32 results about "Phenol oxidase activity" patented technology

The present invention relates to a composition for detecting an infinitesimal quantity of β-1,3-glucan, a preparation method thereof and a diagnostic kit detecting β-1,3-glucan. The composition of the present invention shows phenoloxidase activity by β-1,3-glucan in the presence of calcium ions. Using the composition of the present invention, a sample is gathered from a specimen, the composition of the present invention and calcium ions are added to the sample, and β-1,3-glucan is detected by measuring phenoloxidase activity.

The invention discloses a method for brewing cherry fruit wine. The method is characterized by comprising the following steps of: after selecting, washing and stoning cherries serving as raw materials, obtaining primary pulp by juicing; adding the following substances such as pectinase, calcium citrate, vitamin C, and g / L kojic acid into the primary pulp; standing for 5 to 24 hours; preparing clear juice by performing centrifugal separation; inoculating 0.05 to 2 percent of fruit wine yeast based on the weight of the clear juice; controlling a temperature and fermenting, wherein the primary fermentation temperature is between 16 and 28 DEG C, the fermentation time is 7 to 15 days, the secondary fermentation temperature is between 15 and 20 DEG C, and the fermentation time is 20 to 40 days; aging for 3 to 6 months; clarifying and filtering by adopting an ultra-filtration method; and filling, sterilizing and packaging to obtain finished cherry fruit wine. The method effectively improves a production process of cherry wine, reduces the content of polyphenol fundamentally, inhibits the activity of polyphenol oxidase, reduces harsh taste and improves stability. Compared with the prior art, the cherry fruit wine produced by the method maintains effective nutrient components, reduces the harsh taste and has a pure flavor.

The invention relates to the technical field of enzymatic activity, in particular to an apple polyphenol oxidase accelerative activator and application thereof. The apple polyphenol oxidase accelerative activator comprises a mixture of dodecyltrimethylammonium bromide or hexadecyl trimethyl ammonium bromide and a Gemini surfactant according to the weight ratio of 10-2,000: 1-4; and the Gemini surfactant is dibromo-bis(dimethyl dodecyl) pentyl diammonium or dibromo-bis(dimethyl hexadecyl) pentyl diammonium. The invention further provides application of the apple polyphenol oxidase accelerative activator in improvement of apple polyphenol oxidase activity. The apple polyphenol oxidase activity can be increased to be 240%, and the use amount level of the apple polyphenol oxidase accelerative activator is low.

The invention relates to a preparation method and application of a prawn non-specific immunity reinforcing agent oligopeptide. The preparation method comprises the following steps of: mincing pacific alaskapollack dogmeat by a meat mincer and then placing in an enzyme reactor to heat and sterilize; triturating and then heating to 90 DEG C to preserve the temperature; sterilizing for 20 minutes in a constant temperature water bath; adding water and protease preparations to mix; carrying out enzymolysis under the condition of optimized enzymolysis process parameters; then heating enzymolysis liquid so that protease is inactivated; roughly filtering fishbones and centrifugalizing the enzymolysis liquid to obtain a soluble protein oligopeptide; ultrafiltering, purifying, freezing and drying and then obtaining protein oligopeptide powder. The protease preparations comprise Alcalase and Flavourzyme, and the enzymolysis process parameters comprise the enzymolysis temperature of 55 DEG C, the initial pH value of 8.5, the material and water ratio of 1:1 and the enzymolysis time of 3 hours. A macromolecule substance is intercepted by using a method for combining 8,000rpm high-speed centrifugalization and 100kDa ultrafiltration membrane ultrafiltrating purification, and 30 percent of protein oligopeptide with the molecular weight of smaller than 100Da is obtained. The oligopeptide powder replaces 5 percent of fish meal in prawn feed and can obviously improve the blood corpuscle density, the phenoloxidase activity, the respiratory burst activity, the SOD activity, the NOS activity and the specific growing rate of a prawn.

