CN102838757B - Method of extracting gutta-percha through enzymolysis - Google Patents
Method of extracting gutta-percha through enzymolysis Download PDFInfo
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Abstract
Provided is a method of extracting gutta-percha through enzymolysis. One of natural eucommia skin or eucommia leaves or eucommia wing skin is placed in clean water to be cleaned and dried, and moisture content of the eucommia skin, the eucommia leaves, or the eucommia wing skin is enabled to be less than 5%. A compound enzyme which comprises, by mass, 0.27% of cellulose, 0.01% of pectinase, and 0.02% of protease is added, enzymolysis undergoes 1-3 days under stirring at a speed of 500 r/min under 50 DEG C. Mixture after the enzymolysis is filtered, dregs of the filtered mixture are sent to an ultrasonic extraction tank for ultrasound extraction, and glue mixed liquor is produced. The glue mixed liquor is filtered in a grading mode, impurities are eliminated, and the glue mixed liquor with the impurities eliminated is sent to a separation tank. Eventually the gutta-percha is extracted through the method that gutta-percha crystals are precipitated at a low temperature. The method is simple in process, little in pollution, high in yield rate, low in cost, and ideal in effect; the extraction rate reaches more than 95%, and the gutta-percha content reaches 99.9%; and the produced gutta-percha is long in gelatin silk, high in tensile and tearing-proofing physical strength, and capable of being applied to wide fields.
Description
Technical field
The present invention relates to a kind of extracting method of natural product, is a kind of method of enzymolysis and extraction gutta-percha specifically.
Background technology
Chinese gutta percha be uniquely have except para rubber (natural gum) in the world at present huge
The high-quality natural rubber resource of DEVELOPMENT PROSPECT, and China accounts for cosmopolitic species in the cultivated area of the bark of eucommia
Plant more than 99% of area, there is monopolistic resources advantage.More external developed countries, as Japan, give a long price for the Folium Eucommiae of China in a large number, accelerate the research and development to gutta-percha novel material.Therefore, the applicating and exploitation problem Dou Shi various countries of Chinese gutta percha concentrate the brainstorm subject of investigation.
But, the development prospect cognition degree of current domestic industry to bark of eucommia industry is not high, do not form the industrial chain of upstream and downstream Application and Development, therefore under this overall situation both domestic and external, the paces of China's eucommia resource comprehensive utilization need quickening badly, change Natural Rubber in The World Distribution Pattern, strengthen the right of speech of China's natural rubber resource.China is that high-quality eucommia resource concentrates growing area, the exploitation of gutta-percha, effectively will promote that this superior resources of China utilize, particularly in fields such as China's war industry, tyre production industry.This project main body, by striving the bark of eucommia plant husbandry driving the Changjiang river, In The Middle And Lower Reaches, the Yellow River, makes it develop the new natural rubber treasure-house of formation one, Substitute For Partial natural rubber, and part solves the phenomenon of natural rubber scarcity.The integrated application of natural Chinese gutta percha and the tool of setting up of information-based key equipment Showcase Production Line are of great significance.Compared with the applied research of Chinese gutta percha aspect, the extraction process of natural Chinese gutta percha and the research and development of corresponding Showcase Production Line are still in the comparatively primary stage.There is a lot of drawback in traditional extraction, such as, extraction time is many, productive rate is low, pollution is heavy, high in cost of production, and therefore these factors are unfavorable for extraction, the development and apply of natural Chinese gutta percha, and become the bottleneck of the natural Chinese gutta percha development of restriction.
Through the scientific research and development of decades, although the extracting method of Chinese gutta percha has breakthrough, these extracting method all also exist drawback and are difficult to industrialization, and the patent that current China is all and document mainly report following natural Chinese gutta percha extracting method:
A. tight auspicious fragrant patent (Chinese Academy of Sciences chemistry institute): first, mill and expose collodion silk, toluene, benzene, ethylene dichloride extract, and add alcohol, ketone analyses thick glue; Secondly, the dense NaOH infusion hydrocellulose of raw material, pectin, the thick glue of organic solvent extraction.The shortcoming of this method is: collodion silk damage is serious, and physicals is poor, and solvent toxicity is large, and cannot reclaim, complex technical process, seriously polluted, cost is high.
