CN108220361A - A kind of extracting method of julid chitosan and its application - Google Patents
A kind of extracting method of julid chitosan and its application Download PDFInfo
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- CN108220361A CN108220361A CN201810213357.6A CN201810213357A CN108220361A CN 108220361 A CN108220361 A CN 108220361A CN 201810213357 A CN201810213357 A CN 201810213357A CN 108220361 A CN108220361 A CN 108220361A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
- C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
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Abstract
Extracting method and its application the invention discloses a kind of julid chitosan, additionally provide a kind of shrimp feed formula.The preparation process of the julid chitosan is substantially：Dead, crushed after being dried is scalded after cleaning worm living, and julid powder is air-dried, then remove protein with sodium hydroxide with being cleaned after HCl treatment, is cleaned again, distilled water is eventually adding and julid chitosan is obtained with chitin deacetylase cooperation ultrasonication.The present invention is by with the preparation of ultrasonic wave added chitin deacetylase progress chitosan, being made free of contamination pure chitosan, can play the role of adjusting body's immunity.
Description
Technical field
The present invention relates to a kind of extracting methods of chitosan, and in particular to a kind of method that chitosan is extracted from julid.
Background technology：
Julid is also named thousand-legger, thousand foot worms, arm beam of steelyard worm etc..Belong to the insect of arthropods, polypody subphylum, Diplopoda.
Chitosan is also known as chitosan, is to be obtained by the chitin that nature is widely present by deacetylation
, chemical name is Chitosan (1-4) -2- amino-B-D glucose.With biological functionality and compatibility, blood compatibility
The excellent performances such as property, safety, microbic resolvability.
Extraction about julid chitosan is reported less at present, and extraction effect and quality have deficiency.
Invention content：
The object of the present invention is to provide a kind of method that chitosan is extracted from julid, to solve above-mentioned technical problem.
The present invention is by with the preparation of ultrasonic wave added chitin deacetylase progress chitosan, being made free of contamination pure julid shell and gathering
Sugar.
To achieve the above object, the present invention provides following scheme：
The present invention provides a kind of method that chitosan is extracted from julid, includes the following steps：
(1) julid is cleaned up, is then scalded extremely with 65~70 DEG C of hot water, after removing content, dry, pulverize 60
~80 mesh sieve, and obtain julid dry powder；
(2) take the julid dry powder of step (1), add in 5%HCl, at 40~45 DEG C constant temperature impregnate 20~for 24 hours, and interval
Stirring, until bubble-free generates, raw material suction filtration is washed to neutrality, and air-dry, and must pre-process julid dry powder；
(3) it takes the pretreatment julid dry powder of step (2), adds in 8%~10% NaOH solution, constant temperature at 80~100 DEG C
Extraction, and intermittent stirring after removing the protein in julid skin, are washed to neutrality, dry at 60~85 DEG C, obtain dry
Chitin；
(4) chitin of step (3) be crushed into 40~60 mesh sieve, according to solid-liquid ratio 1：(15~20) distilled water is added in,
After ultrasonication, the chitin deacetylase of addition 8~12% is digested, and is inactivated with 90~100 DEG C of water-baths, and fast
Speed is cooled to room temperature, filters to take precipitation, and distilled water is washed to neutrality repeatedly, and drying obtains julid chitosan.
Further, the mass volume ratio of julid dry powder and HCl are 1 in the step (2)：(10~15).
Further, the temperature air-dried in the step (2) is 60~80 DEG C.
Further, the mass volume ratio of julid dry powder and NaOH are 1 in the step (3)：(6~10).
Further, in the step (4) ultrasonication condition：Ultrasonic power is 300~450W, and ultrasonic time is
40~60min.
Further, enzymatic hydrolysis condition in the step (4)：Hydrolysis temperature is 45~50 DEG C, and enzymolysis time is 5~8h.
Further, julid chitosan can be applied in feed made from this method.
Further, the feed includes shrimp feed.
Further, the present invention also provides a kind of shrimp feeds, include the ingredient of following parts by weight：15~35 parts of fish meal, elder brother
It is 5~15 parts of worm albumen powder, 5~15 parts of carrot meal, 5~15 parts of flour, 5~10 parts of earthworm, 1~5 part of julid chitosan, compound
0.1~1 part of vitamin, 0.1~1 part of composite mineral matter.
