KR101452612B1 - Feed composition for abalone - Google Patents

Abstract

The present invention relates to an abalone feed composition, and more particularly, to an abalone feed composition that utilizes surplus resources and has nutrients capable of being degraded smoothly by an enzyme source of abalone.
The abalone feed composition according to the present invention can provide a nutrient source capable of replacing seaweed and sea tangle by utilizing microalgae and polysaccharide which can be decomposed smoothly into an abalone enzyme and allowing abalone to be ingested smoothly.

Description

{Feed composition for abalone}

The present invention relates to an abalone feed composition, and more particularly, to an abalone feed composition that utilizes surplus resources and has nutrients capable of being degraded smoothly by an enzyme source of abalone.

Abalone has been cultivated in the domestic market since the 1970s by releasing the seedling to the sea. Especially in recent years, production and proportion of production by abalone in domestic abalone production has been greatly increased. In the case of food used in abalone farming, the natural marine algae (seaweed) and sea tangle are mainly used as natural feed, because they contain more ash than other seaweeds, which promotes the growth of abalone shells. However, in the case of a supply shortage or difficulty to supply, the use of dried seaweeds or kelp, or imports of expensive mixed feeds from foreign countries, is a hindrance to the systematic development of abalone.

In order to solve the above problems, it is necessary to develop domestic formulated feeds. However, mixed feeds using seaweed, kelp, parasites and the like, which have high palatability of Korean people, may cause the same problems in the future.

Korean Patent Publication No. 10-2002-0066892 describes compound feeds for abalone farming including fish meal, starch, seaweed and kelp powder as conventional compound feeds for abalone farming to replace natural feeds. However, there is a problem that it contains seaweed and sea tangle used as a natural feed as a basic substance.

Korean Patent No. 10-0758934 also discloses a compound feed for abalone cultivation using cabbage and parasites as a carbohydrate source. However, it is considered that this is a method of cultivating cabbage, There is a problem of high price formation.

Korean Patent Publication No. 10-2002-0003430 discloses a compound feed for abalone cultivation which further includes leafy vegetables in soybean meal, fish meal and seaweed, but also contains seaweed and kelp used in natural feeds as they are, There is a problem in that it can not provide concrete grounds on whether the enzyme is decomposed smoothly by the enzyme source of abalone to help growth.

Therefore, in order to overcome the problems of the prior art as described above, the present inventors have found that the use of surplus resources as a result of using microalgae and various polysaccharides in place of seaweed and kelp, which were used as conventional natural feeds, Thereby completing the present invention.

An object of the present invention is to provide an abalone feed composition capable of replacing seaweed and sea tangle by utilizing surplus resources and capable of decomposing smoothly into an abalone enzyme.

It is still another object of the present invention to provide a method of cultivating an abalone by treating the abalone feed composition with abalone.

In order to solve the above-mentioned problems, the present invention includes one or more microalgae selected from the group consisting of microalgae of Dunaliella tertiolecta and Botryococcus , ≪ / RTI >

As used herein, the term " microalgae or microphytes "refers to algae of small size that can be observed under a microscope and is generally found in freshwater or marine systems. It is a single cell type that exists individually or in the form of a chain or cluster. It has a size of several micrometers to several hundreds of micrometers depending on the species and unlike higher plants, it has no roots, stems and leaves. The microalgae are capable of photosynthesis, and when cultivated while irradiating light, they can consume carbon dioxide, accumulate energy while discharging oxygen, and have high growth rate and growth rate.

The microalgae can supply various nutrient sources including abundant carbohydrates, proteins and fats.

In the present invention, the microalgae may be any one or more selected from the group consisting of Dunaliella tertiolecta and Botryococcus microalgae. The microalgae may be any one or more selected from the group consisting of microalgae cell wall polysaccharides and abalone enzymes Considering the activity, it is more preferable to use a duplexer.

The present invention is characterized in that the cell wall polysaccharide of microalgae DUALI ELETTIRETTA is composed of amylose-like glucan and that they have a high substrate specificity for the carbohydrate digestive enzyme.

The monosaccharide composition of cell wall polysaccharides of microalgae of Dunaliella genus is known to be very diverse and species dependent. For example, D. salina contains galactose, glucose, xylose, and fructose as its major components.