B. old increasing ripple patent (chemistry institute of the Chinese Academy of Sciences): raw material destroys cell walls through fermentation, uses
2%NaOH boiling, hydraulic engine is smashed, and dissociate collodion silk.The shortcoming of this method is: boiling temperature is high, and collodion silk is destroyed, and pollute large, collodion silk purity is not high, and the production cycle is long.
C. Yang Zhen hall patent (special product institute of the Chinese Academy of Agricultural Sciences): cultivate wound healing tissue, dries and makes powder, benzene, trichloromethane lixiviate, methanol extraction, ether purification and recover.The shortcoming of this method is: mixed solvent cannot utilize, and cost is high, and wound healing tissue's regeneration period is long, and leather toxicity is large,
Collodion silk physicals is poor.
D. horse Berlin method: 10%NaOH boiling at 90 DEG C, at 40 DEG C, concentrated hydrochloric acid process obtains thick glue.The shortcoming of this method is: concentrated acid, concentrated base are seriously polluted, and blowdown flow rate is large, and alkali destroys serious to gutta-percha, physicals declines, and product is thick glue.
Draw, no matter be which kind of extracting method above in technique, all there is the defects such as extraction time is many, environmental pollution heavy, extraction efficiency is low according to above As-Is analysis.Therefore, the bottleneck realizing Chinese gutta percha extraction industrialization at present mainly processes the problem that extraction cost is high, extract poor accuracy.Therefore, the new extraction process of development is needed in the development of natural Chinese gutta percha industry badly, realizes the industrialization production that natural Chinese gutta percha extracts application, forms one and extract the Showcase Production Line driving its comprehensive utilization with natural Chinese gutta percha.
Summary of the invention
The invention provides a kind of method that Cortex Eucommiae prepares gutta-percha, the method technique is simple, and pollute little, extraction rate reached is to more than 95%, and the content of gutta-percha reaches 99.9%.
A method for enzymolysis and extraction gutta-percha, adopt natural Cortex Eucommiae, Folium Eucommiae or Eucommia skin to make, the molecular weight 200000-300000 of gutta-percha, the foreign matter content in gutta-percha is less than 0.5%, and concrete preparation process is as follows:
Step one, the one chosen in natural Cortex Eucommiae, Folium Eucommiae or Eucommia skin, put into clear water and clean, dry, make the water content of Cortex Eucommiae, Folium Eucommiae or Eucommia intracutaneous be less than 5%, for subsequent use;
Step 2, employing enzymolysis process carry out pre-treatment
A, preparation enzyme solution: get 0.27% cellulase, 0.01% polygalacturonase and 0.02% proteolytic enzyme by mass percentage respectively and be dissolved in water, be mixed with the complex enzyme liquid of cellulase, polygalacturonase and proteolytic enzyme;
B, the Cortex Eucommiae cleaned up in above-mentioned steps one, Folium Eucommiae or Eucommia skin to be joined by solid-liquid ratio 1:1-2 in the complex enzyme liquid of the cellulase, polygalacturonase and the proteolytic enzyme that prepare; 50 DEG C, the rotating speed of 500r/min stir under enzymolysis 2-3 days;
The mixture of step 3, Cortex Eucommiae, Folium Eucommiae or Eucommia skin by after step 2 enzymolysis filters, slag after filtering is sent into ultrasonic wave mixing tank, first in ultrasonic wave mixing tank, put into petroleum ether solution, the slag after filtration and the weight ratio of petroleum ether solution are: 1:6-10; Then carry out ultrasonic, power is 2.4-3.6KW, and temperature is 70-80 DEG C, ultrasonic vibration time 30-60 minute, make the liquid in ultrasonic wave mixing tank form colloid mixed solution;
Dregs and liquor separation device sent into by step 4, colloid mixed solution step 3 formed, carry out classified filtering, its temperature is made to remain on 50 DEG C, first cross 1000 order filter sieve, then be 500 order filter sieve, be then 200 order filter sieve, carry out filtration treatment, deimpurity colloid mixed solution will be gone to send into and to carry glue tank, and reclaim sherwood oil for new bark of eucommia raw material continuation extraction gutta-percha.
