CN102719510A - Amino acid fermentation bacteria utilization method - Google Patents
Amino acid fermentation bacteria utilization method Download PDFInfo
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- CN102719510A CN102719510A CN2012102113999A CN201210211399A CN102719510A CN 102719510 A CN102719510 A CN 102719510A CN 2012102113999 A CN2012102113999 A CN 2012102113999A CN 201210211399 A CN201210211399 A CN 201210211399A CN 102719510 A CN102719510 A CN 102719510A
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- tropina
- enzymolysis
- mycoprotein
- fermentation
- glutamic acid
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Abstract
The invention discloses an amino acid fermentation bacteria utilization method, which belongs to the technical field of amino acid production. The method comprises the steps that by taking glutamic acid fermentation as an example and adopting the isoelectric point of fermentation broth, a disc separator is utilized to separate mycoprotein in the fermentation broth, and a moderate amount of compound enzyme is added into the mycoprotein serum, so that the walls of bacteria are broken and the bacteria are enzymetically hydrolyzed. The enzymatic hydrolysate is separated by the high-speed disc separator, and the cell walls are removed, the obtained clarified enzymatic hydrolysate is concentrated under low temperature to produce the enzymetically hydrolyzed mycoprotein extract, or the clarified enzymatic hydrolysate is dried by a gunite granulation fluidized bed drier, a spray granulation drier and the like to produce the enzymetically hydrolyzed mycoprotein powder. In the method, the utilization value of the mycoprotein is greatly increased, so that the mycoprotein is changed from cheap feed into high value-added yeast extract and yeast powder substitute, the product can be used as fermentation ingredient and in other fields, the resources can be sufficiently utilized in the whole process, the purposes of energy saving and consumption reduction are achieved, and the invention has a broad application prospect.
Description
Technical field
The invention belongs to the amino acid production technical field, relate to a kind of amino acid fermentation thalline and utilize method, be specially a kind of compound enzymic preparation that utilizes the glutamic acid fermentation thalline is carried out the method that enzymolysis prepares enzymolysis protein cream, protein powder.
Background technology
Aminoglutaminic acid thalline is the sub product in the fermentative Production monosodium glutamate process; Be that glutamate producing bacterium generates the discarded thalline of L-glutamic acid after extracting refining gourmet powder; It is a kind of single cell protein; Contain rich in protein, the chemical ingredients of tropina after the drying is analyzed found that Protein content is 78.77% up to 85.8% total aminoacid content in the aminoglutaminic acid thalline, be higher than present protein zymolyte raw material dregs of beans commonly used, yeast etc.Its amino acid kind and proportioning are all more complete, and contain other nutritive substances such as abundant VITAMINs, nucleic acid, polysaccharide.
In the L-glutamic acid production process, the most factories of process method in time do not handle and directly it are drained at present, have both polluted environment, cause great waste again; Also some producer waits the glutamic acid fermentation thalline after fermentation ends; Through from handing over post in waste liquid after the remaining L-glutamic acid absorption; Residue waste water adds the flocculation sediment that quantitative flocculation agent carries out tropina, behind Plate Filtration, obtains tropina, and dry back is as feed.This kind feed obtains through the flocculation agent depositing technology, because the introducing of macromole flocculation agent causes the nutritive value of feed to descend significantly, causes its utility value to reduce, and influences the performance of enterprises.
In recent years because the development part thalline of thalline recovery technology is recovered utilization; It is reported the Japan and the U.S. thereof consumption seasonings in; The protein digestion thing is because of its rich in amino acid content and higher nutritive value, and the actual measurement consumption has been equivalent to surpass the consumption of monosodium glutamate.In China, the consumption of this respect is not a lot of at present, makes cost recovery higher because technology is immature, but the Along with people's growth in the living standard, the protein digestion thing will have vast market.The method of at present glu thalline protein being carried out enzymolysis mainly contains chemistry and learns and enzyme process.Though chemical process is simple, the reaction requirement condition is high, seriously polluted, thereby very restricted; The enzyme process enzymolysis is to utilize protease hydrolyzed protein, and reaction conditions is gentle, and energy consumption is low, and side reaction is few, uses comparatively extensive.
