CN1714863A - Method for extracting curcin and its use - Google Patents
Method for extracting curcin and its use Download PDFInfo
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- CN1714863A CN1714863A CN 200510021004 CN200510021004A CN1714863A CN 1714863 A CN1714863 A CN 1714863A CN 200510021004 CN200510021004 CN 200510021004 CN 200510021004 A CN200510021004 A CN 200510021004A CN 1714863 A CN1714863 A CN 1714863A
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Abstract
The present invention discloses the use and extraction process of curcin. The extraction process includes two steps, extraction and purification. In the extraction step, raw curcin liquid is prepared through homogenizing, stirring, dialysis, etc.; and in the purification step, raw curcin liquid is chromatographic separated in gel column, and the collected solution, after ultraviolet detection and SDS-PAGE detection, is freeze dried and preserved. The curcin is applied in preparing medicine for inhibiting tumor or resisting fungal virus of plant. When it is used in inhibiting tumor, curcin has powerful effect of inhibiting the growth of stomach cancer cell. When it is used in resisting plant viral diseases, curcin has wide action spectrum to inhibit the growth of several kinds of fungi in crops and forest.
Description
Technical field
The present invention relates to a kind of protein extracting method and uses thereof, particularly method for extracting curcin and uses thereof.
Background technology
Cortex jatrophae resource in the world is extremely abundant, Cortex jatrophae (Jatropha curcas) is Euphorbiaceae (Euphorbiaceae) oilseed plant, abundant Cortex jatrophae resource is arranged in the southwest of China, its oil content is up to 40%, also contain rich in protein in the Cortex jatrophae kernel, experimental results show that the curcin (curcin) that separation obtains from the Cortex jatrophae kernel is a kind of I type ribosome inactivating protein, cytotoxicity is extremely low, to almost not influence of Normocellular growth.
Because ribosome inactivating protein (RIPs) is a kind of cytotoxin, and it is combined with monoclonal antibody, makes immunotoxin (immunotoxic), can improve the specificity of its effect greatly, is used for the single-minded cancerous cell that kills and wounds, and very big potentiality are arranged in treatment of cancer.At the beginning of the sixties, Paul Ehrlich has just proposed with antibody as pharmaceutical carrier, improves the optionally imagination of medicine.Up to the mid-1970s, just there is the people that RIPs and antibody coupling are made the target medicine, show good specificity, but toxicity decreases.Rise the eighties, and people have caused extensive interest to the medicine that deeply presses for the single-minded kill cancer cell of development of cancer research with RIPs and the bonded imagination of monoclonal antibody.Plant ribosome inactivating protein RIPs has the broad-spectrum anti-tumor activity, this may be since tumor cell surface have more phytotoxin receptor or its receptor and toxin combine after easily by endocytosis.1970, Lin etc. reported that ricin and abrin have therapeutical effect to mouse ascites tumor, Yoshida sarcoma, experimental leukemia, Bl melanoma etc.The Lin laboratory directly with ricin and abrin local injection with the treatment human tumor, in many cases, observe tumor regression, have in addition cure fully.But because toxin also has lethal effect to normal cell, so side effect is bigger.
All higher plants all are subjected to the infringement of multiple fungus.Pathogenic fungi often causes a large amount of underproduction of crops, and mycotoxin also causes the severe contamination of food.In recent years, the report of searching and the various antifungal protein of developmental research is increasing in plant, and this is because show tempting prospect gradually in the plant ribosome inactivating protein RIPs plant resistance to fungal disease engineering.
The extract of Cortex jatrophae has stronger inhibitory action to the B.Haptosporus and the B.Ranarum of the mould genus of frog excrement, and this albumen is as the class in the plant antifungal protein family, and up to now, its antifungal activity yet there are no report.Based on curcin development and utilization is widely needed, we have carried out the extracorporeal antifungal activity test to curcin, plan is assessed this proteic antifungal activity, for the realization of next step transgenic engineering of curcin provides experimental basis, for the application on agricultural lays the first stone.