Step 5, control at 10 DEG C by the temperature carrying glue tank, time 1-12 hour, makes the colloid in colloid mixed solution precipitate completely;
Step 6, propose in glue tank colloidal precipitation after, release the liquid on colloid top layer, take out colloid maybe by carry glue tank greenhouse cooling to 10 DEG C below, take out after making colloid crystallization, i.e. obtained a kind of natural gutta-percha.
Beneficial effect
1, pre-treatment of the present invention adopts enzymolysis process, select cellulase, be equipped with a small amount of polygalacturonase and proteolytic enzyme, three's cooperative cooperating, micelle cell and impeller structure cell is destroyed through biochemical action, owing to forming fiber, the pectin of cell walls, and the starch of easily gelatinization and the albumen that easily solidifies are degraded, and improve the permeability of Gutta_containing cell, impel the destruction of Folium Eucommiae weave construction, gutta-percha is free out, for the dissolving of next step organic dissolution is ready, makes extraction effect better.And enzymolysis process is the producing and manufacturing technique of environmental protection, can reach the requirement of scale operation.
2, leaching process adopt single ether as solvent, the feature of this solvent low toxicity, environmental protection, uses this solvent to extract without discharge cycle, capable of reducing energy consumption, does not relate to separated from solvent problem, can improve solvent utilising efficiency, reduce production cost;
3, adopt classified filtering, avoid blocking, make filter effect better, improve filtration efficiency, save processing operation time;
4, have employed advanced ultrasound countercurrent extraction technique, realize high efficiency extraction, make its extraction yield more than 90%, the content of gutta-percha reaches 99.9%, whole separation carry glue process have pollute low, the feature that productive rate is high, realizes the classification of gutta-percha, rapid extraction.
5. obtain gutta-percha collodion silk long, tension, the physical property intensity of anti tear is high, can Application Areas wide.
Accompanying drawing explanation
Fig. 1 is Eucommia ulmoides Oliv. leaves before Folium Eucommiae enzymolysis;
Fig. 2 is the gutta-percha residue of Folium Eucommiae after complex enzyme zymohydrolysis;
Fig. 3 is air-dry rear gutta-percha long filament;
Fig. 4 is Eucommia pericarp photo before complex enzyme zymohydrolysis;
Fig. 5 is Eucommia pericarp photo after complex enzyme zymohydrolysis;
Fig. 6 is the Photomicrograph of Eucommia pericarp after complex enzyme zymohydrolysis;
Fig. 7 is bark of eucommia Du stem dry hide photo before Stem of Eucommia ulmoides Oliv dry hide enzymolysis;
Fig. 8 is the photo that Stem of Eucommia ulmoides Oliv dry hide causes bark of eucommia Du stem skin structure destroyed after complex enzyme zymohydrolysis;
Fig. 9 is the gutta-percha photo of bark of eucommia Du's stem dry hide after complex enzyme zymohydrolysis;
Figure 10 is the nmr spectrum of extracted natural rubber.
