CN105249434A - Method for preparing natural alginate oligosaccharide water-retaining agent by utilizing abalone visceral enzyme liquid - Google Patents
Method for preparing natural alginate oligosaccharide water-retaining agent by utilizing abalone visceral enzyme liquid Download PDFInfo
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- CN105249434A CN105249434A CN201510757644.XA CN201510757644A CN105249434A CN 105249434 A CN105249434 A CN 105249434A CN 201510757644 A CN201510757644 A CN 201510757644A CN 105249434 A CN105249434 A CN 105249434A
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Abstract
The invention discloses a method for preparing a natural alginate oligosaccharide water-retaining agent by utilizing abalone visceral enzyme liquid. The method for preparing the natural alginate oligosaccharide water-retaining agent by utilizing the abalone visceral enzyme liquid comprises the following steps: deshelling fresh abalone, and taking out viscera; mixing and mashing abalone viscera, and adding distilled water; uniformly stirring, regulating pH to 8.0-9.0, and continuously stirring, so as to obtain crude enzyme liquid; filtering the crude enzyme liquid by adopting a coarse filtration device to take filtrate; carrying out membrane filtration on the obtained filtrate for removing impurities, wherein liquid outside a membrane is the prepared enzyme solution; regulating pH value of 1 mass% of alginate aqueous solution to be 8.5, adding the alginate aqueous solution into the obtained enzyme solution, and reacting, so as to obtain alginate oligosaccharide solution; and carrying out rotary evaporation and concentration until solid content is 15-25%, and carrying out spray drying to obtain powder, thus obtaining the natural alginate oligosaccharide water-retaining agent, wherein the molecular weight of the prepared alginate oligosaccharide is 300-1000Da. The method for preparing the natural alginate oligosaccharide water-retaining agent by utilizing the abalone visceral enzyme liquid has the advantages that raw materials are available, the cost is low, the product additive value of alginate is improved, and the environmental pollution is reduced; meanwhile, the technique is simple, the source is natural, and the production cost is reduced; besides, the water-retaining property of an aquatic production is effectively improved, and the colour and lustre of the natural alginate oligosaccharide water-retaining agent product can not be changed.
Description
Technical field
The invention belongs to field of processing of aquatic products, particularly a kind of method preparing natural brown alga oligose water-loss reducer with abalone visceral enzym liquid.
Background technology
Aquatic products are in processing and storage transportation, and the use with the additive of water retaining function can improve the hold facility of moisture in aquatic products to improve the stability of moisture.The principle active component of traditional aquatic products water-loss reducer is polyphosphate, but excessive use phosphate not only causes harmful effect to environment, also can work the mischief to the health of human body.Therefore, European Union from 1 day January in 2006 nonimportation use the aquatic products crossed of phosphate treated.China also must make adjustment in time to meet international change demand.Appearance along with China's " the law of food safety " in 2009 and people's improving constantly food safety requirements, use safety, reliably food additives become inexorable trend.So the Development and Production of novel, efficient, safe aquatic food antistaling agent, has huge market potential.
Algin is the important composition composition of brown alga cell membrane, and being mainly derived from the brown algas such as sea-tangle, bulk kelp, sargassum, is the straight chain acidic polysaccharose be formed by connecting by 1 → 4 glycosidic bond by D-MANNOSE aldehydic acid (M) and L-guluronic acid (G).Food industry, textile industry and the industry such as medical and health, daily-use chemical industry are widely used in.But algin molecular weight is large, and dissolubility is low, viscosity is large, thus makes its application in every profession and trade be subject to a definite limitation.At present, main employing enzyme edman degradation Edman and chemical degradation method degraded algin, obtain low-molecular-weight brown alga oligose.The advantages such as there are some researches show, it is strong that brown alga oligose not only has dissolubility, the high and safety non-toxic of stability, also have multiple biologically active, as: Promoting plant growth, antitumor, neuroprotective etc.Algin wide material sources, utilize it to produce brown alga oligose, have important researching value and huge potentiality to be exploited.
