CN104404094A - Method for extracting taurine by use of enzymatic conversion method on the basis of clams - Google Patents

Abstract

The invention discloses a method for extracting taurine based on clams using an enzyme conversion method, comprising the following steps: 1. Pretreatment of clams or their processed boiled liquid: separating clam meat, cleaning, and homogenizing; or collecting and processing clams Remove the discarded clam boiled liquid in the water and filter to remove impurities. 2. Enzyme conversion: Add enzymes under appropriate pH conditions to carry out enzyme conversion reactions; 3. Removal of proteins: Precipitate and centrifuge under appropriate pH conditions to remove acidic and alkaline proteins. 4. Ultrafiltration equipment to remove macromolecular substances: the macromolecular polysaccharide polypeptide is cut off by ultrafiltration equipment to retain taurine. 5. Electrodialysis removes salt ions. 6. Ion-exchange chromatography: collect taurine by adsorption on strongly acidic cation-exchange resin. 7. Ethanol recrystallization: add a certain volume of ethanol to precipitate taurine crystals. The method of the invention makes full use of waste raw materials, and has the advantages of simple operation, safety, low cost, environmental protection, continuous production, high yield of taurine, and the like.

Description

Method for extracting taurine based on clam using enzyme conversion method

technical field

The invention relates to an industrialized method for continuously preparing taurine in clams or their boiled liquid through an enzyme conversion method.

Background technique

Clams, such as Philippine clams, commonly known as sand clams, are a type of bivalve molluscs belonging to the clam family, and are widely distributed in the north and south seas of my country. Its main characteristics are rapid growth, short reproduction cycle, strong adaptability to salt and temperature, delicious taste, rich nutrition and low price. It is the second largest shellfish farming product in my country, so the deep development and utilization of clams and their processing by-products has extremely important economic value. Fresh clam meat is rich in protein, fat, calcium, phosphorus, iron and other minerals, vitamins, amino acids, and taurine.

The scientific name of taurine is 2-aminoethanesulfonic acid. Because it was isolated from ox bile for the first time, it is commonly known as taurine and ox bile. Taurine is a sulfur-containing non-protein structural amino acid, one of the essential amino acids for the human body, and has strong biological activity. Taurine is very rich in fish and shellfish. Through in-depth research, it is found that taurine can increase the ability of cells to resist oxidation, free radical damage and anti-virus damage. It is also a good liver protection agent, which can enhance vision, promote brain development, relieve fatigue, improve work efficiency, and lower cholesterol. , Inhibition of gallstones, anti-inflammatory, analgesic effects, and has a certain anti-tumor activity. Clinically, taurine can be used to treat diseases such as acute and chronic hepatitis, colds, hypertension, arteriosclerosis and cancer.

The demand for taurine has increased significantly. Our country has a potential market of 20,000 tons per year. The annual consumption in the United States is more than 6,000 tons, and the output in Japan is nearly 2,000 tons.

In response to the growing demand, more technologies for extracting taurine are being updated. At present, taurine is mainly extracted by chemical synthesis at home and abroad, and methods such as ethyl chloride method, ethanolamine method, and ethyleneimine method are mostly used. However, these chemical synthesis methods have problems such as high toxicity of raw materials, complicated process operation, difficult recovery, and serious environmental pollution.

The current comprehensive utilization of marine resources, on the one hand, is to obtain relatively high value-added products from low-value aquatic products; on the other hand, it is to recycle the waste generated during the processing of aquatic products. Since the production waste brings great pressure to the environment, its utilization will undoubtedly bring higher environmental benefits. Extracting taurine from the leftovers of marine product processing can not only improve the utilization rate of marine resources, but also reduce environmental pollution. my country is rich in marine biological resources, and extracting taurine from fish and shellfish and their leftovers has huge economic benefits. Therefore, it is of great practical significance to study how to efficiently extract taurine from marine organisms.

Contents of the invention

In view of the above-mentioned problems, the present invention relates to a technology that can replace the traditional chemical synthesis method to extract taurine or adopt the boiling method to extract taurine, that is, add proteases to the clam and its boiling liquid, and use the enzyme conversion method to extract taurine. , extracting taurine, the invention has the advantages of low cost, environmental protection, simple operation, high extraction efficiency, fully uses waste raw materials and can effectively improve the yield of taurine through the effect of enzymatic conversion.

