CN108713558A - The growth-promoting preparation of general flavone content in a kind of promotion ginkgo leaf - Google Patents
The growth-promoting preparation of general flavone content in a kind of promotion ginkgo leaf Download PDFInfo
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Abstract
The present invention relates to biotechnology, specifically a kind of growth-promoting preparation for promoting general flavone content in ginkgo leaf is made of the raw material of following parts by weight：15.0-20.0 parts of 1.5-3.5 parts of bacillus licheniformis, 5.6-7.8 parts of bacillus subtilis, 4.1-6.3 parts of bacillus pumilus, 2.5-5.7 parts of Burkholderia and distilled water.The invention further relates to a kind of preparation methods of the growth-promoting preparation of general flavone content in promotion ginkgo leaf, include the following steps：(1) distilled water is added according to the ratio into container, at room temperature sealing a period of time；(2) at room temperature, bacillus licheniformis, bacillus subtilis, bacillus pumilus and Burkholderia are then added according to the ratio successively, concussion is uniform, you can.Growth-promoting preparation through the invention, general flavone content is promoted by original 0.85% to 0.98% or so in ginkgo leaf, greatly improves economic value and medical value.
Description
Technical field
The present invention relates to biotechnology, specifically a kind of growth-promoting preparation for promoting general flavone content in ginkgo leaf.
The invention further relates to a kind of preparation method and applications of the growth-promoting preparation of general flavone content in promotion ginkgo leaf.
Background technology
GINKGO BILOBA EXTRACT, that is, ginkgo biloba p.e, it can increase cerebrovascular blood flow, improve blood brain vascular circulation work(
Can, protect brain cell, coronary artery dilator, pre- angina pectoris and myocardial infarction, antithrombotic to improve immunity of organism energy
Power, it is highly beneficial per capita to coronary heart diseases and angina pectoris, cerebral arteriovenous malformation, senile dementia, high blood pressure, have very high medicinal
Value.Therefore, how to promote general flavone content in ginkgo leaf is our urgent problems.
Invention content
Technical problem to be solved by the present invention lies in overcome the deficiencies of the prior art and provide in a kind of promotion ginkgo leaf
The growth-promoting preparation of general flavone content.
In order to solve the above technical problems, the present invention now proposes following technical scheme：General flavone contains in a kind of promotion ginkgo leaf
The growth-promoting preparation of amount, is made of the raw material of following parts by weight：1.5-3.5 parts of bacillus licheniformis, bacillus subtilis 5.6-7.8
15.0-20.0 parts of part, 4.1-6.3 parts of bacillus pumilus, 2.5-5.7 parts of Burkholderia and distilled water.
Further, it is made of the raw material of following parts by weight：2.5 parts of bacillus licheniformis, 6.6 parts of bacillus subtilis,
18 parts of 5.2 parts of bacillus pumilus, 4.1 parts of Burkholderia and distilled water.
Further, the Bacillus licheniformis falls that edge is irregular, and rough surface is creamy white；Thalline rod-short,
It is arranged in heap, it is raw in gemma.The bacterium is used for detaching licoflavone element, and the preparation method of the bacillus licheniformis is：8% inoculation
The bacillus licheniformis is stirring while adding in fluid nutrient medium, after addition, controls certain speed of agitator, aerobic
Culture keeps certain temperature, fermentation pH to be not less than 6.5.
Further, the Liquid Culture based component is：0.5 weight % of starch, 0.5 weight % of urea, 1 weight of bean cake powder
Measure %, 0.6 weight % of dipotassium hydrogen phosphate, 0.3 weight % of potassium dihydrogen phosphate, 0.05 weight % of yeast extract, 0.1 weight of magnesium sulfate
Measure %, 0.02 weight % of manganese sulfate.
Further, the bacillus subtilis colony edge is neat, and surface is smooth, and protuberance is creamy white, thalline quarter butt
Shape arranges, central spore in heap.For detaching licoflavone element and similar metabolin, the preparation side of the bacillus subtilis
Method is：Include the following steps：(1)) Bacillus subtilis strain is inoculated on slant medium, carries out inclined-plane culture；(2) tiltedly
Bacillus subtilis after the culture of face is inoculated on seed culture medium, carries out seed culture；(3) by the seed liquor through seed culture
Be inoculated on fermentation medium, carry out fermented and cultured, since fermentation by 5~6 hours not control ph, at 5~6 hours to 8
Between 6.0-6.5, pH is controlled between 7.0-7.5 to fermentation ends after 8 hours, obtains withered grass bud for pH value control between hour
Spore bacillus product, viable count 200-400cfu/ml in producing bacillus subtilis product, spore forming rate 87~92%.
