CN108997511A - One main laminaria active polysaccharide and preparation method thereof - Google Patents

One main laminaria active polysaccharide and preparation method thereof Download PDF

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CN108997511A
CN108997511A CN201811045230.4A CN201811045230A CN108997511A CN 108997511 A CN108997511 A CN 108997511A CN 201811045230 A CN201811045230 A CN 201811045230A CN 108997511 A CN108997511 A CN 108997511A
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active polysaccharide
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于海彦
张莉
刘尚军
仝飞飞
张希
臧晋
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Nanyang Institute of Technology
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    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof

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Abstract

The invention discloses a main laminaria active polysaccharides, including raw material below: Fresh Laminaria Japonica, dehydrated alcohol, cellulase, pectase, zytase and deionized water.The invention also discloses the preparation methods of the kelp active polysaccharide.Kelp active polysaccharide prepared by the present invention have reducing blood lipid, it is hypoglycemic, adjust immune, anticoagulation, be antitumor, lead discharging removing toxic substances and a variety of biological functions such as anti-oxidant, by repeatedly extracting and repeatedly digesting, effectively increase extraction effect, pass through ultrasound, centrifugation and repeatedly washing simultaneously, be conducive to improve the purity of product, it can be used for solving the problems, such as due to lacking corresponding catabolic enzyme in human body and can not directly absorb laminarin by edible kelps, have a vast market foreground.

Description

One main laminaria active polysaccharide and preparation method thereof
Technical field
The present invention relates to field of biotechnology, specifically a kind of kelp active polysaccharide and preparation method thereof.
Background technique
Active polysaccharide refers to the active compound of polysaccharide of certain special physiological, has bidirectional modulation Human Physiology rhythm Function, be widely present in plant and microorganism wall, small toxicity, highly-safe, function is extensive, have it is extremely important with Special physiological activity is the high molecular polymer connected with carbonyl by glycosidic bond by aldehyde radical, and constitutes four big bases of life One of this substance.
Kelp is a kind of raw brown alga plant in large size sea grown in low temperature seawater, belongs to seaweed plant, be rich in algin With iodine matter, can be adapted for the cooking methods such as mixing, burn, stew, boiling in a covered pot over a slow fire.Laminarin refers to the polysaccharide component purified from kelp, tool Have reducing blood lipid, it is hypoglycemic, adjust immune, anticoagulation, be antitumor, lead discharging removing toxic substances and a variety of biological functions such as anti-oxidant.But by In lacking corresponding catabolic enzyme in human body, by edible kelps, human body can not directly absorb laminarin, it is therefore desirable to from kelp Middle extraction laminarin.But present extracting method, it cannot be guaranteed that the purity and effect extracted, obtained product cannot expire , there is poor product quality, while can not make by human body good absorption, last laminarin in the demand of foot society Health-care effect cannot guarantee well.
Summary of the invention
The purpose of the present invention is to provide main laminaria active polysaccharides and preparation method thereof, to solve in above-mentioned background technique The problem of proposition.
To achieve the above object, the invention provides the following technical scheme:
One main laminaria active polysaccharide, including below according to the raw material of parts by weight: 17-25 parts of Fresh Laminaria Japonica, dehydrated alcohol 140-160 parts, 0.038 ‰ -0.05 ‰ parts of cellulase, 0.01 ‰ -0.03 ‰ parts of pectase, zytase 0.012 ‰ - 0.016 ‰ parts and suitable deionized water.
As a further solution of the present invention: including below according to the raw material of parts by weight: 20 parts of Fresh Laminaria Japonica, dehydrated alcohol 150 parts, 0.045 ‰ parts of cellulase, 0.02 ‰ parts of pectase, 0.015 ‰ parts of zytase and suitable deionized water.
