CN102719510B - Amino acid fermentation bacteria utilization method - Google Patents
Amino acid fermentation bacteria utilization method Download PDFInfo
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- CN102719510B CN102719510B CN 201210211399 CN201210211399A CN102719510B CN 102719510 B CN102719510 B CN 102719510B CN 201210211399 CN201210211399 CN 201210211399 CN 201210211399 A CN201210211399 A CN 201210211399A CN 102719510 B CN102719510 B CN 102719510B
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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 acids 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 to the method that enzymolysis prepares enzymolysis protein cream, protein powder.
Background technology
Aminoglutaminic acid thalline is the byproduct in fermentative Production monosodium glutamate process, 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, it is 78.77% up to 85.8% total amino acid content that the chemical composition of tropina after drying is analyzed to the content of finding protein in aminoglutaminic acid thalline, higher than protein zymolyte is commonly used at present raw material dregs of beans, yeast etc.Its amino acid kind and proportioning are all more complete, and contain other nutritive substances such as abundant VITAMIN, nucleic acid, polysaccharide.
In the L-glutamic acid production process, the most factories of processing method do not process in time and directly it are drained at present, have both polluted environment, cause again great waste; Also some producer waits the glutamic acid fermentation thalline for electricity after fermentation ends, after the L-glutamic acid from handing over post by remnants in waste liquid adsorbs, the residue waste water add quantitative flocculation agent to carry out the flocculation sediment of tropina, obtain tropina after Plate Filtration, after drying as feed.This kind of feed obtains through the flocculation agent depositing technology, because the introducing of macromole flocculation agent causes the nutritive value of feed significantly to descend, causes its utility value to reduce, and affects the performance of enterprises.
Development part thalline due to the thalline recovery technology is recovered utilization in recent years, it is reported in the seasonings of Japan and the U.S. thereof consumption, the protein digestion thing is because of its abundant aminoacids 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, because technology is immature, makes cost recovery higher, but, along with the raising of people's living standard, the protein digestion thing will have wide market.The method of at present glu thalline protein being carried out to enzymolysis mainly contains Science of Chemistry and enzyme process.Although 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, the reaction conditions gentleness, and energy consumption is low, and side reaction is few, applies comparatively extensive.
Therefore, study a kind of system to the utilization of glu thalline protein enzymolysis, reducing contaminated wastewater, reducing introducing due to flocculation agent, to cause the tropina nutritive value to descend be the technical problem that this area is needed solution badly.
Summary of the invention
The contriver is through a series of a kind of researched and developed glutamic acid fermentation thalline methods of utilizing, the method is passed through the tropina in disc separator separate fermentation liquid before having adopted the electricity such as fermented liquid, adds appropriate compound enzymic preparation to make the tropina enzymolysis in above-mentioned tropina slurries.This enzymolysis solution separates and removes cell walls through high-speed dish piece powder machine, and gained clarification enzymolysis solution is made enzymolysis tropina cream after cryoconcentration, or, after spray granulating fluid bed dryer, Spray Grain-make Drier etc. carry out drying, makes enzymolysis tropina powder.The method has increased substantially the tropina utility value, thereby make it by cheap feed, change high added value yeast extract paste, yeast powder surrogate into, this product is owing to containing 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 is concentrated through Multi-effect plate type vaporizer low-temperature evaporation, the secondary steam water of condensation produced, for the glutamic acid fermentation batching, L-glutamic acid concentrated solution after evaporation is by waiting continuously electricity to extract L-glutamic acid, supernatant liquor is by adsorbing L-glutamic acid from the friendship post, the L-glutamic acid of resolving goes to wait electricity again to extract, hc effluent removes fertilizer Workshop Production fertilizer, heavy phase (tropina) is squeezed rear granulation by sheet frame, by fluidised bed drying, produces high protein feed.
Above-mentioned enzymolysis tropina cream or protein powder are owing to being 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 the feed amino acids such as part Methionin, Threonine, methionine(Met) and make the anino acid feed pre-mixture, this product cost of decrease.
For achieving the above object, a kind of glutamic acid fermentation thalline of the present invention utilizes the concrete operation 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 fermented liquid is separated with tropina, the rotating speed of high-speed dish piece machine separating thallus is 4000～5000r/min, by above condition, the L-glutamic acid feed liquid is separated with tropina.