In order to achieve the above object, the present invention provides a method for extracting taurine based on clams using enzyme conversion method, comprising the steps of:

S1, using clams to prepare a reaction solution;

S2, take one of pepsin, papain, trypsin, and alkaline protease, adjust the pH value suitable for the enzyme conversion reaction to the reaction solution according to the selected enzyme, and add the selected enzyme to the reaction solution Carrying out an enzyme conversion reaction; after that, inactivating the enzyme activity to obtain an enzyme conversion solution;

S3, the enzyme conversion solution removes acidic and basic proteins;

S4, remove macromolecular substances through ultrafiltration equipment, and retain taurine;

S5, electrodialysis to remove salt ions;

S6, collect taurine effluent by strong acidic cation exchange resin adsorption;

S7, adding a certain volume of ethanol to the effluent to precipitate taurine crystals.

In a preferred manner, step S2 specifically includes the steps of:

S21. Add one of pepsin, papain, trypsin, and alkaline protease with a dry matter mass fraction of 0.1% to 10% in the reaction solution; adjust the pH to a suitable environment for the enzyme used (suitable The environment is: pepsin pH 1-3, papain pH 5-9, trypsin pH 7-9, alkaline protease pH 8-11), enzymolysis at 20-50°C for 0.5-10 hours;

S22, cooling, adjusting the pH to 6-8 (this step can effectively guarantee the final yield of the product);

S23, inactivate enzyme activity at 80-100°C for 5-30 minutes.

In addition, in step S1, clam meat or clam boiled liquid can be used to prepare the reaction liquid, specifically: the clam meat is added with 1:1-1:100 water according to its volume ratio and then homogenized to obtain the reaction liquid; or, The clam boiled liquid is concentrated until the dry matter content is between 1% and 30%, and the water-insoluble impurities are removed by centrifugation to obtain the reaction liquid. The boiled liquid can be the waste liquid in the clam processing process (which is discarded in the prior art), so as to further increase the product value and increase the profit.

Step S3 can refer to the mode of prior art, specifically can refer to " taurine in meretrix " [Gong Lifen, Huang Weisheng, Xie Xiaolan, Zheng Zhifu, Hu Donghong. Extraction of Taurine in Meretrix (I). Fine Chemical Industry. 2003, 20 (7 ), 393-395], first adjust the acidity of the solution, if there is acidic protein precipitation, it can be removed by centrifugation, and then the solution is adjusted to be alkaline, if there is alkaline protein precipitation, it can be removed by centrifugation. The order of adjusting the acid or base can be varied. The present invention provides a preferred method, specifically step S3 includes the process:

S31. Add 1mol/L-6mol/L HCl to the treated enzyme conversion solution to adjust the pH to 2-4, that is, acidic protein precipitates, centrifuge, and take the supernatant:

S32. Use 1mol/L-6mol/L NaOH to adjust the pH of the supernatant to 8-10, then the alkaline protein precipitates, centrifuge, and take the supernatant.

Step S4: Ultrafiltration is performed on the protein-removed solution, the macromolecular polypeptide polysaccharides are cut off by the ultrafiltration membrane, and the filtrate is collected. Suitable electrodialysis process parameters in step S5 are: voltage 1V-20V, flow rate 0.5mL/min-4L/min, sample concentration 1mg/mL-100mg/mL.

The above steps S6 and S7 can be realized by referring to the existing technology, such as "Extraction of natural taurine in mussel waste liquid" [Cui Zhongai, Li Pingping, Wang Daobo, Fan Zhendi, Du Yanbing. Mussel waste liquid Extraction of natural taurine. Anhui Agricultural Sciences. 2010,38(16):8671-8673]. Wherein, step S6 includes the following operations:

S61, passing the extract through a strongly acidic cation exchange resin column, eluting with distilled water, and taking the effluent whose conductivity begins to drop to a stable period;

S62. Put the extract on a 6×60 cm Na ⁺ type strong acid resin column with a loading volume of 300 mL, elute with distilled water at an elution rate of 20 mL/min, and collect the effluent when the conductivity begins to decrease until the conductivity is stable.