The preparation method of the kelp active polysaccharide, steps are as follows:
1) Fresh Laminaria Japonica is weighed according to parts by weight, is cut into the sheet that length and width are 1cm, then weighed according to parts by weight anhydrous Ethyl alcohol is mixed with the aforementioned Fresh Laminaria Japonica for being cut into sheet, then is ultrasonically treated, and 20 mesh filter to get filtrate A and filter residue B；
2) filter residue B obtained in step 1) is added to the deionized water of 10 times of volumes, 200min, 40 mesh are stirred at 42 DEG C Filter to get filtrate C and filter residue D；
3) filter residue D obtained in step 2) is added to the deionized water of 8 times of volumes, with the stirring of 100r/min at 38 DEG C Rate is stirred 100min, and 80 mesh filter to get filtrate E and filter residue F；
4) filtrate F obtained in liquor C obtained in filtrate A obtained in step 1) and step 2) and step 3) is mixed It closes uniformly, obtains mixed liquor G；By repeatedly extracting, extraction effect is effectively increased；
5) deionized water of 9 times of weight will be added in filter residue F obtained in step 3), be warming up to 70 DEG C after impregnating 40min Continue to impregnate 50min, be then cooled to 48 DEG C, cellulase is added and reacts 40min, then is warming up to 55 DEG C, zytase is added 40min is reacted, is then cooled to 50 DEG C, pectin enzyme reaction 30min is added, obtains mixture H；By repeatedly digesting, effectively improve Extraction effect；
6) mixture H obtained in step 5) is warming up to 105 DEG C, keeps 3 minutes progress destroy the enzyme treatments, is then added 12 The deionized water of times weight impregnates 140min, and the filtering of 100 mesh obtains filtrate I；
7) filtrate I obtained in step 6) is uniformly mixed with mixed liquor G obtained in step 4), is concentrated into The 10% of original volume carries out centrifugation 15min with the rate of 5000rpm, takes supernatant to carry out ultrafiltration using ultrafiltration membrane and leaves and takes super Filtrate obtains mixture J；
8) mixture J obtained in step 7) is concentrated under reduced pressure into 1/3 volume, the positive fourth of isometric chloroform-is then added Alcohol mixed liquor carries out uniformly mixed, then carries out ultrasonic treatment 5min, stands and carries out centrifugation 15min with the rate of 5000rpm after 2h, Upper layer centrifugate is taken, in aforementioned upper layer centrifugate plus the concentration of 3.5 times of volumes is 98% ethanol water, is cooled to 4 DEG C Standing 12h is carried out, centrifugation 15min is then carried out with the rate of 4000rpm and obtains sediment, is replaced with anhydrous propanone and anhydrous ether Washing precipitate 6 times, then carry out vacuum freeze drying to get；Ultrasound and centrifugation are carried out by the way that chloroform-n-butanol mixed liquor is added Processing effectively increases extraction effect, and then be conducive to improve the purity of product also by repeatedly washing.
As further scheme of the invention: in step 1), the ultrasonic power of the ultrasonic treatment is 200W；It is described super The time of sonication is 20min.
As further scheme of the invention: in step 2), the stirring rate of the stirring is 400r/min.
As further scheme of the invention: in step 7), the molecular cut off of the ultrafiltration membrane is 20000- 40000。
As further scheme of the invention: in step 8), the chloroform-n-butanol mixed liquor is chloroform and n-butanol It is mixed according to the ratio that volume ratio is 4:1；The ultrasonic power of the ultrasonic treatment is 180W.
As further scheme of the invention: in step 8), the temperature of the vacuum freeze drying is -18 DEG C, pressure For 10Pa, time 12h.
The preparation method of the kelp active polysaccharide is preparing the purposes in active polysaccharide product.
Compared with prior art, the beneficial effects of the present invention are:
Kelp active polysaccharide prepared by the present invention have reducing blood lipid, it is hypoglycemic, adjust immune, anticoagulation, antitumor, lead discharging Removing toxic substances and a variety of biological functions such as anti-oxidant, by repeatedly extracting and repeatedly digesting, compared to more single extracting mode, extraction Effect greatly improves, by experimental verification, it is ensured that the quality of extraction, and repeatedly digest, the effect that can have been obtained, together When by ultrasound, centrifugation and repeatedly washing, be conducive to the purity for improving product, can be used for solving corresponding due to lacking in human body Catabolic enzyme and the problem of can not directly absorb laminarin by edible kelps, have a vast market foreground.
Specific embodiment
Technical solution of the present invention is described in more detail With reference to embodiment.
Embodiment 1
One main laminaria active polysaccharide, including below according to the raw material of parts by weight: 17 parts of Fresh Laminaria Japonica, 140 parts of dehydrated alcohol, 0.038 ‰ parts of cellulase, 0.01 ‰ parts of pectase, 0.012 ‰ parts of zytase and suitable deionized water.
