CN104404094A - Method for extracting taurine by use of enzymatic conversion method on the basis of clams - Google Patents
Method for extracting taurine by use of enzymatic conversion method on the basis of clams Download PDFInfo
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Abstract
The invention discloses a method for extracting taurine by use of enzymatic conversion method on the basis of clams, and the method comprises the following steps: 1 pretreatment of the clams or a clam processing water boiling liquid, to be more specific, separating clam meat, cleaning, homogenizing; or collecting waste clam water boiling liquid in the processing process of the clams, and filtering to remove impurities; 2, enzymatic conversion, to be more specific, adding enzyme in appropriate pH conditions for enzymatic conversion reaction; 3, removal of protein, to be more specific, precipitating in appropriate pH conditions, and centrifuging for removal of acidic and alkaline proteins; 4, use of an ultrafiltration device for removal of macromolecules, to be more specific, blocking macromolecular polysaccharides and peptides to retain the taurine by the ultrafiltration device; 5, electrodialysis for removal of salt ions; 6, ion exchange chromatography, to be more specific, using strong acidic cation exchange resin for adsorption and collection of the taurine; and 7, alcohol recrystallization, to be more specific, adding a certain volume of ethanol for precipitation of taurine crystals. The method fully utilizes the waste material, and has the advantages of simple operation, safety, low cost, green environmental protection, continuous production, high taurine yield and the like.
Description
Technical field
The present invention relates to the method for taurine in a kind of industrialized utilization enzymatic methods of conversion continuous production clam or its water cooking liquid.
Background technology
Clam class, such as philippine clam whelp, be commonly called as husky a species of small clam living in fresh water, is a class bivalve shellfish, is under the jurisdiction of Veneridae, is distributed widely in the marine site, north and south of China.Its principal feature be growth rapidly, the breeding cycle is short, to the adaptable of salt and temperature and flavour delicious, nutritious, low price.Be the second largest shellfish culture product of China, therefore have extremely important economic worth to the deep development utilization of clam and processing byproduct thereof.Containing mineral substance, VITAMIN, amino acid, taurines such as rich in protein, fat, calcium, phosphorus, iron in fresh clam meat.
Taurine (Taurine) formal name used at school 2-aminoethyl sulfonic acid, because separating from oxgall first, therefore is commonly called as taurocholic acid, Taurine.Taurine is a kind of nonprotein structural amino acid of sulfur-bearing, is one of amino acid of needed by human, has very strong biological activity.Taurine content in fish, shellfish is very abundant.Found by further investigation; taurine can increase cellular anti-oxidant, Green Tea Extract damages and the ability of antiviral infringement; it or good liver-protecting preparation; there is vision enhancing, promote brain development, relieving fatigue, increase work efficiency, reduce cholesterol, suppression gallbladdergallstonecholetithiasis, anti-inflammatory, analgesic effect, there is certain anti-tumor activity simultaneously.Taurine can be used for the diseases such as treatment acute, chronic hepatitis, flu, hypertension, arteriosclerosis and cancer clinically.
The demand of taurine significantly increases, and there is the potential market of 20000 tons/year in China, and U.S.'s year consumption reaches more than 6000 tons, nearly 2000 tons of Japanese output.
For growing demand, the more technology extracting taurine upgrades day by day.At present, the main chemical synthesis that relies on extracts taurine both at home and abroad, adopts the methods such as second monochloroethane method, girbotol process, ethyleneimine method more.But the problems such as material toxicity is large, complex operation, recovery is difficult, environmental pollution is serious that these chemical synthesiss exist.
At present for the comprehensive utilization of oceanic resources, be obtain relative high value-added product from the fishery products of low value on the one hand; That the waste produced in processing of aquatic products process is recycled on the other hand.Bring larger pressure owing to producing waste to environment, higher environmental benefit can be brought undoubtedly to its utilization.The tankage utilizing marine prods to process extract taurine, both can improve oceanic resources utilization ratio, and can reduce environmental pollution again.Chinese Marine Biology Resources is enriched, and extracts taurine, have great economic benefit from fish and shellfish and tankage thereof.So how research extracts taurine efficiently from marine organisms have important practical significance.