Therefore, study a kind of system to the utilization of glu thalline protein enzymolysis, to reduce contaminated wastewater, to reduce because the introducing of flocculation agent causes the tropina nutritive value to descend is the technical problem that this area is needed solution badly.
Summary of the invention
The contriver is through a series of methods of utilizing of having researched and developed a kind of glutamic acid fermentation thalline; This method is passed through the tropina in the disc separator separate fermentation liquid before having adopted electricity such as fermented liquid, above-mentioned tropina slurries is added an amount of compound enzymic preparation make the tropina enzymolysis.This enzymolysis solution separate to be removed cell walls through high-speed dish piece powder machine, and gained clarification enzymolysis solution is processed enzymolysis tropina cream behind cryoconcentration, or after carrying out drying through spray granulating fluid bed dryer, spray granulating and drying machine etc., processes enzymolysis tropina powder.This method has increased substantially the tropina utility value; Thereby make it change high added value yeast extract paste, yeast powder surrogate into by cheap feed; This product is owing to contain a large amount of each anthropoid and necessary amino acid of animal; Thereby can be used as anino acid feed Preblend raw material, it is turned waste into wealth.
Utilize high-speed dish piece to separate and remove thalline and insolubles; L-glutamic acid feed liquid through after separating concentrates through multiple-effect plate-type evaporator low-temperature evaporation, and the secondary steam water of condensation of generation is used for the glutamic acid fermentation batching; L-glutamic acid liquid concentrator after the evaporation extracts L-glutamic acid through waiting electricity continuously; Supernatant is through adsorbing L-glutamic acid from the friendship post, and the L-glutamic acid of parsing goes to wait electricity to extract once more, and hc effluent removes fertilizer Workshop Production fertilizer; Heavy phase (tropina) is produced high protein feed through sheet frame squeezing back granulation through fluidised bed drying.
Above-mentioned enzymolysis tropina cream or protein powder are owing to be rich in a large amount of human bodies and animal institute indispensable amino acid; Can its as the compounded amino acid starting material; And suitably add feed amino acids such as part Methionin, Threonine, methionine(Met) and process the anino acid feed pre-mixture, reduce this product cost significantly.
For realizing above-mentioned purpose, a kind of glutamic acid fermentation thalline of the present invention utilizes the concrete operations step of method to be:
(1) glutamic acid fermentation bacterium liquid utilizes the high-speed dish piece separating machine that the L-glutamic acid feed liquid in the fermented liquid is separated with tropina, and the rotating speed of high-speed dish piece machine separating thallus is 4000～5000r/min, through above condition, the L-glutamic acid feed liquid is separated with tropina.
(2) separation is obtained using prozyme that it is carried out enzymolysis after tropina is diluted with water to contents on dry basis 5%～10%; Obtain enzymolysis solution, enzymatic hydrolysis condition is: 55 ℃ of temperature, pH6.5; Time 6h; Wherein prozyme is: N,O-Diacetylmuramidase, aspartic protease, beta-glucanase, three's ratio are N,O-Diacetylmuramidase: aspartic protease: beta-glucanase is 2: 5: 1, and prozyme addition (weight ratio of enzyme and tropina (substrate) dry weight) is: 5-7%;
(3) adopt disc separator to separate to above-mentioned enzymolysis solution and remove cell walls; The gained supernatant is through multiple-effect plate-type evaporator low-temperature evaporation; Said vaporizer one is imitated feeding temperature＜80 ℃; Drop temperature＜45 ℃ are imitated at the end, gained paste contents on dry basis＞65%, with above-mentioned paste directly packing process commercially available yeast extract paste substitute products enzymolysis tropina cream; Perhaps above-mentioned paste is processed powdery or granular material through spray granulating fluidized-bed or spray granulating and drying, after packing, process commercially available yeast powder surrogate enzymolysis tropina powder.