However, the cell wall polysaccharides obtained from the microalgae of the present invention, It was confirmed by FT-IR analysis that the starch-like glucagon was composed of glucose, which is a monosaccharide having a structure similar to starch, unlike the genus.

Therefore, when the microalgae of the present invention is used, the microalgae of the present invention are superior to other microalgae composed of polysaccharides including various kinds of monosaccharides, in that they are superior in the conversion efficiency into glucose and the degrading activity of turnip enzymes.

In addition, in one embodiment of the present invention, degreasing process and protein removal process are performed on microalgae dunaliella tritorerata to obtain a defatted cell wall, and the defatted cell wall is hydrolyzed under specific conditions, thereby obtaining 90% or more of glucose And it was confirmed that it can be produced from microalgae, Dunali elateriotera. As a result, it was confirmed that the microalgae dunaliella tritororetta would be easy to use as a carbon source for abalone. (Examples 3, 4 and 5)

Furthermore, it was confirmed that the substrate specificity of the overturning enzyme against the microalgae dunaliella tritoretta was excellent in one embodiment of the present invention. (Example 11)

In the present invention, the microalga of the Botryococcus is a genus of a small and irregular lumpy clustering green alga generally floating in a pond or the like. Although only three species are known globally, the species is not specifically limited in the present invention.

In the present invention, the microalgae botryokocus is different from the dinal eletertiorerator in that the cell wall polysaccharide is a peptidoglycan, and the monosaccharide composition is more diverse.

The term " top extract " used in the present invention refers to a material obtained by dissolving active ingredients using various solvents including water.

In order to obtain the top extract of the present invention, methods for extracting and processing by hot water extraction, alkali, acid or enzyme treatment generally used in extracting and using useful components of seaweeds can be carried out. Specifically, in the case of the hot water extraction method, water of 17 times as much as the weight of the bottom weight is added, and the hot water can be extracted by heating at 120 to 125 ° C. for 10 minutes to 20 minutes. At this time, the fucoidan ingredient content accounts for 15% of the dry weight of the extract and can be utilized as a nutrient source of abalone. Therefore, the above hot water extraction method is preferable, but is not limited thereto.

As used herein, the term "top-milled material" means a material obtained by milling tops into small-sized materials using a general blender.

It is a perennial seaweed that exists only in the metamorphic age and lives in the lower part of the intertidal zone. It is used for food because it has good taste, especially calcium and iron.

In the present invention, the saccharide component of the substance extracted from the root is contained about 29% of alginic acid (Alginic acid or alginate) and fucoidan, and can replace seaweed and sea tangle used as natural feed of abalone. It can be a great help for growth.

Alginic acid is a polysaccharide that constitutes the cell membrane of brown algae such as seaweed, kelp, and tortoise. The nature of the acid is due to the carboxy group of uronic acid. Its structure is a linear uronic acid polymer composed of? (1? 4) -D-mannuronic acid and? (1? 4) -L-glutaric acid residue. Its chemical formula is (C ₆ H ₈₀ ) n and its typical molecular weight is 20,000 -240,000. Because mammals do not have enzymes that degrade alginic acid, they can not use alginic acid for nutrition. However, marine molluscs (abalones) have degrading enzymes, which are known to be related to abalone feeding seaweed, such as seaweed and kelp.

Fucoidan is a sulfated polysaccharide of sticky, viscous structure contained in brown algae such as seaweed, kelp, and tortoise. The average molecular weight is 20KDa, which is a combination of a basic sugar and a sulfate group called fucose.

In the present invention, the abalone feed composition may further include chitosan and beta-glucan.

The term " Chitosan "used in the present invention is a substance obtained by deacetylation of chitin contained in crustaceans such as crabs, crawfishes and shrimp shells, and processed to facilitate absorption into the body of an organism.

In the present invention, the chitosan uses chitin obtained from crab shells, which may be purchased commercially or used in a conventional manner.

The term " beta-glucan "used in the present invention is a polysaccharide, and is a substance existing in cell walls, mushrooms, cereals, etc. of yeast. Beta Glucan is known to have immunoenhancing action. As a glucose polymer, the glucose unit has a basic structure of β-glycoside bonds at positions 1 and 3, and its structure and physicochemical properties are different depending on the position where glucose is bonded.

In the present invention, commercially available beta-glucan may be purchased and used, or may be prepared and used by a conventional method.

