cause concern, the novel material becoming trophotherapy is expected to.Because L-glutamine not only has important scientific research, actual production and using value, L-glutamine is in the effect of physiological function importance simultaneously, and present domestic conventional production methods still seriously polluted, the production capacity output value is lower, therefore, through State Council and deputy to the National People's Congress's approval, Production of L-Glutamine by Microbial Fermentation lists national Eleventh Five-Year Plan emphasis tackling key problem science and technology item in.
Fermented liquid → ceramic membrane filter → flocculation, decolouring+ceramic membrane filter → condensing crystal → centrifugal → drying → product.
Summary of the invention
In order to solve above-mentioned technical problem, the present invention utilizes ceramic membrane, reverse osmosis, nanofiltration membrane and two Crystallization Procedure to carry out the method for purifying L-glutaminate; Its yield of product adopting the method to obtain and qualification rate improve greatly.
L-glutaminate of the present invention extracts, process for purification solves above technical problem by following technical scheme:
The preparation method of L-glutaminate, the method comprises following step:
(1) preparation of fermented liquid:
With brevibacterium flavum MQG121 bacterial strain for fermentation strain;
(1) slant culture: peace roasting pipe bacterial classification is accessed aseptic slant medium 28-32 DEG C and cultivates 20-24 hour;
(2) shaking flask bacterial classification: by cultured slant strains access sterile flasks seed culture medium, shaking speed 100rpm, shaking table stroke 65cm, cultivate 16-20 hour for 28-32 DEG C;
(3) first order seed is cultivated: cultured shaking flask kind liquid is accessed aseptic primary-seed medium, under 1:0.5 ventilation condition, cultivates 12-16 hour for mixing speed 240rpm, 28-32 DEG C;
(4) secondary seed is cultivated: cultured one-level kind liquid is accessed aseptic secondary seed medium, under 1:0.5 ventilation condition, cultivates 2-6 hour for mixing speed 220rpm, 28-32 DEG C;
(5) cultured secondary seed is inoculated in the fermention medium of sterilizing, under 1:0.3 ventilation condition, mixing speed 140rpm, fermenting process is by liquefied ammonia adjust ph to 7.0-7.2, and through 0-12 hours yeast culture, then adjust ph was to 5.5-5.7, product synthesis in 12-44 hours is cultivated, process liquefied ammonia control PH, stream sugaring controls sugared content 2.0-0%, obtains the fermented liquid containing glutamine product;
Above-mentioned fermention medium is:
Glucose 12-16% corn steep liquor 1-6%
Molasses 0.5-2% (NH 4) 2sO 43-7%
VITMAIN B1 1-4mg/L KH 2pO 42-5%,
MgSO 4 0.05-0.08%, MnSO 48-12mg/ml
ZnSO 40.5-2 mg/ml CuSO 40.8-2mg/ml
Above-mentioned brevibacterium flavum bacterial strain ( brevibacterium flavum) being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 12nd, 2013, deposit number is CGMCC NO.8468;
(2) leavened prod Hydrolysis kinetics:
Ceramic membrane filter: the fermented liquid under normal temperature state is sent into ceramic membrane circulating filtration and obtains fermentation clear liquid, use recirculated water for the cooling of ceramic membrane circulation feed liquid in working cycle, make the temperature of circulating fermentation liquid remain on less than 70 DEG C, pottery filter concentration liquid adds reverse osmosis water and cleans out residual product; By the thalline in ceramic membrane filter removing fermented liquid and macromole impurity, obtained pottery cleaner liquid.
Primary crystallization: by the feed liquid after concentrated at 50 DEG C, vacuum concentration under 80rpm condition, concentrated solution product content is 60%, vacuum tightness is-0.098Mpa, 15 DEG C are cooled to the cooling rate of 10-15 DEG C/h with recirculated water, growing the grain 4 hours again, growing the grain mixing speed is 20rpm, obtains wet product crystallization through centrifuge;
Crude product back dissolving: first reverse osmosis water is warming up to 70 DEG C, mixing speed is 40rpm, by wet product dissolving crystallized, the amount being 10% by product content drops into crude product, is adjusted to pH3.5, crystallization is dissolved completely with reagent hydrochloric acid, obtained dissolving crude product liquid.
