CN1297190C - Superoxide dismutase apple, its prodn. method and special use biological prepn. therefrom - Google Patents
Superoxide dismutase apple, its prodn. method and special use biological prepn. therefrom Download PDFInfo
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- CN1297190C CN1297190C CNB2004100065483A CN200410006548A CN1297190C CN 1297190 C CN1297190 C CN 1297190C CN B2004100065483 A CNB2004100065483 A CN B2004100065483A CN 200410006548 A CN200410006548 A CN 200410006548A CN 1297190 C CN1297190 C CN 1297190C
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Abstract
The present invention discloses a super oxide dimutase (SOD) apple, a production method thereof and a special biological preparation thereof. The active component of the special biological preparation is 3.0*10<10>cfu/g of bacillus CGMCC0175. The super oxide dimutase (SOD) apple contains 20 to 50 IU/g of super oxide dimutase (SOD). The method for producing the super oxide dimutase (SOD) apple comprises the step of spraying the special apple SOD preparation to a tree crown or/and injecting the special apple SOD preparation to a trunk. After the special apple SOD preparation is used, the produced SOD enzyme has stronger heat resistance and stress tolerance capability, and the enzyme activity is 37% of the original enzyme activity. The resistance to ozone, high temperature and high acid is very strong. The SOD apple keeps the favorable performance and shape of the original apple, and the content of SOD is obviously enhanced. The super oxide dimutase (SOD) apple has the functions of enhancing the human body's immunity and delaying senescence. In the deep processing course, higher SOD enzyme activity can still be kept.
Description
Technical field
The present invention relates to a kind of method and special biological agent thereof of producing the apple of high-content biological protective enzyme, particularly relate to the method and the special biological agent thereof of the apple of a kind of production high-load superoxide dismutase (SOD).
Background technology
The SOD enzyme is one of biological important protective enzyme, has enhancing body immunity, anti-aging function.The patent No. is 93103774.3, denomination of invention discloses a bacillus in the Chinese invention patent of " a kind of bacillus and production method thereof that produces superoxide dismutase ", the bacterial strain preserving number is CGMCC 0175, and this bacterial strain has the ability of stronger generation superoxide dismutase.
Apple is the fruit that is loved by the people, and also is few in numberly after the China's WTO to have one of agricultural product of obvious international competitiveness.World's apple gross annual output amount is nearly 6,000 ten thousand tons at present, the apple occupancy volume is about 10 kilograms per capita, and China is 15 kilograms, is higher than 5 kilograms of world's per capita shares, therefore apple is carried out quality improvement, it is very great particularly to make apple have some special medical care effect meanings.
Summary of the invention
The purpose of this invention is to provide the special biological agent that a kind of production has high-load superoxide dismutase (SOD) apple.
The active component of this special biological agent is 3.0 * 10
10The bacillus CGMCC 0175 of cfu/g.
Also contain precipitated calcium carbonate or the peat composed of rotten mosses or the humic acid of described thalline 5-7 times ratio of weight and number in the described preparation, the fineness of the precipitated calcium carbonate or the peat composed of rotten mosses or humic acid is generally 0.08-0.1mm, preferably 0.09mm.
Second purpose of the present invention provides the method for a kind of production high-load superoxide dismutase (SOD) apple.
The method of production superoxide dismutase provided by the present invention (SOD) apple is to the tree crown spraying or/and the special-purpose SOD preparation of trunk injection apple.
In actual applications, described method to the special-purpose SOD preparation of tree crown spraying apple is that the every mu of each special-purpose SOD biologic product of apple 40-60 gram of using is converted water 100-200 kilogram, sprays 5-7 time each 25-30 days at interval altogether., wherein even more preferably every mu each restrain with 50, spray altogether 6 times,
The special-purpose SOD preparation of described trunk injection apple may further comprise the steps: 1) boring, and in the boring of rhizome portion, hole depth reaches heart;
2) injection, the special-purpose SOD preparation of Xiang Kongzhong injection apple, every mu at every turn with the 65-85 gram, is dissolved in the 1-3 kg of water, inject 3-5 time altogether, wherein even more preferably every mu restrain with 75 at every turn, be dissolved in 2 kg of water, Apple with Fruit-bagged Treatment is injected 4 times altogether.
In the said process, boring should be within the 50cm of rhizome portion, and boring and trunk are the angle of 45-80 degree, aperture 5-5.5mm, drilling hole amount 2-3.