In the present invention, the chitosan may be 15 to 35 parts by weight, the beta-glucan may be 15 to 35 parts by weight, and the top extract may be 200 to 300 parts by weight based on 100 parts by weight of the microalgae.

In the present invention, 15 to 35 parts by weight of the chitosan, 15 to 35 parts by weight of the beta-glucan, and 50 to 150 parts by weight of the top-crushed product may be included in 100 parts by weight of the microalgae.

In one embodiment of the present invention, the substrate specificity of the microalgae, beta-glucan, and top extract was checked based on the abalone lipid vesicles containing the enzyme. As a result, the best specificity for the amylose derived from microalgae, And the blending ratio of the microalgae was determined as a main constituent.

Furthermore, in the case of chitosan or beta-glucan, if the weight ratio is out of the above range, there is a disadvantage that the abundant nutrition of abalone can not be provided due to the relatively low substrate specificity.

In the case of the top extract or the top crumbs, since the seaweed used in the abalone and the brown algae polysaccharide in the kelp are contained as they are, they are used as the main nutrient source together with the microalgae and determined as the above weight parts. However, because the nutrient sources are more dense than that of the top extract, the weight of the top extract is determined to be about half of the weight of the top extract.

In the present invention, the malt extract may be added to the overturned feed composition comprising microalgae, chitosan, beta-glucan, top extract or top powder.

The term "malt extract " used in the present invention refers to a solution obtained by adding malt powder (malt flour) into water and keeping it at 50 캜 to 70 캜 for several hours to allow glycation to take place.

Malt is rich in amylase, an enzyme that degrades sugars. Therefore, when maltose is added to the abalone feed composition, it can effectively decompose polysaccharides and thus it can be a great help for digestion of abalone. Respectively.

In the present invention, the abalone feed composition may further include vitamins and nutrients. This may provide additional nutrient supplements to the abalone feed composition.

The vitamin may include vitamins A, B, and C, and the nutritional agent may include casein, EPA, DHA and various inorganic nutrients, but is not particularly limited thereto.

In one embodiment of the present invention, digestive enzymes are contained in each of the alpha amylase derived from dunaliolaterothorecta, peptidoglycan derived from botarium cocus, chitosan, beta glucan and top extract, which constitute the abalone feed composition The degradation activity of the abalone extract was measured. As a result, it was found that the above components were decomposed by enzymes of abalone, and as a result, it was confirmed that abalone could ingest them and decompose them and use them as a nutrient source. (Examples 6 to 10)

The present invention also provides a method of cultivating an abalone, comprising the step of treating the abalone feed composition in an abalone.

A method of treating the composition with abalone is to allow the abalone to ingest the feed composition.

Since the abalone feed composition contains microalgae diner eletertiorerator composed of amylose-like glucan consisting only of glucose as a monosaccharide, the conversion efficiency to glucose and the degrading activity of abalone enzyme are superior to those of conventional abalone feed compositions. Therefore, such a composition can be usefully used in abalone farming business.

The abalone feed composition according to the present invention can provide a nutrient source capable of replacing seaweed and sea tangle by utilizing microalgae and polysaccharide which can be decomposed smoothly into an abalone enzyme and allowing abalone to be ingested smoothly.

FIG. 1 shows the results of HPLC analysis of the monosaccharide composition of the degreased cell wall. FIG. (a) is the result of analysis for a standard substance, Fuc is fructose, Ara is arabinose, Rha is rhamnose, GlcN is glucosamine, Gal is galactose, Man represents mannose and Xyl represents xylose. (b) represents a single peak resulting from the trifluoroacetic acid hydrolyzate of the microalgae D. tertiolecta .
2 is a graph showing the FT-IR spectrum of defatted cell wall (solid line), corn starch (string), and cereal protein gluten (dotted line).
Fig. 3 is a graph showing the results of HPLC analysis of monosaccharides produced by hydrolysis of cell wall polysaccharides into? -Amylase under optimal conditions. (a) shows 10 nmole of standard monosaccharide, (b) shows 10 μmole of cell wall polysaccharide hydrolyzate, (c) shows 1.0 μmole of cell wall polysaccharide hydrolyzate, and (d) shows 0.4 μmole of cell wall polysaccharide hydrolyzate .
Fig. 4 shows the results of α-amylase reducing sugar analysis of dunaliella tritoreta by abalone-derived mucilaginous juice. This means the amount of reducing sugar according to the decomposition time by the abalone enzyme.
FIG. 5 shows the results of peptidoglycan reducing sugar analysis of bortoluccus by abalone-derived mucilage.
Fig. 6 shows the results of reducing sugar analysis of fucoidan derived from the extract of Top extract with abalone-derived mucilage.
Fig. 7 shows the result of the reduction sugar analysis of chitosan by the abalone-derived vaginal epithelium.
Fig. 8 shows the results of analysis of reducing sugar of beta-glucan by abalone-derived mucilaginous juice.
9 is a graph showing the results of measurement of the concentration of α-amylose (Du) derived from dunaliolaterioterecta, peptidoglycan (Bo) derived from bovarium cocus, fucoidan (Fu) derived from top extract, (To) and beta glucan ([beta] -).

