CN106086002A - A kind of supersonic induced promotion micro algae growth and the method for astaxanthin accumulation - Google Patents
A kind of supersonic induced promotion micro algae growth and the method for astaxanthin accumulation Download PDFInfo
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Abstract
The invention discloses a kind of supersonic induced promotion micro algae growth and the method for astaxanthin accumulation, described method is by two stages of production of astaxanthin algae incubation: travelling stage and motionless stage carry out the supersound process of suitable intensity, can be effectively improved the cell growth rate in the travelling stage and the accumulation at motionless stage astaxanthin.The method that the present invention provides can shorten the cultivation cycle of production of astaxanthin algae effectively, improves the astaxanthin productivity in incubation, is effectively reduced the production cost of algae base astaxanthin.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of supersonic induced promotion micro algae growth and astaxanthin accumulation
Method.
Background technology
Astaxanthin (molecular formula: C40H52O4) it is a kind of carotenoid with extremely strong antioxidant activity, also it is a kind of simultaneously
Excellent natural pigment, is widely used in the industries such as food, medicine, health product, aquaculture, has the most wide city
Field prospect, it is estimated that the market value that astaxanthin is annual in the world at present is up to 200,000,000 dollars.Currently, the astaxanthin master on market
Chemical synthesis to be depended on produce, relative to natural astaxanthin, synthetic astaxanthin owing to there is substantial amounts of mesomer,
Thus activity is well below natural astaxanthin, and the astaxanthin of chemosynthesis is because the problems such as safety are in the industries such as food and medicine
There is great limitation, therefore, the exploitation of natural astaxanthin is extremely important.Microorganism is natural astaxanthin
Best source, especially some microalgae, such as Haematocoocus Pluvialls (Haematococcus pluvialis), intracellular containing higher
Astaxanthin, its content up to the 3～5% of dry cell weight, further, since in microalgae accumulation astaxanthin be pure S-S type structure,
Activity be better than other source astaxanthin, therefore, microalgae be considered as natural astaxanthin most preferably produce body.
Its growth course of microalgae for production of astaxanthin is generally divided into two stages, i.e. moves about stage and motionless stage,
When cell is under suitable growth conditions, cell is in the travelling stage, and in this stage, cell number accumulates rapidly；Work as cultivation
Nutrient substance in base is depleted, or when cell is under other adverse environmental factors, cell wall is the most thickening, and cellular morphology becomes
Greatly, losing flagellum, cell is in motionless state, and now astaxanthin starts to accumulate in a large number, and cellular colours gradually deepens.Currently, base
The highest in the production cost of the astaxanthin of microalgae, it is difficult to compete mutually with the product of chemosynthesis, this is mainly due to the life of microalgae
Long relatively slower, cause toxigenic capacity higher, it addition, the content astaxanthin in cell is relatively low, cause final product yield relatively
Low.Therefore, during microdisk electrode, how to improve the growth rate of microalgae, shorten cultivation cycle, improve intracellular shrimp blue or green
Cellulose content, being effectively reduced the production cost of algae base astaxanthin is present stage technical barrier to be solved.
Summary of the invention
During utilizing microalgae to produce astaxanthin, slow for micro algae growth speed, and need to be right in the motionless stage
It carries out the problem of stress-inducing astaxanthin accumulation, it is an object of the invention to provide a kind of supersonic induced promotion micro algae growth and
The method of astaxanthin accumulation.Described method is moved about the stage at microalgae, can effectively facilitate the growth of microalgae cell, shortens and cultivates week
Phase；When cell is in the motionless stage, can the effectively accumulation of astaxanthin in inducing cell, be greatly improved astaxanthin in incubation
Productivity.
For achieving the above object, a kind of supersonic induced promotion micro algae growth of the present invention and the method for astaxanthin accumulation
Comprise the following steps:
(1) in conventional micro-algae culture medium, the algae that can accumulate astaxanthin is accessed with certain inoculum concentration；
(2) cultivate at suitable temperature and light intensity, in the middle of incubation at regular intervals, with certain frequency
The regular hour is processed with power ultrasonic；
(3) after cell goes to the motionless stage, adjust to suitable temperature and light intensity, at regular intervals, with necessarily
Frequency and power ultrasonic process the regular hour；
(4), after cultivation terminates, collect cell, measure and extract the astaxanthin of intracellular.