The invention relates to a method for enhancing immunity of Fenneropenaeus chinensis. The method is characterized in that Fenneropenaeus chinensis is fed with single Fenneropenaeus chinensis basal feed and Fenneropenaeus chinensis basal feed mixed with 1.4% (by weight) of hydrolyzed fish protein powder alternately, i.e. Fenneropenaeus chinensis is firstly fed with Fenneropenaeus chinensis basal feed mixed with 1.4% (by weight) of hydrolyzed fish protein powder for 7 days, and then fed with single Fenneropenaeus chinensis basal feed for 7 days, over and over again. The measured serum acid phosphatase activity, serum phenol oxidase activity, serum lysozyme activity, peroxidase activity change and serum superoxide dismutase activity of Fenneropenaeus chinensis fed by using the method provided by the invention are not significantly different from those of a Fenneropenaeus chinensis group continuously fed with hydrolyzed fish protein powder and a Fenneropenaeus chinensis group continuously fed with peptidoglycans; and the survival rate and weight increase rate are not significantly different from those of the Fenneropenaeus chinensis group continuously fed with hydrolyzed fish protein powder and the Fenneropenaeus chinensis group continuously fed with peptidoglycans. Therefore, compared with the method in which Fenneropenaeus chinensis is continuously fed with Fenneropenaeus chinensis basal feed mixed with 1.4% (by weight) of hydrolyzed fish protein powder, the method provided by the invention can obtain the same immunity effect but can reduce half of economic cost.

There is provided a process for enhancing the activity of a phenol oxidizing enzyme, comprising adding to the enzyme, as an enhancer for the activity of the enzyme, one or more compounds having the formula:wherein:A is an optionally substituted electron-withdrawing group, chosen from the group consisting of nitrile, carboxyl, and esters and salts thereof, amides, aldehydes, ketones, sulfoxides, sulfones, or sulphonates, andB is an optionally substituted electron-donating group, chosen from the group consisting of the oxide anion, sulfide anion, oxides, amines, imines, hydroxides, sulfides, ethers, carboxylic acids, and halogen substituents, andR1-R5 may each independently represent hydrogen, hydroxy, halogen, nitroso, formyl, carboxyl, and esters and salts thereof, carbamoyl, sulfo, and esters and salts hereof, sulfamoyl, nitro, amino, phenyl, C1-C20 alkyl, C1-C8 alkoxy, carbonyl-C1-C6-alkoxy, aryl-C1-C6-alkyl, and R2 and R5 together may form an alkylene group or an alkenylene group.

It is intended to provide a method for producing a non-browning apple. Apples in which total polyphenol content is not more than 40 mg / 100 g fresh weight in terms of catechin when extracted from apple flesh using methanol and assayed by the Folin-Denis method are used as cross parents and by crossing and breeding them, using a low level of the browning degree of fruit flesh as an index and performing selection based on the index, the non-browning apple is produced. The non-browning apple has a non-browning characteristic that browning of flesh is less likely to occur and a polyphenol oxidase activity is significantly lower than that of a conventional cultivar.

The present invention relates to a method for producing an acidified milk drink using an enzyme having phenoloxidase activity.

The invention discloses a preparation method and application of a browning inhibitor solution for alcoholization of flue-cured tobacco. The browning inhibitor solution for the alcoholization of the flue-cured tobacco is prepared from 5-30 g / L of sodium sulfite, 10-30 g / L of sodium citrate, 10-30 g / L of EDTA, 6-12 g / L of ascorbic acid and 1-10 g / L of Amadori compounds. Sodium nitrite and ascorbic acid in the browning inhibitor solution for the alcoholization of the flue-cured tobacco are good bio-antioxidants, the Maillard reaction can be effectively retarded, and the polyphenol oxidative condensation reaction caused by the oxidation effect is prevented; sodium citrate, EDTA and the Amadori compounds can effectively inhibit the activity of polyphenol oxidase, enzymatic browning is inhibited, and browning in the flue-cured tobacco alcoholization process is effectively inhibited. Meanwhile, the alcoholization quality of tobacco leaves can be improved, and the aroma of the alcoholized flue-cured tobacco is better and richer.

The invention belongs to the technical field of processing of traditional Chinese medicines, and particularly relates to a sulfur-free processing method for Bulbus Fritillariae Thunbergii. The sulfur-free processing method for Bulbus Fritillariae Thunbergii comprises the following steps: A, liquid preparation: adding vitamin C, sodium chloride and citric acid in purified water to obtain a polyphenol oxidase sulfur-free inhibitor for subsequent use; B, soaking: soaking screened Bulbus Fritillariae Thunbergii in the polyphenol oxidase sulfur-free inhibitor for 15-60 minutes, wherein the polyphenol oxidase sulfur-free inhibitor completely covers the Bulbus Fritillariae Thunbergii during soaking; C, air-drying: draining water of the soaked Bulbus Fritillariae Thunbergii, and drying the Bulbus Fritillariae Thunbergii in an oven by hot air at the temperature of 50-70 DEG C for 100-120 minutes; and D, storing: storing the dried Bulbus Fritillariae Thunbergii in a sealing bag. The sulfur-free processing method for Bulbus Fritillariae Thunbergii has the advantages that the vitamin C, the sodium chloride, the citric acid and the like are used as main enzyme inhibition ingredients to inhibit activity of the polyphenol oxidase, activity of PPO is inhibited to prevent the circumstance that the appearance quality is affected by browning of raw herbal materials, after being soaked in the inhibiting liquid, the Bulbus Fritillariae Thunbergii is dried,, and therefore, good internal quality and appearance quality can be achieved.