Ion-exchange: be 10% by content, pH be 3.5, temperature be 45 DEG C feed liquid with 1 times of resin volume/hour flow velocity by 732 cationic exchange coloums, 330 anion-exchange columns are flowed into identical speed by the feed liquid of cationic exchange coloum, by the organic and inorganic impurity in ion exchange resin removing feed liquid, obtain from friendship liquid;
Nanofiltration: enter nanofiltration equipment circulating filtration by under the feed liquid normal temperature state after the removal of impurities of cation and anion exchange post; Use recirculated water for the cooling of nanofiltration circulation feed liquid in working cycle, make the temperature of circulation feed liquid remain on less than 50 DEG C, nanofiltration concentrated solution adds reverse osmosis water and cleans out residual product; By the organic molecule impurity in nanofiltration removing feed liquid, obtain nanofiltration dialysis feed liquid.
Decolouring removal of impurities process: decolouring removal of impurities process is carried out to nanofiltration dialysis feed liquid with gac, feed liquid is warming up to 70 DEG C, add 0.3% medicinal 767 gacs, be incubated 30 minutes, mixing speed is 60rpm, reach by the gac in fragrant special strainer circulating filtration removing feed liquid after more than 95% until transmitance, obtained destainer.
Reverse osmosis concentration: to the feed liquid reverse osmosis concentration under normal temperature, make material liquid pH be 7.0, in feed liquid, product content is 10%, makes product content in concentrated rear feed liquid be 15%; Obtained reverse osmosis concentrated liquid.Feed liquid PH to 6.0-6.5 are regulated with H type 732 Zeo-karb.
Secondary crystal: reverse osmosis concentrated liquid is squeezed into single-action condensing crystal tank, thickening temperature is kept to be≤50 DEG C, vacuum tightness is-0.098Mpa, mixing speed keeps 60rpm, when concentrated solution product content is more than 60%, keep mixing speed 20rpm, be down to 15 DEG C by the cooling rate of recirculated water 15-15 DEG C /h, growing the grain 4 hours, obtains wet product crystallization through rotating speed 1200rpm centrifuge;
Dry: wet product crystalline product is dry through double cone dryer, and its condition is: vacuum tightness-0.098Mpa, use temperature is that the hot water circulation of 80 DEG C keeps dry temperature≤50 DEG C, dry 6h, treat product temperature≤40 DEG C packaging, pack environment condition is: temperature 18 DEG C-25 DEG C, humidity≤70%.
Preferably, in the preparation process of above-mentioned fermented liquid:
With brevibacterium flavum MQG121 bacterial strain for fermentation strain;
(1) slant culture: peace roasting pipe bacterial classification is accessed aseptic slant medium 28-32 DEG C and cultivates 20-24 hour;
(2) shaking flask bacterial classification: by cultured slant strains access sterile flasks seed culture medium, shaking speed 100rpm, shaking table stroke 65cm, cultivate 16-20 hour for 28-32 DEG C;
(3) first order seed is cultivated: cultured shaking flask kind liquid is accessed aseptic primary-seed medium, under 1:0.5 ventilation condition, cultivates 12-16 hour for mixing speed 240rpm, 28-32 DEG C;
(4) secondary seed is cultivated: cultured one-level kind liquid is accessed aseptic secondary seed medium, under 1:0.5 ventilation condition, cultivates 2-6 hour for mixing speed 220rpm, 28-32 DEG C;
(5) cultured secondary seed is inoculated in the fermention medium of sterilizing, under 1:0.3 ventilation condition, mixing speed 140rpm, fermenting process is by liquefied ammonia adjust ph to 7.0-7.2, and through 0-12 hours yeast culture, then adjust ph was to 5.5-5.7, product synthesis in 12-44 hours is cultivated, process liquefied ammonia control PH, stream sugaring controls sugared content 2.0-0%, obtains the fermented liquid containing glutamine product;
Preferred, above-mentioned fermention medium is:
Glucose 12% corn steep liquor 1%
Molasses 0.5% (NH 4) 2sO 44%
VITMAIN B1 1mg/L KH 2pO 43%,
MgSO 4 0.05%, MnSO 48mg/ml
ZnSO 41mg/ml CuSO 41mg/ml
Brevibacterium flavum bacterial strain of the present invention ( brevibacterium flavum) being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 12nd, 2013, deposit number is CGMCC NO.8468; Preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, postcode: 100101.