Summary of the invention
For above-mentioned Problems existing; the present invention relates to a kind of conventional chemical synthesis that can substitute to extract taurine or adopt water-boiling method to extract the technology of taurine; namely in clam and water cooking liquid thereof, protease is added; utilize enzyme transforming process; extract taurine; the present invention is with low cost, environmental protection, simple to operate, extraction efficiency is high, fully uses scrap feed material and effectively improves the yield of taurine by the effect of enzymatic conversion.
In order to achieve the above object, the invention provides a kind of method utilizing enzyme transforming process extraction taurine based on clam, comprise the steps:
S1, utilize clam preparation feedback liquid;
S2, the one of getting in stomach en-, papoid, trypsinase, Sumizyme MP, the pH value suitable to the conversion reaction of described reaction solution regulatory enzyme according to selected enzyme, adds selected enzyme in described reaction solution and carries out enzymatic conversion reaction; Afterwards, the enzyme that goes out is lived, and obtains enzymatic conversion solution;
S3, described enzymatic conversion solution removal are acid, basic protein;
S4, remove macromolecular substance by ultrafiltration apparatus, retain taurine;
S5, electrodialysis remove salt ion;
S6, by storng-acid cation exchange resin absorptive collection taurine effluent liquid;
Add the ethanol of certain volume in S7, effluent liquid, separate out taurine crystal.
Under optimal way, step S2 specifically comprises operation:
S21, in described reaction solution, add stomach en-, papoid, trypsinase, one in Sumizyme MP that dry biomass number in solution is 0.1% ~ 10%; Regulate pH to the control environment (control environment is: stomach en-pH1 ~ 3, papoid pH5 ~ 9, trypsinase pH7 ~ 9, Sumizyme MP pH8 ~ 11) of enzyme used, 20 ~ 50 DEG C of enzymolysis 0.5 ~ 10 hour;
S22, cooling, adjust pH to 6 ~ 8 (this step effectively can ensure the last yield of product);
S23,80 ~ 100 DEG C of enzymes that go out are lived 5 ~ 30 minutes.
In addition, step S1 can select clam meat or clam water cooking liquid preparation feedback liquid, is specially: clam meat carries out homogenized after adding the water of 1:1 ~ 1:100 by its volume ratio, obtains described reaction solution; Or it is between 1% ~ 30% that clam water cooking liquid is concentrated into dry matter content, the water-fast impurity of centrifugal segregation, obtains described reaction solution.Described water cooking liquid can be the waste liquid (prior art is all abandoned) in the clam course of processing, thus improves value of the product further, promotes profit.
Step S3 can with reference to the mode of prior art, specifically can with reference to " in clam taurine " [Gong Lifen, Huang Weisheng; Xie Xiaolan, Zheng Zhifu, Hu Donghong. the extraction (I) of taurine in clam. fine chemistry industry .2003; 20 (7); 393-395], first regulator solution is acid, has acidic protein to precipitate; can centrifugally remove; rear regulator solution alkalescence, has basic protein to precipitate, can centrifugally remove.The order of acid or alkali is regulated to change.The invention provides a kind of optimal way, be specially step S3 and comprise operation:
S31, to process after enzymatic conversion solution in add 1mol/L ~ 6mol/L HCl adjust pH to 2 ~ 4, namely have acidic protein to precipitate, centrifugation, get supernatant liquor:
S32, supernatant liquor 1mol/L ~ 6mol/L NaOH adjusts pH to 8 ~ 10, and namely have basic protein to precipitate, centrifugation, gets supernatant liquor.
Step S4: the solution after removing protein is carried out uf processing, clips macromolecular polypeptide polysaccharose substance with ultra-filtration membrane, collects filtrate.Electrodialysis process parameter suitable in step S5 is: voltage 1V ~ 20V, flow velocity 0.5mL/min ~ 4L/min, sample concentration 1mg/mL ~ 100mg/mL.