Summary of the invention
Purpose of the present invention promptly is to overcome the shortcoming of prior art, a kind of method for extracting curcin and uses thereof is provided, curcin (curcin) belongs to I type ribosome inactivating protein (RIPs), it stops intracellular protein synthetic as a kind of N-glycosidase, thereby the propagation of cell is suppressed, and curcin may bring into play antitumor action by number of mechanisms.Ribosome inactivating protein RIPs, it specifically hydrolysis ribosomal RNA 3 '-end loop-stem structure the adenine residue and cause the ribosome inactivation, and then Profilin is synthetic, but it does not make the ribosome inactivation of self, only other species ribosome is shown high degree of specificity, this obviously has the function that prevents that the exotic disease substance from infecting, because this proteinoid can be used for the plant resistance to fungal disease.Extraction step of the present invention is simple, can reduce extraction cost, and omnidistance 3～5 ℃, can preserve activity and the fine solubility of toxalbumin, suitable salt ionic concentration and pH value can guarantee the yield of toxalbumin.
Purpose of the present invention is achieved through the following technical solutions: a kind of purposes of curcin, this curcin are used to prepare the medicine that suppresses tumor or are used for plant epiphyte resisting, virus.The extracting method of curcin, it comprises two big steps, slightly carry and purification, slightly carry is that jatropha curcas seed is added in the phosphate buffer, temperature is 3～5 ℃, after homogenate on the plant refiner, stirs 4～6 hours, temperature is 3～5 ℃ and left standstill 20～30 hours, homogenate is got supernatant with the centrifugal 15～30min of 8000～12000g, and supernatant moves into clean container, adding ammonium sulfate solids to final concentration is 60%, left standstill 3～6 hours, and, preserved supernatant with the centrifugal 8～12min of 14000～18000g, precipitation is dissolved in the phosphate buffer of 30-40ml, the supernatant of preserving is continued to add ammonium sulfate solids to final concentration reach 80%,, abandon supernatant with the centrifugal 15～30min of 14000～18000g, collecting precipitation, and be dissolved in the phosphate buffer of 30-40ml.Above-mentioned protein solution is merged, and low temperature dialysis 45～50 hours with the centrifugal 15～30min of 14000～18000g, is abandoned precipitation with bag filter, gets supernatant, and-15～-30 ℃ of preservations are standby, are the toxalbumin crude extract;
Purification is that the toxalbumin crude extract is carried out gel filtration chromatography, last sample front pillar is used earlier the phosphate buffer balance, use same buffer solution elution behind the last sample, elution speed is 12～20ml/h, collect then, the solution of collecting is carried out ultraviolet detection, and SDS-PAGE detects then, and last lyophilization is preserved.
By top narration as can be seen, the present invention has the following advantages:
1, antineoplastic advantage:
● to the strong inhibitory action of stomach cancer cell growth;
● toxicity is low: curcin belongs to I type ribosome inactivating protein, and cytotoxicity is extremely low, to almost not influence of Normocellular growth;
● output height, cost are low: Cortex jatrophae resource in the world is extremely abundant, and endosperm toxalbumin content height can be developed jointly with biodiesel, reduces integrated cost.
2, be used for the plant epiphyte resisting disease,
● action spectrum is wide: under certain concentration range, the growth of the fungal disease of multiple staple crops and forest species is had inhibitory action;
● can with other antimycotic resource coordinating effect, improve action intensity and increase action spectrum;
● the effect specificity is strong, and is little to plant self influence;
● catabolite is nontoxic, and is little to environmental hazard.
3, toxalbumin is extracted advantage
● reduce extraction step, reduced extraction cost
● omnidistance 3～5 ℃, preserved activity and the fine solubility of toxalbumin.
● suitable salt ionic concentration and pH value have guaranteed the yield of toxalbumin.
