CN102242100A - Method for using compound biological enzyme in plant extraction process, and conditions thereof - Google Patents
Method for using compound biological enzyme in plant extraction process, and conditions thereof Download PDFInfo
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- CN102242100A CN102242100A CN2011100987255A CN201110098725A CN102242100A CN 102242100 A CN102242100 A CN 102242100A CN 2011100987255 A CN2011100987255 A CN 2011100987255A CN 201110098725 A CN201110098725 A CN 201110098725A CN 102242100 A CN102242100 A CN 102242100A
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Abstract
The invention relates to a method for using compound biological enzyme in a plant extraction process, and conditions thereof. The compound biological enzyme comprises 5-60% of cellulose, 5-30% of hemicellulase, 5-50% of diastase, 5-50% of pectinase and 1-25% of protease, wherein the sum of the weight percentages of the components is 100%, and activity unit of the compound biological enzyme is 200000 U/g-1200000 U/g. The method for using the compound biological enzyme comprises: completely crushing a substrate until the substrate can passes a sieve having 80-200 meshes; adding the substrate to a water bath solution according to a ratio of material to liquid of 1:5-1:8; adjusting a pH value of the water bath solution to 4.0-5.0 through a acid (for example, acetic acid, oxalic acid, citric acid and the like); adding the compound biological enzyme to the water bath solution according to a certain ratio based on different substrates, followed by stirring completely and uniformly. A water bath temperature is controlled between 15-60 DEG C. The extraction time is controlled to 1-72 hours. The compound biological enzyme addition comprises the following conditions that: when the substrates are flowers, leaves or fruits of land plants, the compound biological enzyme addition is 0.1-0.5% of the substrate mass; when the substrates are roots and stems of the land plants, the compound biological enzyme addition is 0.3-0.8% of the substrate mass; when the substrates are marine plants, the compound biological enzyme addition is 0.8-3% of the substrate mass.
Description
Technical field
The present invention relates to method and condition that a kind of compound biological enzyme is used for plant extract technology.
Background technology
The contained effective constituent of plant is wrapped in the cell walls mostly, traditional hot water, acid, alkali, organic solvent extraction, because of being subjected to divide cellulosic obstruction in the cell walls, often the extraction time long, solvent load is many, extraction efficiency is lower, and the interior impurities of cell is (as albumen, pectin, tannin, ash content and viscous substance etc.) also disturb the extraction of effective constituent.After plant was handled with alcohol simultaneously, change in various degree may take place or lose in its composition, so that directly influence the pharmacological action and the curative effect of Chinese medicine preparation.Therefore traditional extracting method has not only increased the production cost height, has also wasted valuable plant resources.
Enzyme is produced by the living organisms cell, is subjected to multiple factor to regulate the biological catalyst with catalytic capability of control.Compared its common point with general catalyzer, but outstanding feature is arranged, the catalytic efficiency height of enzyme has the specificity of height.Therefore, utilize enzyme can change the permeability of cell walls, help the effective component stripping in the wall, thereby improve the extraction yield of effective component.
Summary of the invention
The invention provides method and condition that a kind of compound biological enzyme is used for plant extract, extraction efficiency height not only, and also simple to operate, production cost is low, has improved the utilization ratio of plant.
The technical solution adopted in the present invention is: compound biological enzyme is made up of cellulase, hemicellulase, amylase, polygalacturonase and proteolytic enzyme, wherein cellulase 5%~60%, hemicellulase 5%~30%, amylase 5%~50%, polygalacturonase 5%~50%, proteolytic enzyme 1%~25%, total mass consists of 100%.The activity unit of enzyme is 200,000 U/g~1,200,000 U/g.
Using method: substrate is fully pulverized, make it can pass through 80 orders~200 mesh sieves, be to put into water-bath in 1: 5~1: 8 by solid-liquid ratio then, regulate pH value to 4.0~5.0 of water-bath with acid (as acetate, oxalic acid, citric acid etc.) after, in proportion compound biological enzyme is poured in the water-bath according to different substrates, fully stirred evenly.The temperature of water-bath is controlled between 15 ℃~60 ℃, extraction time 1h~72h.When extracting, need suitable stirring water-bath, help the hydrolysis effect of enzyme like this, shorten extraction time.
The dosage of compound biological enzyme: with the flower of land plant, leaf, dosage was the 0.1%-0.5% of substrate quality when fruit was substrate.With the root of land plant, dosage was the 0.3%-0.8% of substrate when stem was substrate; Dosage is the 0.8%-3% of substrate when being substrate with the marine plant.