Compared with prior art, the present invention has following advantageous effect：
The present invention prepares julid chitosan product using the method for supersonic synergic chitin deacetylase, has deacetylation
High, the advantages of acetylation is uniform, relative rate is high, the narrow high quality julid chitosan of range of molecular weight distributions, Er Qiegong can be obtained
Skill is environmentally friendly, pollution-free；The julid chitosan of the present invention also can apply in shrimp feed, can play the work for adjusting body's immunity
With.
Specific embodiment
With reference to specific embodiment, the present invention is described in further detail.
Embodiment 1
A kind of method that chitosan is extracted from julid, includes the following steps：
(1) julid is cleaned up, is then scalded extremely with 68 DEG C of hot water, after removing content, dry, pulverize 70 mesh
Sieve, obtains julid dry powder；
(2) the julid dry powder of step (1) is taken, adds in the 5%HCl of 12 mass volume ratios, constant temperature impregnates 22h at 43 DEG C,
And intermittent stirring, until bubble-free generates, raw material suction filtration is washed to neutrality, and is air-dried at 70 DEG C, must pre-process julid dry powder；
(3) it takes the pretreatment julid dry powder of step (2), adds in the NaOH solution of 8 mass volume ratios 9%, constant temperature at 90 DEG C
Extraction, and intermittent stirring after removing the protein in julid skin, are washed to neutrality, dry at 75 DEG C, obtain dry crust
Matter；
(4) chitin of step (3) be crushed into 50 mesh sieve, according to solid-liquid ratio 1：18 add in distilled water, are through power
After 400W ultrasonications 50min, the enzymolysis that 10% chitin deacetylase carries out 6.5h at 48 DEG C is added in, with 95 DEG C of water
Bath inactivation, and room temperature is quickly cooled to, precipitation is filtered to take, distilled water is washed to neutrality repeatedly, and drying obtains julid chitosan.
Embodiment 2
A kind of method that chitosan is extracted from julid, includes the following steps：
(1) julid is cleaned up, is then scalded extremely with 70 DEG C of hot water, after removing content, dry, pulverize 80 mesh
Sieve, obtains julid dry powder；
(2) the julid dry powder of step (1) is taken, adds in the 5%HCl of 15 mass volume ratios, constant temperature impregnates for 24 hours at 45 DEG C,
And intermittent stirring, until bubble-free generates, raw material suction filtration is washed to neutrality, and is air-dried at 80 DEG C, must pre-process julid dry powder；
(3) it takes the pretreatment julid dry powder of step (2), adds in the NaOH solution of 10 mass volume ratios 10%, it is permanent at 100 DEG C
Temperature extraction, and intermittent stirring, after removing the protein in julid skin, are washed to neutrality, dry at 85 DEG C, obtain dry first
Chitin；
(4) chitin of step (3) be crushed into 60 mesh sieve, according to solid-liquid ratio 1：20 add in distilled water, are through power
After 450W ultrasonications 40min, the enzymolysis that 12% chitin deacetylase carries out 5h at 50 DEG C is added in, with 100 DEG C of water
Bath inactivation, and room temperature is quickly cooled to, precipitation is filtered to take, distilled water is washed to neutrality repeatedly, and drying obtains julid chitosan.
Embodiment 3
A kind of method that chitosan is extracted from julid, includes the following steps：
(1) julid is cleaned up, is then scalded extremely with 65 DEG C of hot water, after removing content, dry, pulverize 60 mesh
Sieve, obtains julid dry powder；
(2) the julid dry powder of step (1) is taken, adds in the 5%HCl of 10 mass volume ratios, constant temperature impregnates 20h at 40 DEG C,
And intermittent stirring, until bubble-free generates, raw material suction filtration is washed to neutrality, and is air-dried at 60 DEG C, must pre-process julid dry powder；
(3) it takes the pretreatment julid dry powder of step (2), adds in the NaOH solution of 6 mass volume ratios 8%, constant temperature at 80 DEG C
Extraction, and intermittent stirring after removing the protein in julid skin, are washed to neutrality, dry at 60 DEG C, obtain dry crust
Matter；
(4) chitin of step (3) be crushed into 40 mesh sieve, according to solid-liquid ratio 1：15 add in distilled water, are through power
After 300W ultrasonications 60min, the enzymolysis that 8% chitin deacetylase carries out 8h at 45 DEG C is added in, with 90 DEG C of water-baths
Inactivation, and room temperature is quickly cooled to, precipitation is filtered to take, distilled water is washed to neutrality repeatedly, and drying obtains julid chitosan.