Step S7 includes the following operations:

S71. The above effluent was concentrated under reduced pressure, added 3 times the volume of 95% ethanol, and placed at 4°C to precipitate taurine.

S72. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain taurine crystals with high purity.

In an optimal mode, the solution is freeze-dried or vacuum-dried to obtain taurine.

Taurine in aquatic products exists in free form and is easily soluble in hot water but insoluble in ethanol. Therefore, white crystals can be obtained by hot water extraction, ion exchange purification, ethanol precipitation and other methods. The invention uses an enzymatic conversion method to extract taurine in clam boiled liquid, and has the advantages of simple operation, low cost, high yield, environmental protection, safety and continuous production. The main steps include: 1. Collect the waste clam boiled liquid produced by the enterprise, and filter to remove water-insoluble impurities; 2. Add protease to the boiled liquid to carry out the enzyme conversion reaction under the appropriate pH condition; 3. Precipitate under the appropriate pH value and remove it by centrifugation Acidic and basic proteins; 4. Remove macromolecular substances through ultrafiltration equipment and retain taurine; 5. Remove salt ions by electrodialysis; 6. Collect taurine effluent through strong acid cation exchange resin adsorption; 7. Outflow A certain volume of ethanol is added to the solution to precipitate taurine crystals.

The present invention has the following advantages;

1. The entire extraction process does not use any organic solvents, which reduces environmental pollution and ensures the safety of the staff.

2. The obtained purified taurine does not contain any toxic reagents, and is safe and reliable.

3. It can make full use of low-value clams or the waste boiling liquid produced after processing, and improve the utilization rate of waste liquid.

4. The taurine is extracted by enzymatic conversion, which can effectively increase the yield.

5. Using ultrafiltration treatment and electrodialysis technology, while effectively improving the purity of taurine, it can also remove heavy metal elements present in aquatic products.

6. Fully automatic continuous production can be realized, the equipment is simple, the operation is easy, and the labor intensity is reduced.

Description of drawings

Fig. 1 is the changing situation of taurine content in the product before and after enzymatic conversion.

Detailed ways

Example one

Shell clams, wash, add water at a volume ratio of 1:10, and homogenate; adopt enzyme-assisted extraction method, add alkaline protease powder to the homogenate, the amount added is 2.5% of the substrate amount, and the pH of the solution is adjusted with 1mol/L NaOH To 10, react at 37°C for 3.5 hours, when the reaction is over, adjust the pH to 6, inactivate the enzyme in a boiling water bath for 10 minutes, and cool down.

Adjust the pH of the solution to 3 with 6mol/L HCl, that is, acidic protein precipitates, centrifuge, take the supernatant, and then adjust the pH to 10 with 4mol/L NaOH, that is, alkaline protein precipitates, centrifuge, take the supernatant Serum.

The supernatant was subjected to ultrafiltration treatment. The ultrafiltration membrane intercepted macromolecular polysaccharide polypeptide substances, collected the filtrate, and then removed salt ions through electrodialysis. The electrodialysis voltage was 1V, the flow rate was 4L/min, and the sample concentration was 1mg/mL. The filtrate was adjusted to pH 3 with 6mol/L HCl, and the solution was put on the column.

Take 300mL of the filtrate to pass through the Na ion-type acidic resin column, elute with distilled water at a rate of 10mL/min, take the effluent whose conductivity begins to drop to a stable period, concentrate the above effluent under reduced pressure, and add 3 times the volume 95% ethanol, placed in 4 ° C environment, precipitated taurine. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain solid taurine. The yield of taurine products measured by colorimetry with acetylacetone and formaldehyde can reach 2.4%.

Example two

Clams were shelled, washed, added water at a volume ratio of 1:10, and homogenized; an enzyme-assisted extraction method was adopted, and acid protease powder was added to the homogenate in an amount of 2.5% of the substrate amount, and the pH of the solution was adjusted to 1 with 4mol/L HCl , react at 45°C for 3.5 hours, after completion, adjust the pH to 7, inactivate the enzyme in a boiling water bath for 10 minutes, and cool down.

Adjust the pH of the solution to 3 with 6mol/L HCl, that is, acidic protein precipitates, centrifuge, take the supernatant, and then adjust the pH to 8 with 4mol/L NaOH, that is, alkaline protein precipitates, centrifuge, take the supernatant liquid.