The invention discloses a method and specific primers of detecting the character of the activity of wheat polyphenol oxidase. The primers provided by the invention for detecting the character of the activity of wheat polyphenol oxidase is a pair of primers consisting of the nucleotide sequences of the sequence 1 and the sequence 2 in the sequence table. The method of detecting the character of the activity of wheat polyphenol oxidase is that PCR amplification can be conducted by taking the DNA of a to-be-detected gene group as a templet and taking the nucleotide sequences of the sequence 1 and the sequence 2 in the sequence table as primers; and then the amplified outcomes can be detected to see whether 750 to 1000 bp of bands exist. The method and specific primers can play an important role both in the detection of the character of the activity of wheat polyphenol oxidase and in the breeding.

The invention discloses a processing method of a sulfur-free konjak slice. The processing method comprises the following steps of (1) washing the konjak; (2) rubbing the peel of the konjak bulb; (3) slicing and collecting the konjak bulb; (4) blanching the konjak slice by steam, deactivating enzyme, and inhibiting the activity of polyphenol oxidase; (5) treating the konjak slice by konjak powder,so as to coat one layer of konjak powder onto the surface of the konjak slice; (6) drying the konjak slice by microwaves. The processing method of the sulfur-free konjak slice has the advantages thatthe fresh konjak is used as the raw material; the special processing method is adopted, so as to obtain the sulfur-free konjak slice.

The invention provides a processing method of a hypsizygus marmoreus quick-frozen product, and belongs to the technical field of food processing. The processing method of the hypsizygus marmoreus quick-frozen product comprises the steps of pre-freezing, soaking, blanching, draining, quick-freezing, ice glaze plating and the like. According to the method, the storage time of the hypsizygus marmoreus can be remarkably prolonged, and the inhibition effect on the activity of polyphenol oxidase in the hypsizygus marmoreus reaches 95%. The quick-frozen hypsizygus marmoreus product prepared by adopting the technical scheme not only keeps good appearance, but also keeps the special flavor and nutritional ingredients of hypsizygus marmoreus, and the method has the characteristics of simple process and low cost, can realize large-scale industrial production, and has good economic benefits.

The invention discloses a preparation method of a phenoloxidase active protein. The preparation method comprises the following steps: salting out proteins from a raw material waste blood plasma residual on Tachypleus Amebocyte Lysate by a saturated ammonium sulfate solution, performing refrigerating centrifugation to obtain limulus hemocyanin, performing dialysis desalting on a PBS solution of thehemocyanin through an MW 3500 Da dialysis bag, and carrying out Sephacryl S-100 allyl dextran gel column purification and concentrating treatment to obtain high-efficiency phenoloxidase active hemocyanin, wherein the initial reaction rate Vi of the high-efficiency phenoloxidase active hemocyanin is 74.52 nmol.(min.mg). The product obtained in the invention has a high purity and a high activity, and can be used for preparing a phenolic pollutant micro-detector with the advantages of high efficiency, simplicity and low cost; and the preparation method is simple, and simultaneously achieves desalting, separating, purifying and concentrating effects by cooperating the MW 3500 Da dialysis bag with the Sephacryl S-100 allyl dextran gel column.