Further, the slant medium includes following components：Peptone 10g/L, yeast extract powder 5g/L, grape
Sugared 5g/L, sodium chloride 10g/L, agar 20g/L.
Further, the bacillus pumilus colony edge is wavy, and surface is smooth, and concave mesa-shaped is creamy white, semi-transparent
It is bright；Thalline thin rod shape, fence type arrangement.The bacterium is used for detaching licoflavone element, and the preparation method of the bacillus pumilus is：
Include the following steps：(1) actication of culture, bacillus pumilus preserve bacterium and line on plating medium, and 37 DEG C of cultures are for 24 hours；(2)
Strain fermentation culture enters the strain transfer after activation in 10ml seed fermentation culture mediums, 37 DEG C of culture 48h；(3) fermentation is trained
The strain supported is inoculated in 1000ml triangular flasks, 37 DEG C of culture 48h；(4) 10L kinds will be inoculated in the cultured strain of triangular flask
Sub- fermentation tank, 37 DEG C of cultures 44h, rotating speed 150r/min, ventilatory capacity 0.4vvm.
Further, step (1) the middle plateform culture medium be by 5g peptones, 1g yeast extracts, 0.1g high ferric phosphates,
20g agar, appropriate seawater are configured through 121 DEG C of sterilizing 30min, pH 7.6.
Further, the Burkholderia colony edge is neat, and surface is smooth, and protuberance is creamy white, bacterium
Body rod-short arranges, central spore in heap.The bacterium is used for detaching licoflavone element, first isolated secondary metabolite.
It is another object of the present invention to additionally provide a kind of preparation promoting the growth-promoting preparation of general flavone content in ginkgo leaf
Method includes the following steps：(1) distilled water is added according to the ratio into container, at room temperature sealing a period of time；(2) at room temperature,
Then bacillus licheniformis, bacillus subtilis, bacillus pumilus and Burkholderia are added according to the ratio successively,
Concussion is uniform, you can.
Another object of the present invention is application of the growth-promoting preparation in promoting ginkgo leaf in general flavone content.
Compared with prior art, the beneficial effects of the present invention are：1, growth-promoting preparation through the invention, educates in ginkgo
It is applied on root or blade during seedling or cultivation, the general flavone content in ginkgo leaf is promoted to increase general flavone in ginkgo leaf
Content is promoted by original 0.85% to 0.98%, greatly improves economic value and medical value；
2, growth-promoting preparation production method of the invention is simple, highly practical.
Specific implementation mode
, there are further understanding and understanding in the effect of to make to structure feature of the invention and being reached, to preferable
Embodiment is described in detail, and is described as follows：
Embodiment 1
The growth-promoting preparation of general flavone content, is made of the raw material of following parts by weight in a kind of promotion ginkgo leaf：Lichens gemma
1.5 parts of bacillus, 5.6 parts of bacillus subtilis, 4.1 parts of bacillus pumilus, 2.5 parts of Burkholderia and distilled water
15.0 parts.
Embodiment 2
The growth-promoting preparation of general flavone content, is made of the raw material of following parts by weight in a kind of promotion ginkgo leaf：Lichens gemma
2.5 parts of bacillus, 6.6 parts of bacillus subtilis, 5.2 parts of bacillus pumilus, 4.1 parts of Burkholderia and distilled water
18 parts.
Embodiment 3
The growth-promoting preparation of general flavone content, is made of the raw material of following parts by weight in a kind of promotion ginkgo leaf：Lichens gemma
3.5 parts of bacillus, 7.8 parts of bacillus subtilis, 6.3 parts of bacillus pumilus, 5.7 parts of Burkholderia and distilled water
20.0 parts.
Embodiment 4
The preparation method of the growth-promoting preparation of general flavone content, includes the following steps in a kind of promotion ginkgo leaf：(1) to container
In add distilled water according to the ratio, at room temperature sealing a period of time；(2) at room temperature, lichens gemma is then added according to the ratio successively
Bacillus, bacillus subtilis, bacillus pumilus and Burkholderia, concussion are uniform, you can.