The supernatant was subjected to ultrafiltration treatment. The ultrafiltration membrane intercepted macromolecular polysaccharide polypeptide substances, collected the filtrate, and then removed salt ions through electrodialysis. The electrodialysis voltage was 15V, the flow rate was 100mL/min, and the sample concentration was 8g/mL. The filtrate was adjusted to pH 3 with 6 mol/L HCl, and the filtrate was put on the column.

Take 300mL of the filtrate to pass through the Na ion-type acidic resin column, elute with distilled water at a rate of 10mL/min, take the effluent whose conductivity begins to drop to a stable period, concentrate the above effluent under reduced pressure, and add 3 times the volume 95% ethanol, placed in 4 ° C environment, precipitated taurine. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain solid taurine. The yield of taurine products measured by colorimetry with acetylacetone and formaldehyde can reach 1.8%.

Example three

Collect the clam boiled liquid produced by the enterprise to remove insoluble impurities; adjust the pH of the solution to 3 with 6mol/L HCl, that is, acidic protein precipitates, centrifuge, take the supernatant, and then adjust the pH to 9 with 4mol/L NaOH, That is, the basic protein precipitated, centrifuged, and the supernatant was taken. The supernatant was subjected to ultrafiltration treatment. The ultrafiltration membrane intercepted macromolecular polysaccharide polypeptide substances, collected the filtrate, and then removed salt ions through electrodialysis. The electrodialysis voltage was 1V, the flow rate was 0.5mL/min, and the sample concentration was 1mg/mL. The filtrate was adjusted to pH 3 with 6mol/L HCl, and the solution was put on the column.

Take 300mL of the filtrate to pass through the Na ion-type acidic resin column, elute with distilled water at a rate of 10mL/min, take the effluent whose conductivity begins to drop to a stable period, concentrate the above effluent under reduced pressure, and add 3 times the volume 95% ethanol, placed in 4 ° C environment, precipitated taurine. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain solid taurine. The yield of taurine products measured by colorimetry with acetylacetone and formaldehyde can reach 0.182g/L.

Example four

Collect the boiled clam liquid produced by the enterprise to remove insoluble impurities; adopt the enzyme-assisted extraction method, add alkaline protease powder to the boiled liquid, the amount added is 2.5% of the substrate amount, and adjust the pH of the solution to 10 with 1mol/L NaOH at 37°C After reacting for 3.5 hours, when the reaction is over, adjust the pH to 8, inactivate the enzyme in a boiling water bath for 10 minutes, and cool down.

Adjust the pH of the solution to 3 with 6mol/L HCl, that is, acidic protein precipitates, centrifuge, take the supernatant, and then adjust the pH to 10 with 6mol/L NaOH, that is, alkaline protein precipitates, centrifuge, take the supernatant Serum.

The supernatant is subjected to ultrafiltration treatment, the ultrafiltration membrane intercepts macromolecular polysaccharide polypeptide substances, the filtrate is collected, and then the salt ions are removed by electrodialysis, the electrodialysis voltage is 1V, the flow rate is 1L/min, and the sample concentration is 1mg/mL. The filtrate was adjusted to pH 3 with 6mol/L HCl, and the solution was put on the column.

Take 300mL of the filtrate to pass through the Na ion-type acidic resin column, elute with distilled water at a rate of 10mL/min, take the effluent whose conductivity begins to drop to a stable period, concentrate the above effluent under reduced pressure, and add 3 times the volume 95% ethanol, placed in 4 ° C environment, precipitated taurine. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain solid taurine. The yield of taurine products measured by colorimetry with acetylacetone and formaldehyde can reach 0.316g/L.

Example five

Collect clam boiled liquid produced by enterprises to remove water-insoluble impurities. Adopt the enzyme-assisted extraction method, add pepsin to the boiled liquid, the addition amount is 2.5% of the substrate amount, adjust the pH of the solution to 1 with 1mol/L HCl, react at 20°C for 3.5 hours, after the reaction is completed, adjust the pH to 7, Inactivate the enzyme in a boiling water bath for 10 minutes and cool down.

Adjust the pH of the solution to 3 with 6mol/L HCl, that is, acidic protein precipitates, centrifuge, take the supernatant, and then adjust the pH to 8 with 4mol/L NaOH, that is, alkaline protein precipitates, centrifuge, take the supernatant liquid.