According to China Fisheries yearbook statistics, within 2013, China's abalone output is 11.03 ten thousand tons, and wherein Fujian Province's output is 8.84 ten thousand tons, and in the trend increased year by year.Abalone take seaweeds as food, containing abundant polysaccharide hydrolysis enzyme in its alimentary canal, as: cellulase, algin catenase, agarase etc.In abalone process, internal organ, as processing fent, often by reject or be prepared into the low value products such as fish meal, are difficult to realize higher value application.Therefore, with the accessory substance internal organ after abalone processing for raw material, algin is degraded, reduce its viscosity, obtain low-molecular-weight brown alga oligose, be applied in the exploitation of aquatic products water-loss reducer, effectively can improve the added value of abalone and algin processing, the comprehensive high-efficiency for China's marine resources utilizes makes contributions.
Summary of the invention
The object of the present invention is to provide a kind of preparation method utilizing natural enzyme efficiently to prepare the algin oligosaccharide improving retention ability.
For achieving the above object, the invention provides a kind of preparation method of natural brown alga oligose water-loss reducer, it is characterized in that, concrete steps are as follows:
Prepared by crude enzyme liquid: by fresh abalone decladding, get internal organ; Abalone internal organ are blended to without block solid content, adds distilled water; Stir, regulate pH to 8.0-9.0 with food-grade NaOH, Keep agitation; Preferred stirring 30-60min; Gained is crude enzyme liquid;
The preparation of enzyme liquid: adopt coarse filter device to filter crude enzyme liquid, get filtrate; By gained filtrate through membrane filtration removal of impurities, the outer liquid of film is the enzyme liquid of preparation.
The preparation of algin oligosaccharide solution: by soluble in water for commercial algin, regulates pH to 8.5, and preparation mass percentage concentration is the alginate solution of 1%, reacts, obtain algin oligosaccharide solution after adding gained enzyme liquid;
Water-loss reducer powder preparation: be 15-25% to solid content by described algin oligosaccharide solution rotating evaporation and concentration, is spray dried to powder; Prepared algin oligosaccharide molecular weight is 300-1000Da.
Further, in described crude enzyme liquid preparation process, the mass ratio of described abalone internal organ and distilled water is 1:4-1:10;
Optional, carry out at described crude enzyme liquid is prepared in 4 DEG C.
Further, in the preparation process of described enzyme liquid, coarse filter device is nylon filter bag or silk or 3 layers of gauze.
Further, in the preparation process of described enzyme liquid, described film is ceramic membrane or hollow polyester film, and ceramic membrane aperture is 0.1-0.3mm, and hollow polyester membrane aperture is 0.2-0.8mm.
Further, in the preparation of described algin oligosaccharide solution, the volume ratio of the alginate solution of described enzyme liquid and 1 quality % is 1:20-1:50.
Further, in the preparation of described algin oligosaccharide solution, reaction condition is Keep agitation reaction 30min-3h under 37 DEG C of conditions.
Further, in described water-loss reducer powder preparation, in rotary evaporation process, 50-70 DEG C is maintained the temperature at; Spraying dry inlet temperature is at 160-185 DEG C, and charging rate is 200-400mL/h.
The natural brown alga oligose water-loss reducer that the present invention also protects the preparation method of described natural brown alga oligose water-loss reducer to prepare.
The natural brown alga oligose water-loss reducer that the present invention also protects the preparation method of described natural brown alga oligose water-loss reducer to prepare is for the water conservation purposes of aquatic products;
Preferably, described purposes is that water intaking product eats the natural brown alga oligose water-loss reducer solution 0.01-0.1 % by weight for preparing of preparation method that part is placed in described natural brown alga oligose water-loss reducer and soaks 30-120min;
Preferably, the natural brown alga oligose water-loss reducer solution of every kg water product interpolation 1L-4L carries out complete submergence immersion.
Further, aquatic products are fresh-water fishes, fish, prawn and crab.
The present invention's key make use of natural enzyme and efficiently prepares algin oligosaccharide, and the algin oligosaccharide suitable molecular weight of preparation is used for fresh-keeping.Not every algin oligosaccharide can improve retention ability, and molecular weight is that the algin oligosaccharide of 300-1000Da is just passable.The oligosaccharides that the present invention identifies preparation is not only effective but also identify molecular weight.
Beneficial effect of the present invention:
(1) with abalone internal organ for raw material, the abundant hydrolase utilizing it to contain, preparation technology is simple, and processing ease, originates natural, has saved production cost simultaneously.