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.

Example  One: Dünaliera Tertiorector  Strain and culture

Dunaliella , a green halophilic alga that accumulates β-carotene, tertiolecta , UTRX LB 999) from the University of Texas (UTEX) in Austin. Compressed air containing 2% carbon dioxide was supplied to 0.1 _VVM and cells were incubated in 5 L of artificial seawater added with f / 2 medium for 10 days at 25 캜 for 7 days while being fluorescently irradiated with 150 μE / (m ² s). Culture flasks were irradiated with external illumination that automatically turned on and off mimicking the biological cycle without adjusting the pH. The effects of various lighting on the growth rate of the birds were confirmed in advance. If necessary, initial illumination intensities of 100, 200 and 350 μE / (m ² s) were achieved using one, two and four fluorescent lights, respectively.

Example  2: Identification of cell growth rate

All experiments were performed with seed culture during logarithmic growth period. Bird cells were cultured at 4 ° C under a continuous illumination of 15 μE / (m ² s) intensity using a white fluorescent lamp. The number of alveolar cells and the mean cell size were measured with a Coulter counter (Model Z2; Coulter Electronics, Inc., Hialeah, FL, USA). Data from the Coulter counter was collected with AccuComp software and transmitted to an Excel spreadsheet to calculate not only the average cell size but also the cell number and size distribution.

Example  3: Dünaliera Tertiorecta  Degreasing cell wall separation and analysis

To obtain a defatted cell wall of a dunaliolateritorector, lipid extraction was performed based on the method of Bligh and Dyer and the mass of defatted biomass was measured after lyophilizer (Ilshin Co., Korea) . Briefly, cells were recovered from the culture broth by centrifugation at 10,000 rpm for 30 minutes and washed twice with 10 volumes of distilled water. The cells were lyophilized and the lipids were extracted twice from the dried cells using a 10-fold volume of extraction solvent (chloroform: methanol = 1: 2, v / v). The cells obtained from the solvent extraction procedure were used as crude defatted biomass. The protein was removed from the degreased, dried biomass by washing with 0.1 M NaOH solution to obtain a partially purified defatted cell wall. The total weight of carbohydrates in the defatted cell walls was determined by a modified phenol-sulfuric acid method. The degreasing biomass was sonicated at 50 ° C for 30 minutes (Power Sonic 405, Whashin Co., Korea) to disperse all the dry ingredients in distilled water and determine the protein concentration in the degreased biomass using the Bradford method. The ash content was determined as the amount of mineral matter remaining as an incombustible residue of the degreased cell wall. The ash burning of the degreased cell walls was performed at 700 ° C and was completed when the cooled residue was near white. The ash quantity of defatted cell walls was expressed as dried matter.

The monosaccharide composition of the degreased cell wall was measured using a Bio-LC system (Dionex, USA) equipped with a pulsed amperometric detector (PAD) and a high performance anion of the hydrolyzate obtained under strong acid conditions Was determined by high performance anion-exchange chromatography (HPAEC). Briefly, the degreased cell wall was resuspended in 1 ml of distilled water and mixed with an equal volume of 4.0 M trifluoroacetic acid (TFA). The sample was placed at 100 ° C for 6 hours to allow acid-hydrolysis to take place. Filtered through a 0.45 μm syringe filter, and vacuum dried using a Speed-Vac (Biotron, Korea). Vacuum drying was repeated to remove residual acid and the dried sample was analyzed on a CarboPac PA-1 column (Dionex, USA) according to the manufacturer's instructions for conditions for monosaccharide analysis by HPAEC-PAD.