The supernatant was subjected to ultrafiltration treatment. The ultrafiltration membrane intercepted macromolecular polysaccharide polypeptide substances, collected the filtrate, and then removed salt ions through electrodialysis. The electrodialysis voltage was 5V, the flow rate was 100mL/min, and the sample concentration was 2mg/mL. The filtrate was adjusted to pH 3 with 6 mol/L HCl, and the filtrate was put on the column.

Take 300mL of the filtrate to pass through the Na ion-type acidic resin column, elute with distilled water at a rate of 10mL/min, take the effluent whose conductivity begins to drop to a stable period, concentrate the above effluent under reduced pressure, and add 3 times the volume 95% ethanol, placed in 4 ° C environment, precipitated taurine. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain solid taurine. The yield of taurine products measured by colorimetry with acetylacetone and formaldehyde can reach 0.284g/L.

Example six

Collect clam boiled liquid produced by enterprises to remove water-insoluble impurities. Adopt the enzyme-assisted extraction method, add papain in the boiled liquid, the addition amount is 2.5% of the substrate amount, adjust the pH of the solution to 8 with 6mol/L NaOH, react at 50°C for 3.5 hours, after completion, adjust the pH to 8, and extinguish in a boiling water bath Enzyme activity 20min, cooling.

Adjust the pH to 10 with 1mol/L NaOH, that is, the alkaline protein precipitates, centrifuge, and take the supernatant. Then use 6mol/L HCl to adjust the pH of the solution to 4, that is, acidic protein precipitates, centrifuge, and take the supernatant.

The supernatant was subjected to ultrafiltration treatment. The ultrafiltration membrane intercepted macromolecular polysaccharide polypeptide substances, collected the filtrate, and then removed salt ions through electrodialysis. The electrodialysis voltage was 10V, the flow rate was 3L/min, and the sample concentration was 4mg/mL. The filtrate was adjusted to pH 3 with 6 mol/L HCl, and the filtrate was put on the column.

Take 300mL of the filtrate to pass through the Na ion-type acidic resin column, elute with distilled water at a rate of 10mL/min, take the effluent whose conductivity begins to drop to a stable period, concentrate the above effluent under reduced pressure, and add 3 times the volume 95% ethanol, placed in 4 ° C environment, precipitated taurine. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain solid taurine. The yield of taurine products measured by colorimetry with acetylacetone and formaldehyde can reach 0.332g/L.

Example seven

Collect clam boiled liquid produced by enterprises to remove water-insoluble impurities. Take the enzyme-assisted extraction method, add trypsin powder to the boiled liquid, the amount added is 2.5% of the substrate amount, adjust the pH of the solution to 9 with 4mol/L NaOH, and react at 30°C for 3.5 hours. After completion, adjust the pH to 6 and place in a boiling water bath Inactivate the enzyme for 20 minutes and cool down.

Adjust the pH of the solution to 3 with 6mol/L HCl, that is, acidic protein precipitates, centrifuge, take the supernatant, and then adjust the pH to 10 with 4mol/L, that is, alkaline protein precipitates, centrifuge, take the supernatant .

The supernatant was subjected to ultrafiltration treatment. The ultrafiltration membrane intercepted macromolecular polysaccharide polypeptide substances, collected the filtrate, and then removed salt ions through electrodialysis. The electrodialysis voltage was 15V, the flow rate was 60mL/min, and the sample concentration was 6mg/mL. The filtrate was adjusted to pH 3 with 6 mol/L HCl, and the filtrate was put on the column.

Take 300mL of the filtrate to pass through the Na ion-type acidic resin column, elute with distilled water at a rate of 10mL/min, take the effluent whose conductivity begins to drop to a stable period, concentrate the above effluent under reduced pressure, and add 3 times the volume 95% ethanol, placed in 4 ° C environment, precipitated taurine. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain solid taurine. The yield of taurine products measured by colorimetry with acetylacetone and formaldehyde can reach 0.276g/L.