Description of drawings
Fig. 1 is the inhibition curve chart of different purification phase curcins (curcin) liquid to stomach cancer cell
Fig. 2 is the inhibition curve chart of curcin (curcin) to different tumor cells
Fig. 3 is the inhibitory action sketch map of curcin (curcin) to the pathogenic fungi mycelial growth
The specific embodiment
The invention will be further described below in conjunction with drawings and Examples
Embodiment one:
The extracting method of curcin comprises two big steps, slightly carry and purification, slightly carry is that jatropha curcas seed is added in the phosphate buffer, temperature is 3 ℃, after homogenate on the plant refiner, with magnetic stirrer 4 hours, temperature is 3 ℃ and left standstill 30 hours that homogenate is with the centrifugal 15min of 10 000g, and supernatant moves into clean container, adding ammonium sulfate solids to final concentration is 60%, left standstill 3 hours, and, preserved supernatant with the centrifugal 12min of 15000g, precipitation is dissolved in the phosphate buffer of 30ml, the supernatant of preserving is continued to add ammonium sulfate solids to final concentration reach 80%,, abandon supernatant with the centrifugal 15min of 15000g, collecting precipitation, and be dissolved in the phosphate buffer of 30ml.Above-mentioned protein solution is merged, and low temperature dialysis 45 hours with the centrifugal 30min of 14000g, is abandoned precipitation with bag filter, gets supernatant, and-15 ℃ of preservations are standby, are the toxalbumin crude extract;
Purification is that the toxalbumin crude extract is carried out gel filtration chromatography, and last sample front pillar is used the phosphate buffer balance earlier, uses same buffer solution elution behind the last sample, elution speed is 20ml/h, collects then, and the solution of collecting is carried out ultraviolet detection, SDS-PAGE detects then, and last lyophilizing is preserved.
Above-mentioned ammonium sulfate is faintly acid, and need slowly to add, and neutralize with NaOH (1M), thus when adding ammonium sulfate, need magnetic agitation, and detect the solution pH value at any time with the PH reagent paper, thereby adding ammonium sulfate, decision still drips NaOH (1M).
During dialysis, outer liquid is phosphate buffer, needs magnetic agitation, is interrupted and changes phosphate buffer, stirs 50 hours in 4 ℃.
Discover that the curcin of different purification phase (curcin) is to the suppression ratio difference of stomach cancer cell, experimental result is listed in table 1.Protein crude extract administration can suppress vitro growth of gastric cancer cell and survival as can be seen from Table 1, and increase along with concentration, cell survival rate reduces gradually, and be further purified along with proteinic, protein concentration under the identical suppression ratio also reduces accordingly, show from Fig. 1 electrophoretogram, having only molecular weight at peak II is 28.2kDa albumen one band, can illustrate thus, the composition of the inhibition stomach cancer cell growth effect that has in the barbadosnut protein crude extract is this albumen, just our curcin (curcin) of being purified into.
The different purification phase curcins of table 1 (curcin) are to the influence of the survival rate of stomach cancer cell
| Protein concentration (μ g/ml) | Survival rate % (X ± S) | ||
| Protein crude extract | Peak I | Peak II | |
| 750 187.50 46.88 11.72 2.93 0.73 0.18 | 24.28±7.6 30.61±3.5 39.88±4.7 53.13±2.4 60.99±3.6 70.38±3.5 79.98±6.3 | 39.31±1.2 52.56±4.8 63.72±3.2 69.41±6.4 85.21±5.3 87.44±9.1 94.64±7.8 | 3.33±0.1 11.11±0.6 18.76±1.6 20.68±0.8 31.50±1.8 42.56±3.5 54.53±2.1 |
Because there is linear relationship in the logarithm of cell inhibiting rate and protein concentration, therefore the logarithm each point of cell inhibitory rate and corresponding proteins concentration is carried out rectilinear regression and statistical disposition.With the cell inhibiting rate is vertical coordinate, the logarithm of protein concentration is that abscissa carries out linear mapping, as Fig. 1, according to rectilinear calculate Cortex jatrophae curcin lixiviating solution, peak I, peak II are respectively 14.27 ± 4.4 μ g/ml, 187.80 ± 6.8 μ g/ml, 0.23 ± 0.004 μ g/ml to the IC50 of stomach cancer cell.
Experiment in vitro shows, curcin (curcin) can suppress the growth of murine myeloma cell (Sp2/0), human liver cancer cell (Human hepatoma), growth effect to HeLa cell, normal human embryonic lung diploid fibroblast (MRC) is very little, this can illustrate that curcin do not have a cytotoxicity, similar performance with I type RIPs, also shown curcin to the selective effect of tumor cell, various tumor cells are also different to its sensitivity simultaneously.