Above-mentioned steps S6 and S7 can realize with reference to the mode of prior art; as " in mussel waste liquid, the extraction of natural taurine is studied " [Cui Zhongai; Li Pingping; kingly way ripple; Fan Zhendi; Du Yanbing. the extraction research of natural taurine in mussel waste liquid. Agriculture of Anhui science .2010,38 (16): 8671-8673].Wherein, step S6 comprises following operation:
S61, by extracting solution by strong acid cation exchange resin column, distilled water wash-out, power taking conductance starts the effluent liquid dropped between stationary phase;
S62, by 6 × 60cmNa on extracting solution
+type highly acidic resin post, loading volume is 300mL, distilled water wash-out, and elution speed is 20mL/min, accesses the effluent liquid during specific conductivity starts to drop to stable conductivity.
Step S7 comprises following operation:
S71, above-mentioned effluent liquid, through concentrating under reduced pressure, add 3 times of volume 95% ethanol, under being placed in 4 DEG C of environment, separate out taurine.
S72, crude product taurine add suitable quantity of water and dissolve, then use 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain the taurine crystal that purity is higher.
Under optimum way, solution obtains taurine through lyophilize or vacuum drying method.
Taurine in fishery products exists in a free form, and is soluble in hot water, is insoluble to ethanol, so can adopt hot water extraction, ion exchange purification, the methods such as alcohol settling obtain white crystals.The present invention utilizes enzyme transforming process to extract taurine in clam water cooking liquid, have simple to operate, cost is low, yield is high, environmental protection, safety can continuous seepage.Its key step comprises: 1, collect enterprise and produce discarded clam water cooking liquid, filter and remove water-insoluble impurity; 2, enzymatic conversion reaction is carried out under adding proteolytic enzyme appropriate pH condition in water cooking liquid; 3, precipitation also centrifugal segregation acidity, basic protein under proper pH value condition; 4, remove macromolecular substance by ultrafiltration apparatus, retain taurine; 5, electrodialysis removes salt ion; 6, by storng-acid cation exchange resin absorptive collection taurine effluent liquid; 7, add the ethanol of certain volume in effluent liquid, separate out taurine crystal.
Tool of the present invention has the following advantages;
1, whole leaching process does not use any organic solvent, decreases environmental pollution, ensure that the inherently safe of staff.
2, gained taurine purification product being mixed into without any toxic reagent, safe and reliable.
3, the discarded water cooking liquid produced after can making full use of low value clam or its processing, improves the utilization ratio of waste liquid.
4, adopt enzyme transforming process to extract taurine, effectively can improve yield by the method for enzymatic conversion.
5, adopt uf processing and electrodialytic technique, while the purity effectively improving taurine, also can get rid of the heavy metal element existed in fishery products.
6, can realize full-automatic continuous prodution, equipment is simple, operates easy, reduces labour intensity.
Accompanying drawing explanation
Fig. 1 is the changing conditions of taurine content in the product before and after enzymatic conversion.
Embodiment
Example one
Clam is shelled, cleaning, and 1:10 adds water by volume, homogenate; Take enzyme assisted extraction method, add Sumizyme MP powder in homogenate, add-on is 2.5% of amount of substrate, and with 1mol/LNaOH regulator solution pH to 10,37 DEG C are reacted 3.5 hours, and reaction terminates, and adjust pH6, and the enzyme that goes out in boiling water bath 10min alive, cools.
With the HCl regulator solution pH to 3 of 6mol/L, namely have acidic protein to precipitate, centrifugation, gets supernatant liquor, then to be adjusted to pH with 4mol/L NaOH be l0, and namely have basic protein to precipitate, centrifugation, gets supernatant liquor.
Supernatant liquor is carried out uf processing, and ultra-filtration membrane retains macromolecular polysaccharide peptide material, collects filtrate, then removes salt ion through electrodialysis, electrodialysis voltage 1V, flow velocity 4L/min, sample concentration 1mg/mL.Filtrate, with 6mol/LHCl regulator solution pH to 3, treats upper prop.
