CN108624507A - A kind of chlorella W7 that can generate astaxanthin and its application - Google Patents
A kind of chlorella W7 that can generate astaxanthin and its application Download PDFInfo
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- CN108624507A CN108624507A CN201810480849.1A CN201810480849A CN108624507A CN 108624507 A CN108624507 A CN 108624507A CN 201810480849 A CN201810480849 A CN 201810480849A CN 108624507 A CN108624507 A CN 108624507A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12N1/125—Unicellular algae isolates
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- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
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Abstract
The present invention relates to a kind of chlorella that can generate astaxanthin, the chlorella is preserved in China typical culture collection center on December 25th, 2017, and preserving number is CCTCC NO:M2017837；Further relate to application of the above-mentioned chlorella in astaxanthin preparation；Further relate to the cultural method of above-mentioned chlorella.The chlorella W7 speeds of growth of the present invention are fast, and the glucose that can be absorbed while photoautotrophy is grown in environment carries out autotrophy+heterotrophism mixed growth.Under inductive condition, chlorella W7 of the invention can largely generate astaxanthin and lipid, thus, astaxanthin and algae oil can be extracted from frustule, or directly acquire algae powder for food addition or feed, generate high economic benefit.
Description
Technical field
The present invention relates to the production fields of high added value nutrients, more specifically it relates to which a kind of can generate the small of astaxanthin
Ball algae and its application.
Background technology
Carotenoid type is various, has identified more than 600 Carotenoids.In this numerous carotenoid
In, astaxanthin (astaxanthin) is increasingly paid close attention to by researchers due to its anti-oxidation characteristics.The chemical name of astaxanthin
Referred to as 3,3 '-dihydroxy -4,4 '-diketone-β, β '-carrotene, molecular formula：C40H52O4, molecular weight 596.86, because of two terminal hydroxy groups
Optically-active sex differernce has 3 kinds of isomers, and there are a conjugated polyene structures in molecular structure, and the both ends of ring are respectively containing there are one oxygen
Functional group, therefore it is classified as lutein race.
Some researches show that the ability of astaxanthin quenching activity oxygen radical is 2-10 times stronger than beta carotene, compares vitamin E
It is 80-550 times strong, with extremely strong inoxidizability, phosphatide on biomembrane and other lipid molecules can be protected not oxidized,
Protecting effect is good more than beta carotene and lutein, so this antioxygenic property of astaxanthin can be in the anti-of many diseases
It can play a crucial role during controlling.
Astaxanthin is principally found in marine animal such as shrimp, crab, fish, is main carotenoid in marine organisms body
One of.However, content astaxanthin is not high in animal body, and the culture-cycle is long, is not easy to extract.It is therefore desirable to screen shrimp
Green cellulose content is high, and is easy to the microalgae of culture to produce astaxanthin.
Invention content
Inventor obtains the one plant of algae that can generate astaxanthin strain from field acquisition algae sample through culture of isolated.Its cell at
Circle, green, atrichia have core, and the generation that can induce astaxanthin is cultivated under the conditions of nitrogen stress.According to cellular morphology and life
Habit and 18S rDNA sequencing displays, belong to Chlorophyta, Chlorella.
It is found based on above, the present invention provides a kind of chlorellas that can generate astaxanthin, and the chlorella was in 2017
It was preserved in China typical culture collection center on December 25, preserving number is CCTCC NO:M2017837.
The present invention also provides application of the above-mentioned chlorella in astaxanthin preparation.
The present invention also provides a kind of methods preparing astaxanthin, include the following steps：
S1：Above-mentioned chlorella is cultivated, chlorella culture is obtained；
S2：Astaxanthin is extracted from the chlorella culture.
In a preferred embodiment, S1 includes the following steps：
S11：The chlorella is cultivated in full culture medium, accumulates biomass；
S12：Collect the chlorella algae cell of S11 cultures；
S13：The chlorella algae cell is forwarded in the culture medium of nitrogen stress and is cultivated, the chlorella culture is obtained.
In a preferred embodiment, the full culture medium described in S11 is BG11 culture mediums.