The special-purpose SOD preparation of apple is bred in the apple tree body, is shifted by bacillus, improves SOD activity and content in the apple body.
The SOD enzyme that the special-purpose SOD preparation of apple bacterial strain produces has stronger heatproof heat resistance and anti-adversity ability, under 70 ℃ of conditions, handled 1 hour, enzyme live for protoenzyme alive 47%; Handled 30 minutes under 100 ℃ of conditions, enzyme work is 37% of protoenzyme work; Patience to ozone, high temperature peracid is very strong.Superoxide dismutase (SOD) apple of producing with method of the present invention has not only kept the merit of original apple, and SOD content obviously improves, and contains the superoxide dismutase of 20-50IU/g.The soluble solid of apple is more than or equal to 14.7%; Titratable acid is smaller or equal to 0.27g/100ml; Hardness is 12-18kg/cm
2Have the effect that strengthens body immunity and delay senility, in deep-processing process, still can keep higher SOD enzyme to live.
The special-purpose SOD preparation of apple can combine enforcement with other cultivation management measure, do not increase man-hour, simultaneously can mix use, not influence result of use with farming materials such as other insecticide, bactericide (except the bactericide), plant growth regulator, foliage fertilizer, chemical fertilizer.
Embodiment
The production of embodiment 1, the special-purpose SOD biologic product of apple
One, the fermenting and producing of bacterial strain
1, the technical indicator in the production
Sterilising conditions: temperature 121-125 ℃, pressure 0.11-0.14mpa, time 30min
Condition of culture: temperature 28-34 ℃, pressure 0.05mpa, ventilation values 1: 1, pH value 7-7.5
Put a jar index: bacteria containing amount 〉=5,000,000,000/ml, gemma forms 〉=70%, assorted bacterium rate≤0.3%
2, bacterial classification production
From the bacillus CGMCC 0175 former strain inclined plane picking a little in standby slant tube, rule, place 28-30 ℃ of incubator to cultivate 24h, take out and check, confirm no living contaminants, when thalli growth is neat, continue to cultivate 24-48h again, smear, dyeing then, and under oily mirror, observe no living contaminants, when spore is neat, as the bacterial classification of fermenting and producing.
3, fermenting and producing
Fermenting and producing branch seed tank culture and fermentation tank culture two parts carry out, and in time put jar when zymotic fluid reaches set quota.
Two, preparation biologic product
1, adds adsorbent
In the zymotic fluid that obtains, drop into 6 times of precipitated calcium carbonate or the peat composed of rotten mosses or humic acids that the thalline weight fineness is 0.09mm, mechanical agitation 3-6 hour then, make the abundant suppressed by vector absorption of bacillus.
2, press filtration
Adopt open delivery or closed delivery plate to shoulder filter press elimination excessive moisture.
3, drying
Low temperature drying (being no more than 70 ℃) makes moisture content≤5%.
4, pulverize
Pulverize with the ultrafine cracker, fineness requirement is 100% by the 0.09mm aperture sieve, and the purpose of control fineness is to stop up shower nozzle when preventing to use.
5, packing
Embodiment 2, employing tree crown spray-on process are produced the SOD apple
Select for use the apple tree growing way vigorous, the Fuji apple orchard that has entered campaign.
Every mu of each special-purpose SOD biologic product 50 of apple of using restrains, and generally converts water 100-200 kilogram, and mixed dilution is even, looks tree body size, planting density adjustment water consumption.Bagging Fuji sprays 6 times altogether, sprays squaring period 1 time, and the young fruit period spray is 1 time before the bagging, and spray is 3 times behind the bagging, and each 25-30 days at interval, spray was 1 time after bag taking; Sprinkling should reach position dilutions such as blade, trunk, fruit evenly adheres to, generally should be before at 10 in the morning or at 3 in afternoon after spray.Normal cultivation management is carried out in the orchard, and the apple that obtains during results is the superoxide dismutase apple, and the superoxide dismutase of this kind apple is 20-50IU/g, and the soluble solid of apple is more than or equal to 14.7%; Titratable acid is smaller or equal to 0.27g/100ml; Hardness is 12-18kg/cm
2
The method of this embodiment also is applicable to kinds such as new Red Star, corolla,, sprays 6 times at least at interval 25 days at every turn altogether for these kinds in squaring period to picking time.
Embodiment 3, employing trunk injection method are produced the SOD apple
Select for use the apple tree growing way vigorous, the orchard that has entered campaign.
