CN101724012A - Deep processing method for cultivated gardon asparagus in gracilaria - Google Patents

Deep processing method for cultivated gardon asparagus in gracilaria Download PDF

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CN101724012A
CN101724012A CN200810228026A CN200810228026A CN101724012A CN 101724012 A CN101724012 A CN 101724012A CN 200810228026 A CN200810228026 A CN 200810228026A CN 200810228026 A CN200810228026 A CN 200810228026A CN 101724012 A CN101724012 A CN 101724012A
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protein solution
frond
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agar
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CN101724012B (en
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张卫
李海燕
虞星炬
曹旭鹏
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Dalian Institute of Chemical Physics of CAS
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Dalian Institute of Chemical Physics of CAS
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Abstract

The invention relates to a set of economic and feasible deep processing method for cultivating seaweed, gardon asparagus in large scale in China's sea areas, and high quality agaragars, reagent pure phycoerythrins, halogen peroxide enzyme products, trace element feed additives and trace element fertilizers can be produced simultaneously. The deep processing method is characterized in that protein crude extract can be produced via pretreatment of the gardon asparagus fronds, crude extract preparation and fractionated precipitation of the ammonium sulphate; the electrophoresis pure halogen peroxide enzyme products and the phycoerythrins crude extract are produced via the negative ion exchange column chromatography; after the phycoerythrins is purified through hydroxylapatite and the molecular sieve column chromatography, pure phycoerythrins products are obtained; after leaching the protein, the remaining gardon asparagus leavings is used for extracting the high-quality agaragars; and then the leavings is dried and smashed to get trace element feed additives and trace element fertilizers. The invention has economic process and high efficiency, is environment-friendly, opens up a new path for cultivating the gardon asparagus products in large quantity, and becomes the important technique basis for realizing the gardon asparagus comprehensive utilization industrialization.

Description

A kind of deep processing method of cultivated gardon asparagus in gracilaria
Technical field
The present invention relates to halobiontic deep processing, specifically a kind of deep processing technology of cultivated gardon asparagus can obtain agar, the pure phycoerythrin of reagent, haloperoxidase and microelement feed addictive and trace element fertilizer simultaneously.
Background technology
Thallus Gracilariae as a peculiar kind of Gracilaria, because of its adaptability is fast, the growth rate advantages of higher, has become the breed kelp of Chinese output the 3rd.Cultivated gardon asparagus can absorb nitrogen, phosphorus and the carbonic acid gas in the seawater in a large number, improves the ecotope of coastal ocean, promotes fish and shellfish procreation growth, makes the breeding ecological environment reach benign cycle.The most important thing is that Thallus Gracilariae has very important economic value: be one of important source material of agar, agar content accounts for 20%～30% of its dry weight, and the agar-agar quality of Thallus Gracilariae is good and content is high, also has antitumous effect.Contain abundant phycoerythrin and functional enzyme component in the frond, as haloperoxidase.Phycoerythrin is widely used in foods and cosmetics industry, with aspects such as fluorescent mark and biomedicines very big application prospect is arranged in fluorescence immunoassay, diagnosis, in addition, also is familiar with by people in application prospect pharmaceutically; Haloperoxidase has the effect of the oxidation of catalysis sulphur, epoxidation, indoles oxidation and other reaction, more and more draws attention aspect synthetic and high added value compound synthesizes at medicine.Residue behind the comprehensive utilization process still contains abundant trace element, as sodium, magnesium, aluminium, silicon, phosphorus, sulphur, potassium, calcium, manganese, iron, nickel, zinc, strontium, chlorine, bromine, iodine etc., can be used for trace element fertilizer in the microelement feed addictive auxiliary material of domestic animal and the agriculture production.Therefore, no matter Thallus Gracilariae is the raw materials for production of staple commodities agar, still all has good economic worth as the phycoerythrin of high added value and the raw material of functional enzyme etc., very necessary to comprehensive utilization and the high-quality Products Development of Thallus Gracilariae.
