CN106305785B - It is a kind of to improve the composite bacteria agent of content beta-carotene and its application in carrot - Google Patents
It is a kind of to improve the composite bacteria agent of content beta-carotene and its application in carrot Download PDFInfo
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- CN106305785B CN106305785B CN201610585714.2A CN201610585714A CN106305785B CN 106305785 B CN106305785 B CN 106305785B CN 201610585714 A CN201610585714 A CN 201610585714A CN 106305785 B CN106305785 B CN 106305785B
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Abstract
The invention discloses a kind of composite bacteria agents of bata-carotene content in raising carrot and its application, the microbial inoculum to be made of lactobacillus plantarum (Lactobacillus plantarum) P 8 and Lactobacillus casei (Lactobacillus casei) Zhang.Both bacterium have apparent influence to the growth metabolism of carrot, the content for improving bata-carotene in carrot can especially be cooperateed with, to provide ideal raw material for the preparation of natural Beta-carotene, the production cost of bata-carotene is eventually reduced, improves yield and benefit.
Description
Technical field
The present invention relates to a kind of microbial bacterial agent more particularly to a kind of it can significantly improve content beta-carotene in carrot
Microbial inoculum belongs to microorganisms technical field.
Background technology
With the development of economy with the continuous quickening of social progress and rhythm of life, present people are in work, life
The pressure of various aspects is all bigger, and while normal exercise and health care, more and more consumers are also in daily life
It is appropriate to supplement the nutrients.
Beta carotene is one of carotenoid (carotenoids) and crocus fat-soluble compound, it is nature
Most generally existing is also most stable of natural pigment in boundary.In many natural foods for example：Green vegetables, sweet potato, carrot, spinach
Dish, pawpaw, mango etc. all have abundant beta carotene, but exist in most still carrots.Beta carotene is one
Kind antioxidant, has very strong inoxidizability, can prevent, delay and treat certain diseases, especially cancer, while
The immune function of body can be improved.Meanwhile beta carotene has detoxication, is to safeguard the indispensable nutrition of health
Element, prevention of cardiovascular disease, cataract and it is anti-oxidant on have significant function, and prevent in turn more caused by aging and aging
Kind of degenerative disorders, are widely used in food, drug, cosmetics and health products trade at present, be FAO (Food and Agriculture Organization of the United Nation) and
The outstanding nourishing food additive of A classes that the food additives joint specialist committee of the World Health Organization is assert, the whole world is
There are 52 countries, area approval to use, at 1000 tons or more, annual growth has wide demand up to 10%~15% in the world
Wealthy market prospects.
Currently, the method for production beta carotene mainly has plant extraction method, chemical synthesis, microbe fermentation method.Naturally
Carrotene distorts with good anti-dye body, protective effect on cancer risk and stronger physiological activity, and carrotene production in the market
Most of product are composite prepared by chemical synthesis, therefore physiological activity is relatively low.Plant extraction method refers to being rich in from carrot etc.
Beta carotene is extracted in the plant of beta carotene, and it has been main trend that beta carotene is extracted in carrot.But by carrot
The limitation of middle content beta-carotene, the price of current natural Beta-carotene are twice of chemicals, and the cost is relatively high.If energy
The content for enough improving beta carotene in carrot, then can reduce cost, improve yield and benefit.
Invention content
In view of the universal relatively low problem of content beta-carotene in current carrot, the object of the present invention is to provide one kind
Improve the composite bacteria agent of content beta-carotene and its application in carrot.
In order to achieve the object of the present invention, inventor is finally obtained as follows by a large number of experiments research and unremitting effort
Technical solution：
A kind of composite bacteria agent, the microbial inoculum is by lactobacillus plantarum (Lactobacillus plantarum) P-8 and cheese breast bar
Bacterium (Lactobacillus casei) Zhang is formed.
It should be noted that lactobacillus plantarum (Lactobacillus plantarum) P-8 of the present invention, point
In fermenting yoghurt from Inner Mongol traditional natural, there is excellent anti-gastrointestinal tract digestive juice tolerance, can survive in dynamic
In object enteron aisle；The bacterial strain has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and address is China
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, preservation time are on 06 28th, 2012, and preserving number is CGMCC No.6312.