In a preferred embodiment, the also glucose containing 1g/L in the BG11 culture mediums.Adding glucose can
Make micro algae growth faster.
In a preferred embodiment, the culture environment of the chlorella is：22-26 DEG C of temperature, illumination 60-80 μ E,
Incubation time is 5-7 days.
In a preferred embodiment, in S13, culture environment is：28 DEG C of temperature, illumination 150-200 μ E, incubation time
It is 7-9 days.It is highly preferred that the culture medium of this step is the BG11 culture mediums containing 30g/L glucose.It is shone by nitrogen stress, bloom
Oxdative stress can be generated with the glucose that high concentration is added, astaxanthin is generated to induce in chlorella W7.
The chlorella W7 speeds of growth of the present invention are fast, and the glucose in environment can be absorbed while photoautotrophy is grown
Carry out autotrophy+heterotrophism mixed growth.Under inductive condition, chlorella W7 of the invention can largely generate astaxanthin and lipid, because
And astaxanthin and algae oil can be extracted from frustule, or algae powder is directly acquired for food addition or feed, it generates high economical
Benefit.
Microbial preservation
The water sample of the microalgae being related to from field acquisition of the present invention is isolated, reflects through 18S rDNA sequencings and morphology
Fixed, which belongs to Chlorophyta Chlorella.The microalgae has been preserved in Wuhan City of Hubei China province force on December 25th, 2017
The China typical culture collection center (CCTCC) of prosperous No. 299 Wuhan Universitys of area's Bayi Road, preserving number are CCTCC NO:
M2017837, is named as chlorella W7, and Classification system is Chlorella sp.W7.
Description of the drawings
Fig. 1 is the light micrograph after the chlorella W7 of the present invention is cultivated 7 days in the BG11 culture mediums containing 1g/L；
Fig. 2 is the light micrograph after chlorella W7 is cultivated 7 days in the nitrogen stress BG11 culture mediums containing 30g/L；
Fig. 3 is the growth curve that W7 is cultivated in the BG11 culture mediums containing 1g/L.
Specific implementation mode
Principles and features of the present invention are described below in conjunction with example, the given examples are served only to explain the present invention, and
It is non-to be used to limit the scope of the present invention.
1. the acquisition and preservation of chlorella W7
The chlorella W7 (Chlorella sp.W7) of the present invention is detached by the algae-containing water sample acquired from Wuhan City East Lake
It obtains, for cellular morphology as shown in Figure 1, under normal condition, cell is spherical in shape, and yellow green, atrichia has core.Through 18S rDNA
Sequencing identification, the microalgae belong to Chlorophyta Chlorella.After environmental induction, which largely generates astaxanthin, makes frustule
In yellow, and cell size before induction than increasing 5-10 times.The microalgae is preserved in Chinese lake on December 25th, 2017
The China typical culture collection center (CCTCC) of Wuhan University of the Wuhan City Bei Sheng, preserving number are CCTCC NO:M2017837,
It is named as chlorella W7, Classification system Chlorella sp.W7.
2. the culture of chlorella W7
Grope experiment display according to conditions such as temperature, illumination, the preference temperature for being conducive to chlorella W7 growths is 22-26
DEG C, it is 60-80 μ E suitable for illumination.
Chlorella W7 is seeded to equipped in BG11 (containing 1g/L glucose) triangular flask, illumination is carried out at 24 ± 1 DEG C
It cultivates, in oscillation (100rpm/min) or 2-6%CO on shaking table in incubation2Ventilation.Test frustule number daily, as a result such as
Shown in Fig. 3, chlorella W7 reaches maximum algae cell density for 5-7 days in
BG11 culture mediums are prepared by the following method：Take Stock II, Stock III, Stock IV, Stock
VI, Stock VII, each 1ml of Stock VIII, add 1.5g NaNO3, water is added to be settled to 1000ml.Each liquid storage ingredient such as 1 institute of table
Show, the ingredient of A5 is as shown in table 2.