In the present embodiment, the preparation method of the kelp active polysaccharide, steps are as follows:
1) Fresh Laminaria Japonica is weighed according to parts by weight, is cut into the sheet that length and width are 1cm, then weighed according to parts by weight anhydrous Ethyl alcohol is mixed with the aforementioned Fresh Laminaria Japonica for being cut into sheet, then is ultrasonically treated, and 20 mesh filter to get filtrate A and filter residue B；
Wherein, the ultrasonic power of the ultrasonic treatment is 200W；The time of the ultrasonic treatment is 20min；
2) filter residue B obtained in step 1) is added to the deionized water of 10 times of volumes, the stirring with 400r/min at 42 DEG C It mixes rate and is stirred 200min, 40 mesh filter to get filtrate C and filter residue D；
3) filter residue D obtained in step 2) is added to the deionized water of 8 times of volumes, with the stirring of 100r/min at 38 DEG C Rate is stirred 100min, and 80 mesh filter to get filtrate E and filter residue F；
4) filtrate F obtained in liquor C obtained in filtrate A obtained in step 1) and step 2) and step 3) is mixed It closes uniformly, obtains mixed liquor G；By repeatedly extracting, extraction effect is effectively increased；
5) deionized water of 9 times of weight will be added in filter residue F obtained in step 3), be warming up to 70 DEG C after impregnating 40min Continue to impregnate 50min, be then cooled to 48 DEG C, cellulase is added and reacts 40min, then is warming up to 55 DEG C, zytase is added 40min is reacted, is then cooled to 50 DEG C, pectin enzyme reaction 30min is added, obtains mixture H；By repeatedly digesting, effectively improve Extraction effect；
6) mixture H obtained in step 5) is warming up to 105 DEG C, keeps 3 minutes progress destroy the enzyme treatments, is then added 12 The deionized water of times weight impregnates 140min, and the filtering of 100 mesh obtains filtrate I；
7) filtrate I obtained in step 6) is uniformly mixed with mixed liquor G obtained in step 4), is concentrated into The 10% of original volume carries out centrifugation 15min with the rate of 5000rpm, takes supernatant to carry out ultrafiltration using ultrafiltration membrane and leaves and takes super Filtrate obtains mixture J；The molecular cut off of the ultrafiltration membrane is 30000；
8) mixture J obtained in step 7) is concentrated under reduced pressure into 1/3 volume, the positive fourth of isometric chloroform-is then added Alcohol mixed liquor carries out uniformly mixed, then carries out ultrasonic treatment 5min, stands and carries out centrifugation 15min with the rate of 5000rpm after 2h, Upper layer centrifugate is taken, in aforementioned upper layer centrifugate plus the concentration of 3.5 times of volumes is 98% ethanol water, is cooled to 4 DEG C Standing 12h is carried out, centrifugation 15min is then carried out with the rate of 4000rpm and obtains sediment, is replaced with anhydrous propanone and anhydrous ether Washing precipitate 6 times, then carry out vacuum freeze drying to get；
Wherein, the chloroform-n-butanol mixed liquor be chloroform and n-butanol according to the ratio mixing that volume ratio is 4:1 and At；The ultrasonic power of the ultrasonic treatment is 180W；The temperature of the vacuum freeze drying is -18 DEG C, pressure 10Pa, the time For 12h；Ultrasound and centrifugal treating are carried out by the way that chloroform-n-butanol mixed liquor is added, also by repeatedly washing, effectively increases and mentions Effect is taken, and then is conducive to improve the purity of product.
Embodiment 2
One main laminaria active polysaccharide, including below according to the raw material of parts by weight: 25 parts of Fresh Laminaria Japonica, 160 parts of dehydrated alcohol, 0.05 ‰ parts of cellulase, 0.03 ‰ parts of pectase, 0.016 ‰ parts of zytase and suitable deionized water.
In the present embodiment, the preparation method of the kelp active polysaccharide, steps are as follows:
1) Fresh Laminaria Japonica is weighed according to parts by weight, is cut into the sheet that length and width are 1cm, then weighed according to parts by weight anhydrous Ethyl alcohol is mixed with the aforementioned Fresh Laminaria Japonica for being cut into sheet, then is ultrasonically treated, and 20 mesh filter to get filtrate A and filter residue B；
Wherein, the ultrasonic power of the ultrasonic treatment is 200W；The time of the ultrasonic treatment is 20min；
2) filter residue B obtained in step 1) is added to the deionized water of 10 times of volumes, the stirring with 400r/min at 42 DEG C It mixes rate and is stirred 200min, 40 mesh filter to get filtrate C and filter residue D；
3) filter residue D obtained in step 2) is added to the deionized water of 8 times of volumes, with the stirring of 100r/min at 38 DEG C Rate is stirred 100min, and 80 mesh filter to get filtrate E and filter residue F；
4) filtrate F obtained in liquor C obtained in filtrate A obtained in step 1) and step 2) and step 3) is mixed It closes uniformly, obtains mixed liquor G；By repeatedly extracting, extraction effect is effectively increased；
5) deionized water of 9 times of weight will be added in filter residue F obtained in step 3), be warming up to 70 DEG C after impregnating 40min Continue to impregnate 50min, be then cooled to 48 DEG C, cellulase is added and reacts 40min, then is warming up to 55 DEG C, zytase is added 40min is reacted, is then cooled to 50 DEG C, pectin enzyme reaction 30min is added, obtains mixture H；By repeatedly digesting, effectively improve Extraction effect；