In addition, Lactobacillus casei (Lactobacillus casei) Zhang of the present invention, be detached from koumiss it is resistance to
The probiotics strain of sour and resistance to bile acid；The bacterial strain is preserved in Chinese microorganism strain preservation management committee on April 21st, 2006
Member's meeting General Microbiological Culture collection, preserving number CGMCC No.1697.
Preferably, composite bacteria agent as described above, the microbial inoculum is by lactobacillus plantarum (Lactobacillus plantarum
P-8) and Lactobacillus casei (Lactobacillus casei) Zhang presses 1: the number of viable of (0.5~3) is than composition.
The present inventor has been surprisingly found that when screening inhibits the Antagonistic Fungi of carrot pathogen, contains lactobacillus plantarum
The composite bacteria liquid of (Lactobacillus plantarum) P-8 and Lactobacillus casei (Lactobacillus casei) Zhang
There is no obvious inhibiting effect to pathogen, but have apparent influence to the growth metabolism of carrot, it is particularly possible to improve in carrot
The content of beta carotene.Based on this discovery, the present inventor is by being inoculated in cultivation and fermentation in MRS culture solutions, by both bacterium systems
For at composite bacteria agent.Therefore, the present invention provides the applications of above-mentioned composite bacteria agent, i.e.,：Above-mentioned composite bacteria agent is improving recklessly
Application in radish in content beta-carotene.And application of the above-mentioned composite bacteria agent in preparing Carrot liquid.
Composite bacteria agent of the present invention may be used following method and prepare：
(1) by the lactobacillus casei bacterial strain (Lactobacillus casei) Zhang and lactobacillus plantarum strain
(Lactobacillus plantarum) P-8 is inoculated in MRS culture solutions, is cultivated 12-24 hours under the conditions of 37 DEG C, through centrifugation
After washing, 8%-12% sterilized non-fat cow's milk liquid is added on bacterial sediment, and it is 2.0 × 10 to adjust its bacterium number10Cfu/mL is mixed
Culture is poured into after even；
(2) strain culture that step (1) obtains is inoculated in MRS culture solutions, it is small that 12-24 is cultivated under the conditions of 37 DEG C
When, inoculate the sterilized non-fat cow's milk liquid of 8%-12%, cultivated 6-10 hours under the conditions of 37 DEG C, adjust its bacterium number be 3 ×
109Cfu/mL, pH value are 4.4~4.6.
Compared with prior art, composite bacteria agent of the present invention is by lactobacillus plantarum (Lactobacillus
Plantarum) P-8 and Lactobacillus casei (Lactobacillus casei) Zhang compositions, growth of both bacterium to carrot
Metabolism has apparent influence, it is particularly possible to which collaboration improves the content of beta carotene in carrot, to the system for natural Beta-carotene
It is standby to provide ideal raw material, the production cost of beta carotene is eventually reduced, yield and benefit are improved.
Description of the drawings
Fig. 1 is standard curve, and abscissa is content beta-carotene (μ g/ml), and ordinate is light absorption value.
Specific implementation mode
Form is described in further detail the above of the present invention again by the following examples, but should not manage this
Solution is only limitted to embodiment below for the range of the above-mentioned theme of the present invention, and all technologies realized based on the above of the present invention are equal
Belong to the scope of the present invention.In addition, the experimental method in following embodiments is unless otherwise instructed conventional method.
Embodiment 1：Lactobacillus plantarum (Lactobacillus plantarum P-8) and Lactobacillus casei
The experiment of (Lactobacillus casei) Zhang content beta-carotenes in improving carrot
Test place：Laixi Dian Bu Hou Tun (lactobacillus plantarum group, Lactobacillus casei group, compound bacteria group and blank control group
Each 1.7 mu)
Cultivate administration：Canopy specification long 115m, wide 10m, per 13 ridge of canopy ridging, plantation 2 row of radish on every ridge, spacing in the rows 5-6cm,
Per canopy basal dressing：300 jin of compound potassium sulfate fertilizer (N: P: K 15: 15: 15), 600-700 jins of organic fertilizer, later stage not top dressing.