Summary of the invention
The objective of the invention is to set up a kind of deep processing method that from cultivated gardon asparagus, obtains agar, phycoerythrin, haloperoxidase and microelement feed addictive and trace element fertilizer simultaneously, realize the deep development utilization of cultivated gardon asparagus resource.
For achieving the above object, the technical scheme taked of the present invention is:
Integrated by technology and product, realize the thought of the complete utilization of Thallus Gracilariae, the present invention has adopted a kind of deep processing method that obtains agar, phycoerythrin, haloperoxidase and microelement feed addictive and trace element fertilizer from cultivated gardon asparagus, and specific operation process is as follows:
A kind of deep processing method of cultivated gardon asparagus in gracilaria, culturing the Gracilaria Thallus Gracilariae with Chinese marine site is raw material,
A. extract the pure phycoerythrin of reagent
(1) pre-treatment: clean and remove silt and other foreign material in the Thallus Gracilariae, control water purification branch, lyophilize, liquid nitrogen grinding becomes gardon asparagus powder;
(2) crude extract preparation: in gardon asparagus powder, add the phosphate buffered saline buffer of 0.01～0.2M of 10～30 times of Thallus Gracilariae frond dry weights, 0～4 ℃, soak 24～72h, filtered through gauze gets algae-residue and filtrate; Filtrate is in 0～4 ℃, the centrifugal 10～50min of 3000g～8000g, and supernatant liquor is a protein crude extract;
(3) ammonium sulfate precipitation: crude extract adds solid ammonium sulfate, and to make the mass concentration of ammonium sulfate in crude extract be 15～40%, 0～4 ℃, the centrifugal 10～50min of 6000～15000g; Abandon precipitation, supernatant adds solid ammonium sulfate once more, and to make the mass concentration of ammonium sulfate in solution be 55～80%, 0～4 ℃, the centrifugal 10～50min of 6000～15000g; Collecting precipitation, the centrifuging and taking throw out with 0.01～0.2M phosphoric acid buffer dialysis 10～48 hours, obtains protein solution A; The dialysis membrane molecular weight cut-off is 14000;
(4) anion exchange chromatography separates: with the 0.01～0.2M phosphoric acid buffer linear gradient elution protein solution A that contains NaCl, the concentration of NaCl in phosphoric acid buffer is from 0.05～2.0M linear change during wash-out, protein solution A is 1: 20～40 with the cumulative volume ratio of elutriant, the wash-out component was collected by the timed interval, collected a pipe in per 3～10 minutes, measure every pipe respectively and collect liquid in the light absorption value at 280nm and 565nm place, according to elution peak and OD 565/ OD 280The ratio size is collected the interior protein solution B of purer collection tube of ratio 〉=1;
(5) hydroxyapatite column chromatography for separation: adopt concentration to make linear gradient elution protein solution B from the phosphoric acid buffer that 0.01～0.4M changes, protein solution B is 1: 10～30 with the cumulative volume ratio of elutriant, the wash-out component was collected by the timed interval, collected a pipe in per 4～12 minutes, measure every pipe respectively and collect the light absorption value at liquid 280nm and 565nm place, according to elution peak and OD 565/ OD 280The ratio size is collected the interior protein solution C of purer collection tube of ratio 〉=2.6;
(6) gel-filtration molecular sieve column chromatography: adopt the 0.01～0.2M phosphoric acid buffer elutriant eluted protein solution C that contains 0.05～0.5M NaCl, the wash-out component was collected by the timed interval, collected a pipe in per 4～10 minutes, measure every pipe respectively and collect the 280nm of liquid and the light absorption value at 565nm place, according to OD 565/ OD 280The ratio size is collected the pure phycoerythrin of reagent of purity 〉=4.0;
B. extract the pure haloperoxidase of electrophoresis:
Every pipe is collected the haloperoxidase vigor of protein solution B and the light absorption value at 280nm place among the determination step A (4), collects the solution that is associated with enzyme activity, obtains haloperoxidase; Through 0.01～0.1M, pH7～9Tris-Cl damping fluid dialysed overnight, the dialysis membrane molecular weight cut-off is 14000, improves enzyme activity with the solution collected;
C. agar extracts:
Lixiviate in the steps A (2) is got crude protein algae-residue afterwards, handle in mass concentration 0.5～9%NaOH solution, system solid-liquid mass volume ratio is 1: 10～50, and treatment temp is 50～100 ℃, and the time is 0.5～6 hour; The reaction back is extremely neutral with fresh water flushing frond; Frond is soaked in the water, and the water surface did not have frond, and loose distribution fully contacts illumination, and intensity of illumination is 0.5 ten thousand～150,000 Lux conditions 2～72 hours, and pale brown look frond becomes off-white color, finishes bleaching process;
30～40 times of water yields that add the frond dry weight, normal pressure boiling water are carried glue or carry glue under gauge pressure 0.05～0.07MPa, and the time is 1～4 hour, and 80～500 eye mesh screens filter;
Filter residue is carried glue once more, adds 15～20 times of water yields of frond dry weight, and normal pressure boiling water is carried glue or carry glue under gauge pressure 0.05～0.07MPa, and the time is 0.5～2 hour, and 80～500 eye mesh screens filter, and twice filtrate merges, and collects waste residue; After solidifying, the filtrate naturally cooling gets product;
Solidify the back product and be cut into elongated side's bar, be put in 2～6 ℃ of precoolings, the time is 4～6 hours; Put into again under-20～-25 ℃ and freezed 24～36 hours; Dehydration is soaked, is cleaned adhesive tape, is drying to obtain strip agar; Drying is pulverized, and crosses 100～300 mesh sieves and can be made into powder-like product;
D. microelement feed addictive and trace element fertilizer preparation:
Waste residue after step C agar extracts is put 40～80 ℃ of oven dry, weighs, and pulverizes, and crosses 60～120 mesh sieves and promptly gets product;
The product that more than shows journey and obtained is high-quality agar, the pure phycoerythrin of reagent, haloperoxidase and microelement feed addictive and trace element fertilizer, and detailed process is seen Fig. 1.
Described anion exchange filler is DE52 or DEAE Sepharose; Described gel-filtration molecular sieve filling is Sephacryl S-300 or Sephadex G-200.
The pH=7.0 of described phosphoric acid buffer.
Described Tris-Cl damping fluid contains 1mM VO 4 3-
Advantage of the present invention:
1. the present invention can obtain phycoerythrin, haloperoxidase, agar and microelement feed addictive and trace element fertilizer simultaneously from cultivated gardon asparagus, and the phycoerythrin purity behind the purifying is greater than 4.0, and productive rate is 4.03mg/g phycoerythrin (dry weight); The haloperoxidase productive rate is a 5.67U/ gram (dry weight); Gained agar productive rate is 19.1% (dry weight), and is superior in quality, suitable with direct extraction agar method productive rate; The productive rate of trace mineral supplement and trace element fertilizer is 48.2%.Can improve the utilization ratio of Thallus Gracilariae greatly by the present invention, realize the low-cost high production of Thallus Gracilariae.
2. the applied range of the multiple seaweed bio product of separation and Extraction.
3. method of the present invention is applicable to all Gracilarias and Gelidium marine alga.
In a word, the present invention is low, the efficient feasible cultivated gardon asparagus in gracilaria deep processing method of a kind of environmental pollution, has wide application prospect.
Description of drawings
Fig. 1 is a cultivated gardon asparagus in gracilaria comprehensive utilization process schema.
Fig. 2 is the inorganic elements analytical results of Thallus Gracilariae raw material and comprehensive utilization back residue thereof.