Beneficial effect of the present invention is:
(1) in the extraction, treating process of L-glutaminate, utilize the production bacterium in ceramic membrane equipment removing fermented liquid and high molecular weight protein, replace traditional sheet frame sterilization technique, decrease floor space and labour intensity; Adopt ceramic membrane to carry out the degerming of glutamine ferment liquid, not only can reclaim albumen, due to the raising of filtered liquid quality, significantly can also reduce consumption (100 kg/ms original by the exchange capacity of resin to product of ion exchange resin 3bring up to 150 kg/m 3), two Crystallization Procedure significantly can reduce thalline, organism and inorganic contaminants content in glutamine subsequent extracted refined liquid, improves the yield of glutamine;
(3) adopt the organic pigment in sodium membrane filtration technology removing feed liquid and small organic molecule, organic impurity in feed liquid is reduced greatly, improves the transmittance of feed liquid, reduce the consumption of gac;
(4) reverse osmosis is adopted to carry out concentrated pre-treatment, decrease high temperature and concentrate destruction to product, decrease the use of steam, reduce energy consumption, decrease sewage discharge, ensure that the quality into ion exchange column feed liquid, increase the adsorptive power of ion exchange column to product, decrease the consumption of resin; Ion-exchange is resolved liquid glutamine content and is brought up to 8% by traditional 5.5%, is greatly enhanced, improves quality product, reduce production cost, significantly reduce the labour intensity of workman from friendship liquid purity.
By above-mentioned treatment process, finished product can reach more than 85% (the clean L-glutaminate cubage relative to removing thalline) for total recovery fermented liquid, finished product first-time qualification rate reaches 100%, and traditional technology can only reach the total recovery of 50%, and first-time qualification rate only has 98%.
Accompanying drawing explanation
Fig. 1 is process flow sheet of the present invention.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is further described, so that those skilled in the art more understands the present invention, but does not limit the present invention with this.
Extraction production technology
The preparation method of L-glutaminate, the method comprises following step:
(1) preparation of fermented liquid:
With brevibacterium flavum MQG121 bacterial strain for fermentation strain;
(1) slant culture: peace roasting pipe bacterial classification is accessed aseptic slant medium 28-32 DEG C and cultivates 20-24 hour;
(2) shaking flask bacterial classification: by cultured slant strains access sterile flasks seed culture medium, shaking speed 100rpm, shaking table stroke 65cm, cultivate 16-20 hour for 28-32 DEG C;
(3) first order seed is cultivated: cultured shaking flask kind liquid is accessed aseptic primary-seed medium, under 1:0.5 ventilation condition, cultivates 12-16 hour for mixing speed 240rpm, 28-32 DEG C;
(4) secondary seed is cultivated: cultured one-level kind liquid is accessed aseptic secondary seed medium, under 1:0.5 ventilation condition, cultivates 2-6 hour for mixing speed 220rpm, 28-32 DEG C;
(5) cultured secondary seed is inoculated in the fermention medium of sterilizing, under 1:0.3 ventilation condition, mixing speed 140rpm, fermenting process is by liquefied ammonia adjust ph to 7.0-7.2, and through 0-12 hours yeast culture, then adjust ph was to 5.5-5.7, product synthesis in 12-44 hours is cultivated, process liquefied ammonia control PH, stream sugaring controls sugared content 2.0-0%, obtains the fermented liquid containing glutamine product;
Above-mentioned fermention medium is:
Glucose 12% corn steep liquor 1%
Molasses 0.5% (NH 4) 2sO 44%
VITMAIN B1 1mg/L KH 2pO 43%,
MgSO 4 0.05%, MnSO 48mg/ml
ZnSO 41mg/ml CuSO 41mg/ml
Above-mentioned brevibacterium flavum bacterial strain ( brevibacterium flavum) being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 12nd, 2013, deposit number is CGMCC NO.8468;
(2) leavened prod Hydrolysis kinetics:
Ceramic membrane filter: the fermented liquid under normal temperature state is sent into ceramic membrane circulating filtration and obtains fermentation clear liquid, use recirculated water for the cooling of ceramic membrane circulation feed liquid in working cycle, make the temperature of circulating fermentation liquid remain on less than 70 DEG C, pottery filter concentration liquid adds reverse osmosis water and cleans out residual product; By the thalline in ceramic membrane filter removing fermented liquid and macromole impurity, obtained pottery cleaner liquid.