6) mixture H obtained in step 5) is warming up to 105 DEG C, keeps 3 minutes progress destroy the enzyme treatments, is then added 12 The deionized water of times weight impregnates 140min, and the filtering of 100 mesh obtains filtrate I；
7) filtrate I obtained in step 6) is uniformly mixed with mixed liquor G obtained in step 4), is concentrated into The 10% of original volume carries out centrifugation 15min with the rate of 5000rpm, takes supernatant to carry out ultrafiltration using ultrafiltration membrane and leaves and takes super Filtrate obtains mixture J；The molecular cut off of the ultrafiltration membrane is 30000；
8) mixture J obtained in step 7) is concentrated under reduced pressure into 1/3 volume, the positive fourth of isometric chloroform-is then added Alcohol mixed liquor carries out uniformly mixed, then carries out ultrasonic treatment 5min, stands and carries out centrifugation 15min with the rate of 5000rpm after 2h, Upper layer centrifugate is taken, in aforementioned upper layer centrifugate plus the concentration of 3.5 times of volumes is 98% ethanol water, is cooled to 4 DEG C Standing 12h is carried out, centrifugation 15min is then carried out with the rate of 4000rpm and obtains sediment, is replaced with anhydrous propanone and anhydrous ether Washing precipitate 6 times, then carry out vacuum freeze drying to get；
Wherein, the chloroform-n-butanol mixed liquor be chloroform and n-butanol according to the ratio mixing that volume ratio is 4:1 and At；The ultrasonic power of the ultrasonic treatment is 180W；The temperature of the vacuum freeze drying is -18 DEG C, pressure 10Pa, the time For 12h；Ultrasound and centrifugal treating are carried out by the way that chloroform-n-butanol mixed liquor is added, also by repeatedly washing, effectively increases and mentions Effect is taken, and then is conducive to improve the purity of product.
Embodiment 3
One main laminaria active polysaccharide, including below according to the raw material of parts by weight: 21 parts of Fresh Laminaria Japonica, 150 parts of dehydrated alcohol, 0.044 ‰ parts of cellulase, 0.02 ‰ parts of pectase, 0.014 ‰ parts of zytase and suitable deionized water.
In the present embodiment, the preparation method of the kelp active polysaccharide, steps are as follows:
1) Fresh Laminaria Japonica is weighed according to parts by weight, is cut into the sheet that length and width are 1cm, then weighed according to parts by weight anhydrous Ethyl alcohol is mixed with the aforementioned Fresh Laminaria Japonica for being cut into sheet, then is ultrasonically treated, and 20 mesh filter to get filtrate A and filter residue B；
Wherein, the ultrasonic power of the ultrasonic treatment is 200W；The time of the ultrasonic treatment is 20min；
2) filter residue B obtained in step 1) is added to the deionized water of 10 times of volumes, the stirring with 400r/min at 42 DEG C It mixes rate and is stirred 200min, 40 mesh filter to get filtrate C and filter residue D；
3) filter residue D obtained in step 2) is added to the deionized water of 8 times of volumes, with the stirring of 100r/min at 38 DEG C Rate is stirred 100min, and 80 mesh filter to get filtrate E and filter residue F；
4) filtrate F obtained in liquor C obtained in filtrate A obtained in step 1) and step 2) and step 3) is mixed It closes uniformly, obtains mixed liquor G；By repeatedly extracting, extraction effect is effectively increased；
5) deionized water of 9 times of weight will be added in filter residue F obtained in step 3), be warming up to 70 DEG C after impregnating 40min Continue to impregnate 50min, be then cooled to 48 DEG C, cellulase is added and reacts 40min, then is warming up to 55 DEG C, zytase is added 40min is reacted, is then cooled to 50 DEG C, pectin enzyme reaction 30min is added, obtains mixture H；By repeatedly digesting, effectively improve Extraction effect；
6) mixture H obtained in step 5) is warming up to 105 DEG C, keeps 3 minutes progress destroy the enzyme treatments, is then added 12 The deionized water of times weight impregnates 140min, and the filtering of 100 mesh obtains filtrate I；
7) filtrate I obtained in step 6) is uniformly mixed with mixed liquor G obtained in step 4), is concentrated into The 10% of original volume carries out centrifugation 15min with the rate of 5000rpm, takes supernatant to carry out ultrafiltration using ultrafiltration membrane and leaves and takes super Filtrate obtains mixture J；The molecular cut off of the ultrafiltration membrane is 30000；
8) mixture J obtained in step 7) is concentrated under reduced pressure into 1/3 volume, the positive fourth of isometric chloroform-is then added Alcohol mixed liquor carries out uniformly mixed, then carries out ultrasonic treatment 5min, stands and carries out centrifugation 15min with the rate of 5000rpm after 2h, Upper layer centrifugate is taken, in aforementioned upper layer centrifugate plus the concentration of 3.5 times of volumes is 98% ethanol water, is cooled to 4 DEG C Standing 12h is carried out, centrifugation 15min is then carried out with the rate of 4000rpm and obtains sediment, is replaced with anhydrous propanone and anhydrous ether Washing precipitate 6 times, then carry out vacuum freeze drying to get；
Wherein, the chloroform-n-butanol mixed liquor be chloroform and n-butanol according to the ratio mixing that volume ratio is 4:1 and At；The ultrasonic power of the ultrasonic treatment is 180W；The temperature of the vacuum freeze drying is -18 DEG C, pressure 10Pa, the time For 12h；Ultrasound and centrifugal treating are carried out by the way that chloroform-n-butanol mixed liquor is added, also by repeatedly washing, effectively increases and mentions Effect is taken, and then is conducive to improve the purity of product.