Table 2 curcin (curcin) is to the influence of the survival rate of different tumor cells
| Protein concentration (μ g/ml) | Survival rate % (X ± S) | |||
| Human hepatoma | Sp2/0 | HeLa | MRC | |
| 750 187.50 46.88 11.72 2.93 0.73 0.18 | 20.59±4.6 26.81±8.9 36.61±6.3 41.93±1.8 50.84±3.1 58.65±5.5 66.90±1.6 | 6.91±5.5 16.04±1.7 22.98±1.1 31.01±3.5 40.23±0.8 49.86±1.3 58.83±2.7 | 54.67±2.6 69.05±1.1 99.08±1.9 98.23±6.3 98.56±8.4 97.88±2.6 99.55±4.3 | 53.11±2.1 68.88±5.6 99.33±5.3 99.62±1.9 98.77±3.8 98.46±6.7 98.11±4.8 |
With the cell inhibiting rate is vertical coordinate, the logarithm of protein concentration is that abscissa carries out linear mapping, see Fig. 2, calculate Cortex jatrophae curcin according to rectilinear the IC50 of the growth of murine myeloma cell (Sp2/0), human liver cancer cell (Human hepatoma) is respectively 0.66 ± 0.015 μ g/ml, 3.16 ± 0.031 μ g/ml.But the suppression ratio to HeLa cell, normal human embryonic lung diploid fibroblast growth under Cmax 750 μ g/ml is respectively 45.33% and 46.89%.(SGC-7901, Sp2/0 Humanhepatoma) present lethal effect to curcin, but its sensitivity difference to some extent to three kinds of tumor cells.Sensitivity is SGC-7901＞Sp2/0＞Human hepatoma, three kinds of sensitive cells strains of the suppression ratio of different curcin dosage groups and this difference not significantly (p＞0.05) of comparing.We it can also be seen that from figure, rising along with concentration increases curcin to the suppression ratio of tumor cell, and linearly relevant between the concentration logarithm value of curcin and the suppression ratio (the r value is respectively 0.98511,0.99897,0.99871, p＜0.001), illustrates that curcin concentration logarithm value and its suppression ratio are proportionate.We also can see from figure, curcin is very weak to the suppression ratio of HeLa cell, normal human embryonic lung diploid fibroblast growth, and (the r value is respectively 0.80228 not show the concentration logarithm value of curcin and the rectilinear correlation between the suppression ratio, 0.78449, p=0.03), illustrate between curcin concentration logarithm value and its suppression ratio not have dependency.
Embodiment two:
The extracting method of curcin, it comprises two big steps, slightly carries and purification, and slightly carrying is that jatropha curcas seed is added in the phosphate buffer, 5 ℃, after homogenate on the plant refiner, stirred 6 hours, temperature is 5 ℃ and left standstill 30 hours, homogenate is with the centrifugal 15min of 10000g, get supernatant, it is 60% that supernatant adds ammonium sulfate solids to final concentration, leaves standstill 6 hours, with the centrifugal 8min of 15000g, preserve supernatant, precipitation is dissolved in the phosphate buffer of 30ml, the supernatant of preserving is continued to add ammonium sulfate solids to final concentration reach 80%, with the centrifugal 15min of 15000g, abandon supernatant, collecting precipitation, and be dissolved in the phosphate buffer of 30ml.With above-mentioned protein solution dialysis 45 hours, bag filter with the centrifugal 15min of 15000g, is abandoned precipitation, get supernatant ,-15 ℃ of preservations are standby, are the toxalbumin crude extract;
Purification is that the toxalbumin crude extract is carried out gel filtration chromatography, and last sample front pillar is used the phosphate buffer balance earlier, uses same buffer solution elution behind the last sample, elution speed is 12ml/h, collects then, and the solution of collecting is carried out ultraviolet detection, SDS-PAGE detects then, and last lyophilizing is preserved.Above homogenate need be interrupted homogenate.Ammonium sulfate is faintly acid, and need slowly to add, and neutralize with NaOH (1M), thus when adding ammonium sulfate, need magnetic agitation, and detect the solution pH value at any time with the PH reagent paper, thereby adding ammonium sulfate, decision still drips NaOH (1M).During dialysis, outer liquid is phosphate buffer, needs magnetic agitation, is interrupted and changes phosphate buffer, stirs 45 hours in 5 ℃.