The invention relates to a kirschwasser preparation method. The kirschwasser is prepared by using medicinal and edible cherries as a main raw material and using papayas having an effect of reinforcing spleen to promote digestion as well as bitter gourds, shrub althea flowers, red dates and lichees having an effect of beautifying the face as auxiliary materials through brewage. The kirschwasser preparation method is characterized in that high-pressure steam instantaneous treatment is performed for the first time to deactivate enzymes of the cherries, so that the activity of polyphenol oxidase is basically inhibited; and meanwhile, proteinase in the papayas hydrolyzes proteins and other phenol substances in the fruit juice of the raw materials, and the hydrolyzate can provide a necessary nitrogen source for yeast growth. According to the invention, proportioning of cherries, papayas, bitter gourds, shrub althea flowers, red dates and lichees, high-pressure steam instantaneous treatment parameters and other factors are optimized through a uniform design method, and a regression equation model is optimized. The kirschwasser produced by the invention keeps active nutritional ingredients of cherries, and also has the nutrition and food therapy functions of papayas, bitter gourds, shrub althea flowers, red dates and lichees. Besides, the kirschwasser has a mellow flavor, a high nutrition function and a health care function.

The invention discloses a color protection agent for preparing vegetable paper and a color protection method thereof. Added polyphenol oxidase activity inhibition agent can be chelated with surface layer metal ions to prevent the color changing and fading of the metal ions for natural pigments; an antioxidant can reduce the oxidized color changing; polyvinylpyrrolidone can be associated with polyphenol substance hydroxyls to form strong hydrogen bonds, so that the polyphenol substances can be selectively absorbed, and the enzyme-accelerating browning can be prevented; saponin can synergistically inhibit the activity of polyphenol oxidase; and citric acid has an effect of chelating the metal ions, adjusting PH and preventing the acidic corrosion, so that the color protection agent providedby the invention has a complex effect, can retard the browning, can reduce the browning rate, can play a good color protection effect, and can solve the problem that the vegetable is likely to oxidizeand brown in the preparation process of the vegetable paper; and moreover, the preparation process is simple, the cost is low, and the effect is apparent.

The invention discloses a method for brewing cherry fruit wine. The method is characterized by comprising the following steps of: after selecting, washing and stoning cherries serving as raw materials, obtaining primary pulp by juicing; adding the following substances such as pectinase, calcium citrate, vitamin C, and g / L kojic acid into the primary pulp; standing for 5 to 24 hours; preparing clear juice by performing centrifugal separation; inoculating 0.05 to 2 percent of fruit wine yeast based on the weight of the clear juice; controlling a temperature and fermenting, wherein the primary fermentation temperature is between 16 and 28 DEG C, the fermentation time is 7 to 15 days, the secondary fermentation temperature is between 15 and 20 DEG C, and the fermentation time is 20 to 40 days; aging for 3 to 6 months; clarifying and filtering by adopting an ultra-filtration method; and filling, sterilizing and packaging to obtain finished cherry fruit wine. The method effectively improvesa production process of cherry wine, reduces the content of polyphenol fundamentally, inhibits the activity of polyphenol oxidase, reduces harsh taste and improves stability. Compared with the prior art, the cherry fruit wine produced by the method maintains effective nutrient components, reduces the harsh taste and has a pure flavor.

The invention presents a measuring method of the activation of wheat polyphenol oxidase and its special primer. The primer provided by this invention, which is used to measure the activation of wheat polyphenol oxidase, is actually a pair of v formed by the nucleotide sequence of the sequence 1 and nucleotide sequence of the sequence 2 in the sequence list. The way of measuring the activation of wheat polyphenol oxidase is to make PCR amplification by taking the genome DNA of wheat waiting for measuring as the templet and the nucleotide sequence of the sequence 1 and nucleotide sequence of the sequence 2 in the sequence list as primer, and then test whether there is a stripe with the size of 198bp in the amplification amplified products. The method of this invention and its special primer will have an important role in the measuring of the activation of wheat polyphenol oxidase as well as in breeding.

The invention relates to the technical field of feed additives, in particular to a compound microbial preparation and application thereof in aquatic feed after spraying. The compound microbial preparation is prepared from enterococcus faecalis, clostridium butyricum, bacillus licheniformis and a compound protecting agent. By means of an after spraying technology, the compound microbial preparation is added into aquatic feed; thus, microorganism activity loss in a feed processing and feeding process is effectively reduced; furthermore, a survival rate and a weight gain rate of turbot are both obviously improved, the weight gain rate is improved by 19.8%, a feed coefficient is reduced by 14.3%, and phenol oxidase activity and lysozyme activity in blood of experimental-group turbot are respectively improved by 76.5% and 64.4%; thus, feed cost is beneficial to being reduced, immunity of aquatic livestock is improved, culture benefit is improved, and an effect is obvious.