Primary crystallization: by the feed liquid after concentrated at 50 DEG C,, vacuum concentration under 80rpm condition, concentrated solution product content is 60%, vacuum tightness is-0.098Mpa, 15 DEG C are cooled to the cooling rate of 10-15 DEG C/h with recirculated water, growing the grain 4 hours again, growing the grain mixing speed is 20rpm, obtains wet product crystallization through centrifuge;
Crude product back dissolving: first reverse osmosis water is warming up to 70 DEG C, mixing speed is 40rpm, by wet product dissolving crystallized, the amount being 10% by product content drops into crude product, is adjusted to pH3.5, crystallization is dissolved completely with reagent hydrochloric acid; Obtained dissolving crude product liquid.
Ion-exchange: be 10% by content, pH be 3.5, temperature be 45 DEG C feed liquid with 1 times of resin volume/hour flow velocity by 732 cationic exchange coloums, 330 anion-exchange columns are flowed into identical speed by the feed liquid of cationic exchange coloum, by the organic and inorganic impurity in ion exchange resin removing feed liquid, obtain from friendship liquid;
Nanofiltration: enter nanofiltration equipment circulating filtration by under the feed liquid normal temperature state after the removal of impurities of cation and anion exchange post; Use recirculated water for the cooling of nanofiltration circulation feed liquid in working cycle, make the temperature of circulation feed liquid remain on less than 50 DEG C, nanofiltration concentrated solution adds reverse osmosis water and cleans out residual product; By the organic molecule impurity in nanofiltration removing feed liquid, obtain nanofiltration dialysis feed liquid.
Decolouring removal of impurities process: decolouring removal of impurities process is carried out to nanofiltration dialysis feed liquid with gac, feed liquid is warming up to 70 DEG C, add 0.3% medicinal 767 gacs, be incubated 30 minutes, mixing speed is 60rpm, reach by the gac in fragrant special strainer circulating filtration removing feed liquid after more than 95% until transmitance, obtained destainer.
Reverse osmosis concentration: the feed liquid after decolouring is cooled to 40 DEG C, enters the circulation of reverse osmosis concentration device and is concentrated into content and reaches 15%; Use recirculated water to lower the temperature for circulation fluid in working cycle, keep feed temperature≤50 DEG C, obtained reverse osmosis concentrated liquid.Feed liquid PH to 6.0-6.5 are regulated with H type 732 Zeo-karb.
Secondary crystal: reverse osmosis concentrated liquid is squeezed into single-action condensing crystal tank, thickening temperature is kept to be≤50 DEG C, vacuum tightness is-0.098Mpa, mixing speed keeps 60rpm, when concentrated solution product content is more than 60%, keep mixing speed 20rpm, be down to 15 DEG C by the cooling rate of recirculated water 15-15 DEG C /h, growing the grain 4 hours, obtains wet product crystallization through centrifuge;
Dry: wet product crystalline product is dry through double cone dryer, and its condition is: vacuum tightness-0.098Mpa, use temperature is that the hot water circulation of 80 DEG C keeps dry temperature≤50 DEG C, dry 6h, treat product temperature≤40 DEG C packaging, pack environment condition is: temperature 18 DEG C-25 DEG C, humidity≤70%.