Embodiment 4
A kind of shrimp feed includes the ingredient of following parts by weight：35 parts of fish meal, 15 parts of insect albumen powder, 15 parts of carrot meal,
5 parts of flour, 5 parts of earthworm, 1 part of julid chitosan, 0.1 part of multi-vitamins, 0.1 part of composite mineral matter.
Embodiment 5
A kind of shrimp feed includes the ingredient of following parts by weight：15 parts of fish meal, 5 parts of insect albumen powder, 5 parts of carrot meal, face
15 parts of powder, 10 parts of earthworm, 5 parts of julid chitosan, 1 part of multi-vitamins, 1 part of composite mineral matter.
Comparative example 1
A kind of method that chitosan is extracted from julid, includes the following steps：
(1) julid is cleaned up, is then scalded extremely with 68 DEG C of hot water, after removing content, dry, pulverize 70 mesh
Sieve, obtains julid dry powder；
(2) the julid dry powder of step (1) is taken, adds in the 5%HCl of 12 mass volume ratios, constant temperature impregnates 22h at 43 DEG C,
And intermittent stirring, until bubble-free generates, raw material suction filtration is washed to neutrality, and is air-dried at 70 DEG C, must pre-process julid dry powder；
(3) it takes the pretreatment julid dry powder of step (2), adds in the NaOH solution of 8 mass volume ratios 9%, constant temperature at 90 DEG C
Extraction, and intermittent stirring after removing the protein in julid skin, are washed to neutrality, dry at 75 DEG C, obtain dry crust
Matter；
(4) chitin of step (3) be crushed into 50 mesh sieve, according to solid-liquid ratio 1：18 add in distilled water, are through power
After 400W ultrasonications 50min, precipitation is filtered to take, distilled water is washed to neutrality repeatedly, and drying obtains julid chitosan.
Comparative example 2
A kind of method that chitosan is extracted from julid, includes the following steps：
(1) julid is cleaned up, is then scalded extremely with 68 DEG C of hot water, after removing content, dry, pulverize 70 mesh
Sieve, obtains julid dry powder；
(2) the julid dry powder of step (1) is taken, adds in the 5%HCl of 12 mass volume ratios, constant temperature impregnates 22h at 43 DEG C,
And intermittent stirring, until bubble-free generates, raw material suction filtration is washed to neutrality, and is air-dried at 70 DEG C, must pre-process julid dry powder；
(3) it takes the pretreatment julid dry powder of step (2), adds in the NaOH solution of 8 mass volume ratios 9%, constant temperature at 90 DEG C
Extraction, and intermittent stirring after removing the protein in julid skin, are washed to neutrality, dry at 75 DEG C, obtain dry crust
Matter；
(4) chitin of step (3) be crushed into 50 mesh sieve, according to solid-liquid ratio 1：18 add in distilled water, add in 10%
Chitin deacetylase carries out the enzymolysis of 6.5h at 48 DEG C, is inactivated with 95 DEG C of water-baths, and be quickly cooled to room temperature, it is heavy to filter to take
It forms sediment, distilled water is washed to neutrality repeatedly, is dried, is obtained julid chitosan.
Comparative example 3
A kind of method that chitosan is extracted from julid, includes the following steps：
(1) julid is cleaned up, is then scalded extremely with 68 DEG C of hot water, after removing content, dry, pulverize 70 mesh
Sieve, obtains julid dry powder；
(2) the julid dry powder of step (1) is taken, adds in the 5%HCl of 12 mass volume ratios, constant temperature impregnates 22h at 43 DEG C,
And intermittent stirring, until bubble-free generates, raw material suction filtration is washed to neutrality, and is air-dried at 70 DEG C, must pre-process julid dry powder；
(3) it takes the pretreatment julid dry powder of step (2), adds in the NaOH solution of 8 mass volume ratios 9%, constant temperature at 90 DEG C
Extraction, and intermittent stirring after removing the protein in julid skin, are washed to neutrality, dry at 75 DEG C, obtain dry crust
Matter；
(4) chitin of step (3) be crushed into 50 mesh sieve, according to solid-liquid ratio 1：18 add in distilled water, filter to take precipitation,
Distilled water is washed to neutrality repeatedly, and drying obtains julid chitosan.
Julid chitosan extracts Content evaluation
Preferably to prove that a kind of julid chitosan extracting method provided by the present invention has excellent effect, choose real
It applies example 1,2,3 and the extracting method of comparative example 1,2,3 extracts julid chitosan respectively, julid chitosan extraction content is carried out down
Measure described in table.