Above-mentioned zymolyte is through measuring aminoacids content, and after adding feed a part amino acid adjustment prescription in right amount, can be made into anino acid feed Preblend.
The present invention at first uses the high-speed dish piece separating machine that the L-glutamic acid feed liquid in the fermented liquid is separated with tropina, removes the fermentation thalline, and the rotating speed of high-speed dish piece machine separating thallus is 4000～5000r/min; Through above condition, high in efficiency and convenience the L-glutamic acid feed liquid is separated with tropina, this uses the high-speed dish piece separating machine to initiate as the applicant; Disc separator is a kind of in the settling centrifuge; Be used to separate difficult isolating material, it is more than extraction rate reached to 97% to tropina, and it is low that the membrane filtration technique in compared to prior art has a cost; The advantage of simple operation is more suitable for large-scale industrial production.
The application need not to add chemical reagent such as flocculation agent; The feed safety hidden danger of having avoided the flocculation agent monomer to exist, the easy enzymolysis of gained tropina has improved quality product; This method technology is simple, can be widely used in the extraction of L-glutamic acid and other amino acid fermentation tropina.
The inventor tests the enzymatic hydrolysis system of enzyme digestion reaction, addition, PH, temperature etc. through creative work and factorial experiment in a large number; Draw best enzyme digestion reaction parameter system, confirm that from a large amount of enzymolysis protein enzymes the best complex enzyme is combined as N,O-Diacetylmuramidase, aspartic protease, beta-glucanase, and confirmed its best adding proportion; This combination enzyme enzyme digestion reaction mild condition; Enzymolysis solution nutrition is kept perfectly, and the mutual induction of more cooperating improves reaction efficiency and transformation efficiency;
Significant variation takes place with the change of pH value in the enzyme reaction result, and each enzymatic reaction is all deposited and all had an optimum action pH value, when being lower than optimum pH; The protein digestion rate increases with the increase of pH value; After reaching optimum pH, with the increase of pH value, enzymatic hydrolyzation descends on the contrary.The present invention confirms that the best enzymolysis PH of prozyme is 6.5; The increase of enzyme amount can improve the ability to function of enzyme to substrate, can cause the raising of enzymatic hydrolyzation thus, when the enzyme dosage increases to certain numerical value; Protein molecule reaches capacity to the demand of enzyme; Enzyme no longer is the restrictive factor that improves the protein digestion rate, so enzymatic hydrolyzation becomes gently further increase along with the increase of enzyme dosage; Its enzymatic hydrolyzation descends on the contrary, and originally determining best complex enzyme addition is 5-7%; Enzymatic reaction is also very responsive to temperature, and under the situation that keeps enzyme activity, the protein digestion rate raises with temperature, reach the climax after, the continuation elevated temperature, enzyme activity begins to reduce, enzymolysis speed descends rapidly, the protein digestion rate descends thereupon.Originally determining optimal reaction temperature is 55 ℃;
The present invention need not to add chemical reagent such as flocculation agent with existing production technique; The feed safety hidden danger of having avoided the flocculation agent monomer to exist, the egg white icing of its preparation can directly be packed and process commercially available yeast extract paste substitute products, and gained enzymolysis protein cream substitutes commercially available yeast powder as nitrogenous source; Carry out the Threonine fermentation test; Continuous 5 jars of average fermentation parameters are respectively: with the fermented liquid product acid 13% of enzymolysis protein cream as nitrogenous source, fermentation period 28h, fermentation glucose acid invert ratio 68%.With the fermented liquid product acid 14% of commercially available yeast powder as nitrogenous source, fermentation period 28h, fermentation glucose acid invert ratio 67%.Be used for the alternative commercially available yeast powder product of Threonine fermentation finished thoroughly1 through above-mentioned relatively products made thereby of the present invention, thereby reduce the Threonine fermentation costs significantly.
The whole simple operation of process of the present invention, production cost is lower, and remarkable in economical benefits very is suitable for suitability for industrialized production.