Embodiment 5
The preparation method of bacillus subtilis is：Include the following steps：(1)) Bacillus subtilis strain is inoculated in tiltedly
On the culture medium of face, inclined-plane culture is carried out；(2) bacillus subtilis after inclined-plane culture is inoculated on seed culture medium, is planted
Son culture；(3) seed liquor through seed culture is inoculated on fermentation medium, carries out fermented and cultured, it is small to 6 since fermentation
When not control ph, between 6 hours to 8 hours pH value control between 6.3, after 8 hours pH controls between 7.2 to fermenting
Terminate, obtains producing bacillus subtilis product, viable count 300cfu/ml in producing bacillus subtilis product, spore forming rate 90%.
Further, in the present embodiment, slant medium includes following components：Peptone 10g/L, yeast extract powder 5g/
L, glucose 5g/L, sodium chloride 10g/L, agar 20g/L.
Embodiment 6
The preparation method of bacillus pumilus is：Include the following steps：(1) actication of culture, bacillus pumilus preserve bacterium
It lines on plating medium, 37 DEG C of cultures are for 24 hours；(2) strain transfer after activation is entered 10ml seeds hair by strain fermentation culture
In ferment culture medium, 37 DEG C of culture 48h；(3) the good strain of fermented and cultured is inoculated in 1000ml triangular flasks, 37 DEG C of culture 48h；
(4) 10L seed fermentation tanks, 37 DEG C of cultures 44h, rotating speed 150r/min, ventilation will be inoculated in the cultured strain of triangular flask
Amount is 0.4vvm.
Further, in the present embodiment, plating medium is by 5g peptones, 1g yeast extracts, 0.1g high ferric phosphates, 20g
Agar, appropriate seawater are configured through 121 DEG C of sterilizing 30min, pH 7.6.
Test example
By test example as can be seen that each ingredient of the growth-promoting preparation of the present invention has promotion to make general flavone content
With, but it is more preferable to combine effect after the promotion preparation for forming the present invention, and can obtain each ingredient of embodiment 2
Proportioning facilitation is best, and general flavone content can reach 0.98%.
Finally it should be noted that：The above embodiments are merely illustrative of the technical solutions of the present invention, rather than to limit it；
Although the present invention is described in detail referring to the foregoing embodiments, it will be understood by those of skill in the art that it still may be used
To modify to the technical solution that previous embodiment is recorded or equivalent replacement of some of the technical features；And this
A little modifications or substitutions, the spirit and scope for various embodiments of the present invention technical solution that it does not separate the essence of the corresponding technical solution.
Claims (10)
1. a kind of growth-promoting preparation promoting general flavone content in ginkgo leaf, which is characterized in that be made of the raw material of following parts by weight：
1.5-3.5 parts of bacillus licheniformis, 5.6-7.8 parts of bacillus subtilis, 4.1-6.3 parts of bacillus pumilus, onion Bai Kehuo
15.0-20.0 parts of 2.5-5.7 parts of moral Salmonella of that and distilled water.
2. it is according to claim 1 it is a kind of promoted ginkgo leaf in general flavone content growth-promoting preparation, which is characterized in that by with
The raw material of lower parts by weight forms：2.5 parts of bacillus licheniformis, 6.6 parts of bacillus subtilis, 5.2 parts of bacillus pumilus, onion
18 parts of 4.1 parts of bulkholderia cepasea and distilled water.
3. a kind of growth-promoting preparation promoting general flavone content in ginkgo leaf according to claim 1, which is characterized in that described
The preparation method of bacillus licheniformis is：The 8% inoculation bacillus licheniformis is stirring while adding in fluid nutrient medium, adds
After adding, certain speed of agitator is controlled, aerobic culture keeps certain temperature, fermentation pH to be not less than 6.5.
4. a kind of growth-promoting preparation promoting general flavone content in ginkgo leaf according to claim 3, which is characterized in that described
Liquid Culture based component is：0.5 weight % of starch, 0.5 weight % of urea, 1 weight % of bean cake powder, 0.6 weight of dipotassium hydrogen phosphate
Measure %, 0.3 weight % of potassium dihydrogen phosphate, 0.05 weight % of yeast extract, 0.1 weight % of magnesium sulfate, 0.02 weight % of manganese sulfate.