3.1. Dünaliera Tertiorecta  Degreasing cell wall separation

Since algal biomass contains many other ingredients in addition to polysaccharides, it is important to remove the ballast compound to separate and purify the cell walls. Lipids and other hydrophobic molecules were separated from the algal biomass by extraction with dry chloroform: methanol mixture (1: 2, v / v) to obtain a defatted material . In the present invention, a 69.4% content of degreasing biomass was obtained from total algal cell mass. The degreasing biomass consisted of the four components observed in the initial composition shown in Table 1.

Since it is difficult to separate proteins and polysaccharides due to the complexation between biopolymers, an alkali reagent (0.1 M NaOH) is used as a subsequent purification step to remove most of the protein from the degreased cell wall Respectively. The degreased cell walls were then washed twice more with ethanol and lyophilized with powder to obtain a purified defatted cell wall. This includes 91.94% total carbohydrate, 0.28% w / w protein, 6.50% ash and 1.28% other non-identified impurities, respectively. The impurities may be phenol-based molecules that are firmly attached to the degreased cell wall.

3.2. Dünaliera Tertiorecta  Analysis of defatted cell wall composition

The specific analysis of the defatted cell walls used in the present invention confirmed some general characteristics of industrially important polysaccharides. Defatted cell walls isolated and purified from D. tertiolecta degreasing biomass, unlike reported for other industrial microalgae and giant algae, were identified by thin-layer chromatography (TLC) and HPAEC-PAD analysis and were predominantly composed of glucose It was confirmed to be a homopolysaccharide, that is, glucan (Fig. 1). The results support that degreased cell walls can be used as a major carbon source for glucose production and further upturns. The monosaccharide composition of defatted cell wall polysaccharides in microalgae of Dunaliella genus is very diverse and species dependent. For example, by HPLC analysis, four major constituent monosaccharides (galactose, glucose, xylose, and fructose) were detected against D. salina . The presence of pentose sugar (i.e., xylose) that is not present in the polysaccharide of the prokaryotic origin has been confirmed. The present inventors have been able to obtain a monosaccharide composition consisting only of glucose from the microalgae D. tertiolecta and to identify its primary structure (see FT-IR analysis). This means that it can be a suitable carbon source for abalone.

Example  4: FT - IR  Spectroscopy

KBr pellets were prepared using a Nicolet 6700 FT-IR spectrometer (Nicolet Analytical Instruments, USA) and the absorbed FT-IR spectrum (400-4000 cm ^-1 ) of the degreased cell wall was recorded. Sixty-four scans were accumulated with a spectral resolution of 4.0 cm- ¹ . The spectrum was smoothed and the baseline corrected using Omnic 8.0 (Nicolet Analytical Instruments, USA) software. The calibrated spectrum was then converted to comma-separated values (CSV) format to produce graphs and sent to Origin 6.0 software (Microcal Origin, USA).

FT-IR spectra of defatted cell walls were also shown using starch and gluten (cereal protein) as a reference material (FIG. 2). Analysis of structurally sensitive characteristic IR bands showed that the degreased cell walls of D. tertiolecta were amylose- or starch-like α-glucan rather than cellulose-like β-glucan, glucan. Some IR bands of degreased cell walls appearing at 1154, 1081, 1022, 928, 862, 765, 708, 612, 578 and 532 cm ^-1 are typical for starch and amylose. Furthermore, the presence of protein impurities was confirmed by IR bands at 1649 and 1536 cm ^-1 (amide I and amide II vibrations). The protein composition is much less clear than the polysaccharide composition because the above-mentioned protein bands are not stronger than those at the predominant sugar region (950-1200 cm ^-1 ) of CO and CC stretching vibration. Thus, the defatted cell wall of a dunaliella tritoretta is structurally different from other polysaccharides, such as heterozygosity from plants and other microalgae-derived? - (1 → 2) -D-glucans. Although more accurate structural analysis is required to determine the exact structure of the degreased cell wall, the monosaccharide composition and FT-IR analysis disclosed in the present invention are sufficient to confirm that the polysaccharide is? - (1 → 4) -D-glucan. Therefore, the enzyme of abalone can be converted into glucose by hydrolyzing it, and can be used as a useful carbon source of abalone.