1, boring, within the 50cm of rhizome portion, the hole of brill and trunk angle 45-80 degree, aperture 5-5.5mm, drilling hole amount 2-3, the degree of depth reaches heart.
2. injection, every mu each generally is dissolved in 2 kg of water with special-purpose SOD preparation 75 grams of apple, and Apple with Fruit-bagged Treatment is injected 4 times altogether, and squaring period (by the end of April) 1 time, behind the bagging 3 times, the time is respectively by the end of June, by the end of July, by the end of August.With the special-purpose SOD preparation of the required apple of the individual plant dilution bag that instils of packing into, vertically be hung on about the 1.5 meters high center branches in ground, and discharge ducted gas, then the instillation syringe needle is inserted boring and can inject.Injection finishes, and orifice cuts flat with bark with dried branch jam-pack, and mound protection orifice.Be generally 2-12 hour inject time when fine.During injection, the dissolving in 4 hours at least in advance of the special-purpose SOD preparation of apple.Normal cultivation management is carried out in the orchard, and the apple that obtains during results is the superoxide dismutase apple, and the superoxide dismutase of this kind apple is 20-50IU/g, and the soluble solid of apple is more than or equal to 14.7%; Titratable acid is smaller or equal to 0.27g/100ml; Hardness is 12-18kg/cm
2
The mensuration that embodiment 4, superoxide dismutase apple SOD enzyme of the present invention are lived
One, SOD enzyme mensuration alive
The mensuration that the SOD enzyme is lived adopts the NBT photochemical method
1, the preparation of NBT reactant liquor: preparation pH7.8, the phosphate buffer 1000ml of 50mmol/L, the L-methionine (L-Met) that adds 1.933mg, after treating to dissolve fully, put into 45.8mg chlorination nitrate blue tetrazolium (NBT), 0.47mg vitamin b3 (be made into 0.47mg/ml solution, draw 1ml and add) and 29.0mg ethylenediamine tetra-acetic acid (EDTA), solution final concentration: Met is 13mmol/L; NBT is 6.3 * l0
-3Mmol/L; Vitamin b3 (VB
2) be 1.3 * 10
-3Mmol/L; EDTA is 0.1mmol/L, places brown reagent bottle, with the shading of black cloth cover, can preserve 1 month in 4 ℃ of refrigerators.
2, the preliminary treatment of material
Apple pulp grinds with mortar, and according to pulp: the ratio of 50mM phosphate buffer=1: 2 adds buffer solution, and centrifugal 10 minutes of 10000rpm gets supernatant and carries out the SOD enzyme activity determination.
3, assay method
Draw 30 μ l samples, put into 10ml clear glass small beaker, add the 3mlNBT reactant liquor, mixing is in 28 ℃ of SOD photochemical reaction chambers, 2 * 15W fluorescent lamp illumination 20 minutes (illumination is 4000 luxs), measure absorbance at the 560nm place, make blank, establish 3 repetitions for every group with the NBT reactant liquor, whole mensuration process is all carried out under lucifuge or low light condition except that photochemical reaction.Contrast as test with the standard enzyme of the U.S. " Sigma " company, Bao'an, Shanghai company and the SOD oral liquid of Laolaifu Pharmaceutical Co., Guizhou's production in the test process simultaneously.
4, calculate
SOD enzyme (IU/g)=(OD alive
Contrast-OD
Sample)/(1/2OD
Contrast) * extension rate
Two, superoxide dismutase apple SOD enzyme of the present invention mensuration alive
SOD enzymic activity in the SOD apple that mensuration embodiment 1 and embodiment 2 produce, the result is as shown in table 1, shows that the work of SOD enzyme is significantly higher than common apple in the SOD apple, and remarkable at the a=0.05 level difference.Explanation can be produced by this technology has the SOD health care apple that height contains the work of SOD enzyme.
SOD enzyme activity determination result in table 1, the SOD apple
| Project | Number of repetition | SOD enzyme (IU/g) alive |
| The SOD apple of embodiment 1 | 3 | 22.5B |
| The SOD apple of embodiment 2 | 3 | 35.0A |
| Contrast | 3 | 2.8C |
The heat resistance of embodiment 5, superoxide dismutase of the present invention (SOD) apple SOD is measured
SOD apple sample was kept 20 minutes in 100 ℃, measure the SOD enzymic activity of SOD apple and contrast apple.