Experimental period：2016.04.26-2016.6.23
Test crop：(kind is carrot：Mendel 808)
Usage amount and method：After carrot seeding terminates, the microbial bacterial agent (bacterium that experimental group uses is filled with water droplet immediately
Total viable count content is more than 10 in agent9Cfu/mL, the quantity ratio of two kinds of bacterium is 1 in compound bacteria group: 1) 2 liters/canopy, later stage are not dripping
Infected water is filled, and not infected water is irrigated according to crop growing state, ensures that each processing irrigation volume is identical.
Content beta-carotene assay method (high-efficient liquid phase technique)：
Sample pre-treatments：It takes the tissue 4.0g grindings of different carrot different parts to be slurried, is transferred in 100ml conical flasks,
It is separately added into 0.5g vitamin Cs, 30ml ethyl alcohol, 10ml is saturated potassium hydroxide solution, and saponification 30min is shaken in 53 DEG C of water-baths,
Then the extraction and separation repeatedly of 75ml petroleum ethers, extracting solution ether layer is used to be washed with water repeatedly to weakly acidic pH (pH6.0 or more), distillation and concentration
It is used in combination nitrogen to dry up, is then settled to 25ml with 50% chloroform (liquid phase is special), 50% petroleum ether mixed liquor, draw 1ml mistakes
HPLC measurement is carried out after 0.45 μm of filter membrane.
Chromatographic condition：C18 chromatographic columns, mobile phase methanol-acetonitrile-chloroform, Detection wavelength 448nm.
Standard curve is prepared：It weighs beta carotene standard items 10mg (amount weighed when measurement is 10.8mg) and then uses
50% chloroform and 50% petroleum ether mixed liquor constant volume 100ml, are equivalent to 108 μ g/ml, are then diluted to 72,54,36,
21.6,13.5 μ g/ml draw after 1ml crosses 0.45 μm of filter membrane and carry out HPLC measurement.The standard curve of making is as shown in Figure 1.
Calculation formula：
The carotene carotene content that c-- is checked in from mark song, μ g/ml；
25-- sample constant volume total volumes
M-- example weights
1 embodiment 1 of table respectively handles content beta-carotene
Compared with blank control group,▲P ＜ 0.01；Compared with lactobacillus plantarum group,★P ＜ 0.01；With Lactobacillus casei group
Compare,■P ＜ 0.01.
Embodiment 2：Lactobacillus plantarum (Lactobacillus plantarum P-8) and Lactobacillus casei
The experiment of (Lactobacillus casei) Zhang content beta-carotenes in improving carrot
Test place：Laixi Dian Bu Hou Tun, (experimental group and each 1.7 mu of control group)
Cultivate administration：Respectively about 1.7 mu, canopy specification long 115m, wide 10m of experimental group and control group, per 13 ridge of canopy ridging, per ridge
Upper 2 row of plantation radish, spacing in the rows 5-6cm, per canopy basal dressing：300 jin of compound potassium sulfate fertilizer (N: P: K 15: 15: 15), organic fertilizer
600-700 jins, later stage not top dressing.
Experimental period：2016.04.26-2016.6.23
Test crop：(kind is carrot：Promise profit is red)
Usage amount and method：After carrot seeding, filling microbial bacterial agent with water droplet, (the quantity ratio of two kinds of bacterium is 1: 1, living
Bacterium sum content is more than 109Cfu/mL) 2 liters/canopy, later stage no longer trickle irrigation infected water, and not infected water is irrigated according to crop growing state,
Ensure that each processing irrigation volume is identical.
Content beta-carotene assay method (high-efficient liquid phase technique)：With embodiment 1
2 embodiment 2 of table respectively handles content beta-carotene
Compared with blank control group,▲P ＜ 0.01；Compared with lactobacillus plantarum group,★P ＜ 0.01；With Lactobacillus casei group
Compare,■P ＜ 0.01.