Example  5: Dünaliera Tertiorecta  Enzymatic hydrolysis of cell wall polysaccharides enzymatic hidrolysis )

To confirm the conversion of the cell wall polysaccharide to glucose, the degreasing biomass was sonicated at 50 DEG C for 30 minutes to disperse all the dry ingredients in distilled water. Highly soluble proteins and other non-glucan substances were removed by centrifugation at 13,000 rpm for 30 minutes to isolate the cell wall polysaccharide. Approximately 89% of the purified cell wall polysaccharide after 24 hours of enzymatic hydrolysis was converted to glucose (2.67 g glucose / 3.0 g purified cell wall polysaccharide) in excess of the enzyme (? -Amylase (Sigma, St. Louis, USA) . This amount almost corresponds to the total carbohydrate amount in the defatted cell wall (see Table 1), indicating that most of the glucan has been converted to glucose. TLC and HPAEC analysis of the enzyme hydrolyzate showed no other monosaccharide (FIG. 3). This indicates that the cell wall polysaccharide was completely converted to glucose by the? -Amylase treatment. As a result, it has been confirmed that the present invention has a remarkable potential as a feed component as an excellent carbon source for the abalone through the use of the microalgae, Dunaliolateriotorecta.

Example  6: Abalone derived In the vaginal fluid  by Dünaliera Tertiorecta  α-amylase reducing sugar analysis

In the above example, it was confirmed that the cell wall polysaccharide of Dulneriella tritoretta was composed of amylose-like glucan. In order to confirm the hydrolysis of the abalone enzyme to the α-amylose derived from the dunaliella tritorecta, reducing sugar assay.

The abalone enzyme was contained in abalone - derived vaginal epithelium extracted from abalone so that the vaginal vaginal fluid was extracted from abalone. Specifically, 10 fresh abalones were placed in a beaker containing 800 ml of distilled water, and the temperature was raised to 50 DEG C and the mixture was heated for 10 minutes. The abalone was immediately recovered and the temperature was lowered by ice or cold water to prevent the enzyme activity from being lowered. Finally, a crude extract as an abalone-derived mucilaginous juice was obtained.

The α-amylose derived from the dunaliella triteracta was obtained by separating the cell wall polysaccharide of the dunaliella triteractata according to the method of Example 3.1. Above.

Thereafter, 1 ml of the abalone-derived vaginal epithelium was added to the above-mentioned α-amylose derived from the diluent. Through repeated experiments, it was confirmed that pH 6.0 was the optimum reaction condition and acetate buffer (pH 6.07) was additionally applied. Finally, a mixture was prepared so as to have 0.5% by weight of? -Amylose derived from the dunaliella tetra lecta and subjected to a hydrolysis reaction.

The mixture was incubated at 37 DEG C for 0 to 24 hours, and then the sample was boiled at 90 DEG C for 10 minutes in order to quench the reaction. The solid residue and the supernatant were separated by centrifugation at 9,000 x g for 15 minutes. The supernatant was subjected to a reducing sugar assay and the results are shown in FIG.

From the above results, it was confirmed that α-amylose of dunaliolateritiora was decomposed smoothly by digestive enzymes of abalone, indicating that abalone can be ingested and utilized as a nutrient source.

Example  7: Abalone derived In the vaginal fluid  by Boat Rio Cocus  Peptidoglycan ( Peptidoglycan ) Reducing sugar analysis

Reducing sugar analysis was carried out to confirm the hydrolysis of the abalone enzyme against peptidoglycan derived from microalgae, bovioccocus.

Peptidoglycan derived from a microalga, bovinioccus, was obtained by isolating the cell wall polysaccharide of bortriocococcus by the method of Example 3.1. Above.

The specific analysis method is the same as in Example 6, and the results are shown in Fig.

From the above results, it was confirmed that peptidoglycan derived from boranioccus was degraded smoothly by digestive enzymes of abalone, indicating that abalone can be ingested and utilized as a nutrient source.

Example  8: Abalone derived In the vaginal fluid  Derived from the extract Fucoidan  Reducing sugar analysis

Reducing sugar analysis was carried out to investigate the hydrolysis of the abalone enzyme to fucoidan, one of the major components of the extract.

In the case of fucoidan derived from the extract, 30 g of starch was added to 500 ml of distilled water and heated at 120 to 125 ° C for 10 minutes to 20 minutes to obtain an extract. Alginic acid reacted with calcium chloride to precipitate, and centrifuged (at 13,000 rpm for 10 minutes to 30 minutes) to remove the precipitate. The extract from which the precipitate was removed was frozen at -80 ° C, dried using a freeze dryer, and finally fucoidan derived from the extract was obtained.