As shown in Figure 3, after 7 kinds of tests, find that curcin (curcin) has had strong inhibitory effects to 5 kinds of pathogenic fungi mycelial growths for the examination strain.Minimum inhibitory concentration to rice blast fungus (P.oryzae Cav.), loose pestalotiopsis funerea bacterium (P.funerea), Sclerotinia sclerotiorum [S.sclerotiorum (Lib) de Bary] is 5 μ g/mL.Suppression ratio all can reach 100% when curcin concentration is 100 μ g/mL; To the inhibition effect of corn sheath blight fungus (R.solani Kuha) slightly a little less than, its minimum inhibitory concentration is at 15 μ g/mL, when curcin concentration reached 100 μ g/mL, suppression ratio was 89.2%; To glue born of the same parents anthrax bacillus [C.gloeosporioides (Perz.) sacc.] minimum inhibitory concentration is 5 μ g/mL, when curcin (curcin) concentration is 100 μ g/mL, the growth inhibited of mycelia is reached 80%.
Curcin (curcin) is to the tangible inhibition of being formed with of fungal spore.Experimental result shows, curcin (curcin) is the strongest to the inhibitory action of rice blast fungus (P.oryzae Cav.) Sporulation, in 120h, when curcin (curcin) concentration during greater than 50 μ g/mL, pathogenic fungi can not produce spore basically, and suppression ratio is 100%; Curcin to the inhibition effect of loose pestalotiopsis funerea bacterium (P.funerea) Sporulation slightly a little less than, but curcin (curcin) concentration is during greater than 75 μ g/mL, suppression ratio all can reach 100%; When curcin (curcin) concentration during, in 120 hours, can suppress the formation (Fig.3.4C) of Sclerotinia sclerotiorum [S.sclerotiorum (Lib) de Bary] sclerotium greater than 15 μ g/mL; At viewing duration, the contrast and the treatment samples of corn sheath blight fungus (R.solani Kuha) all do not produce spore; The effect of glue born of the same parents anthrax bacillus [C.gloeosporioides (Perz.) sacc.] shown as when curcin concentration is 15 μ g/mL can suppress Sporulation, suppression ratio reaches 100% when curcin concentration is 75 μ g/mL.
To produce more loose pestalotiopsis funerea bacterium (P.funerea), rice blast fungus (P.oryzae Cav.) and the glue born of the same parents anthrax bacillus [C.gloeosporioides (Perz.) sacc.] of spore is experimental bacteria, measures the inhibitory action of curcin to the pathogenic fungi spore germination.Observe behind 16h and find, curcin is also not obvious to the inhibition effect of these 3 kinds of fungus spore germinations, almost is not affected.
Curcin (curcin) with 50 μ g/mL is handled plant pathogenic fungi.Behind 28 ℃ of cultivation 24h, light microscopic is found loose pestalotiopsis funerea bacterium (P.funerea), corn sheath blight fungus (R.solani Kuha), Sclerotinia sclerotiorum [S.sclerotiorum (Lib) de Bary], unusual, wherein obvious with corn sheath blight fungus (R.solani Kuha) and Pyricularia oryzae (P.oryzae Cav.) with the growthform generation of rice blast fungus (P.oryzae Cav.), glue born of the same parents anthrax bacillus 5 kinds of pathogenic fungi mycelia such as [C.gloeosporioides (Perz.) sacc.] down.Most of mycelia distortion, clustering phenomena takes place in the protoplasm skewness.Mycelia obviously diminishes.