Wet product crystalline product is dry through bipyramid, and its condition is: vacuum tightness-0.098Mpa, temperature 80 DEG C, and packaging, condition is: temperature 18 DEG C-25 DEG C, humidity≤60%, transmittance >=82%, DE value≤2.5%, obtains finished product.
(2) L-glutaminate is at 50m 3fermentor tank extracts in producing, upgrading result statistics
production leadtime result is as follows
(3) inspection after construction result: pilot product is through inspection, and indices meets L-glutaminate FCC V standard, and main performance index refers to following table:
The present invention extracts workshop section and adopts ceramic membrane device processes, and the production bacterium utilizing ceramic membrane equipment to remove in fermented liquid replaces traditional sheet frame sterilization technique, and impurity in feed liquid is reduced greatly; But also decrease sewage discharge, ensure that the quality into ion exchange column feed liquid, increase the adsorptive power of ion exchange column, decrease the consumption of resin and gac, ion-exchange is crossed flow liquid glutamine content and is reached 8%, and extraction, refining total recovery reach more than 85%, improve quality product, reduce production cost, significantly reduce the labour intensity of workman.Membrane concentration separating technology general is in the world adopted to replace traditional concentrating under reduced pressure technique at product enriching stage, namely the destruction to product is decreased, in turn save steam consumption, decrease coal quantity of boiler, the resource consumptions such as water, electricity, coal reduce greatly, be conducive to saving energy and reduce the cost, production cost is low.And have employed sodium filter membrane concentration technology, improve the quality of product, the resin demand of the traditional extraction concentration technique of fermentation method of also reducing over, greatly reduces the quantity discharged of the waste water such as water, acid, alkali, reduces the pollution to environment.
Embodiment 2
The preparation method of L-glutaminate, the method comprises following step:
(1) preparation of fermented liquid:
With brevibacterium flavum MQG121 bacterial strain for fermentation strain;
Slant culture: peace roasting pipe bacterial classification access slant medium 28-32 DEG C is cultivated 20-24 hour;
Shaking flask bacterial classification: cultured slant strains access shake-flask seed substratum 28-32 DEG C is cultivated 16-20 hour;
First order seed is cultivated: cultivate 12-16 hour by primary-seed medium 28-32 DEG C of cultured shaking flask kind liquid access sterilization;
Secondary seed is cultivated: cultured one-level kind liquid is accessed the secondary seed medium 28-32 DEG C of cultivation 2-6 hour sterilized; Be inoculated in the fermention medium of sterilizing by cultured secondary seed, pass into sterile wind, adjust ph, to 6.2-6.8, cultivated through 40-48 hour the fermented liquid obtained containing glutamine product;
Above-mentioned fermention medium is:
Extractum carnis 2.2%, glucose 13.5%,
Sucrose 3.6% wort 4.8%
CSL 4.0%， （NH 4） 2SO 4 2.5%,
KH 2PO 4 0.2%, MgSO 4 0.04%,
MnSO 410 mg/L， CuSO 41 mg/L，
Vitamins B 11mg/L, NaH 2pO 41.0mg/L,
Yeast extract 5.5% CaCO 32%;
Above-mentioned brevibacterium flavum bacterial strain ( brevibacterium flavum) being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 12nd, 2013, deposit number is CGMCC NO.8468;
(2) leavened prod Hydrolysis kinetics:
Ceramic membrane filter: the fermented liquid under normal temperature state is sent into ceramic membrane filter, adopt recirculated water to be the cooling of circulation feed liquid in treating processes, make the temperature of fermented liquid remain on less than 70 DEG C, concentrated solution adds reverse osmosis water and cleans out residual product;
Reverse osmosis concentration: to the feed liquid reverse osmosis concentration under normal temperature, make material liquid pH be 7.0, in feed liquid, product content is 10%, makes product content in concentrated rear feed liquid be 15%;