(2) separation being obtained to tropina is diluted with water to the rear use prozyme of contents on dry basis 5%～10% it is carried out to enzymolysis, obtain enzymolysis solution, enzymatic hydrolysis condition is: 55 ℃ of temperature, pH6.5, time 6h, wherein prozyme is: N,O-Diacetylmuramidase, aspartic protease, beta-glucanase, and three's ratio is N,O-Diacetylmuramidase: aspartic protease: beta-glucanase is 2: 5: 1, prozyme addition (weight ratio of enzyme and tropina (substrate) dry weight) is: 5-7%;
(3) to above-mentioned enzymolysis solution, adopt disc separator to separate and remove cell walls, the gained supernatant liquor is through Multi-effect plate type vaporizer low-temperature evaporation, described vaporizer one effect feeding temperature<80 ℃, end effect drop temperature<45 ℃, gained paste contents on dry basis>65%, above-mentioned paste is directly packed and made commercially available yeast extract paste substitute products enzymolysis tropina cream, perhaps above-mentioned paste is made to powdery or granular material through spray granulating fluidized-bed or spray granulating and drying, after packing, make commercially available yeast powder surrogate enzymolysis tropina powder.
Above-mentioned zymolyte is aminoacids content after measured, and, after adding in right amount feed a part amino acid adjustment formula, can be made into anino acid feed Preblend.
At first the present invention uses the high-speed dish piece separating machine that the L-glutamic acid feed liquid in fermented liquid is separated with tropina, remove the fermentation thalline, the rotating speed of high-speed dish piece machine separating thallus is 4000～5000r/min, by above condition, efficiently easily the L-glutamic acid feed liquid is separated with tropina, this uses the high-speed dish piece separating machine pioneering for the applicant, disc separator is a kind of in settling centrifuge, the material separated for separating of difficulty, it is more than extraction rate reached to 97% to tropina, it is low that membrane filtration technique compared to prior art has a cost, the advantage of simple operation, be more suitable for large-scale industrial production.
The application is without adding the 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, improved quality product, this method technique is simple, can be widely used in the extraction of L-glutamic acid and other amino acid fermentation tropina.
The inventor carries out great many of experiments by creative work and factorial experiment to the enzymatic hydrolysis system of enzyme digestion reaction, addition, PH, temperature etc., draw best enzyme digestion reaction parameter system, determine that from a large amount of enzymolysis protein enzymes the best complex enzyme is combined as N,O-Diacetylmuramidase, aspartic protease, beta-glucanase, and determined its best adding proportion, this combination enzyme reaction conditions gentleness, it is complete that enzymolysis solution nutrition keeps, and the mutual induction that more can cooperate improves reaction efficiency and transformation efficiency;
Significant variation occurs 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, lower than optimum pH the time, 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 determines 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 is no longer the restrictive factor that improves the protein digestion rate, thus enzymatic hydrolyzation along with the increase of enzyme dosage, become gently, further increase, 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, in the situation that keep enzyme activity, the protein digestion rate raises with temperature, after reaching climax, continues the rising temperature, and enzyme activity starts to reduce, and enzymolysis speed descends rapidly, and the protein digestion rate descends thereupon.Originally determining optimal reaction temperature is 55 ℃;
The present invention with existing production technique without adding the chemical reagent such as flocculation agent, the feed safety hidden danger of having avoided the flocculation agent monomer to exist, its egg white icing prepared can directly be packed and make commercially available yeast extract paste substitute products, gained enzymolysis protein cream substitutes commercially available yeast powder as nitrogenous source, carry out the Threonine Fermentation test, continuous 5 tank average fermentation parameters are respectively: the fermented liquid product acid 13% with enzymolysis protein cream as nitrogenous source, fermentation period 28h, fermentation glucose acid invert ratio 68%.Fermented liquid product acid 14% with commercially available yeast powder as nitrogenous source, fermentation period 28h, fermentation glucose acid invert ratio 67%.Be used for Threonine Fermentation alternative commercially available yeast powder product fully by above-mentioned relatively products made thereby of the present invention, thus decrease Threonine Fermentation cost.
The whole simple operation of process of the present invention, production cost is lower, and remarkable in economical benefits extremely is suitable for suitability for industrialized production.
Embodiment
Embodiment 1
(1) at first by 10 cubes of glutami acid fermentation liquors, by disc separator, the fermentation thalline is separated, the rotating speed of high-speed dish piece machine separating thallus is 4000～5000r/min, by above condition, the L-glutamic acid feed liquid is separated with tropina, obtain tropina 6500kg.