The invention belongs to the technical field of traditional Chinese medicine processing, and in particular relates to a polyphenol oxidase sulfur-free inhibitor for bulbus fritillariae thunbergii as well as a formula and a preparation method of the polyphenol oxidase sulfur-free inhibitor. The inhibitor is prepared from the following optimized components in parts by weight: 1-5 parts of vitamin C, 8-12 parts of sodium chloride, and 10-14 parts of citric acid. The polyphenol oxidase sulfur-free inhibitor has the advantages that the activity of polyphenol oxidase is inhibited by the main enzyme inhibition components including vitamin C, sodium chloride and citric acid, the activity of PPO (polyphenol oxidase) is inhibited, so that the browning of the raw medicinal material can be prevented, then the appearance quality is not influenced, therefore, the effect of preventing the appearance quality decrease of bulbus fritillariae thunbergii is achieved, meanwhile, the real sulfur-free processing of the overall process is realized, and the integral design is reasonable.

The present invention discloses a shrimp meat separation device using a vibration and separation method and belongs to the technical field of shrimp meat processing. The device comprises a box body and a vibration screen arranged inside the box body; a clamping block is connected with one side of the vibration screen; a sifting spring is connected with the other side of the vibration screen; a material suction pipe is connected with the inner wall of the box body at the lower end of the vibration screen; the material suction pipe is connected with a water pump through a support pipe; the waterpump is connected with a material outlet pipe; connection springs are connected with the outer wall of the head end of the material suction pipe; and the connection springs are connected with a movable cover distributed with cover holes. The device is high in working reliability, rapid in shrimp meat separation speed and good in effects, enables the shrimp meat to be compact in texture, inhibitsshrimp polyphenol oxidase activity in the shrimp meat body, effectively extends shelf life of the shrimp meat and improves mouthfeel of the shrimp meat.

The invention provides a potato pulp preparation method and potato pulp prepared by the same, and relates to the field of processing of fresh potatoes. The preparation method comprises the following steps: pouring a solution which contains KOH with mass concentration of 6-8% and ethanol with volume percentage of 70-90% in an ultrasonic washing machine; stirring for 5-10 min at the temperature of 65-75 DEG C to obtain peeled potatoes; soaking the peeled potatoes by using an aqueous solution of a polyphenol oxidase inhibitor which contains citric acid with mass concentration of 3-7%, ascorbic acid with mass concentration of 3-7% and glutathione with mass concentration of 0.5-1% to dealkalize the potatoes so as to obtain the dealkalized potatoes; and grinding the dealkalized potatoes into the potato pulp. In the preparation method, peeling efficiency is high, peel residues are less, alkaloid in the potatoes is removed, activity of the polyphenol oxidase is inhibited, and freshness of the potato pulp is guaranteed.

The invention discloses a method and specific primers of detecting the character of the activity of wheat polyphenol oxidase. The primers provided by the invention for detecting the character of the activity of wheat polyphenol oxidase is a pair of primers consisting of the nucleotide sequences of the sequence 1 and the sequence 2 in the sequence table. The method of detecting the character of the activity of wheat polyphenol oxidase is that PCR amplification can be conducted by taking the DNA of a to-be-detected gene group as a templet and taking the nucleotide sequences of the sequence 1 and the sequence 2 in the sequence table as primers; and then the amplified outcomes can be detected to see whether 750 to 1000 bp of bands exist. The method and specific primers can play an important role both in the detection of the character of the activity of wheat polyphenol oxidase and in the breeding.

The invention discloses a preparation method of antioxidant soybean oil containing phospholipid, which comprises the following steps: step 1, optimizing the addition amount of the phospholipid in the soybean oil, respectively adding 0.05%, 0.1%, 0.2% and 0.5% of the phospholipid into 100g of soybean oil according to the mass of the soybean oil, simultaneously adding a blank experiment as a contrast, accelerating oxidation, and detecting by referring to the national standard GB 5009.227-2016; the oxidized soybean oil phospholipid can be combined with free radicals in the body, so that the SOD activity and the serum phenol oxidase activity are enhanced, the oxidation resistance in the human body is improved, the oxidation stability of the soybean oil is enhanced, and the process is low in cost, high in efficiency, environmentally friendly and easy to realize industrial production.

The invention discloses a processing method of sulfur-free konjac chips, which comprises the following steps: 1) washing the konjac; 2) rubbing the konjac corms; 3) slicing and collecting the konjac corms; 4) 1. Use steam to blanch the konjac chips to inactivate enzymes to inhibit the activity of polyphenol oxidase; 5) process the konjac chips with konjac flour so that the surface of the konjac chips is coated with a layer of konjac flour; 6) konjac chips Carrying out microwave drying The object of the present invention is to provide a kind of processing method of sulfur-free konjac slices, use fresh konjac as raw material, through specific processing method, obtain sulfur-free konjac slices.