Fig. 3 is the absorption and the fluorescence emission spectrum spectrogram of phycoerythrin
Embodiment
Below by specific embodiment method of the present invention and result are described:
Embodiment 1
Clean and remove silt and other foreign material in the Thallus Gracilariae, control water purification branch, lyophilize, liquid nitrogen grinding powdered; Get 10 gram algae powder, (0.01M pH7.0), 2 ℃, soaks 30h, filtered through gauze to add 100ml potassium phosphate salt damping fluid.2 ℃, the centrifugal 15min of 4000g gets supernatant liquor.Crude extract adds solid (NH 4) 2SO 4To 25%, 2 ℃ of final quality concentration, the centrifugal 20min of 7000g.Abandon precipitation, supernatant is used (NH once more 4) 2SO 4Precipitation (mass concentration 60%), 2 ℃, the centrifugal 30min of 9000g.Collecting precipitation was dialysed 15 hours with 0.02M, pH7.0 potassium phosphate salt damping fluid, obtained protein crude extract A; The dialysis membrane molecular weight cut-off is 14000;
Protein solution A after the dialysis goes up the DEAE-sepharose post and separates, with the 0.02M that contains NaCl, pH7.0 potassium phosphate salt damping fluid linear gradient elution, the concentration of NaCl in phosphoric acid buffer is from 0.1～1.0M linear change during wash-out, protein solution A is 1: 35 with the cumulative volume ratio of elutriant, the wash-out component was collected by the timed interval, collected a pipe in per 5 minutes, measure respectively that every pipe collects that the enzyme of liquid is lived and 280, the light absorption value at 565nm place, be associated with the component that enzyme is lived, obtain haloperoxidase; Collect OD simultaneously 565/ OD 280Protein solution B in the purer collection tube of ratio 〉=1, protein solution B is through 0.02M, pH7.0Tris-Cl damping fluid dialysed overnight, and the dialysis membrane molecular weight cut-off is 14000, improves enzyme activity.
Protein solution B, last hydroxyapatite column chromatography for separation, adopt concentration to make linear gradient elution from the potassium phosphate salt damping fluid that 0.01～0.2M changes, protein solution B is 1: 15 with the cumulative volume ratio of elutriant, the wash-out component was collected by the timed interval, collected a pipe in per 6 minutes, measure the light absorption value that every pipe is collected liquid 280nm and 565nm place respectively, according to elution peak and OD 565/ OD 280The ratio size is collected the interior protein solution C of purer collection tube of ratio 〉=2.6.Protein solution C goes up the gel-filtration molecular sieve column chromatography, employing contains the 0.05M potassium phosphate salt buffer solution elution liquid wash-out of 0.1M NaCl, and the wash-out component was collected by the timed interval, collects one and manages in per 6 minutes, measure every pipe respectively and collect the 280nm of liquid and the light absorption value at 565nm place, according to OD 565/ OD 280The ratio size is collected the pure phycoerythrin of reagent of purity 〉=4.0.
Preparation 9%NaOH (mass percent) solution, solid-to-liquid ratio is 1: 50, is heated to 90 ℃, adds lixiviate again and gets crude protein algae-residue afterwards, isothermal reaction 1h; 80 eye mesh screens filter, and reclaim alkali lye, and frond is fully washed near neutral; The frond freshwater soaking, liquid level did not have frond, loose distribution, intensity of illumination is under the 120000 Lux conditions, and substantial light was according to 3 hours, and frond becomes off-white color; Algae weighs 40 times water and is added in the frond that filters after cleaning, and the normal pressure poach was carried glue 3 hours, 500 eye mesh screen filtered while hot; Filter residue adds water and carries out carrying the second time glue, and time and water consumption all reduce by half, 500 eye mesh screen filtered while hot; Twice filtering filtrate merges, and after naturally cooling solidifies, is cut into elongated side's bar.The strip gel is placed 3 ℃ of precoolings 4 hours; Put under-20 ℃ and freezed 36 hours, adhesive tape is freezed fully; Thaw adhesive tape dehydration of clear water, ice crystal melts, with clear water soak, the rinsing adhesive tape, filter and moisture dried; Adhesive tape is arranged arrangement evenly, under daylight, dry, get the strip agar-agar; Dried agar-agar is pulverized, and crosses 200 eye mesh screens, promptly gets the agar-agar powder.
Waste residue after agar extracts is put 45 ℃ of oven for drying, weighs, and pulverizes, and crosses 100 mesh sieves, obtains microelement feed addictive and trace element fertilizer.The micronutrient levels comparative result is seen Fig. 2 before and after the Thallus Gracilariae comprehensive utilization process.