Get 300mL filtrate by Na ionic acidic resins post, distilled water wash-out, elution speed is 10mL/min; access the effluent liquid that specific conductivity starts to drop between stationary phase, above-mentioned effluent liquid, through concentrating under reduced pressure, adds 3 times of volume 95% ethanol; under being placed in 4 DEG C of environment, separate out taurine.Crude product taurine adds suitable quantity of water and dissolves, then uses 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain taurine solid, and the yield that warp and methyl ethyl diketone and formaldehyde colorimetry record taurine product can reach 2.4%.
Example two
Clam is shelled, cleaning, and 1:10 adds water by volume, homogenate; Take enzyme assisted extraction method, add aspartic protease powder in homogenate, add-on is 2.5% of amount of substrate, and with 4mol/LHCl regulator solution pH Isosorbide-5-Nitrae 5 DEG C reaction 3.5 hours, after completing, adjust pH7, go out in boiling water bath enzyme 10min, cooling.
With the HCl regulator solution pH to 3 of 6mol/L, namely have acidic protein to precipitate, centrifugation, gets supernatant liquor, then adjusts pH to 8 with 4mol/L NaOH, and namely have basic protein to precipitate, centrifugation, gets supernatant liquor.
Supernatant liquor is carried out uf processing, and ultra-filtration membrane retains macromolecular polysaccharide peptide material, collects filtrate, then removes salt ion through electrodialysis, electrodialysis voltage 15V, flow velocity 100mL/min, sample concentration 8g/mL.Filtrate, with the HCl regulator solution pH to 3 of 6mol/L, treats upper prop.
Get 300mL filtrate by Na ionic acidic resins post, distilled water wash-out, elution speed is 10mL/min; access the effluent liquid that specific conductivity starts to drop between stationary phase, above-mentioned effluent liquid, through concentrating under reduced pressure, adds 3 times of volume 95% ethanol; under being placed in 4 DEG C of environment, separate out taurine.Crude product taurine adds suitable quantity of water and dissolves, then uses 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain taurine solid, and the yield that warp and methyl ethyl diketone and formaldehyde colorimetry record taurine product can reach 1.8%.
Example three
Collect enterprise and produce clam water cooking liquid, remove insoluble impurities; With the HCl regulator solution pH to 3 of 6mol/L, namely have acidic protein to precipitate, centrifugation, gets supernatant liquor, then to be adjusted to pH with 4mol/L NaOH be 9, and namely have basic protein to precipitate, centrifugation, gets supernatant liquor.Supernatant liquor is carried out uf processing, and ultra-filtration membrane retains macromolecular polysaccharide peptide material, collects filtrate, then removes salt ion through electrodialysis, electrodialysis voltage 1V, flow velocity 0.5mL/min, sample concentration 1mg/mL.Filtrate, with 6mol/LHCl regulator solution pH to 3, treats upper prop.
Get 300mL filtrate by Na ionic acidic resins post, distilled water wash-out, elution speed is 10mL/min; access the effluent liquid that specific conductivity starts to drop between stationary phase, above-mentioned effluent liquid, through concentrating under reduced pressure, adds 3 times of volume 95% ethanol; under being placed in 4 DEG C of environment, separate out taurine.Crude product taurine adds suitable quantity of water and dissolves, then uses 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain taurine solid, and the yield that warp and methyl ethyl diketone and formaldehyde colorimetry record taurine product can reach 0.182g/L.
Example four
Collect enterprise and produce clam water cooking liquid, remove insoluble impurities; Take enzyme assisted extraction method, add Sumizyme MP powder in water cooking liquid, add-on is 2.5% of amount of substrate, and with 1mol/LNaOH regulator solution pH to 10,37 DEG C are reacted 3.5 hours, and reaction terminates, and adjust pH8, and the enzyme that goes out in boiling water bath 10min alive, cools.
With the HCl regulator solution pH to 3 of 6mol/L, namely have acidic protein to precipitate, centrifugation, gets supernatant liquor, then to be adjusted to pH with 6mol/L NaOH be l0, and namely have basic protein to precipitate, centrifugation, gets supernatant liquor.