Table 3：The performance test results of embodiment 1-3 and comparative example 1-3：
The result shows that：
From table 3 it is observed that contained by the julid chitosan that the julid chitosan extracting method of the present invention is extracted
Amount is higher, and better than comparative example.Without chitin deacetylase comparative example 1 compared with Example 1, julid chitosan carries
It takes content relatively low, shows the method using chitin deacetylase, the julid chitosan of high-content can be obtained；At ultrasound
Compared with Example 1, julid chitosan extraction content is relatively low for the comparative example 2 of reason, and showing to be ultrasonically treated can help to extract high-content
Julid chitosan；Without chitin deacetylase and the comparative example 3 that is not ultrasonically treated compared with Example 1, julid shell gathers
Sugar extraction content is significant lower, shows that the method using supersonic synergic chitin deacetylase prepares julid chitosan product, energy
Obtain high-content julid chitosan.
The present invention is described by way of example above, but the present invention is not limited to above-mentioned specific embodiment, all to be based on
Any changes or modifications that the present invention is done belong to the scope of protection of present invention.
Claims (9)
1. a kind of extracting method of julid chitosan, which is characterized in that include the following steps：
(1) julid is cleaned up, is then scalded extremely with 65~70 DEG C of hot water, after removing content, dry, pulverize 60~80
Mesh sieves, and obtains julid dry powder；
(2) take the julid dry powder of step (1), add in 5%HCl, at 40~45 DEG C constant temperature impregnate 20~for 24 hours, and intermittent stirring,
Until bubble-free generates, raw material suction filtration is washed to neutrality, and air-dry, and must pre-process julid dry powder；
(3) it takes the pretreatment julid dry powder of step (2), adds in 8%~10% NaOH solution, extracting at constant temperature at 80~100 DEG C,
And intermittent stirring after removing the protein in julid skin, is washed to neutrality, dry at 60~85 DEG C, obtains dry crust
Matter；
(4) chitin of step (3) be crushed into 40~60 mesh sieve, according to solid-liquid ratio 1：(15~20) distilled water is added in, through super
After sonicated, the chitin deacetylase of addition 8~12% is digested, and is inactivated with 90~100 DEG C of water-baths, and rapid cold
But room temperature is arrived, filters to take precipitation, distilled water is washed to neutrality repeatedly, and drying obtains julid chitosan.
2. extracting method according to claim 1, which is characterized in that the quality of julid dry powder and HCl in the step (2)
Volume ratio is 1：(10~15).
3. extracting method according to claim 1, which is characterized in that the temperature air-dried in the step (2) is 60~80
℃。
4. extracting method according to claim 1, which is characterized in that the matter of julid dry powder and NaOH in the step (3)
It is 1 to measure volume ratio：(6~10).
5. extracting method according to claim 1, which is characterized in that the condition of ultrasonication in the step (4)：It is super
Acoustical power is 300~450W, and ultrasonic time is 40~60min.
6. extracting method according to claim 1, which is characterized in that enzymatic hydrolysis condition in the step (4)：Hydrolysis temperature is
45~50 DEG C, enzymolysis time is 5~8h.
7. according to application of the julid chitosan in feed made from claim 1~6 any one of them extracting method.
8. application according to claim 7, which is characterized in that the feed is shrimp feed.
9. a kind of shrimp feed, which is characterized in that include the ingredient of following parts by weight：15~35 parts of fish meal, insect albumen powder 5~15
Part, 5~15 parts of carrot meal, 5~15 parts of flour, 5~10 parts of earthworm, 1~5 part of julid chitosan, multi-vitamins 0.1~1
Part, 0.1~1 part of composite mineral matter.
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2018
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| CN101144097A (en) * | 2007-09-18 | 2008-03-19 | 重庆百奥帝克微生态科技有限公司 | Method for preparing chitin and its chitosan and chitosan oligosaccharide |
| CN102558387A (en) * | 2010-12-13 | 2012-07-11 | 青岛中仁药业有限公司 | Method for extracting chitin and antibacterial peptide from fly larvae |
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| CN104262507A (en) * | 2014-10-31 | 2015-01-07 | 江苏财经职业技术学院 | Method for preparing lobster shell chitosan through cooperation of ultrasonic wave and CDA (Chitin Deacetylase) |
| CN105249019A (en) * | 2015-11-10 | 2016-01-20 | 厦门市海林生物科技有限公司 | Young shrimp feed and preparation method thereof |
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