Embodiment 4
One main laminaria active polysaccharide, including below according to the raw material of parts by weight: 20 parts of Fresh Laminaria Japonica, 150 parts of dehydrated alcohol, 0.045 ‰ parts of cellulase, 0.02 ‰ parts of pectase, 0.015 ‰ parts of zytase and suitable deionized water.
In the present embodiment, the preparation method of the kelp active polysaccharide, steps are as follows:
1) Fresh Laminaria Japonica is weighed according to parts by weight, is cut into the sheet that length and width are 1cm, then weighed according to parts by weight anhydrous Ethyl alcohol is mixed with the aforementioned Fresh Laminaria Japonica for being cut into sheet, then is ultrasonically treated, and 20 mesh filter to get filtrate A and filter residue B；
Wherein, the ultrasonic power of the ultrasonic treatment is 200W；The time of the ultrasonic treatment is 20min；
2) filter residue B obtained in step 1) is added to the deionized water of 10 times of volumes, the stirring with 400r/min at 42 DEG C It mixes rate and is stirred 200min, 40 mesh filter to get filtrate C and filter residue D；
3) filter residue D obtained in step 2) is added to the deionized water of 8 times of volumes, with the stirring of 100r/min at 38 DEG C Rate is stirred 100min, and 80 mesh filter to get filtrate E and filter residue F；
4) filtrate F obtained in liquor C obtained in filtrate A obtained in step 1) and step 2) and step 3) is mixed It closes uniformly, obtains mixed liquor G；By repeatedly extracting, extraction effect is effectively increased；
5) deionized water of 9 times of weight will be added in filter residue F obtained in step 3), be warming up to 70 DEG C after impregnating 40min Continue to impregnate 50min, be then cooled to 48 DEG C, cellulase is added and reacts 40min, then is warming up to 55 DEG C, zytase is added 40min is reacted, is then cooled to 50 DEG C, pectin enzyme reaction 30min is added, obtains mixture H；By repeatedly digesting, effectively improve Extraction effect；
6) mixture H obtained in step 5) is warming up to 105 DEG C, keeps 3 minutes progress destroy the enzyme treatments, is then added 12 The deionized water of times weight impregnates 140min, and the filtering of 100 mesh obtains filtrate I；
7) filtrate I obtained in step 6) is uniformly mixed with mixed liquor G obtained in step 4), is concentrated into The 10% of original volume carries out centrifugation 15min with the rate of 5000rpm, takes supernatant to carry out ultrafiltration using ultrafiltration membrane and leaves and takes super Filtrate obtains mixture J；The molecular cut off of the ultrafiltration membrane is 30000；
8) mixture J obtained in step 7) is concentrated under reduced pressure into 1/3 volume, the positive fourth of isometric chloroform-is then added Alcohol mixed liquor carries out uniformly mixed, then carries out ultrasonic treatment 5min, stands and carries out centrifugation 15min with the rate of 5000rpm after 2h, Upper layer centrifugate is taken, in aforementioned upper layer centrifugate plus the concentration of 3.5 times of volumes is 98% ethanol water, is cooled to 4 DEG C Standing 12h is carried out, centrifugation 15min is then carried out with the rate of 4000rpm and obtains sediment, is replaced with anhydrous propanone and anhydrous ether Washing precipitate 6 times, then carry out vacuum freeze drying to get；
Wherein, the chloroform-n-butanol mixed liquor be chloroform and n-butanol according to the ratio mixing that volume ratio is 4:1 and At；The ultrasonic power of the ultrasonic treatment is 180W；The temperature of the vacuum freeze drying is -18 DEG C, pressure 10Pa, the time For 12h；Ultrasound and centrifugal treating are carried out by the way that chloroform-n-butanol mixed liquor is added, also by repeatedly washing, effectively increases and mentions Effect is taken, and then is conducive to improve the purity of product.