(2) utilize abalone visceral enzym to be hydrolyzed to algin and prepare brown alga oligose, raw material sources are extensive, cheap, improve the added value of product of algin, reduce the use amount of traditional phosphate water-loss reducer simultaneously, solve the problem that phosphate exceeds standard, decrease problem of environmental pollution.
(3) this water-loss reducer effectively can improve the water-retaining property of aquatic products, decreases phosphatic use, and can not cause the change of product color, improves the phenomenon that the product that uses phosphate water-loss reducer to bring turns white.
Accompanying drawing explanation
Fig. 1 is that oligosaccharides electron spray Plasma-Mass Spectroscopy figure prepared by abalone visceral enzym liquid enzymolysis algin;
Fig. 2 is the content of reducing sugar variation relation figure in time that abalone visceral enzym liquid enzymolysis algin different time produces;
Fig. 3 is enzymolysis different time alginate solution viscosity variation relation figure in time;
Fig. 4 is the influence curve figure of brown alga oligose to peeled shrimp water-retaining property;
Fig. 5 is the influence curve figure of algin oligosaccharide to Tilapia mossambica fillet water-retaining property.
Detailed description of the invention
Be described below in detail embodiments of the invention, the example of described embodiment is shown in the drawings, and wherein same or similar label represents same or similar element or has element that is identical or similar functions from start to finish.Be exemplary below by the embodiment be described with reference to the drawings, be intended to for explaining the present invention, and can not limitation of the present invention be interpreted as.Unreceipted concrete technology or condition person in embodiment, according to the technology described by the document in this area or condition or carry out according to product description.Agents useful for same or the unreceipted production firm person of instrument, being can by the conventional products of commercial acquisition.
embodiment 1:
(1) buy 10kg abalone, by fresh abalone decladding, get internal organ for subsequent use, altogether 2.5kg;
(2) blended by abalone internal organ, add 15L volume distilled water, tissue mashing, regulate pH to 8.5, Keep agitation 60min with food-grade NaOH, described operating process is all carried out at 4 DEG C.
(3) adopt filter bag to filter crude enzyme liquid, get filtrate.By crude enzyme liquid through hollow polyester (0.6mm) membrane filtration, pressure 1.8MPa before post, after post, pressure is 0.8MPa, gets the outer liquid of film and is obtained abalone visceral enzym liquid, 16L altogether.
(4) prepare the alginate solution 10L that mass percentage concentration is 1%, 1:40 adds enzyme liquid 250mL by volume, under 37 DEG C of conditions, react 1h, obtains algin oligosaccharide solution.Get the differential responses time, the content of reducing sugar of assaying reaction product and viscosity number (the results are shown in Figure 2 and Fig. 3).As can be seen from the figure, along with the reaction time extends, product content of reducing sugar constantly increases, and solution viscosity constantly declines.
(5) be 20% by brown alga oligose solution rotating evaporation and concentration to solid content, be spray dried to powder for subsequent use, altogether 92g algin oligosaccharide.
Adopt distilled water to be configured to the solution that concentration is 50 μ g/mL the algin oligosaccharide of acquisition, adopt ESI/MS(MicromassLCTPremierTMXE, Waters, USA) oligosaccharide molecular amount is identified.As shown in Figure 1, after ESI-MS qualification, algin oligosaccharide molecular weight concentrates on about 300-700Da.
embodiment 2
The internal organ that shelled by prawn get peeled shrimp part.Prepare the brown alga oligose solution of variable concentrations in advance, concentration is respectively 0, and 0.01%, 0.05%, 0.1%.In the solution of often kind of concentration, put into 3 peeled shrimps, soak 60min, soak after terminating and dry.Measure the water content of peeled shrimp under variable concentrations with electronics moisture teller, weigh peeled shrimp under variable concentrations centrifugal before with centrifugal after quality, encase peeled shrimp with filter paper time centrifugal, be put into centrifugal 2000 × g, 10min in centrifuge tube.According to the retention ability of peeled shrimp under formulae discovery variable concentrations oligosaccharides.Soaking each 5.0g of peeled shrimp under getting variable concentrations carries out centrifugal, the peeled shrimp weight that variable concentrations is corresponding is 3.2,3.8,4.3,4.4g.