Embodiment
Embodiment 1
(1) at first with 10 cubes of glutami acid fermentation liquor; Through disc separator the fermentation thalline is separated, the rotating speed of high-speed dish piece machine separating thallus is 4000～5000r/min, through above condition; The L-glutamic acid feed liquid is separated with tropina, get tropina 6500kg.
(2) above-mentioned tropina is added stirred reactor, and add an amount of warm water and mix well, adjustment solid content 8%, 55 ℃ of adjustment enzyme digestion reaction temperature add a little sulfuric acid adjustment pH6.5, add N,O-Diacetylmuramidase 10kg/m respectively
3, aspartic protease 25Kg/m
3, beta-glucanase 5Kg/m
3, slowly stirring enzymolysis time is 6 hours, measures all kinds of aminoacids contents of enzymolysis solution with amino acidanalyser.
(3) adopt disc separator to separate to above-mentioned enzymolysis solution and remove cell walls; The gained supernatant is through triple effect plate-type evaporator low-temperature evaporation, and said vaporizer one is imitated 78 ℃ of feeding temperatures, and 43 ℃ of drop temperatures are imitated at the end; Enzymolysis protein cream is processed in the direct barrelling of gained paste, gained egg white icing product 4000kg.
Perhaps above-mentioned paste is processed powdery or granular material through spray granulating fluidized-bed or spray granulating and drying, after packing, process commercially available yeast powder surrogate enzymolysis tropina powder.
Embodiment 2
Through passing through the spray granulating fluidised bed drying, the finished product packing that obtains is processed enzymolysis tropina powder, 130 ℃ of spray granulating fluidized-bed hot blast temperatures with embodiment 1 gained enzymolysis tropina cream.
Embodiment 3
Substitute commercially available yeast powder with embodiment 1 gained enzymolysis protein cream as nitrogenous source, carry out the Threonine fermentation test, continuous 5 jars of average fermentation parameters are respectively:
Produce acid 13%, fermentation period 28h, fermentation glucose acid invert ratio 68% with the enzymolysis protein cream of embodiment 1 preparation as the fermented liquid of nitrogenous source.
With the fermented liquid product acid 14% of commercially available yeast powder as nitrogenous source, fermentation period 28h, fermentation glucose acid invert ratio 67%.
Be used for the alternative commercially available yeast powder product of Threonine fermentation finished thoroughly1 through above-mentioned relatively products made thereby of the present invention, thereby reduce the Threonine fermentation costs significantly.
Embodiment 4
With the yeast powder in the enzymolysis protein powder alternate standard YPD substratum of embodiment 2 preparations, all the other components unchanged are cultivated yeast saccharomyces cerevisiae and pichia spp under the same conditions, estimate the culture effect of product through the growing state that compares cell.
Utilize turbidimetry for Determination OD
600The growing state of characterize cells shows with this product to substitute commercially available yeast powder product, and achieves the desired result.
Producing 200000 tons of L-glutamic acid production plants per year with Hulunbuir Northeast Fufeng Biotechnology Co., Ltd. is example, to adopting the cost analysis of comparing with traditional technology behind the present invention following:
2200 yuan/ton of the tropina prices of extracting through flocculating, annual 36000 tons of tropinas, about 9,000,000 yuan of flocculation agent and the drying cost sold; 7,920 ten thousand yuan of sales revenue after the separation, can be made into 25000 tons/year of enzymolysis protein powders with above-mentioned thalline before waiting electricity; 22000 yuan/ton of present commercially available yeast powder unit prices; Therefore can realize 55,000 ten thousand yuan of sales revenue, 2000 yuan of protein powder consumption zymin per ton and dry processing costs, so can increase 43,000 ten thousand yuan of incomes every year newly.
The present invention compares with common process, stable and reliable product quality, and production cost reduces significantly; For glutamate production enterprise; Not only can reduce environmental pollution, reduce the WWT expense, simultaneously resource made full use of; Reach the purpose of energy-saving consumption-reducing, will produce considerable social benefit and economic benefit.