Haloperoxidase enzyme biopsy survey method is referring to document 1:Itoh N, Izumi Y, Yamada H, 1986,261:5194.The phycoerythrin method for detecting purity is referring to document 2:Wang GC, 2002,56:509.
Calculate with dry weight, the extracted amount that extracts various products from cultivated gardon asparagus sees Table 1.The phycoerythrin that the deep processing method purifying obtains shows fabulous spectroscopic properties, and it absorbs and fluorescence emission spectrum is seen Fig. 3.
Table 1 extracts the extracted amount (embodiment 1) of various products from cultivated gardon asparagus
Product Extracted amount (dry weight)
The pure phycoerythrin of reagent ??4.03mg/g
Haloperoxidase ??5.67U/g
Agar ??19.1％
Microelement feed addictive and trace element fertilizer ??48.2％
The agar that this technology is extracted shows good physicochemical property, sees Table 2.
The physical chemistry spare matter of agar (embodiment 1) in the table 2 Thallus Gracilariae comprehensive utilization process
Physicochemical property Numerical value
Gel-strength (g/cm 2) ??1860
Temperature of solidification (℃) ??41.3
3, acyl galactose content (%) in the 6- ??40.8
Physicochemical property Numerical value
Sulfate group content (%) ??0.81
Embodiment 2
Clean and remove silt and other foreign material in the Thallus Gracilariae, control water purification branch, lyophilize, liquid nitrogen grinding powdered; Get 15 gram algae powder, add 300ml 0.1M sodium phosphate salt damping fluid, 4 ℃ are soaked 72, filtered through gauze.4 ℃, the centrifugal 25min of 6000g gets supernatant liquor.Crude extract adds solid (NH 4) 2SO 4To 30%, 4 ℃ of final quality concentration, the centrifugal 15min of 13000g.Abandon precipitation, supernatant is used (NH once more 4) 2SO 4Precipitation (mass concentration 70%), 6 ℃, the centrifugal 25min of 12000g.Collecting precipitation was dialysed 36 hours with 0.1M sodium phosphate salt damping fluid, got protein crude extract A; The dialysis membrane molecular weight cut-off is 14000.
Protein solution A after the dialysis goes up the DEAE-52 post and separates, with the 0.05M that contains NaCl, pH8.0 sodium phosphate salt damping fluid linear gradient elution, the concentration of NaCl in the sodium phosphate salt damping fluid is from 0.1～2.0M linear change during wash-out, protein solution A is 1: 20 with the cumulative volume ratio of elutriant, the wash-out component is collected by 8 minutes interval, measure every pipe and collect the enzyme work and 280 of liquid, the light absorption value at 565nm place, be associated with the component that enzyme is lived, get haloperoxidase; Collect OD simultaneously 565/ OD 280Protein solution B in the purer collection tube of ratio 〉=1, protein solution B is through 0.1M, pH9.0Tris-Cl damping fluid dialysed overnight, and the dialysis membrane molecular weight cut-off is 14000.
The protein solution B that collects, last hydroxyapatite column chromatography for separation, adopt concentration to make linear gradient elution from the sodium phosphate salt damping fluid that 0.05～0.3M changes, protein solution B is 1: 30 with the cumulative volume ratio of elutriant, the wash-out component was collected by the timed interval, collected a pipe in per 8 minutes, measure the light absorption value that every pipe is collected liquid 280nm and 565nm place respectively, according to elution peak and OD 565/ OD 280The ratio size is collected the interior protein solution C of purer collection tube of ratio 〉=2.6.The protein solution C that collects goes up Sephadex G-200 molecular sieve: with the 0.1M sodium phosphate salt buffer solution elution that contains 0.4M NaCl, the wash-out component is pressed the interval of 8min and is collected, measure every pipe collect liquid 280, the light absorption value at 565nm place, the pure phycoerythrin of reagent of collection purity 〉=4.0.