The invention foundation
Cellulase has decomposition, softening fibre element, destruction cell walls, increases the effect of the stripping quantity of vegetable cell content, it is the general name of the glucogenic one group of enzyme of degraded cellulose, comprises endoglucanase, cellobiohydrolase, 3 components of beta-glucosidase.Optimum pH 4~5,40 ℃~60 ℃ of the best use of temperature.
Hemicellulase is by endo-type enzymes such as 'beta '-mannase, beta-xylanases, circumscribed-type enzyme and compositions such as arabinofuranosidase/xylosidase, tilactase, glucuronidase and acetyl zytase such as beta-glucosidase, beta-Mannosidase, xylobiase.Effect with digestion plant cell wall.
Polygalacturonase is the general name of a class prozyme of glycan lytic enzyme, the pectin substance Acyl-hydrolase of decompose pectin matter.45 ℃-50 ℃ of optimum temperatures, action pH value 3~6.
Amylase is the enzyme general name of hydrolyzed starch and glycogen, is divided into α-Dian Fenmei and beta-amylase usually.
Proteolytic enzyme plays the group attribute of katalysis according to the active centre of enzyme, can be divided into serine/threonine protein enzyme, thiol proteinase, metalloprotease and aspartate protease etc.Proteolytic enzyme can resolve into peptide with protein, is hydrolyzed into amino acid through peptase again.
The advantage that compound biological enzyme of the present invention is used for the effective ingredients in plant extraction has:
1, utilize enzyme reaction to have highly narrow spectrum characteristics, select different enzymes to be mixed in proportion, moiety (as Mierocrystalline cellulose, hemicellulose, pectin substance etc.) hydrolysis or degraded with plant cell wall, destroy cell wall structure, make the fully dissolving of intracellular composition, suspendible or peptization in solvent, thereby reach the extraction purpose, improve extraction efficiency, reduce production costs.Utilize the present invention can make extraction yield increase more than 15%.
2, plant combinative enzyme hydrolysis mild condition has reduced the difficulty of solvent extraction subsequently, makes whole extraction process mild conditionization, helps to keep original character of effective component.
3, can destroy plant cell wall at low temperatures quickly and effectively, the extract at low temperature of pharmaceutically active ingredient in helping to realize, the destruction of reducing effective constituent is reduced energy consumption.
Embodiment
Following examples are used for understanding better the present invention, but are not used for limiting the scope that the present invention protects.
The invention will be further described below in conjunction with embodiment:
Embodiment 1: utilize compound biological enzyme that Japanese Honeysuckle is extracted.
(1) get the Japanese Honeysuckle 2000g of oven dry, micronizing makes it can cross 100 mesh sieves.
(2) extracting cellulose enzyme 4.5g, hemicellulase 2.0g, amylase 1.5g, polygalacturonase 1.5g, proteinase-10 .5g are mixed into the heavy compound biological enzyme of 10g, and the activity unit of enzyme is 500,000 U/g.
(3) the Japanese Honeysuckle fine powder after 2000g is pulverized adds the water of 6 times of amounts, temperature is risen to 48 ℃ then, add pre-configured compound biological enzyme liquid 10g, transferring pH value with oxalic acid is 4.5,48 ℃ of enzymolysis 60min of back maintenance stir, be warming up to 100 ℃ then and carry out quick inactivating, keep naturally cooling cooling behind the temperature lixiviate 1h again.
(4) filter, remove slag, concentrate, dry extractive substance.
(5) compare simultaneously, get the Japanese Honeysuckle 2000g of oven dry, micronizing makes it can cross 100 mesh sieves, adds the water of 8 times of amounts, heating lixiviate 6 hours, filter extractive substance.
Through check relatively, the compound bio enzyme extraction can make the chlorogenic acid yield higher by 24.9% than water extraction.
Embodiment 2: utilize compound bio-enzyme that the coptis is extracted.
(1) get coptis 2000g, micronizing makes it can cross 200 mesh sieves.
(2) extracting cellulose enzyme 7.0g, hemicellulase 3.0g, amylase 2.0g, polygalacturonase 1.0g, proteolytic enzyme 1.0g are mixed into the heavy compound biological enzyme of 14g, and the activity unit of enzyme is 400,000 U/g.
(3) coptis fine powder after the 2000g pulverizing is added 3 times of water gagings, add pre-configured compound biological enzyme liquid 14g, soak with 0.3% sulfuric acid adjust pH to 5 back, at 40 ℃ of following water bath with thermostatic control 90min, the coptis and 0.3% sulfuric acid are placed percolator as solvent, and dipping, diacolation are collected percolate, with milk of lime adjust pH to 10~12, precipitation, suction filtration, filtrate is used concentrated hydrochloric acid adjust pH to 1~2, adding table salt makes saltiness reach 7%, fully stir, leave standstill 24h, filter, dry under 60 ℃, get the berberine hydrochloride crude product.