Supernatant liquor is carried out uf processing, and ultra-filtration membrane retains macromolecular polysaccharide peptide material, collects filtrate, then removes salt ion through electrodialysis, electrodialysis voltage 1V, flow velocity 1L/min, sample concentration 1mg/mL.Filtrate, with 6mol/LHCl regulator solution pH to 3, treats upper prop.
Get 300mL filtrate by Na ionic acidic resins post, distilled water wash-out, elution speed is 10mL/min; access the effluent liquid that specific conductivity starts to drop between stationary phase, above-mentioned effluent liquid, through concentrating under reduced pressure, adds 3 times of volume 95% ethanol; under being placed in 4 DEG C of environment, separate out taurine.Crude product taurine adds suitable quantity of water and dissolves, then uses 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain taurine solid, and the yield that warp and methyl ethyl diketone and formaldehyde colorimetry record taurine product can reach 0.316g/L.
Example five
Collect enterprise and produce clam water cooking liquid, remove water-insoluble impurity.Take enzyme assisted extraction method, add stomach en-in water cooking liquid, add-on is 2.5% of amount of substrate, and with the HCl regulator solution pH to 1 of 1mol/L, 20 DEG C are reacted 3.5 hours, and reaction terminates, and adjust pH to 7, and the enzyme that goes out in boiling water bath 10min alive, cools.
With the HCl regulator solution pH to 3 of 6mol/L, namely have acidic protein to precipitate, centrifugation, gets supernatant liquor, then adjusts pH to 8 with 4mol/L NaOH, and namely have basic protein to precipitate, centrifugation, gets supernatant liquor.
Supernatant liquor is carried out uf processing, and ultra-filtration membrane retains macromolecular polysaccharide peptide material, collects filtrate, then removes salt ion through electrodialysis, electrodialysis voltage 5V, flow velocity 100mL/min, sample concentration 2mg/mL.Filtrate, with the HCl regulator solution pH to 3 of 6mol/L, treats upper prop.
Get 300mL filtrate by Na ionic acidic resins post, distilled water wash-out, elution speed is 10mL/min; access the effluent liquid that specific conductivity starts to drop between stationary phase, above-mentioned effluent liquid, through concentrating under reduced pressure, adds 3 times of volume 95% ethanol; under being placed in 4 DEG C of environment, separate out taurine.Crude product taurine adds suitable quantity of water and dissolves, then uses 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain taurine solid, and the yield that warp and methyl ethyl diketone and formaldehyde colorimetry record taurine product can reach 0.284g/L.
Example six
Collect enterprise and produce clam water cooking liquid, remove water-insoluble impurity.Take enzyme assisted extraction method, add papoid in water cooking liquid, add-on is 2.5% of amount of substrate, is adjusted to pH value of solution 8 with 6mol/LNaOH, and 50 DEG C are reacted 3.5 hours, after completing, adjusts pH8, the enzyme that goes out in boiling water bath 20min alive, cooling.
Be adjusted to pH for l0 with 1mol/L NaOH, namely have basic protein to precipitate, centrifugation, gets supernatant liquor.Use the HCl regulator solution pH to 4 of 6mol/L again, namely have acidic protein to precipitate, centrifugation, gets supernatant liquor.
Supernatant liquor is carried out uf processing, and ultra-filtration membrane retains macromolecular polysaccharide peptide material, collects filtrate, then removes salt ion through electrodialysis, electrodialysis voltage 10V, flow velocity 3L/min, sample concentration 4mg/mL.Filtrate, with the HCl regulator solution pH to 3 of 6mol/L, treats upper prop.
Get 300mL filtrate by Na ionic acidic resins post, distilled water wash-out, elution speed is 10mL/min; access the effluent liquid that specific conductivity starts to drop between stationary phase, above-mentioned effluent liquid, through concentrating under reduced pressure, adds 3 times of volume 95% ethanol; under being placed in 4 DEG C of environment, separate out taurine.Crude product taurine adds suitable quantity of water and dissolves, then uses 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain taurine solid, and the yield that warp and methyl ethyl diketone and formaldehyde colorimetry record taurine product can reach 0.332g/L.