(2) above-mentioned tropina is added to stirred reactor, and add appropriate warm water to mix well, adjust solid content 8%, adjust 55 ℃ of enzyme digestion reaction temperature, add a little sulfuric acid and adjust pH6.5, add respectively N,O-Diacetylmuramidase 10kg/m
3, aspartic protease 25Kg/m
3, beta-glucanase 5Kg/m
3, slowly stirring enzymolysis time is 6 hours, with amino acidanalyser, measures all kinds of aminoacids contents of enzymolysis solution.
(3) to above-mentioned enzymolysis solution, adopt disc separator to separate and remove cell walls, the gained supernatant liquor is through triple effect plate-type evaporator low-temperature evaporation, 78 ℃ of described vaporizer one effect feeding temperatures, 43 ℃ of end effect drop temperatures, enzymolysis protein cream is made in the direct barrelling of gained paste, gained egg white icing product 4000kg.
Perhaps above-mentioned paste is made to powdery or granular material through spray granulating fluidized-bed or spray granulating and drying, after packing, make commercially available yeast powder surrogate enzymolysis tropina powder.
Embodiment 2
By embodiment 1 gained enzymolysis tropina cream, through passing through the spray granulating fluidised bed drying, the finished product packing obtained is made enzymolysis tropina powder, 130 ℃ of spray granulating fluid bed heat air temperatures.
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 tank average fermentation parameters are respectively:
The enzymolysis protein cream prepared with embodiment 1 produces acid 13% as the fermented liquid of nitrogenous source, fermentation period 28h, fermentation glucose acid invert ratio 68%.
Fermented liquid product acid 14% with commercially available yeast powder as nitrogenous source, fermentation period 28h, fermentation glucose acid invert ratio 67%.
Be used for Threonine Fermentation alternative commercially available yeast powder product fully by above-mentioned relatively products made thereby of the present invention, thus decrease Threonine Fermentation cost.
Embodiment 4
With the yeast powder in the enzymolysis protein powder alternate standard YPD substratum of embodiment 2 preparations, all the other components unchanged, cultivate yeast saccharomyces cerevisiae and pichia spp under the same conditions, estimates the culture effect of product by the growing state that compares cell.
Utilize turbidimetry for Determination OD
600the growing state of characterize cells, show can substitute commercially available yeast powder product with this product, and achieve the desired result.
Hulunbuir Northeast Fufeng Biotechnology Co., Ltd.'s 200,000 tons of L-glutamic acid production plants of annual output of take are example, to adopting the cost analysis of comparing with traditional technology after the present invention as follows:
2200 yuan/tons of the tropina prices of extracting by flocculation, annual 36000 tons of the tropinas of selling, approximately 9,000,000 yuan of flocculation agent and drying costs, 7,920 ten thousand yuan of sales revenue, after above-mentioned thalline is separated before waiting electricity, can be made into 25000 ton/years of enzymolysis protein powders, 22000 yuan/tons of current commercially available yeast powder unit prices, but therefore the enforcement sale income is 55,000 ten thousand yuan, 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, the production cost decrease, for glutamate production enterprise, not only can environmental contamination reduction, reduce the sewage disposal expense, resource is taken full advantage of simultaneously, reach the purpose of energy-saving consumption-reducing, will produce considerable Social benefit and economic benefit.
Claims (1)
1. a glutamic acid fermentation thalline utilizes method, it is characterized in that concrete operation step is as follows:
(1) utilize the high-speed dish piece separating machine that the L-glutamic acid feed liquid in glutamic acid fermentation bacterium liquid is separated with tropina, the rotating speed of high-speed dish piece machine separating thallus is 4000～5000r/min, by above condition, the L-glutamic acid feed liquid is separated with tropina;
(2) separation being obtained to tropina is diluted with water to the rear use prozyme of solid content 8% it is carried out to enzymolysis, obtain enzymolysis solution, enzymatic hydrolysis condition is: 55 ℃ of temperature, pH6.5, time 6h, wherein prozyme is: N,O-Diacetylmuramidase, aspartic protease, beta-glucanase, and three's ratio is N,O-Diacetylmuramidase: aspartic protease: beta-glucanase is 2: 5: 1, and the prozyme addition is: 5-7%;
(3) to above-mentioned enzymolysis solution, adopt disc separator to separate and remove cell walls, the gained supernatant liquor is through triple effect plate-type evaporator low-temperature evaporation, and described vaporizer one is imitated 78 ℃ of feeding temperatures, 43 ℃ of end effect drop temperatures, gained paste contents on dry basis reaches 65%, and directly packing makes enzymolysis tropina cream.
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| CN1233619C (en) * | 2004-07-22 | 2005-12-28 | 徐昌洪 | Glutamic acid extraction technology in production of glutamic acid by fermentation method |
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