The specific analysis method is the same as in Example 6, and the results are shown in Fig.

From the above results, it was confirmed that the fucoidan derived from the root extract was decomposed smoothly by the digestive enzymes of abalone, indicating that abalone can be ingested and utilized as a nutrient source.

Example  9: abalone origin In the vaginal fluid  Chitosan Reducing sugar analysis by

Reducing sugar analysis was performed to confirm the hydrolysis of the abalone enzyme to chitosan.

The specific analysis method is the same as in Example 6, and the results are shown in Fig.

From the above results, it was confirmed that chitosan was decomposed smoothly by digestive enzymes of abalone, indicating that abalone can be ingested and utilized as a nutrient source.

Example  10: abalone origin In the vaginal fluid  by Beta Glucan  Reducing sugar analysis

Reducing sugar analysis was performed to confirm the hydrolysis of the overturning enzyme to beta - glucan.

The specific analysis method is the same as in Example 6, and the results are shown in FIG.

From the above results, it was confirmed that beta-glucan was decomposed smoothly by digestive enzymes of abalone, indicating that abalone can be ingested and utilized as a nutrient source.

Example  11: abalone Viscous  Relative substrate specificity

In order to confirm the relative substrate specificity for each substrate of the overturning enzyme, it was confirmed that α-amylose from dunery eletertorecta, peptidoglycan derived from botolio cocus, fucoidan derived from top extract, alginic acid derived from top extract and betaglucan Relative enzyme activity was analyzed.

The substrates were mixed at the same weight ratio and further mixed with abalone-derived mucilage solution and acetate buffer (pH 6.07) to finally prepare a mixture containing 0.5% by weight of the substrates.

The mixture was incubated at 37 DEG C for 24 hours and the relative enzyme activity was analyzed, and the results are shown in Fig.

The results showed that the enzyme activity against α-amylase derived from dunaliella terrortorca was the most excellent, and it could be the most effective nutrient source for abalone.

Example  12: Preparation of abalone feed composition (1)

The mixture was prepared by mixing 1: 1: 0.5: 0.5: 1 chitosan (200 mesh or less): betaglucan: chitosan extract in weight ratio.

The malt extract was immersed in 1.0 L of water per 100 g of malt, kept at 50 캜 for several hours, and glycated was prepared by filtration.

The resulting mixture was further mixed with the above extract and the prepared malt extract in a weight of 1 fold of the weight of the whole mixture, followed by complete lyophilization with a lyophilizer (Ilshin Co., Korea) to prepare an abalone feed composition .

In addition to the prepared feed composition, various vitamins and inorganic nutrients were added. The vitamins and nutrients included vitamins A, B, C, casein, EPA, and DHA.

Example  13: Preparation of abalone feed composition (2)

The mixture was prepared by mixing 1: 1: 0.5: 0.5: 2 chitosan (200 mesh or less): betaglucan: top crumbs in weight ratio.

The top crushing product was prepared by pulverizing the tops with a general household blender.

The malt extract was prepared in the same manner as in Example 12 above.

The prepared mixture was added with the above prepared malt extract in an amount of twice the weight of the whole mixture, mixed, and completely lyophilized with a lyophilizer (Ilshin Co., Korea) to prepare an abalone feed composition.

In addition to the prepared feed composition, various vitamins and inorganic nutrients were added. The vitamins and nutrients included vitamins A, B, C, casein, EPA, and DHA.

Claims (9)

Wherein the composition comprises cell wall polysaccharide and top extract or top powder of one or more microalgae selected from the group consisting of Dunaliella tertiolecta and Botryococcus microalgae.
The feed composition according to claim 1, wherein the microalgae is Dunaliella tertiolecta .
The feed composition of claim 1, further comprising chitosan and beta glucan.
The feed composition according to claim 1, wherein the safflower extract or top-crush comprises alginic acid and fucoidan.
delete delete The feed composition according to claim 1, further comprising a malt extract.
The feed composition according to claim 1, further comprising a vitamin and a nutrient.
A method of growing an abalone, comprising the step of treating the feed composition of any one of claims 1 to 4, 7, and 8 in an abalone.

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