By SDS-PAGE electrophoresis detection curcin (curcin) to the synthetic influence of mycelian protein matter.Corn sheath blight fungus (R.solani Kuha) mycelia after curcin handles has protein band disappearance (line 2) at the 52kD place; Pyricularia oryzae (P.oryzae Cav.) has protein band disappearance (line 4) at 27kD and 40kD place; Pestalotiopsis funerea bacterium (P.funerea) has protein band disappearance (line 6) at about 22kD and 36kD place.Sclerotinia sclerotiorum [S.sclerotiorum (Lib) de Bary] has protein band disappearance (line 7) at the 52kD place after handling through curcin; Glue born of the same parents anthrax bacillus [C.gloeosporioides (Perz.) sacc.] is in 28kD place protein band disappearance (line 9).
Table 3 curcin (Curcin) is to the influence (spore count: * 10 of pathogenic fungi Sporulation
3Individual)
| Concentration (μ g/mL) | Pine pestalotiopsis funerea bacterium (P.funerea) | Pyricularia oryzae (P. oryzae Cav.) | Glue born of the same parents anthrax bacillus [C.gloeosporioides (Perz.) sacc.] | |||
| Spore count | Suppression ratio (%) | Spore count | Suppression ratio (%) | Spore count | Suppression ratio (%) | |
| 0(control) 5 15 25 50 75 100 | 1.85±0.15 1.18±0.08 1.06±0.19 0.83±0.17 0.30±0.10 0 0 | 0 36.2 42.4 55.1 83.8 100 100 | 5.65±0.75 3.15±0.45 3.15±0.25 1.93±0.27 0 0 0 | 0 44.2 44.2 65.8 100 100 100 | 4.33±0.54 4.31±0.41 3.85±0.24 3.45±0.29 1.51±0.24 0 0 | 0 0 11 20 65.4 100 100 |
Claims (5)
1, a kind of purposes of curcin is characterized in that: this curcin is used to prepare the medicine that suppresses tumor or is used for plant antifungal virus.
2, a kind of extracting method of curcin, it is characterized in that: it comprises two big steps, slightly carry and purification, slightly carry is that jatropha curcas seed is added in the phosphate buffer, temperature is 3～5 ℃, after homogenate on the plant refiner, stirs 4～6 hours, temperature is 3～5 ℃ and left standstill 20～30 hours, homogenate is got supernatant with the centrifugal 15～30min of 8000～12000g, and supernatant moves into clean container, adding ammonium sulfate solids to final concentration is 60%, left standstill 3～6 hours, and, preserved supernatant with the centrifugal 8～12min of 14000～18000g, precipitation is dissolved in the phosphate buffer of 30-40ml, the supernatant of preserving is continued to add ammonium sulfate solids to final concentration reach 80%,, abandon supernatant with the centrifugal 15～30min of 14000～18000g, collecting precipitation, and be dissolved in the phosphate buffer of 30-40ml.Above-mentioned protein solution is merged, and low temperature dialysis 45～50 hours with the centrifugal 15～30min of 14000～18000g, is abandoned precipitation with bag filter, gets supernatant, and-15～-30 ℃ of preservations are standby, are the toxalbumin crude extract;
Purification is that the toxalbumin crude extract is carried out gel filtration chromatography, last sample front pillar is used earlier the phosphate buffer balance, use same buffer solution elution behind the last sample, elution speed is 12～20ml/h, collect then, the solution of collecting is carried out ultraviolet detection, and SDS-PAGE detects then, and last lyophilization is preserved.
3, the extracting method of a kind of curcin according to claim 2 is characterized in that: homogenate needs homogenate at low temperatures.
4, the extracting method of a kind of curcin according to claim 2, it is characterized in that: ammonium sulfate is faintly acid, need slowly to add, and neutralize with NaOH (1M), so when adding ammonium sulfate, need stir, and detect the solution pH value at any time, still drip NaOH (1M) thereby decision adds ammonium sulfate with the PH reagent paper.