Embodiment
embodiment 1
A preparation method for gutta-percha, adopt natural Cortex Eucommiae, Folium Eucommiae or Eucommia skin to make, the molecular weight 200000-300000 of gutta-percha, the foreign matter content in gutta-percha is less than 0.5%, and concrete preparation process is as follows:
Step one, choose Cortex Eucommiae, put into clear water and clean, dry, make the water content of Cortex Eucommiae be less than 5%, for subsequent use;
Step 2, employing enzymolysis process carry out pre-treatment
A, preparation enzyme solution: get 0.27% cellulase, 0.01% polygalacturonase and 0.02% proteolytic enzyme by mass percentage respectively and be dissolved in water, be mixed with the complex enzyme liquid of cellulase, polygalacturonase and proteolytic enzyme;
B, the Cortex Eucommiae cleaned up in above-mentioned steps one to be joined by solid-liquid ratio 1:1 in the complex enzyme liquid of the cellulase, polygalacturonase and the proteolytic enzyme that prepare; 50 DEG C, the rotating speed of 500r/min stir under enzymolysis 2 days;
Step 3, filtered by the mixture of the Cortex Eucommiae after step 2 enzymolysis, the slag after filtering is sent into ultrasonic wave mixing tank, and first in ultrasonic wave mixing tank, put into petroleum ether solution, the slag after filtration and the weight ratio of petroleum ether solution are: 1:6; Then carry out ultrasonic, power is 2.4KW, and temperature is 70 DEG C, 30 minutes ultrasonic vibration time, makes the liquid in ultrasonic wave mixing tank form colloid mixed solution;
Dregs and liquor separation device sent into by step 4, colloid mixed solution step 3 formed, carry out classified filtering, its temperature is made to remain on 50 DEG C, first cross 1000 order filter sieve, then be 500 order filter sieve, be then 200 order filter sieve, carry out filtration treatment, deimpurity colloid mixed solution will be gone to send into and to carry glue tank, and reclaim sherwood oil for new bark of eucommia raw material continuation extraction gutta-percha.
Step 5, the temperature carrying glue tank is controlled at 10 DEG C, 1 hour time, the colloid in colloid mixed solution is precipitated completely;
Step 6, propose in glue tank colloidal precipitation after, release the liquid on colloid top layer, take out colloid maybe by carry glue tank greenhouse cooling to 10 DEG C below, take out after making colloid crystallization, i.e. obtained a kind of natural gutta-percha.
embodiment 2
A preparation method for gutta-percha, adopt natural Cortex Eucommiae, Folium Eucommiae or Eucommia skin to make, the molecular weight 200000-300000 of gutta-percha, the foreign matter content in gutta-percha is less than 0.5%, and concrete preparation process is as follows:
Step one, choose Folium Eucommiae, put into clear water and clean, dry, make the water content in Cortex Eucommiae, Folium Eucommiae or seed of bark of eucommia be less than 5%, for subsequent use;
Step 2, employing enzymolysis process carry out pre-treatment
A, preparation enzyme solution: get 0.27% cellulase, 0.01% polygalacturonase and 0.02% proteolytic enzyme by mass percentage respectively and be dissolved in water, be mixed with the complex enzyme liquid of cellulase, polygalacturonase and proteolytic enzyme;
B, the Folium Eucommiae cleaned up in above-mentioned steps one to be joined by solid-liquid ratio 1:2 in the complex enzyme liquid of the cellulase, polygalacturonase and the proteolytic enzyme that prepare; 50 DEG C, the rotating speed of 500r/min stir under enzymolysis 3 days;
Step 3, filtered by the mixture of the Folium Eucommiae after step 2 enzymolysis, the slag after filtering is sent into ultrasonic wave mixing tank, and first in ultrasonic wave mixing tank, put into petroleum ether solution, the slag after filtration and the weight ratio of petroleum ether solution are: 1:8; Then carry out ultrasonic, power is 3.0KW, and temperature is 60 DEG C, 50 minutes ultrasonic vibration time, make the liquid in ultrasonic wave mixing tank form colloid mixed solution;
Dregs and liquor separation device sent into by step 4, colloid mixed solution step 3 formed, carry out classified filtering, its temperature is made to remain on 50 DEG C, first cross 1000 order filter sieve, then be 500 order filter sieve, be then 200 order filter sieve, carry out filtration treatment, deimpurity colloid mixed solution will be gone to send into and to carry glue tank, and reclaim sherwood oil for new bark of eucommia raw material continuation extraction gutta-percha.