5. a kind of growth-promoting preparation promoting general flavone content in ginkgo leaf according to claim 1, which is characterized in that described
The preparation method of bacillus subtilis is：Include the following steps：(1)) Bacillus subtilis strain is inoculated in slant medium
On, carry out inclined-plane culture；(2) bacillus subtilis after inclined-plane culture is inoculated on seed culture medium, carries out seed culture；
(3) seed liquor through seed culture is inoculated on fermentation medium, carries out fermented and cultured, since fermentation not by 5~6 hours
Control ph, between 5~6 hours to 8 hours pH value control between 6.0-6.5, after 8 hours pH control 7.0-7.5 it
Between to fermentation ends, obtain producing bacillus subtilis product, viable count 200-400cfu/ml in producing bacillus subtilis product, gemma
Formation rate 87~92%.
6. a kind of growth-promoting preparation promoting general flavone content in ginkgo leaf according to claim 5, which is characterized in that described
Slant medium include following components：Peptone 10g/L, yeast extract powder 5g/L, glucose 5g/L, sodium chloride 10g/L, fine jade
Fat 20g/L.
7. a kind of growth-promoting preparation promoting general flavone content in ginkgo leaf according to claim 1, which is characterized in that described
The preparation method of bacillus pumilus is：Include the following steps：(1) actication of culture, bacillus pumilus preservation bacterium line flat
On plate culture medium, 37 DEG C of cultures are for 24 hours；(2) strain transfer after activation is entered 10ml seed fermentation culture mediums by strain fermentation culture
In, 37 DEG C of culture 48h；(3) the good strain of fermented and cultured is inoculated in 1000ml triangular flasks, 37 DEG C of culture 48h；It (4) will be three
The angle cultured strain of bottle is inoculated in 10L seed fermentation tanks, and 37 DEG C are cultivated 44h, rotating speed 150r/min, and ventilatory capacity is
0.4vvm。
8. a kind of growth-promoting preparation promoting general flavone content in ginkgo leaf according to claim 7, which is characterized in that described
Step (1) middle plateform culture medium is by 5g peptones, and 1g yeast extracts, 0.1g high ferric phosphates, 20g agar, appropriate seawater is through 121 DEG C
Sterilizing 30min is configured, pH 7.6.
9. a kind of preparation method of growth-promoting preparation described in claim 1, which is characterized in that include the following steps：(1) to container
In add distilled water according to the ratio, at room temperature sealing a period of time；(2) at room temperature, lichens gemma is then added according to the ratio successively
Bacillus, bacillus subtilis, bacillus pumilus and Burkholderia, concussion are uniform, you can.
10. a kind of application of growth-promoting preparation described in claim 1 in promoting ginkgo leaf in general flavone content.
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Cited By (4)
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| CN111363701A (en) * | 2020-03-18 | 2020-07-03 | 泸州正泰生物工程有限公司 | Heat-resistant bacillus licheniformis and application thereof |
| CN112136965A (en) * | 2020-09-08 | 2020-12-29 | 河南惠通天下生物工程有限公司 | Immunity-enhancing and growth-promoting fermented Chinese herbal medicine feed additive and preparation method thereof |
| CN112646731A (en) * | 2020-12-24 | 2021-04-13 | 湖南科技学院 | Growth promoting preparation for increasing content of total flavonoids in ginkgo leaves and preparation method and application thereof |
| CN114410550A (en) * | 2022-03-15 | 2022-04-29 | 中南大学 | Microbial functional flora for increasing flavonoid glycoside content in ginkgo leaves and preparation and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112646731A (en) * | 2020-12-24 | 2021-04-13 | 湖南科技学院 | Growth promoting preparation for increasing content of total flavonoids in ginkgo leaves and preparation method and application thereof |
| CN114410550A (en) * | 2022-03-15 | 2022-04-29 | 中南大学 | Microbial functional flora for increasing flavonoid glycoside content in ginkgo leaves and preparation and application thereof |
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Application publication date: 20181030 Assignee: Xintian County Tianyuan Chunxiao Ecological Agriculture Co.,Ltd. Assignor: HUNAN University OF SCIENCE AND ENGINEERING Contract record no.: X2023980036440 Denomination of invention: A growth promoting agent for increasing the total flavonoid content in Ginkgo biloba leaves Granted publication date: 20200901 License type: Common License Record date: 20230612 |
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