Embodiment 5
One main laminaria active polysaccharide, including below according to the raw material of parts by weight: 20 parts of Fresh Laminaria Japonica, 150 parts of dehydrated alcohol, 0.045 ‰ parts of cellulase, 0.02 ‰ parts of pectase, 0.015 ‰ parts of zytase and suitable deionized water.
In the present embodiment, the preparation method of the kelp active polysaccharide, steps are as follows:
1) Fresh Laminaria Japonica is weighed according to parts by weight, is cut into the sheet that length and width are 1cm, then weighed according to parts by weight anhydrous Ethyl alcohol is mixed with the aforementioned Fresh Laminaria Japonica for being cut into sheet, then is ultrasonically treated, and 20 mesh filter to get filtrate A and filter residue B；
Wherein, the ultrasonic power of the ultrasonic treatment is 200W；The time of the ultrasonic treatment is 20min；
2) filter residue B obtained in step 1) is added to the deionized water of 10 times of volumes, the stirring with 400r/min at 42 DEG C It mixes rate and is stirred 200min, 40 mesh filter to get filtrate C and filter residue D；
3) filter residue D obtained in step 2) is added to the deionized water of 8 times of volumes, with the stirring of 100r/min at 38 DEG C Rate is stirred 100min, and 80 mesh filter to get filtrate E and filter residue F；
4) filtrate F obtained in liquor C obtained in filtrate A obtained in step 1) and step 2) and step 3) is mixed It closes uniformly, obtains mixed liquor G；By repeatedly extracting, extraction effect is effectively increased；
5) deionized water of 9 times of weight will be added in filter residue F obtained in step 3), be warming up to 70 DEG C after impregnating 40min Continue to impregnate 50min, be then cooled to 48 DEG C, cellulase is added and reacts 40min, then is warming up to 55 DEG C, zytase is added 40min is reacted, is then cooled to 50 DEG C, pectin enzyme reaction 30min is added, obtains mixture H；By repeatedly digesting, effectively improve Extraction effect；
6) mixture H obtained in step 5) is warming up to 105 DEG C, keeps 3 minutes progress destroy the enzyme treatments, is then added 12 The deionized water of times weight impregnates 140min, and the filtering of 100 mesh obtains filtrate I；
7) filtrate I obtained in step 6) is uniformly mixed with mixed liquor G obtained in step 4), is concentrated into The 10% of original volume carries out centrifugation 15min with the rate of 5000rpm, takes supernatant to carry out ultrafiltration using ultrafiltration membrane and leaves and takes super Filtrate obtains mixture J；The molecular cut off of the ultrafiltration membrane is 20000；
8) mixture J obtained in step 7) is concentrated under reduced pressure into 1/3 volume, the positive fourth of isometric chloroform-is then added Alcohol mixed liquor carries out uniformly mixed, then carries out ultrasonic treatment 5min, stands and carries out centrifugation 15min with the rate of 5000rpm after 2h, Upper layer centrifugate is taken, in aforementioned upper layer centrifugate plus the concentration of 3.5 times of volumes is 98% ethanol water, is cooled to 4 DEG C Standing 12h is carried out, centrifugation 15min is then carried out with the rate of 4000rpm and obtains sediment, is replaced with anhydrous propanone and anhydrous ether Washing precipitate 6 times, then carry out vacuum freeze drying to get；
Wherein, the chloroform-n-butanol mixed liquor be chloroform and n-butanol according to the ratio mixing that volume ratio is 4:1 and At；The ultrasonic power of the ultrasonic treatment is 180W；The temperature of the vacuum freeze drying is -18 DEG C, pressure 10Pa, the time For 12h；Ultrasound and centrifugal treating are carried out by the way that chloroform-n-butanol mixed liquor is added, also by repeatedly washing, effectively increases and mentions Effect is taken, and then is conducive to improve the purity of product.
Embodiment 6
One main laminaria active polysaccharide, including below according to the raw material of parts by weight: 20 parts of Fresh Laminaria Japonica, 150 parts of dehydrated alcohol, 0.045 ‰ parts of cellulase, 0.02 ‰ parts of pectase, 0.015 ‰ parts of zytase and suitable deionized water.