Example eight

Collect clam boiled liquid produced by enterprises to remove water-insoluble impurities. Adopt the enzyme-assisted extraction method, add acidic protease powder to the boiled liquid, the addition amount is 2.5% of the substrate amount, adjust the pH of the solution to 1 with 4mol/L HCl, react at 45°C for 3.5 hours, after completion, adjust the pH to 7, and extinguish in a boiling water bath Enzyme 10min, cooling.

Adjust the pH of the solution to 3 with 6mol/L HCl, that is, acidic protein precipitates, centrifuge, take the supernatant, and then adjust the pH to 10 with 2mol/L NaOH, that is, alkaline protein precipitates, centrifuge, take the supernatant liquid.

The supernatant was subjected to ultrafiltration treatment. The ultrafiltration membrane intercepted macromolecular polysaccharide polypeptide substances, collected the filtrate, and then removed salt ions through electrodialysis. The electrodialysis voltage was 15V, the flow rate was 100mL/min, and the sample concentration was 8g/mL. The filtrate was adjusted to pH 3 with 6 mol/L HCl, and the filtrate was put on the column.

Take 300mL of the filtrate to pass through the Na ion-type acidic resin column, elute with distilled water at a rate of 10mL/min, take the effluent whose conductivity begins to drop to a stable period, concentrate the above effluent under reduced pressure, and add 3 times the volume 95% ethanol, placed in 4 ° C environment, precipitated taurine. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain solid taurine. The yield of taurine products measured by colorimetry with acetylacetone and formaldehyde can reach 0.304g/L.

Example nine

Collect clam boiled liquid produced by enterprises to remove water-insoluble impurities. Take the enzyme-assisted extraction method, add alkaline protease powder to the boiled liquid, the amount added is 2.5% of the substrate amount, adjust the pH of the solution to 10 with 1mol/L NaOH, react at 37°C for 4 hours, after completion, adjust the pH to 8, and put it in a boiling water bath Inactivate the enzyme activity for 10 minutes and cool down.

Adjust the pH to 10 with 4mol/L NaOH, that is, the alkaline protein precipitates, centrifuge, and take the supernatant. Then use 6mol/L HCl to adjust the pH of the solution to 3, that is, acidic protein precipitates, centrifuge, and take the supernatant.

The supernatant was subjected to ultrafiltration treatment. The ultrafiltration membrane intercepted macromolecular polysaccharide polypeptide substances, collected the filtrate, and then removed salt ions through electrodialysis. The electrodialysis voltage was 15V, the flow rate was 150mL/min, and the sample concentration was 8mg/mL. The filtrate was adjusted to pH 3 with 6 mol/L HCl, and the filtrate was put on the column.

Take 300mL of the filtrate to pass through the Na ion-type acidic resin column, elute with distilled water at a rate of 10mL/min, take the effluent whose conductivity begins to drop to a stable period, concentrate the above effluent under reduced pressure, and add 3 times the volume 95% ethanol, placed in 4 ° C environment, precipitated taurine. Crude taurine is dissolved in an appropriate amount of water, and then recrystallized with 95% ethanol, and repeated 2 to 3 times to obtain solid taurine. The yield of taurine products measured by colorimetry with acetylacetone and formaldehyde can reach 0.312g/L.

The present invention utilizes the enzymatic transformation method to extract the taurine in the clam or its boiled liquid, that is, adds invert protease to the clam or its boiled liquid, utilizes the principle of enzymatic conversion to extract taurine, and the experiment shows that the clam or its boiled liquid is treated with enzyme. After its boiled juice is converted, its taurine content can be increased to more than double (in this step, the taurine content can be increased to 3.3mg/g) compared with before it is not converted, thus it can improve The yield of taurine is shown in accompanying drawing 1; Utilize the method reported in the literature again, carry out cationic resin exchange and ethanol recrystallization method to purify, can reduce the influence of chemical reagent, can improve the yield of taurine again. Accompanying drawing 1 is the changing situation of taurine content in this step product before and after enzymatic transformation; Abscissa 1 among the figure represents the taurine content without enzymatic transformation, and 2,3,4,5 represent to adopt 2 neutral proteases, 3 Alkaline protease, 4 pepsin, 5 papain and other enzymes to change the content of taurine.

The above is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Anyone familiar with the technical field within the technical scope disclosed in the present invention, according to the technical solution of the present invention Any equivalent replacement or change of the inventive concepts thereof shall fall within the 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.