(4) compare simultaneously, get golden cypress 2000g, micronizing makes it can cross 200 mesh sieves, the Diluted Alcohol that adds 8 times of amounts, heating lixiviate 8h, filter extractive substance, use milk of lime adjust pH to 10~12 then, precipitation, suction filtration, filtrate adds table salt and makes saltiness reach 7% with concentrated hydrochloric acid adjust pH to 1~2, fully stirs, leave standstill 24h, filter, dry under 60 ℃, get the berberine hydrochloride crude product.
Through check relatively, the coptis is after Enzymatic Extraction, and the gained content of berberine hydrochloride is 4.3%, and the content of berberine hydrochloride of handling without enzyme is 2.4%, and the compound bio enzyme extraction can make the berberine hydrochloride yield higher by 44.2% than what handle without enzyme.
Embodiment 3: utilize compound bio-enzyme that marine alga is extracted.
(1) get the marine alga 1000g of oven dry, micronizing makes it can cross 200 mesh sieves.
(2) extracting cellulose enzyme 10.5g, hemicellulase 2.5g, amylase 2.5g, polygalacturonase 3.0g, proteolytic enzyme 1.5g are mixed into the heavy compound biological enzyme of 20g, and the activity unit of enzyme is 700,000 U/g.
(3) the marine alga fine powder after the 1000g pulverizing is added 8 times of water gagings, temperature is risen to 50 ℃ then, add pre-configured compound biological enzyme liquid 20g, transferring pH value is 4.5, and the back that stirs keeps 50 ℃ of constant temperature enzymolysis 2h.Be warming up to 90 ℃ then rapidly and carry out quick inactivating, be cooled to 80 ℃ behind the deactivation 1min and extract 1h.
(4) filter, remove slag, concentrate, dry extractive substance.
(5) compare simultaneously, get the marine alga 1000g of oven dry, micronizing makes it can cross 200 mesh sieves, adds 8 times of water gagings, heating lixiviate 8h, filter extractive substance.
Compare through check, but compound bio enzyme extraction chlorogenic acid yield is higher by 35.5% than water extraction.
Claims (5)
1. compound biological enzyme is used for the method and the condition of plant extract technology, it is characterized in that compound biological enzyme is made up of cellulase, hemicellulase, amylase, polygalacturonase and proteolytic enzyme.
2. the described compound biological enzyme of root a tree name claim 1 is used for the method and the condition of plant extract technology, it is characterized in that cellulase 5%~60%, hemicellulase 5%~30%, amylase 5%~50%, polygalacturonase 5%~50%, proteolytic enzyme 1%~25%, total mass consists of 100%.
3. the described compound biological enzyme of root a tree name claim 1 is used for the method and the condition of plant extract technology, and the activity unit that it is characterized in that its enzyme is 200,000 U/g~1,200,000 U/g.
4. the described compound biological enzyme of root a tree name claim 1 is used for the method and the condition of plant extract technology, the using method that it is characterized in that it is that substrate is fully pulverized, make it can pass through 80~200 mesh sieves, be to put into water-bath in 1: 5~1: 8 by solid-liquid ratio then, regulate pH value to 4.0~5.0 of water-bath with acid (as acetate, oxalic acid, citric acid etc.) after, in proportion compound biological enzyme is poured in the water-bath according to different substrates, fully stirred evenly.The temperature of water-bath is controlled between 15 ℃~60 ℃, extraction time 1h~72h.When extracting, need suitable stirring water-bath, help the hydrolysis effect of enzyme like this, shorten extraction time.
5. the described compound biological enzyme of root a tree name claim 1 is used for the method and the condition of plant extract technology, and the dosage of compound biological enzyme that it is characterized in that it is for the flower of land plant, leaf, dosage was the 0.1%-0.5% of substrate quality when fruit was substrate.With the root of land plant, dosage was the 0.3%-0.8% of substrate when stem was substrate; Dosage is the 0.8%-3% of substrate when being substrate with the marine plant.