Measurement result is as shown in table 2, shows, after Overheating Treatment, still keep higher SOD enzyme to live in the SOD apple, but the SOD enzyme that contrast contains is alive extremely low.The SOD apple of producing by this technology is described, is suitable for preserving at normal temperatures and carrying out deep processing, still can keep higher SOD activity.
The heat resistance of SOD is measured in table 2, the SOD apple
| Project | Number of repetition | SOD enzyme (IU/g) alive |
| The SOD apple of embodiment 1 | 3 | 20.5B |
| The SOD apple of embodiment 2 | 3 | 31.0A |
| Contrast | 3 | 0.5C |
Claims (6)
1, a kind of SOD preparation of producing the special use of superoxide dismutase apple, it is characterized in that: its active component is 2.5 * 10
10-3.5 * 10
10The bacillus CGMCC 0175 of cfu/g; The adsorbent of thalline is precipitated calcium carbonate or the peat composed of rotten mosses or the humic acid of described thalline 5-7 times parts by weight.
2, SOD preparation according to claim 1 is characterized in that: the fineness of the described precipitated calcium carbonate or the peat composed of rotten mosses or humic acid is 0.08-0.1mm.
3, a kind of method of producing the superoxide dismutase apple is characterized in that: be to the special-purpose SOD preparation of tree crown spraying apple or/and the special-purpose SOD preparation of trunk injection apple, the active component of said preparation is 2.5 * 10
10-3.5 * 10
10The bacillus CGMCC 0175 of cfu/g, the adsorbent of thalline are precipitated calcium carbonate or the peat composed of rotten mosses or the humic acids of described thalline 5-7 times parts by weight.
4, method according to claim 3 is characterized in that: described method to the special-purpose SOD preparation of tree crown spraying apple is that the every mu of each special-purpose SOD preparation of apple 40-60 gram of using is converted water 100-200 kilogram, sprays 5-7 time each 25-30 days at interval altogether.
5, method according to claim 3 is characterized in that: the special-purpose SOD preparation of described trunk injection apple may further comprise the steps: 1) boring, and in the boring of rhizome portion, hole depth reaches heart;
2) injection, the special-purpose SOD preparation of Xiang Kongzhong injection apple, every mu restrains with 65-85 at every turn, is dissolved in the 1-3 kg of water, and Apple with Fruit-bagged Treatment is injected 3-5 time altogether.
6, method according to claim 5 is characterized in that: described boring is within the 50cm of rhizome portion, and boring and trunk are the angle of 45-80 degree, aperture 5-5.5mm, drilling hole amount 2-3.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100065483A CN1297190C (en) | 2004-03-08 | 2004-03-08 | Superoxide dismutase apple, its prodn. method and special use biological prepn. therefrom |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100065483A CN1297190C (en) | 2004-03-08 | 2004-03-08 | Superoxide dismutase apple, its prodn. method and special use biological prepn. therefrom |
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|---|---|
| CN1559196A CN1559196A (en) | 2005-01-05 |
| CN1297190C true CN1297190C (en) | 2007-01-31 |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100415081C (en) * | 2005-12-28 | 2008-09-03 | 中国农业大学 | SOD-enriched winter date and its production method |
| CN101790952B (en) * | 2010-03-25 | 2012-01-25 | 砀山县科技创业服务中心 | Method for producing superoxide dismutase Dangshan pear |
| CN103461057A (en) * | 2013-09-29 | 2013-12-25 | 大连金州后石实业总公司 | Production method for SOD (superoxide dismutase) apples |
| CN104686270A (en) * | 2015-02-03 | 2015-06-10 | 新疆天物生态科技股份有限公司 | Production method for Korla bergamot pears with SOD enzymatic activity |
| CN105940895A (en) * | 2016-04-27 | 2016-09-21 | 河南省林业科学研究院 | SOD (Superoxide Dismutase) Diospyros kaki fruits and production method therefor |
| CN108782132B (en) * | 2018-06-20 | 2022-03-18 | 五月阳光生物科技(浙江)有限公司 | Cultivation method of fruit rich in SOD |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1082605A (en) * | 1993-04-21 | 1994-02-23 | 北京农业大学 | A kind of genus bacillus and production method thereof that produces superoxide-dismutase |
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2004
- 2004-03-08 CN CNB2004100065483A patent/CN1297190C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1082605A (en) * | 1993-04-21 | 1994-02-23 | 北京农业大学 | A kind of genus bacillus and production method thereof that produces superoxide-dismutase |
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