The ingredient of each liquid storage during 1 BG11 culture mediums of table are prepared
2 A5 ingredients of table
2. astaxanthin induction culture
The chlorella W7 medium centrifugals cultivated through the above steps are collected into frustule, are then forwarded in equal volume
In new triangular flask, culture 7-9 days is carried out in 28 DEG C, light intensity is 150-200 μ E, and used medium is that nitrogen stress BG11 (contains 30g/L
Glucose), that is, other compositions are constant, glucose content 30g/L, and nitrogen content is less than half of BG11.The results show that working as nitrogen
When content is the 1/2-1/5 of BG11, content astaxanthin accounts for dry weight 1.1-2.3%, is secondly canthaxanthin, lipid content up to 40~
60%.When nitrogen content is less than 1/5, cell state is bad, and survival rate reduces.When nitrogen content is higher than 1/2, to generating astaxanthin
Induce unobvious.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention.
Claims (8)
1. a kind of chlorella that can generate astaxanthin, the chlorella was preserved in Chinese Typical Representative culture on December 25th, 2017
Collection, preserving number are CCTCC NO:M2017837.
2. application of the chlorella described in claim 1 in astaxanthin preparation.
3. a method of preparing astaxanthin, which is characterized in that include the following steps：
S1：Chlorella described in claim 1 is cultivated, chlorella culture is obtained；
S2：Astaxanthin is extracted from the chlorella culture.
4. according to the method described in claim 3, it is characterized in that, S1 includes the following steps：
S11：The chlorella is cultivated in full culture medium, accumulates biomass；
S12：Collect the chlorella algae cell of S11 cultures；
S13：The chlorella algae cell is forwarded in the culture medium of nitrogen stress and is cultivated, the chlorella culture is obtained.
5. according to the method described in claim 4, it is characterized in that, the full culture medium described in S11 is BG11 culture mediums.
6. according to the method described in claim 5, it is characterized in that, the also glucose containing 1g/L in the BG11 culture mediums.
7. according to the method described in claim 4, it is characterized in that, in S11, the culture environment of the chlorella is：Temperature 22-
26 DEG C, illumination 60-80 μ E, incubation time is 5-7 days.
8. according to the method described in claim 4, it is characterized in that, in S13, culture environment is：28 DEG C of temperature, illumination 150-
200 μ E, incubation time are 7-9 days.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109439611A (en) * | 2018-11-20 | 2019-03-08 | 华南理工大学 | Improve the method for inducing and cultivating of color green alga astaxanthin and algae oil accumulation intracellular |
| CN114891637A (en) * | 2022-03-16 | 2022-08-12 | 德默特生物科技(珠海)有限公司 | Chlorella zofingiensis mutant capable of producing astaxanthin at high yield and application thereof |
Citations (1)
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| CN104964957A (en) * | 2015-07-02 | 2015-10-07 | 华南理工大学 | Method for rapidly and nondestructively detecting astaxanthin in C.zofingiensis cells |
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2018
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104964957A (en) * | 2015-07-02 | 2015-10-07 | 华南理工大学 | Method for rapidly and nondestructively detecting astaxanthin in C.zofingiensis cells |
Non-Patent Citations (3)
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| 孙妮等: "碳氮比和光强对小球藻合成虾青素的影响", 《微生物学通报》 * |
| 彭娟等: "不同培养基对小球藻Chlorella zofingiensis生长和虾青素产量的影响", 《热带海洋学报》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109439611A (en) * | 2018-11-20 | 2019-03-08 | 华南理工大学 | Improve the method for inducing and cultivating of color green alga astaxanthin and algae oil accumulation intracellular |
| CN109439611B (en) * | 2018-11-20 | 2022-07-12 | 华南理工大学 | Induction culture method for increasing accumulation of astaxanthin and algae oil in chlorella chromosa cells |
| CN114891637A (en) * | 2022-03-16 | 2022-08-12 | 德默特生物科技(珠海)有限公司 | Chlorella zofingiensis mutant capable of producing astaxanthin at high yield and application thereof |
| CN114891637B (en) * | 2022-03-16 | 2023-07-25 | 德默特生物科技(珠海)有限公司 | Chlorella Chlorella zofingiensis mutant with high astaxanthin yield and application thereof |
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