Retention ability/%=[before 1-(m after-m)/water content] * 100
Before m: the quality of centrifugal front peeled shrimp.
After m: the quality of centrifugal rear peeled shrimp.
As can be seen from Figure 4, along with the continuous rising of oligosaccharide concentration, the retention ability of peeled shrimp constantly raises.When water-loss reducer concentration is 0.1%, peeled shrimp retention ability can increase by 30%.
embodiment 3
Fresh Tilapia mossambica is removed the peel and the flesh of fish is cut into about 5g thin slice.Add the brown alga oligose solution of variable concentrations, concentration difference 0,0.01%, 0.05%, 0.1%.Put into 3 in the solution of often kind of concentration to flake a fish, soak 60min, soak after terminating and dry.Measure the water content of fillet under variable concentrations with electronics moisture teller, weigh fillet under variable concentrations centrifugal before with centrifugal after quality, encase fillet with filter paper time centrifugal, be put into centrifugal 2000 × g, 10min in centrifuge tube.According to the retention ability of fillet under formulae discovery variable concentrations oligosaccharides.Get variable concentrations soak fillet 5.9,5.7,5.4,4.9g.Centrifugal rear weight is respectively 4.2,4.4,4.4,4.3g.
Retention ability/%=[before 1-(m after-m)/water content] * 100
Before m: the quality of the flesh of fish before centrifugal
After m: the quality of the centrifugal rear flesh of fish
As can be seen from Figure 5, along with oligosaccharide solution concentration raises, fillet retention ability constantly raises, and proves that oligosaccharide solution can effectively improve fillet water retention.When water-loss reducer concentration reaches 0.1%, fillet retention ability raises about 20%.
Although illustrate and describe embodiments of the invention above, be understandable that, above-described embodiment is exemplary, can not be interpreted as limitation of the present invention, those of ordinary skill in the art can change above-described embodiment within the scope of the invention when not departing from principle of the present invention and aim, revising, replacing and modification.
Claims (10)
1. a preparation method for natural brown alga oligose water-loss reducer, it is characterized in that, concrete steps are as follows:
Prepared by crude enzyme liquid: by fresh abalone decladding, get internal organ; Abalone internal organ are blended to without block solid content, adds distilled water; Stir, regulate pH to 8.0-9.0 with food-grade NaOH, Keep agitation; Preferred stirring 30-60min; Gained is crude enzyme liquid;
The preparation of enzyme liquid: adopt coarse filter device to filter crude enzyme liquid, get filtrate; By gained filtrate through membrane filtration removal of impurities, the outer liquid of film is the enzyme liquid of preparation.
The preparation of algin oligosaccharide solution: by soluble in water for commercial algin, regulates pH to 8.5, and preparation mass percentage concentration is the alginate solution of 1%, reacts, obtain algin oligosaccharide solution after adding gained enzyme liquid;
Water-loss reducer powder preparation: be 15-25% to solid content by described algin oligosaccharide solution rotating evaporation and concentration, is spray dried to powder; Prepared algin oligosaccharide molecular weight is 300-1000Da.
2. the preparation method of natural brown alga oligose water-loss reducer described in claim 1, is characterized in that, in described crude enzyme liquid preparation process, the mass ratio of described abalone internal organ and distilled water is 1:4-1:10;
Optional, carry out at described crude enzyme liquid is prepared in 4 DEG C.
3. the preparation method of natural brown alga oligose water-loss reducer described in claim 1, is characterized in that, in the preparation process of described enzyme liquid, coarse filter device is nylon filter bag or silk or 3 layers of gauze.
4. the preparation method of natural brown alga oligose water-loss reducer described in claim 1, is characterized in that, in the preparation process of described enzyme liquid, described film is ceramic membrane or hollow polyester film, and ceramic membrane aperture is 0.1-0.3mm, and hollow polyester membrane aperture is 0.2-0.8mm.
5. the preparation method of natural brown alga oligose water-loss reducer described in claim 1, is characterized in that, in the preparation of described algin oligosaccharide solution, the volume ratio of the alginate solution of described enzyme liquid and 1 quality % is 1:20-1:50.
6. the preparation method of natural brown alga oligose water-loss reducer described in claim 1, is characterized in that, in the preparation of described algin oligosaccharide solution, reaction condition is Keep agitation reaction 30min-3h under 37 DEG C of conditions.