Claims (5)
1. a glutamic acid fermentation thalline utilizes method, it is characterized in that the concrete operations step is following:
(1) utilize the high-speed dish piece separating machine that the L-glutamic acid feed liquid in the glutamic acid fermentation bacterium liquid is separated with tropina, the rotating speed of high-speed dish piece machine separating thallus is 4000～5000r/min, through above condition, the L-glutamic acid feed liquid is separated with tropina.
(2) separation is obtained using prozyme that it is carried out enzymolysis after tropina is diluted with water to solid content 5%～10%; Obtain enzymolysis solution, enzymatic hydrolysis condition is: 55 ℃ of temperature, pH6.5; Time 6h; Wherein prozyme is: N,O-Diacetylmuramidase, aspartic protease, beta-glucanase, three's ratio are N,O-Diacetylmuramidase: aspartic protease: beta-glucanase is 2: 5: 1, and the prozyme addition is: 5-7%;
(3) adopt disc separator to separate to above-mentioned enzymolysis solution and remove cell walls; The gained supernatant is through triple effect plate-type evaporator low-temperature evaporation; Said vaporizer one is imitated feeding temperature＜80 ℃; Drop temperature＜45 ℃ are imitated at the end, and gained paste contents on dry basis reaches 65%, and directly packing makes enzymolysis tropina cream.
2. method according to claim 1 is characterized in that, solid content is 8% in the step (2), and one imitates 78 ℃ of feeding temperatures in the step (3), and 43 ℃ of drop temperatures are imitated at the end.
3. method according to claim 1 and 2 is characterized in that, egg white icing is processed powdery or granular material through fluidized bed for spraying, granulating or spray granulating and drying in the step (3).
4. the said enzymolysis tropina of claim 1 cream is used for substituting the application of yeast extract paste.
5. claim 1 or 2 said enzymolysis tropina cream and the said powdery of claim 3 or granular material are as the application of amino acid feed pre-mixture raw material.
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| CN104250283A (en) * | 2014-09-19 | 2014-12-31 | 中农颖泰林州生物科园有限公司 | Separation technology for product containing antibacterial peptide |
| CN104726335A (en) * | 2015-04-13 | 2015-06-24 | 呼伦贝尔东北阜丰生物科技有限公司 | Method for manufacturing granule proteins by aid of waste mother liquid of lysine |
| CN104757273A (en) * | 2015-04-12 | 2015-07-08 | 呼伦贝尔东北阜丰生物科技有限公司 | Process for preparing granular type threonine product |
| CN104817376A (en) * | 2015-04-11 | 2015-08-05 | 呼伦贝尔东北阜丰生物科技有限公司 | Bio-organic fertilizer prepared from lysine waste mother liquor |
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| CN106946976A (en) * | 2017-05-19 | 2017-07-14 | 卢松 | Recycle the method that glutamic acid fermentation discards thalline |
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| CN108588139A (en) * | 2018-05-16 | 2018-09-28 | 安徽省芬格欣生物药业有限公司 | A kind of method of spina date seed fermenting and producing glutamic acid and application |
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| CN104250283A (en) * | 2014-09-19 | 2014-12-31 | 中农颖泰林州生物科园有限公司 | Separation technology for product containing antibacterial peptide |
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| CN107164247A (en) * | 2017-06-03 | 2017-09-15 | 秦皇岛华恒生物工程有限公司 | A kind of method of utilization L alanine fermentation wastes culture yeasts |
| CN109987614A (en) * | 2017-12-30 | 2019-07-09 | 许传高 | A method of extracting ammonium sulfate from glutamic acid fermentation tail washings |
| CN109984149A (en) * | 2017-12-30 | 2019-07-09 | 许传高 | A method of biostimulant is prepared using amino acid fermentation tail washings |
| CN108588139A (en) * | 2018-05-16 | 2018-09-28 | 安徽省芬格欣生物药业有限公司 | A kind of method of spina date seed fermenting and producing glutamic acid and application |
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