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Cited By (14)
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CN102626459A (en) * | 2012-03-15 | 2012-08-08 | 重庆市中医院 | Method for compound enzyme-hot water extraction of cortex phellodendri alkaloids |
CN103787885A (en) * | 2014-03-06 | 2014-05-14 | 山东禾本堂生物科技有限公司 | Method of extracting chlorogenic acid with high purity from honeysuckle above ground part |
CN103858945A (en) * | 2014-03-20 | 2014-06-18 | 广州恒广复合材料有限公司 | Natural plant fungicide and preparation method thereof |
CN103919914A (en) * | 2014-04-15 | 2014-07-16 | 河南牧翔动物药业有限公司 | Method for preparing veterinary heat-clearing and detoxifying oral liquid |
CN104258103A (en) * | 2014-09-28 | 2015-01-07 | 神农架神林生物科技有限公司 | Dendrobe buccal tablet and preparation method thereof |
CN104673768A (en) * | 2015-03-20 | 2015-06-03 | 北京农学院 | Process for extracting myrosinase from horseradish |
CN106011112A (en) * | 2016-05-20 | 2016-10-12 | 山东省食品发酵工业研究设计院 | Special complex enzyme system for kelp comprehensive utilization and application thereof |
CN106916021A (en) * | 2017-04-12 | 2017-07-04 | 山东佐田氏生物科技有限公司 | A kind of double source humic acid biological fertilizer and preparation method and application |
CN107875196A (en) * | 2017-11-17 | 2018-04-06 | 成都赛诺联创生物科技有限公司 | Complex enzyme, extract solution system, medicament, extract and preparation method thereof, application |
CN109355343A (en) * | 2018-12-28 | 2019-02-19 | 河北肽都生物科技有限公司 | A kind of preparation method of small molecule honeysuckle chelating peptide |
CN110156797A (en) * | 2019-04-28 | 2019-08-23 | 北海生巴达生物科技有限公司 | A kind of technique for extracting chlorophyll from spirulina |
CN113754626A (en) * | 2021-07-12 | 2021-12-07 | 雅安职业技术学院 | Method for preparing fisetin by enzyme method |
CN113854435A (en) * | 2021-09-30 | 2021-12-31 | 东莞市益得洋科技有限公司 | Agilawood chicken feed additive |
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CN102626459B (en) * | 2012-03-15 | 2014-07-23 | 重庆市中医院 | Method for compound enzyme-hot water extraction of cortex phellodendri alkaloids |
CN102626459A (en) * | 2012-03-15 | 2012-08-08 | 重庆市中医院 | Method for compound enzyme-hot water extraction of cortex phellodendri alkaloids |
CN103787885A (en) * | 2014-03-06 | 2014-05-14 | 山东禾本堂生物科技有限公司 | Method of extracting chlorogenic acid with high purity from honeysuckle above ground part |
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CN103858945B (en) * | 2014-03-20 | 2016-03-23 | 广州恒广复合材料有限公司 | A kind of natural plants bactericide and preparation method thereof |
CN103919914A (en) * | 2014-04-15 | 2014-07-16 | 河南牧翔动物药业有限公司 | Method for preparing veterinary heat-clearing and detoxifying oral liquid |
CN103919914B (en) * | 2014-04-15 | 2016-06-08 | 河南牧翔动物药业有限公司 | A kind of preparation method of antipyretic and detoxicated oral liquid for animals |
CN104258103B (en) * | 2014-09-28 | 2018-01-02 | 神农架神林生物科技有限公司 | A kind of stem of noble dendrobium buccal tablet and preparation method |
CN104258103A (en) * | 2014-09-28 | 2015-01-07 | 神农架神林生物科技有限公司 | Dendrobe buccal tablet and preparation method thereof |
CN104673768A (en) * | 2015-03-20 | 2015-06-03 | 北京农学院 | Process for extracting myrosinase from horseradish |
CN106011112A (en) * | 2016-05-20 | 2016-10-12 | 山东省食品发酵工业研究设计院 | Special complex enzyme system for kelp comprehensive utilization and application thereof |
CN106916021A (en) * | 2017-04-12 | 2017-07-04 | 山东佐田氏生物科技有限公司 | A kind of double source humic acid biological fertilizer and preparation method and application |
CN107875196A (en) * | 2017-11-17 | 2018-04-06 | 成都赛诺联创生物科技有限公司 | Complex enzyme, extract solution system, medicament, extract and preparation method thereof, application |
CN109355343A (en) * | 2018-12-28 | 2019-02-19 | 河北肽都生物科技有限公司 | A kind of preparation method of small molecule honeysuckle chelating peptide |
CN110156797A (en) * | 2019-04-28 | 2019-08-23 | 北海生巴达生物科技有限公司 | A kind of technique for extracting chlorophyll from spirulina |
CN113754626A (en) * | 2021-07-12 | 2021-12-07 | 雅安职业技术学院 | Method for preparing fisetin by enzyme method |
CN113754626B (en) * | 2021-07-12 | 2024-03-15 | 雅安职业技术学院 | Method for preparing fisetin by enzyme method |
CN113854435A (en) * | 2021-09-30 | 2021-12-31 | 东莞市益得洋科技有限公司 | Agilawood chicken feed additive |
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Application publication date: 20111116 |