Example seven
Collect enterprise and produce clam water cooking liquid, remove water-insoluble impurity.Take enzyme assisted extraction method, add trypsinase powder in water cooking liquid, add-on is 2.5% of amount of substrate, and with 4mol/LNaOH regulator solution pH to 9,30 DEG C are reacted 3.5 hours, after completing, adjust pH6, the enzyme that goes out in boiling water bath 20min alive, cooling.
With the HCl regulator solution pH to 3 of 6mol/L, namely have acidic protein to precipitate, centrifugation, gets supernatant liquor, then adjusts pH to l0 with 4mol/L, and namely have basic protein to precipitate, centrifugation, gets supernatant liquor.
Supernatant liquor is carried out uf processing, and ultra-filtration membrane retains macromolecular polysaccharide peptide material, collects filtrate, then removes salt ion through electrodialysis, electrodialysis voltage 15V, flow velocity 60mL/min, sample concentration 6mg/mL.Filtrate, with the HCl regulator solution pH to 3 of 6mol/L, treats upper prop.
Get 300mL filtrate by Na ionic acidic resins post, distilled water wash-out, elution speed is 10mL/min; access the effluent liquid that specific conductivity starts to drop between stationary phase, above-mentioned effluent liquid, through concentrating under reduced pressure, adds 3 times of volume 95% ethanol; under being placed in 4 DEG C of environment, separate out taurine.Crude product taurine adds suitable quantity of water and dissolves, then uses 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain taurine solid, and the yield that warp and methyl ethyl diketone and formaldehyde colorimetry record taurine product can reach 0.276g/L.
Example eight
Collect enterprise and produce clam water cooking liquid, remove water-insoluble impurity.Take enzyme assisted extraction method, add aspartic protease powder in water cooking liquid, add-on is 2.5% of amount of substrate, and with 4mol/LHCl regulator solution pH Isosorbide-5-Nitrae 5 DEG C reaction 3.5 hours, after completing, adjust pH7, go out in boiling water bath enzyme 10min, cooling.
With the HCl regulator solution pH to 3 of 6mol/L, namely have acidic protein to precipitate, centrifugation, gets supernatant liquor, then adjusts pH to l0 with 2mol/L NaOH, and namely have basic protein to precipitate, centrifugation, gets supernatant liquor.
Supernatant liquor is carried out uf processing, and ultra-filtration membrane retains macromolecular polysaccharide peptide material, collects filtrate, then removes salt ion through electrodialysis, electrodialysis voltage 15V, flow velocity 100mL/min, sample concentration 8g/mL.Filtrate, with the HCl regulator solution pH to 3 of 6mol/L, treats upper prop.
Get 300mL filtrate by Na ionic acidic resins post, distilled water wash-out, elution speed is 10mL/min; access the effluent liquid that specific conductivity starts to drop between stationary phase, above-mentioned effluent liquid, through concentrating under reduced pressure, adds 3 times of volume 95% ethanol; under being placed in 4 DEG C of environment, separate out taurine.Crude product taurine adds suitable quantity of water and dissolves, then uses 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain taurine solid, and the yield that warp and methyl ethyl diketone and formaldehyde colorimetry record taurine product can reach 0.304g/L.
Example nine
Collect enterprise and produce clam water cooking liquid, remove water-insoluble impurity.Take enzyme assisted extraction method, add Sumizyme MP powder in water cooking liquid, add-on is 2.5% of amount of substrate, and with 1mol/LNaOH regulator solution pH to 10,37 DEG C are reacted 4 hours, after completing, adjust pH8, the enzyme that goes out in boiling water bath 10min alive, cooling.
Adjust pH to l0 with 4mol/LNaOH, namely have basic protein to precipitate, centrifugation, gets supernatant liquor.Use the HCl regulator solution pH to 3 of 6mol/L again, namely have acidic protein to precipitate, centrifugation, gets supernatant liquor.