5, the extracting method of a kind of curcin according to claim 2 is characterized in that: during dialysis, outer liquid is phosphate buffer, need stir, and is interrupted and changes phosphate buffer, stirs 45～50 hours in 3～5 ℃.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100541185C (en) * | 2007-03-15 | 2009-09-16 | 云南神宇新能源有限公司 | The high-resolution separation method of jatropha curcas seed endosperm protein |
| CN101891798A (en) * | 2010-03-02 | 2010-11-24 | 四川大学 | Method for separating and purifying curcin from seeds of jatropha curcas |
| CN102090512A (en) * | 2011-01-10 | 2011-06-15 | 中国海洋石油总公司 | Detoxication method for jatropha curcas and meal thereof |
| CN101463073B (en) * | 2009-01-08 | 2011-08-31 | 中国科学院西双版纳热带植物园 | Continuous extraction method for Jatrohpa curcas toxic protein |
| CN101623319B (en) * | 2009-07-28 | 2011-08-31 | 四川大学 | Application of antiviral ingredients and barbadosnut phenol extracted from barbadosnut branches and leaves for preparing medicine for influenza C virus and herpes simplex virus I and II |
| CN102260337A (en) * | 2010-05-24 | 2011-11-30 | 丹阳市春润生物能源有限公司 | Preparation method of barbadosnut seed pulp toxic protein |
| CN107058261A (en) * | 2017-04-05 | 2017-08-18 | 四川大学 | Curcin from seeds of Jatropha curcas and its isolation and purification method and application |
| CN110301458A (en) * | 2018-03-27 | 2019-10-08 | 汪顺兴 | A kind of Jatropha curcas biological pesticide technology of preparing and it is used for canker of apple fruit Prevention Technique |
| CN110692653A (en) * | 2019-10-30 | 2020-01-17 | 凉山德农生物能源股份有限公司 | Jatropha curcas source environment disinfectant and preparation method thereof |
| CN114774432A (en) * | 2022-05-12 | 2022-07-22 | 四川大学 | Jatropha curcas ribosome inactivating protein JcRIP12, and coding gene and application thereof |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100541185C (en) * | 2007-03-15 | 2009-09-16 | 云南神宇新能源有限公司 | The high-resolution separation method of jatropha curcas seed endosperm protein |
| CN101463073B (en) * | 2009-01-08 | 2011-08-31 | 中国科学院西双版纳热带植物园 | Continuous extraction method for Jatrohpa curcas toxic protein |
| CN101623319B (en) * | 2009-07-28 | 2011-08-31 | 四川大学 | Application of antiviral ingredients and barbadosnut phenol extracted from barbadosnut branches and leaves for preparing medicine for influenza C virus and herpes simplex virus I and II |
| CN101891798B (en) * | 2010-03-02 | 2012-12-19 | 四川大学 | Method for separating and purifying curcin from seeds of jatropha curcas |
| CN101891798A (en) * | 2010-03-02 | 2010-11-24 | 四川大学 | Method for separating and purifying curcin from seeds of jatropha curcas |
| CN102260337A (en) * | 2010-05-24 | 2011-11-30 | 丹阳市春润生物能源有限公司 | Preparation method of barbadosnut seed pulp toxic protein |
| CN102090512A (en) * | 2011-01-10 | 2011-06-15 | 中国海洋石油总公司 | Detoxication method for jatropha curcas and meal thereof |
| CN107058261A (en) * | 2017-04-05 | 2017-08-18 | 四川大学 | Curcin from seeds of Jatropha curcas and its isolation and purification method and application |
| CN107058261B (en) * | 2017-04-05 | 2020-10-02 | 四川大学 | Jatropha curcas ribosome inactivating protein and separation and purification method and application thereof |
| CN110301458A (en) * | 2018-03-27 | 2019-10-08 | 汪顺兴 | A kind of Jatropha curcas biological pesticide technology of preparing and it is used for canker of apple fruit Prevention Technique |
| CN110692653A (en) * | 2019-10-30 | 2020-01-17 | 凉山德农生物能源股份有限公司 | Jatropha curcas source environment disinfectant and preparation method thereof |
| CN114774432A (en) * | 2022-05-12 | 2022-07-22 | 四川大学 | Jatropha curcas ribosome inactivating protein JcRIP12, and coding gene and application thereof |
| CN114774432B (en) * | 2022-05-12 | 2023-09-29 | 四川大学 | Jatropha curcas ribosome inactivating protein JCRIP12, and encoding gene and application thereof |
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