Step 5, the temperature carrying glue tank is controlled at 10 DEG C, 7 hours time, the colloid in colloid mixed solution is precipitated completely;
Step 6, propose in glue tank colloidal precipitation after, release the liquid on colloid top layer, take out colloid maybe by carry glue tank greenhouse cooling to 10 DEG C below, take out after making colloid crystallization, i.e. obtained a kind of natural gutta-percha.
embodiment 3
A preparation method for gutta-percha, adopt natural Cortex Eucommiae, Folium Eucommiae or Eucommia skin to make, the molecular weight 200000-300000 of gutta-percha, the foreign matter content in gutta-percha is less than 0.5%, and concrete preparation process is as follows:
Step one, the one chosen in Eucommia skin, put into clear water and clean, dry, make the water content of Eucommia intracutaneous be less than 5%, for subsequent use;
Step 2, employing enzymolysis process carry out pre-treatment
A, preparation enzyme solution: get 0.27% cellulase, 0.01% polygalacturonase and 0.02% proteolytic enzyme by mass percentage respectively and be dissolved in water, be mixed with the complex enzyme liquid of cellulase, polygalacturonase and proteolytic enzyme;
B, the Eucommia skin cleaned up in above-mentioned steps one to be joined by solid-liquid ratio 1:2 in the complex enzyme liquid of the cellulase, polygalacturonase and the proteolytic enzyme that prepare; 50 DEG C, the rotating speed of 500r/min stir under enzymolysis 3 days;
Step 3, filtered by the mixture of the Eucommia skin after step 2 enzymolysis, the slag after filtering is sent into ultrasonic wave mixing tank, and first in ultrasonic wave mixing tank, put into petroleum ether solution, the slag after filtration and the weight ratio of petroleum ether solution are: 1:10; Then carry out ultrasonic, power is 3.6KW, and temperature is 80 DEG C, 60 minutes ultrasonic vibration time, makes the liquid in ultrasonic wave mixing tank form colloid mixed solution;
Dregs and liquor separation device sent into by step 4, colloid mixed solution step 3 formed, carry out classified filtering, its temperature is made to remain on 50 DEG C, first cross 1000 order filter sieve, then be 500 order filter sieve, be then 200 order filter sieve, carry out filtration treatment, deimpurity colloid mixed solution will be gone to send into and to carry glue tank, and reclaim sherwood oil for new bark of eucommia raw material continuation extraction gutta-percha.
Step 5, the temperature carrying glue tank is controlled at 10 DEG C, 12 hours time, the colloid in colloid mixed solution is precipitated completely;
Step 6, propose in glue tank colloidal precipitation after, release the liquid on colloid top layer, take out colloid maybe by carry glue tank greenhouse cooling to 10 DEG C below, take out after making colloid crystallization, i.e. obtained a kind of natural gutta-percha.
natural rubber magnetic resonance detection is tested:
Magnetic resonance detection has been carried out to extracted natural rubber, result is as Fig. 4, HNMR collection of illustrative plates display in Fig. 4, at the peak-to-peak signal l.60ppm located than stronger, this peak is trans-1, the proton peak of 4 structure methyl, 1.67ppm place does not almost have signal, shows hardly containing cis-l in natural Chinese gutta percha, the proton peak on 4 structure methyl, also just illustrate in the natural rubber extracted not containing cis-Isosorbide-5-Nitrae polyisoprene.The signal at 2.03 ppm and 1.96 ppm two places is shown to be the proton peak of two methylene radical.The signal at 5.10 ppm places is proton peak of double bond in Isosorbide-5-Nitrae structure.1 is not had, 2 structures and 3, the signal of H in 4 structures in figure.The long-pending calculating by peak, the anti-form-1 characterized in figure, the trans-Isosorbide-5-Nitrae structural content of 4-polyisoprene is 99.9%, and cis-Isosorbide-5-Nitrae-polyisoprene structure is 0.1%.