Supernatant liquor is carried out uf processing, and ultra-filtration membrane retains macromolecular polysaccharide peptide material, collects filtrate, then removes salt ion through electrodialysis, electrodialysis voltage 15V, flow velocity 150mL/min, sample concentration 8mg/mL.Filtrate, with the HCl regulator solution pH to 3 of 6mol/L, treats upper prop.
Get 300mL filtrate by Na ionic acidic resins post, distilled water wash-out, elution speed is 10mL/min; access the effluent liquid that specific conductivity starts to drop between stationary phase, above-mentioned effluent liquid, through concentrating under reduced pressure, adds 3 times of volume 95% ethanol; under being placed in 4 DEG C of environment, separate out taurine.Crude product taurine adds suitable quantity of water and dissolves, then uses 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain taurine solid, and the yield that warp and methyl ethyl diketone and formaldehyde colorimetry record taurine product can reach 0.312g/L.
The present invention utilizes the taurine in enzyme transforming process extraction clam or its water cooking liquid, namely in clam or its water cooking liquid, transforming protein enzyme is added, utilize the principle of enzymatic conversion, extract taurine, experiment show to adopt enzyme to clam or its liquor carry out conversions afterwards its content of taurine compare unconverted before, its content is the highest to be risen to original more than one times (content of taurine can bring up to 3.3mg/g in this step), can improve the yield of taurine thus, see accompanying drawing 1; The method reported in recycling document, carries out resin cation exchange and ethyl alcohol recrystallization method carries out purifying, both can reduce the impact of chemical reagent, can improve again the yield of taurine.Accompanying drawing 1 is the changing conditions of taurine content in this step products before and after enzymatic conversion; In figure, X-coordinate 1 indicates the content of taurine without enzymatic conversion, and 2,3,4,5 represent employing 2 neutral protease, 3 Sumizyme MPs, 4 stomach en-s, the change of content of taurine after the different ferment treatment such as 5 papoids.
The above; be only the present invention's preferably embodiment; but protection scope of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses; be equal to according to technical scheme of the present invention and inventive concept thereof and replace or change, all should be encompassed within protection scope of the present invention.
Claims (9)
1. utilize enzyme transforming process to extract a method for taurine based on clam, it is characterized in that, comprise the steps:
S1, utilize clam preparation feedback liquid;
S2, the one of getting in stomach en-, papoid, trypsinase, Sumizyme MP, the pH value suitable to the conversion reaction of described reaction solution regulatory enzyme according to selected enzyme, adds selected enzyme in described reaction solution and carries out enzymatic conversion reaction; Afterwards, the enzyme that goes out is lived, and obtains enzymatic conversion solution;
S3, described enzymatic conversion solution removal are acid, basic protein;
S4, remove macromolecular substance by ultrafiltration apparatus, retain taurine;
S5, electrodialysis remove salt ion;
S6, by storng-acid cation exchange resin absorptive collection taurine effluent liquid;
Add ethanol in S7, effluent liquid, separate out taurine crystal.
2. utilize enzyme transforming process to extract the method for taurine based on clam according to claim 1, it is characterized in that, S2 step is specially:
S21, in described reaction solution, add stomach en-, papoid, trypsinase, one in Sumizyme MP that dry biomass number in solution is 0.1% ~ 10%; Regulate pH to the control environment of enzyme used, 20 ~ 50 DEG C of enzymolysis 0.5 ~ 10 hour;
Wherein, control environment is: stomach en-pH1 ~ 3, papoid pH5 ~ 9, trypsinase pH7 ~ 9, Sumizyme MP pH8 ~ 11;
S22, cooling, adjust pH to 6 ~ 8;
S23,80 ~ 100 DEG C of enzymes that go out are lived 5 ~ 30 minutes.
3. according to claim 1 or 2, utilize enzyme transforming process to extract the method for taurine based on clam, it is characterized in that, S1 step is specially: clam meat carries out homogenized after adding the water of 1:1 ~ 1:100 by its volume ratio, obtains described reaction solution;
Or it is between 1% ~ 30% that clam water cooking liquid is concentrated into dry matter content, the water-fast impurity of centrifugal segregation, obtains described reaction solution.