In the present embodiment, the preparation method of the kelp active polysaccharide, steps are as follows:
1) Fresh Laminaria Japonica is weighed according to parts by weight, is cut into the sheet that length and width are 1cm, then weighed according to parts by weight anhydrous Ethyl alcohol is mixed with the aforementioned Fresh Laminaria Japonica for being cut into sheet, then is ultrasonically treated, and 20 mesh filter to get filtrate A and filter residue B；
Wherein, the ultrasonic power of the ultrasonic treatment is 200W；The time of the ultrasonic treatment is 20min；
2) filter residue B obtained in step 1) is added to the deionized water of 10 times of volumes, the stirring with 400r/min at 42 DEG C It mixes rate and is stirred 200min, 40 mesh filter to get filtrate C and filter residue D；
3) filter residue D obtained in step 2) is added to the deionized water of 8 times of volumes, with the stirring of 100r/min at 38 DEG C Rate is stirred 100min, and 80 mesh filter to get filtrate E and filter residue F；
4) filtrate F obtained in liquor C obtained in filtrate A obtained in step 1) and step 2) and step 3) is mixed It closes uniformly, obtains mixed liquor G；By repeatedly extracting, extraction effect is effectively increased；
5) deionized water of 9 times of weight will be added in filter residue F obtained in step 3), be warming up to 70 DEG C after impregnating 40min Continue to impregnate 50min, be then cooled to 48 DEG C, cellulase is added and reacts 40min, then is warming up to 55 DEG C, zytase is added 40min is reacted, is then cooled to 50 DEG C, pectin enzyme reaction 30min is added, obtains mixture H；By repeatedly digesting, effectively improve Extraction effect；
6) mixture H obtained in step 5) is warming up to 105 DEG C, keeps 3 minutes progress destroy the enzyme treatments, is then added 12 The deionized water of times weight impregnates 140min, and the filtering of 100 mesh obtains filtrate I；
7) filtrate I obtained in step 6) is uniformly mixed with mixed liquor G obtained in step 4), is concentrated into The 10% of original volume carries out centrifugation 15min with the rate of 5000rpm, takes supernatant to carry out ultrafiltration using ultrafiltration membrane and leaves and takes super Filtrate obtains mixture J；The molecular cut off of the ultrafiltration membrane is 40000；
8) mixture J obtained in step 7) is concentrated under reduced pressure into 1/3 volume, the positive fourth of isometric chloroform-is then added Alcohol mixed liquor carries out uniformly mixed, then carries out ultrasonic treatment 5min, stands and carries out centrifugation 15min with the rate of 5000rpm after 2h, Upper layer centrifugate is taken, in aforementioned upper layer centrifugate plus the concentration of 3.5 times of volumes is 98% ethanol water, is cooled to 4 DEG C Standing 12h is carried out, centrifugation 15min is then carried out with the rate of 4000rpm and obtains sediment, is replaced with anhydrous propanone and anhydrous ether Washing precipitate 6 times, then carry out vacuum freeze drying to get；
Wherein, the chloroform-n-butanol mixed liquor be chloroform and n-butanol according to the ratio mixing that volume ratio is 4:1 and At；The ultrasonic power of the ultrasonic treatment is 180W；The temperature of the vacuum freeze drying is -18 DEG C, pressure 10Pa, the time For 12h；Ultrasound and centrifugal treating are carried out by the way that chloroform-n-butanol mixed liquor is added, also by repeatedly washing, effectively increases and mentions Effect is taken, and then is conducive to improve the purity of product.
Kelp active polysaccharide prepared by the present invention have reducing blood lipid, it is hypoglycemic, adjust immune, anticoagulation, antitumor, lead discharging Removing toxic substances and a variety of biological functions such as anti-oxidant effectively increase extraction effect, pass through simultaneously by repeatedly extracting and repeatedly digesting Ultrasound, centrifugation and repeatedly washing, are conducive to the purity for improving product, can be used for solving due to lacking corresponding catabolic enzyme in human body And the problem of laminarin can not be directly absorbed by edible kelps, it has a vast market foreground.
Better embodiment of the invention is explained in detail above, but the present invention is not limited to above-mentioned embodiment party Formula within the knowledge of one of ordinary skill in the art can also be without departing from the purpose of the present invention Various changes can be made.There is no necessity and possibility to exhaust all the enbodiments.And it thus amplifies out apparent Variation or variation be still in the protection scope of this invention.