Claims (1)
1. a method for enzymolysis and extraction gutta-percha, adopt natural Cortex Eucommiae, Folium Eucommiae or Eucommia skin to make, the molecular weight 200000-300000 of gutta-percha, the foreign matter content in gutta-percha is less than 0.5%, it is characterized in that preparation process is as follows:
Step one, choose the one of Cortex Eucommiae, Folium Eucommiae or Eucommia skin, put into clear water and clean, dry, make the water content of Cortex Eucommiae, Folium Eucommiae or Eucommia intracutaneous be less than 5%, for subsequent use;
Step 2, employing enzymolysis process carry out pre-treatment
A, preparation enzyme solution: get 0.27% cellulase, 0.01% polygalacturonase and 0.02% proteolytic enzyme by mass percentage respectively and be dissolved in water, be mixed with the complex enzyme liquid of cellulase, polygalacturonase and proteolytic enzyme;
B, the Cortex Eucommiae cleaned up in above-mentioned steps one, Folium Eucommiae or Eucommia skin to be joined by solid-liquid ratio 1:1-2 in the complex enzyme liquid of the cellulase, polygalacturonase and the proteolytic enzyme that prepare; 50 DEG C, the rotating speed of 500r/min stir under enzymolysis 2-3 days;
The mixture of step 3, Cortex Eucommiae, Folium Eucommiae or Eucommia skin by after step 2 enzymolysis filters, slag after filtering is sent into ultrasonic wave mixing tank, first in ultrasonic wave mixing tank, put into petroleum ether solution, the slag after filtration and the weight ratio of petroleum ether solution are: 1:6-10; Then carry out ultrasonic, power is 2.4-3.6KW, and temperature is 70 DEG C, ultrasonic vibration time 30-60 minute, makes the liquid in ultrasonic wave mixing tank form colloid mixed solution;
Dregs and liquor separation device sent into by step 4, colloid mixed solution step 3 formed, and makes its temperature remain on 50 DEG C, first crosses 1000 order filter sieve, after 500 order filter sieve, then crosses 200 order filter sieve, and deimpurity colloid mixed solution will be gone to send into separating tank;
Step 5, control at 10 DEG C by the temperature of separating tank, time 1-12 hour, makes the complete crystalline deposit of the colloid in colloid mixed solution;
After colloidal precipitation in step 6, separating tank, release the liquid on colloid top layer, take out colloid maybe by carry glue tank greenhouse cooling to 10 DEG C below, take out after making colloid crystallization, i.e. obtained a kind of natural gutta-percha.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103585421B (en) * | 2013-09-24 | 2016-05-11 | 牛玲 | Bark of eucommia processed product of a kind of liver-kidney tonifying and preparation method thereof |
| CN104288207A (en) * | 2014-10-14 | 2015-01-21 | 南江县百草中药材有限公司 | Method for preparing eucommia ulmoides medicinal slices |
| CN104403113B (en) * | 2014-11-14 | 2020-11-13 | 河南恒瑞源实业有限公司 | Method for extracting gutta-percha from eucommia bark by using enzymatic hydrolysis device |
| CN104356398B (en) * | 2014-11-14 | 2020-11-13 | 河南恒瑞源实业有限公司 | Method for extracting gutta-percha by using full-biological enzyme method |
| CN105566656A (en) * | 2016-01-28 | 2016-05-11 | 张萌萌 | Method for separating residues from eucommia ulmoides gum through enzymolysis |
| CN105832806A (en) * | 2016-03-24 | 2016-08-10 | 浙江旭源杜仲生物科技有限公司 | Production process for eucommia ulmoides leaf extract fine powder oral health-caring tablets |
| CN106864153B (en) * | 2017-01-21 | 2019-08-13 | 云南瑞宝生物科技股份有限公司 | A kind of production method of the colored vein of craftwork |
| CN111826336A (en) * | 2019-04-15 | 2020-10-27 | 北京化工大学 | Method for extracting gutta-percha cells from eucommia ulmoides |
| CN110790946A (en) * | 2019-10-11 | 2020-02-14 | 沈阳化工大学 | Method for extracting gutta-percha by treating eucommia ulmoides nutshells with phosphoric acid/hydrogen peroxide solution |
| CN111011523A (en) * | 2019-12-17 | 2020-04-17 | 顾霆 | Process method for molecular transition and structural recombination |
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