4. utilize enzyme transforming process to extract the method for taurine based on clam according to claim 3, it is characterized in that, described water cooking liquid is the waste liquid in the clam course of processing.
5. according to claim 1 or 2, utilize enzyme transforming process to extract the method for taurine based on clam, it is characterized in that, described step S3 comprises operation:
S31, to process after enzymatic conversion solution in add 1mol/L ~ 6mol/L HCl adjust pH to 2 ~ 4, namely have acidic protein to precipitate, centrifugation, get supernatant liquor:
S32, supernatant liquor 1mol/L ~ 6mol/L NaOH adjusts pH to 8 ~ 10, and namely have basic protein to precipitate, centrifugation, gets supernatant liquor.
6. according to claim 1 or 2, utilize enzyme transforming process to extract the method for taurine based on clam; it is characterized in that; described step S4 comprises the steps: that the solution after by removing protein carries out uf processing, clips macromolecular polypeptide polysaccharose substance with ultra-filtration membrane, collects filtrate.
7. according to claim 1 or 2, utilize enzyme transforming process to extract the method for taurine based on clam; it is characterized in that; electrodialysis process parameter suitable in described step S5 is: voltage 1V ~ 20V, flow velocity 0.5mL/min ~ 4L/min, sample concentration 1mg/mL ~ 100mg/mL.
8. according to claim 1 or 2, utilize enzyme transforming process to extract the method for taurine based on clam, it is characterized in that, described step S6 comprises following operation:
By extracting solution by strong acid cation exchange resin column, distilled water wash-out, power taking conductance starts the effluent liquid dropped between stationary phase;
6 × 60cmNa on extracting solution
+type highly acidic resin post, loading volume is 300mL, distilled water wash-out, and elution speed is 20mL/min, accesses the effluent liquid during specific conductivity starts to drop to stable conductivity.
9. according to claim 1 or 2, utilize enzyme transforming process to extract the method for taurine based on clam, it is characterized in that, it is characterized in that, described step S7 comprises following operation:
S71, above-mentioned effluent liquid, through concentrating under reduced pressure, add 3 times of volume 95% ethanol, under being placed in 4 DEG C of environment, separate out taurine.
S72, crude product taurine add suitable quantity of water and dissolve, then use 95% ethyl alcohol recrystallization, 2 ~ 3 times repeatedly, can obtain the taurine crystal that purity is higher.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104829501A (en) * | 2015-03-24 | 2015-08-12 | 辽宁双增生物工程有限公司 | A method of extracting taurine from a mactra chinensis water decoction |
| CN106986795A (en) * | 2017-05-22 | 2017-07-28 | 南宁学院 | A kind of method that taurine is extracted from cattle liver |
| CN107325027A (en) * | 2017-06-19 | 2017-11-07 | 南宁学院 | A kind of method that taurine is extracted from OX-heart |
| CN107604016A (en) * | 2017-10-31 | 2018-01-19 | 荣成市日鑫水产有限公司 | A kind of method that taurine is extracted in the internal organ from octopus |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104829501A (en) * | 2015-03-24 | 2015-08-12 | 辽宁双增生物工程有限公司 | A method of extracting taurine from a mactra chinensis water decoction |
| CN106986795A (en) * | 2017-05-22 | 2017-07-28 | 南宁学院 | A kind of method that taurine is extracted from cattle liver |
| CN106986795B (en) * | 2017-05-22 | 2018-08-24 | 南宁学院 | A method of extracting taurine from cattle liver |
| CN107325027A (en) * | 2017-06-19 | 2017-11-07 | 南宁学院 | A kind of method that taurine is extracted from OX-heart |
| CN107325027B (en) * | 2017-06-19 | 2019-05-14 | 南宁学院 | A method of extracting taurine from cattle heart |
| CN107604016A (en) * | 2017-10-31 | 2018-01-19 | 荣成市日鑫水产有限公司 | A kind of method that taurine is extracted in the internal organ from octopus |
| CN107604016B (en) * | 2017-10-31 | 2021-05-28 | 荣成市日鑫水产有限公司 | Method for extracting taurine from octopus viscera |
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