The invention relates to a preparation method and application of a prawn non-specific immunity reinforcing agent oligopeptide. The preparation method comprises the following steps of: mincing pacific alaskapollack dogmeat by a meat mincer and then placing in an enzyme reactor to heat and sterilize; triturating and then heating to 90 DEG C to preserve the temperature; sterilizing for 20 minutes in a constant temperature water bath; adding water and protease preparations to mix; carrying out enzymolysis under the condition of optimized enzymolysis process parameters; then heating enzymolysis liquid so that protease is inactivated; roughly filtering fishbones and centrifugalizing the enzymolysis liquid to obtain a soluble protein oligopeptide; ultrafiltering, purifying, freezing and drying and then obtaining protein oligopeptide powder. The protease preparations comprise Alcalase and Flavourzyme, and the enzymolysis process parameters comprise the enzymolysis temperature of 55 DEG C, the initial pH value of 8.5, the material and water ratio of 1:1 and the enzymolysis time of 3 hours. A macromolecule substance is intercepted by using a method for combining 8,000rpm high-speed centrifugalization and 100kDa ultrafiltration membrane ultrafiltrating purification, and 30 percent of protein oligopeptide with the molecular weight of smaller than 100Da is obtained, the further filteration is carried out by 1kDa / 5kDa ultrafiltration membrane to obtain an ultrafiltrate, and the oligopeptide powder is obtained by freezing and drying the ultrafiltrate. The oligopeptide powder replaces 5 percent of fish meal in prawn feed and can obviously improve the blood corpuscle density, the phenoloxidase activity, the respiratory burst activity, the SOD activity, the NOS activity and the specific growing rate of a prawn.

An accurate determination method of activity of shrimp and crab phenol oxidase comprises the steps of extracting hemolymph from an organ for determination, preparing gill tissue phenol oxidase, collecting a gill tissue sample from an ice bag rapidly, adding aseptic normal saline, performing ice-bath homogenization, centrifuging homogenate, sucking supernate for storage for determination, adding a potassium phosphate buffer solution, pretreating in a water bath to allow the temperature of the buffer solution to reach preset temperature, adding L-DOPA (levodopa) and the sample to be determined, mixing intensively, rapidly determining initial absorbance when the wavelength is 490nm, simultaneously starting water bath timing, performing the water bath, determining absorbance (OD (optical density)), further taking two test tubes for each group to measure the absorbance of the substrate L-DOPA and the sample at 490nm to be ODL and OD0 as controls, parallelizing each group for three times, and solving a calculation formula: PO = (OD-ODL-OD0) / t by taking 0.001 of average increase of OD at 490nm per minute (t) as an enzyme activity unit (U).

The invention provides an application of eremothecium in inhibition of polyphenol oxidase activity and belongs to the technical field of polyphenol oxidase activity research. In the application, eremothecium suspension liquid in the logarithmic phase can remarkably inhibit polyphenol oxidase, and the inhibition rate can reach 32%. The inhibition of the polyphenol oxidase by the eremothecium belongs to a mixed inhibition type, the eremothecium can react with not only the free polyphenol oxidase but also a complex of the polyphenol oxidase combined with a substrate, the inhibition ways are multiple, the inhibition rate is high, and the application range is wide.

The invention discloses a preparation method of phenol oxidase active protein. The present invention uses the residual waste plasma of Limulus reagent as raw material, salts out the protein through saturated ammonium sulfate solution, freezes and centrifuges to obtain Limulus hemocyanin, and then desalts the PBS solution of hemocyanin through MW3500Da dialysis bag, and passes through SephacrylS‑ 100 propylene dextran gel column purification, concentrated treatment to obtain high-efficiency phenoloxidase activity of hemocyanin, the reaction initial velocity Vi = 74.52 nmol · (min · mg). The product obtained by the present invention has high purity and high activity, and can be used to prepare efficient, convenient, and low-cost micro-detectors for phenolic pollutants; the preparation method of the present invention is simple, and adopts MW3500Da dialysis bags and SephacrylS-100 acryl-dextran gel The combination of columns can achieve the effects of desalination, separation, purification and concentration at the same time.