Preparation 3%NaOH (mass percent) solution is heated to 75 ℃, adds the algae-residue after the crude protein lixiviate again, solid-to-liquid ratio 1: 20, isothermal reaction 3h; 80 eye mesh screens filter, and reclaim alkali lye, and frond is fully washed near neutral; Under the 50000 Lux intensity of illumination conditions, illumination 24 hours, frond becomes off-white color; Frond and algae weigh 30 times water, carry glue 1 hour under the 0.06MPa gauge pressure, 80 eye mesh screen filtered while hot; Filter residue adds water and carries out carrying the second time glue, and time and water consumption all reduce by half, 80 eye mesh screen filtered while hot; Twice filtering filtrate merges, and after naturally cooling solidifies, is cut into elongated side's bar.The strip gel is placed 5 ℃ of precoolings 6 hours; Put under-25 ℃ again and freezed 24 hours, adhesive tape is freezed fully; Adhesive tape is cleaned in the clear water immersion of thawing repeatedly, turns white until shinny; Row is evenly whole with adhesive tape, dries in 60 ℃ baking oven, gets the strip agar-agar; Dried agar-agar is pulverized, and crosses 200 eye mesh screens, promptly gets agar powder.
Waste residue after agar extracts is put 80 ℃ of oven for drying, weighs, and pulverizes, and crosses 60 mesh sieves, gets microelement feed addictive and trace element fertilizer.

Claims (4)

1. the deep processing method of a cultivated gardon asparagus in gracilaria is characterized in that: culturing the Gracilaria Thallus Gracilariae with Chinese marine site is raw material,
A. extract the pure phycoerythrin of reagent
(1) pre-treatment: clean impurity elimination, lyophilize, liquid nitrogen grinding becomes gardon asparagus powder;
(2) crude extract preparation: in gardon asparagus powder, add the phosphate buffered saline buffer of 0.01～0.2M of 10～30 times of Thallus Gracilariae frond dry weights, 0～4 ℃, soak 24～72h, filtered through gauze gets algae-residue and filtrate; Filtrate is in 0～4 ℃, the centrifugal 10～50min of 3000g～8000g, and supernatant liquor is a protein crude extract;
(3) ammonium sulfate precipitation: crude extract adds solid ammonium sulfate, and to make the mass concentration of ammonium sulfate in crude extract be 15～40%, 0～4 ℃, the centrifugal 10～50min of 6000～15000g; Abandon precipitation, supernatant adds solid ammonium sulfate once more, and to make the mass concentration of ammonium sulfate in solution be 55～80%, 0～4 ℃, the centrifugal 10～50min of 6000～15000g; Collecting precipitation, the centrifuging and taking throw out with 0.01～0.2M phosphoric acid buffer dialysis 10～48 hours, obtains protein solution A; The dialysis membrane molecular weight cut-off is 14000;
(4) anion exchange chromatography separates: with the 0.01～0.2M phosphoric acid buffer linear gradient elution protein solution A that contains NaCl, the concentration of NaCl in phosphoric acid buffer is from 0.05～2.0M linear change during wash-out, protein solution A is 1: 20～40 with the cumulative volume ratio of elutriant, the wash-out component was collected by the timed interval, collected a pipe in per 3～10 minutes, measure every pipe respectively and collect liquid in the light absorption value at 280nm and 565nm place, according to elution peak and OD 565/ OD 280The ratio size is collected the interior protein solution B of purer collection tube of ratio 〉=1;
(5) hydroxyapatite column chromatography for separation: adopt concentration to make linear gradient elution protein solution B from the phosphoric acid buffer that 0.01～0.4M changes, protein solution B is 1: 10～30 with the cumulative volume ratio of elutriant, the wash-out component was collected by the timed interval, collected a pipe in per 4～12 minutes, measure every pipe respectively and collect the light absorption value at liquid 280nm and 565nm place, according to elution peak and OD 565/ OD 280The ratio size is collected the interior protein solution C of purer collection tube of ratio 〉=2.6;