7. the preparation method of natural brown alga oligose water-loss reducer described in claim 1, is characterized in that, in described water-loss reducer powder preparation, maintains the temperature at 50-70 DEG C in rotary evaporation process; Spraying dry inlet temperature is at 160-185 DEG C, and charging rate is 200-400mL/h.
8. the natural brown alga oligose water-loss reducer for preparing of the preparation method of natural brown alga oligose water-loss reducer described in any one of claim 1-7.
9. the natural brown alga oligose water-loss reducer for preparing of the preparation method of natural brown alga oligose water-loss reducer described in any one of claim 1-7 is for the water conservation purposes of aquatic products;
Preferably, the natural brown alga oligose water-loss reducer solution 0.01-0.1 % by weight that the preparation method that described purposes is partly placed in natural brown alga oligose water-loss reducer described in any one of claim 1-8 for water intaking product eats prepares soaks 30-120min;
Preferably, the natural brown alga oligose water-loss reducer solution of every kg water product interpolation 1L-4L carries out complete submergence immersion.
10. described in claim 9 for the water conservation purposes of aquatic products, it is characterized in that, aquatic products are fresh-water fishes, fish, prawn and crab.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107494699A (en) * | 2017-07-05 | 2017-12-22 | 福州大学 | A kind of bio-preservative based on brown alga oligose and its application on prawn is fresh-keeping |
| CN110476851A (en) * | 2019-09-19 | 2019-11-22 | 威海长青海洋科技股份有限公司 | A method of it improving young Bao and turns bait phase survival rate |
| CN110506931A (en) * | 2019-08-22 | 2019-11-29 | 集美大学 | A method of low-viscosity konjac glucomannan is efficiently prepared using the thick enzyme of abalone internal organ |
| CN112870331A (en) * | 2021-02-23 | 2021-06-01 | 广东海洋大学 | Composition for preventing and treating alcoholic liver injury and preparation method and application thereof |
| CN116210814A (en) * | 2022-09-08 | 2023-06-06 | 集美大学 | Application of abalone viscera glycoprotein nano-selenium in preparation of aquatic feed additive |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1341602A (en) * | 2000-09-07 | 2002-03-27 | 青岛海洋大学 | Preparation method of guluronic acid with single degree of polymerization and its application |
| CN104774827A (en) * | 2015-03-23 | 2015-07-15 | 集美大学 | Method for preparing alginate lyase from abalone internal organs |
-
2015
- 2015-11-10 CN CN201510757644.XA patent/CN105249434A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1341602A (en) * | 2000-09-07 | 2002-03-27 | 青岛海洋大学 | Preparation method of guluronic acid with single degree of polymerization and its application |
| CN104774827A (en) * | 2015-03-23 | 2015-07-15 | 集美大学 | Method for preparing alginate lyase from abalone internal organs |
Non-Patent Citations (2)
| Title |
|---|
| 许雷等: "酶解褐藻胶寡糖的吸湿及保湿性能研究", 《日用化学工业》 * |
| 陆云飞等: "褐藻胶寡糖对南美白对虾虾仁品质特性的影响", 《食品科学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107494699A (en) * | 2017-07-05 | 2017-12-22 | 福州大学 | A kind of bio-preservative based on brown alga oligose and its application on prawn is fresh-keeping |
| CN110506931A (en) * | 2019-08-22 | 2019-11-29 | 集美大学 | A method of low-viscosity konjac glucomannan is efficiently prepared using the thick enzyme of abalone internal organ |
| CN110476851A (en) * | 2019-09-19 | 2019-11-22 | 威海长青海洋科技股份有限公司 | A method of it improving young Bao and turns bait phase survival rate |
| CN110476851B (en) * | 2019-09-19 | 2021-06-08 | 威海长青海洋科技股份有限公司 | Method for improving survival rate of juvenile abalone in bait-transferring period |
| CN112870331A (en) * | 2021-02-23 | 2021-06-01 | 广东海洋大学 | Composition for preventing and treating alcoholic liver injury and preparation method and application thereof |
| CN116210814A (en) * | 2022-09-08 | 2023-06-06 | 集美大学 | Application of abalone viscera glycoprotein nano-selenium in preparation of aquatic feed additive |
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