Claims (10)

1. a main laminaria active polysaccharide, which is characterized in that including below according to the raw material of parts by weight: 17-25 parts of Fresh Laminaria Japonica, nothing 140-160 parts of water-ethanol, 0.038 ‰ -0.05 ‰ parts of cellulase, 0.01 ‰ -0.03 ‰ parts of pectase, zytase 0.012 ‰ -0.016 ‰ parts and suitable deionized water.
2. kelp active polysaccharide according to claim 1, which is characterized in that including below according to the raw material of parts by weight: new 20 parts of fresh kelp, 150 parts of dehydrated alcohol, 0.045 ‰ parts of cellulase, 0.02 ‰ parts of pectase, 0.015 ‰ parts of zytase, And suitable deionized water.
3. a kind of preparation method of the kelp active polysaccharide as described in claim 1-2 is any, which is characterized in that steps are as follows:
1) Fresh Laminaria Japonica is weighed according to parts by weight, is cut into the sheet that length and width are 1cm, then weighs dehydrated alcohol according to parts by weight It mixes, then is ultrasonically treated with the aforementioned Fresh Laminaria Japonica for being cut into sheet, 20 mesh filter to get filtrate A and filter residue B；
2) filter residue B obtained in step 1) is added to the deionized water of 10 times of volumes, 200min, the filtering of 40 mesh are stirred at 42 DEG C Obtain liquor C and filter residue D；
3) filter residue D obtained in step 2) is added to the deionized water of 8 times of volumes, with the stirring rate of 100r/min at 38 DEG C It is stirred 100min, 80 mesh filter to get filtrate E and filter residue F；
4) filtrate A obtained in step 1) is mixed with filtrate F obtained in liquor C obtained in step 2) and step 3) It is even, obtain mixed liquor G；
5) deionized water of 9 times of weight will be added in filter residue F obtained in step 3), be warming up to 70 DEG C of continuation after impregnating 40min 50min is impregnated, is then cooled to 48 DEG C, cellulase is added and reacts 40min, then is warming up to 55 DEG C, zytase reaction is added Then 40min is cooled to 50 DEG C, pectin enzyme reaction 30min is added, obtains mixture H；
6) mixture H obtained in step 5) is warming up to 105 DEG C, keeps 3 minutes progress destroy the enzyme treatments, 12 times of weights are then added The deionized water of amount impregnates 140min, and the filtering of 100 mesh obtains filtrate I；
7) filtrate I obtained in step 6) is uniformly mixed with mixed liquor G obtained in step 4), carries out being concentrated into substance Long-pending 10% carries out centrifugation 15min with the rate of 5000rpm, takes supernatant to carry out ultrafiltration using ultrafiltration membrane and leaves and takes ultrafiltrate, Obtain mixture J；
8) mixture J obtained in step 7) is concentrated under reduced pressure into 1/3 volume, it is mixed that isometric chloroform-n-butanol is then added It closes liquid and carries out uniformly mixed, then carry out ultrasonic treatment 5min, stand and centrifugation 15min is carried out with the rate of 5000rpm after 2h, take Layer centrifugate, in aforementioned upper layer centrifugate plus the concentration of 3.5 times of volumes is 98% ethanol water, is cooled to 4 DEG C of progress 12h is stood, centrifugation 15min is then carried out with the rate of 4000rpm and obtains sediment, is alternately washed with anhydrous propanone and anhydrous ether Sediment 6 times, then carry out vacuum freeze drying to get.
4. the preparation method of kelp active polysaccharide according to claim 3, which is characterized in that in step 1), the ultrasound The ultrasonic power of processing is 200W；The time of the ultrasonic treatment is 20min.
5. the preparation method of kelp active polysaccharide according to claim 3, which is characterized in that in step 2), the stirring Stirring rate be 400r/min.
6. the preparation method of kelp active polysaccharide according to claim 3, which is characterized in that in step 7), the ultrafiltration The molecular cut off of film is 20000-40000.
7. the preparation method of kelp active polysaccharide according to claim 6, which is characterized in that in step 8), the chloroform- N-butanol mixed liquor is that chloroform and n-butanol are mixed according to the ratio that volume ratio is 4:1.
8. the preparation method of kelp active polysaccharide according to claim 7, which is characterized in that in step 8), the ultrasound The ultrasonic power of processing is 180W.
9. the preparation method of kelp active polysaccharide according to claim 8, which is characterized in that in step 8), the vacuum The temperature of freeze-drying is -18 DEG C, pressure 10Pa, time 12h.
10. a kind of preparation method of the kelp active polysaccharide as described in claim 3-9 is any is in preparing active polysaccharide product Purposes.
CN201811045230.4A 2018-09-07 2018-09-07 One main laminaria active polysaccharide and preparation method thereof Pending CN108997511A (en)

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