By testing result in table 1 it is found that on carrot variety " Mendel 808 ", compared with blank control group, experimental group
Content (the p that the content of beta carotene in carrot, especially compound bacteria group are remarkably improved beta carotene in carrot can be improved
＜ 0.01)；Relative to blank control, compound bacteria group content beta-carotene improves 112.67%.It can by testing result in table 2
Know, on carrot variety " Nuo Lihong ", compared with blank control group, compound bacteria group also significantly increases β-carrot in carrot
The content (p ＜ 0.01) of element；Relative to blank control, compound bacteria group content beta-carotene improves 67.63%.It is worth noting
, either on carrot variety " Nuo Lihong " or on carrot variety " Mendel 808 ", compound bacteria group is to β-Hu Luo
The raising degree for foretelling cellulose content compares the significant difference (p ＜ 0.01) of single bacterium group.
Claims (4)
1. a kind of application of composite bacteria agent in improving carrot in content beta-carotene, the composite bacteria agent is by plant breast bar
Bacterium（Lactobacillus plantarum）P-8 CGMCC No.6312 and Lactobacillus casei（Lactobacillus casei）
Zhang CGMCC No.1697 compositions.
2. application of the composite bacteria agent in improving carrot in content beta-carotene according to claim 1, which is characterized in that
The composite bacteria agent is by lactobacillus plantarum（Lactobacillus plantarum）P-8 CGMCC No.6312 and cheese breast
Bacillus（Lactobacillus casei）Zhang CGMCC No.1697 press 1：（0.5~3）Number of viable than composition.
3. a kind of application of composite bacteria agent in preparing Carrot liquid, the composite bacteria agent is by lactobacillus plantarum
（Lactobacillus plantarum）P-8 CGMCC No.6312 and Lactobacillus casei（Lactobacillus casei）
Zhang CGMCC No.1697 compositions.
4. application of the composite bacteria agent in preparing Carrot liquid according to claim 3, which is characterized in that described answers
Combined bacteria agent is by lactobacillus plantarum（Lactobacillus plantarum）P-8 CGMCC No.6312 and Lactobacillus casei
（Lactobacillus casei）Zhang CGMCC No.1697 press 1：（0.5~3）Number of viable than composition.
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| JP7088694B2 (en) * | 2018-03-06 | 2022-06-21 | イビデン株式会社 | A method for producing a carrot growth promoter and a functional ingredient content improver in carrots, and a carrot growth promoter and a functional ingredient content improver in carrots. |
| CN108795819B (en) * | 2018-07-03 | 2020-08-07 | 杭州园泰生物科技有限公司 | Compound microorganism culture and application thereof in production of carotenoid |
| CN115251328A (en) * | 2022-06-28 | 2022-11-01 | 中华全国供销合作总社济南果品研究所 | Beta-carotene-rich probiotic fermented carrot powder and preparation method thereof |
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| CN104585506A (en) * | 2015-01-26 | 2015-05-06 | 内蒙古农业大学 | Applications of plant lactobacillus planetarium P-8 |
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| US20030175306A1 (en) * | 2002-01-08 | 2003-09-18 | Garner Bryan E. | Compositions and methods for inhibiting pathogenic growth |
| CN101139557A (en) * | 2006-09-08 | 2008-03-12 | 内蒙古农业大学 | Lactobacillus casei and its application for improvement of blood lipid metabolism and immunological regulation |
| CN101333505A (en) * | 2007-06-26 | 2008-12-31 | 内蒙古农业大学 | Applications of lactobacillus casei for antioxidation |
| CN102994422A (en) * | 2012-11-12 | 2013-03-27 | 北京和美科健生物技术有限责任公司 | Application of lactobacillus plantarum P-8 in improvement of alcoholic liver injury |
| CN104585506A (en) * | 2015-01-26 | 2015-05-06 | 内蒙古农业大学 | Applications of plant lactobacillus planetarium P-8 |
Non-Patent Citations (1)
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