(6) gel-filtration molecular sieve column chromatography: adopt the 0.01～0.2M phosphoric acid buffer elutriant eluted protein solution C that contains 0.05～0.5M NaCl, the wash-out component was collected by the timed interval, collected a pipe in per 4～10 minutes, measure every pipe respectively and collect the 280nm of liquid and the light absorption value at 565nm place, according to OD 565/ OD 280The ratio size is collected the pure phycoerythrin of reagent of purity 〉=4.0;
B. extract the pure haloperoxidase of electrophoresis:
Every pipe is collected the haloperoxidase vigor of protein solution B and the light absorption value at 280nm place among the determination step A (4), collects the solution that is associated with enzyme activity, obtains haloperoxidase; Through 0.01～0.1M, pH7～9 Tris-Cl damping fluid dialysed overnight, the dialysis membrane molecular weight cut-off is 14000, improves enzyme activity with the solution collected;
C. agar extracts:
Lixiviate in the steps A (2) is got crude protein algae-residue afterwards, handle in mass concentration 0.5～9%NaOH solution, system solid-liquid mass volume ratio is 1: 10～50, and treatment temp is 50～100 ℃, and the time is 0.5～6 hour; The reaction back is extremely neutral with fresh water flushing frond; Frond is soaked in the water, and the water surface did not have frond, and loose distribution fully contacts illumination, and intensity of illumination is 0.5 ten thousand～150,000 Lux conditions 2～72 hours, and pale brown look frond becomes off-white color, finishes bleaching process;
30～40 times of water yields that add the frond dry weight, normal pressure boiling water are carried glue or carry glue under gauge pressure 0.05～0.07MPa, and the time is 1～4 hour, and 80～500 eye mesh screens filter;
Filter residue is carried glue once more, adds 15～20 times of water yields of frond dry weight, and normal pressure boiling water is carried glue or carry glue under gauge pressure 0.05～0.07MPa, and the time is 0.5～2 hour, and 80～500 eye mesh screens filter, and twice filtrate merges, and collects waste residue; After solidifying, the filtrate naturally cooling gets product;
Solidify the back product and be cut into elongated side's bar, be put in 2～6 ℃ of precoolings, the time is 4～6 hours; Put into again under-20～-25 ℃ and freezed 24～36 hours; Dehydration is soaked, is cleaned adhesive tape, is drying to obtain strip agar; Drying is pulverized, and crosses 100～300 mesh sieves and can be made into powder-like product;
D. microelement feed addictive and trace element fertilizer preparation:
Waste residue after step C agar extracts is put 40～80 ℃ of oven dry, weighs, and pulverizes, and crosses 60～120 mesh sieves and promptly gets product;
The product that more than shows journey and obtained is high-quality agar, the pure phycoerythrin of reagent, haloperoxidase and microelement feed addictive and trace element fertilizer.
2. according to the described deep processing method of claim 1, it is characterized in that: described anion exchange filler is DE-52 or DEAE-Sepharose; Described gel-filtration molecular sieve is Sephacryl S-300 or Sephadex G-200.
3. according to the described deep processing method of claim 1, it is characterized in that: the pH=7.0 of described phosphoric acid buffer.
4. according to the described deep processing method of claim 1, it is characterized in that: described Tris-C1 damping fluid contains 1mM VO 4 3-
CN2008102280266A 2008-10-10 2008-10-10 Deep processing method for cultivated gardon asparagus in gracilaria Expired - Fee Related CN101724012B (en)

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102150735A (en) * 2011-05-17 2011-08-17 福建省水产研究所 Method for comprehensively utilizing and processing gracilaria
CN102276759A (en) * 2011-06-20 2011-12-14 青岛海康生物有限公司 Preparation technology of agar for high-quality microbial culture medium
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CN104292316B (en) * 2013-07-19 2017-03-29 中国科学院烟台海岸带研究所 The high efficiency separation and purification method of Thallus Gracilariae SILVER REAGENT R type phycoerythrin
CN104336549A (en) * 2014-10-10 2015-02-11 嵊泗县冠岛水产有限公司 Processing method of asparagus
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CN112425503B (en) * 2020-12-10 2022-04-08 江苏海洋大学 Device for full-artificial seedling picking of asparagus

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