Primary crystallization: by the feed liquid primary crystallization after concentrated, in primary crystallization, thickening temperature is 50 DEG C, and concentrated solution product content is 60%, and vacuum tightness is-0.098Mpa, is down to normal temperature growing the grain 4 hours, and mixing speed is 20rpm, obtains wet product crystallization through centrifuge;
Crude product back dissolving: by wet product dissolving crystallized, the amount being 10% by product content drops into crude product reverse osmosis water solvent, adjusts pH3.5, is heated to 50 DEG C, under the rotating speed of 30rpm under agitation condition until dissolve completely;
Ion-exchange: be 10% by content, pH be 3.5, temperature be 45 DEG C feed liquid with 1 times of resin volume/hour flow velocity by 732 cationic exchange coloums, flow into 330 anion-exchange columns by the feed liquid of cationic exchange coloum with identical speed, must from friendship liquid;
Nanofiltration: enter nano-filtration step by under the feed liquid normal temperature state after the removal of impurities of cation and anion exchange post;
Decolouring removal of impurities process: decolouring removal of impurities process is carried out to feed liquid with nanofiltration, by from the organic-inorganic pigment handing over liquid in nanofiltration equipment circulating filtration removing feed liquid, concentrated solution adds after reverse osmosis water cleaning product remains, and makes feed liquid transmittance reach 85%;
Membrane concentration is separated: the feed liquid after decolouring is cooled to 40 DEG C, enters reverse osmosis concentration device and is concentrated into content and reaches 15%;
Secondary crystal: in secondary crystal step, keep thickening temperature to be 50 DEG C, concentrated solution product content is more than 60%, and vacuum tightness is-0.098Mpa, is down to normal temperature growing the grain 4 hours, and mixing speed is 20rpm.Wet product crystallization is obtained through centrifuge;
Wet product crystalline product is dry through bipyramid, and its condition is: vacuum tightness-0.098Mpa, temperature 80 DEG C, and packaging, condition is: temperature 18 DEG C-25 DEG C, humidity≤60%, transmittance >=82%, DE value≤2.5%, obtains finished product.

Claims (2)

  1. The preparation method of 1.L-glutamine, is characterized in that, the method comprises following step:
    (1) preparation of fermented liquid:
    With brevibacterium flavum ( brevibacterium flavum) bacterial strain is fermentation strain;
    (1) slant culture: peace roasting pipe bacterial classification is accessed aseptic slant medium, cultivates 20-24 hour for 28-32 DEG C;
    (2) shaking flask bacterial classification: by cultured slant strains access sterile flasks seed culture medium, shaking speed 100rpm, shaking table stroke 65cm, cultivate 16-20 hour for 28-32 DEG C;
    (3) first order seed cultivate: by cultured shaking flask kind liquid by 10% inoculum size access aseptic primary-seed medium, under 1:0.5 ventilation condition, mixing speed 240rpm, 28-32 DEG C cultivate 12-16 hour;
    (4) secondary seed cultivate: by cultured one-level kind liquid by 10% inoculum size access aseptic secondary seed medium, under 1:0.5 ventilation condition, mixing speed 220rpm, 28-32 DEG C cultivate 2-6 hour;
    (5) by cultured secondary seed solution by 10% inoculum size be inoculated in aseptic fermention medium, under 1:0.3 ventilation condition, mixing speed 140rpm, fermenting process by liquefied ammonia adjust ph to 7.0-7.2, through 0-12 hours yeast culture and adjust ph to 5.5-5.7, the fermented liquid obtained containing glutamine product is cultivated in product synthesis in 12-44 hours;
    Above-mentioned fermention medium is:
    Glucose 12-16% corn steep liquor 1-6%
    Molasses 0.5-2% (NH 4) 2sO 44-8%
    VITMAIN B1 1-4mg/L KH 2pO 42-5%,
    MgSO 4 0.05-0.08%, MnSO 48-12mg/ml
    ZnSO 40.5-2 mg/ml CuSO 40.8-2mg/ml
    Above-mentioned brevibacterium flavum bacterial strain ( brevibacterium flavum) being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 12nd, 2013, deposit number is CGMCC NO.8468;
    (2) leavened prod Hydrolysis kinetics:
    Ceramic membrane filter: the fermented liquid under normal temperature state is sent into ceramic membrane circulating filtration and obtains fermentation clear liquid, use recirculated water for the cooling of ceramic membrane circulation feed liquid in working cycle, make the temperature of circulating fermentation liquid remain on less than 70 DEG C, pottery filter concentration liquid adds reverse osmosis water and cleans out residual product; By the thalline in ceramic membrane filter removing fermented liquid and macromole impurity, obtained pottery cleaner liquid;
    Primary crystallization: by the feed liquid after concentrated at 50 DEG C, vacuum concentration under 80rpm condition, concentrated solution product content is 60%, vacuum tightness is-0.098Mpa, 15 DEG C are cooled to the cooling rate of 10-15 DEG C/h with recirculated water, growing the grain 4 hours again, growing the grain mixing speed is 20rpm, and the centrifuge through 1200rpm rotating speed obtains a wet product crystallization;
    Crude product back dissolving: first reverse osmosis water is warming up to 70 DEG C, mixing speed is 40rpm, by a wet product dissolving crystallized, the amount being 10% by product content drops into crude product, is adjusted to pH3.5, crystallization is dissolved completely with reagent hydrochloric acid; Obtained dissolving crude product liquid;
    Ion-exchange: be 10% by content, pH be 3.5, temperature be 45 DEG C feed liquid with 1 times of resin volume/hour flow velocity by 732 cationic exchange coloums, 330 anion-exchange columns are flowed into identical speed by the feed liquid of cationic exchange coloum, by the organic and inorganic impurity in ion exchange resin removing feed liquid, obtain from friendship liquid;
    Nanofiltration: enter nanofiltration equipment circulating filtration by under the feed liquid normal temperature state after the removal of impurities of cation and anion exchange post; Use recirculated water for the cooling of nanofiltration circulation feed liquid in working cycle, make the temperature of circulation feed liquid remain on less than 50 DEG C, nanofiltration concentrated solution adds reverse osmosis water and cleans out residual product; By the organic molecule impurity in nanofiltration removing feed liquid, obtain nanofiltration dialysis feed liquid;
    Decolouring removal of impurities process: decolouring removal of impurities process is carried out to nanofiltration dialysis feed liquid with gac, feed liquid is warming up to 70 DEG C, add 0.3% medicinal 767 gacs, be incubated 30 minutes, mixing speed is 60rpm, reach by the gac in fragrant special strainer circulating filtration removing feed liquid after more than 95% until transmitance, obtained destainer;
    Reverse osmosis concentration: the feed liquid after decolouring is cooled to 40 DEG C, enters the circulation of reverse osmosis concentration device and is concentrated into content and reaches 15%; Use recirculated water to lower the temperature for circulation fluid in working cycle, keep feed temperature≤40 DEG C, obtained reverse osmosis concentrated liquid;
    Material liquid pH to 6.0-6.5 are regulated with H type 732 Zeo-karb;
    Secondary crystal: reverse osmosis concentrated liquid is squeezed into single-action condensing crystal tank, thickening temperature is kept to be≤50 DEG C, vacuum tightness is-0.098Mpa, mixing speed keeps 60rpm, when concentrated solution product content is more than 60%, keep mixing speed 20rpm, be down to 15 DEG C with recirculated water with the cooling rate of 15-15 DEG C/h, growing the grain 4 hours, obtains the crystallization of secondary wet product through centrifuge;
    Dry: secondary wet product crystalline product is dry through double cone dryer, its condition is: vacuum tightness-0.098Mpa, use temperature is that the hot water circulation of 80 DEG C keeps dry temperature≤50 DEG C, dry 6h, treat product temperature≤40 DEG C packaging, pack environment condition is: temperature 18 DEG C-25 DEG C, humidity≤70%.
  2. 2. the preparation method of a kind of L-glutaminate as claimed in claim 1, is characterized in that, described fermention medium is:
    Glucose 12% corn steep liquor 1%
    Molasses 0.5% (NH 4) 2sO 44%
    VITMAIN B1 1mg/L KH 2pO 43%,
    MgSO 4 0.05%, MnSO 48mg/ml
    ZnSO 41mg/ml CuSO 41mg/ml。
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