Preferably, culture medium preferred BBM culture medium described in step (1), BG 11 culture medium, SE culture medium etc., A9 cultivates
Base, PHM-1 culture medium, Z8 culture medium etc.；Inoculum concentration preferably 3%～20%；The algae producing astaxanthin includes Haematocoocus Pluvialls
(Haematococcus pluvialis), chlorella (Chlorella zofingiensis), Chlorococcum (Chlorococcum
Sp), blood red Euglena (Euglena sanguinea) etc..
Cultivation temperature described in step (2) preferably 20～28 DEG C；Light intensity preferably 30～300 μm ol/m2/s；Described ultrasonic
It is spaced preferably 1～5 day/time, further preferred 3～4 days/time；Supersonic frequency preferably 20～60kHz；Ultrasonic power preferably 500～
6000W/L, further preferred 1000～4000W/L；Sonication treatment time 1～15min, further preferred 3～10min every time.
Cultivation temperature described in step (3) preferably 26～35 DEG C；Light intensity preferably 200～1200 μm ol/m2/s；Described super
Sound intensity be higher than step (2), preferably 1～4 day/time of ultrasonic interval, further preferred 3～4 days/time；Supersonic frequency preferably 40～
80kHz；Ultrasonic power preferably 1000～10000W/L, each sonication treatment time 3～20min, further preferred 5～15min.
A kind of supersonic induced promotion micro algae growth of present invention offer and the method for astaxanthin accumulation, utilize ultrasonic to microalgae
The facilitation of growth, and the coercion that the active oxygen of ultrasonic generation is to microalgae, can be effectively facilitated microalgae travelling
The speed of growth in stage and in the accumulation of motionless stage astaxanthin, makes the production of astaxanthin algae speed of growth in the travelling stage obtain
To being obviously promoted, meanwhile, the accumulation efficiency at motionless stage astaxanthin is significantly improved so that astaxanthin in incubation
Productivity be greatly enhanced.Compared with producing production of astaxanthin process with traditional microalgae, one of the present invention is ultrasonic to lure
Lead and promote that the method for micro algae growth and astaxanthin accumulation has the advantages that
(1) method that the present invention provides effectively promotes the cell growth rate in the travelling stage so that cultivation cycle
Notable shortening；
(2) the alternative traditional high temperature of method of present invention offer, high salt etc. are coerced, and can enter in combination with traditional coercing
One step strengthen intracellular astaxanthin accumulation, can effectively inducing cell in the synthesis of motionless stage astaxanthin；
(3) method that the present invention provides is easy to use, and simple to operate, energy consumption is low, can be effectively reduced the production of astaxanthin
Cost.
Detailed description of the invention
In order to the present invention is better described, it is simple to understand technical scheme, the typical case of the present invention but unrestriced
Embodiment is as follows:
Embodiment 1
(1) in BBM culture medium, the inoculum concentration with 5% accesses Haematocoocus Pluvialls；
(2) at 28 DEG C, 100 μm ol/m2Under/s, it is cultivated, incubation every 4 days, with the frequency of 25kHz and
The power of 2000W/L is to culture fluid supersound process 3min；
(3) after cell goes to the motionless stage, adjustment temperature is to 35 DEG C, and light intensity is to 800 μm ol/m2/ s, every 2 days with
The frequency of 50kHz and the power of 5000W/L are to culture fluid supersound process 5min；
(4), after cultivation terminates, collect cell, measure and extract the astaxanthin of intracellular.
Treatment effect is tested:
In incubation, the most ultrasonic group under astaxanthin productivity ratio the same terms improves 28%.
Embodiment 2
(1) in BG-11 culture medium, the inoculum concentration with 18% accesses chlorella；
(2) at 22 DEG C, 150 μm ol/m2Under/s, it is cultivated, incubation every 1 day, with the frequency of 50kHz and
The power of 3000W/L is to culture fluid supersound process 5min；
(3) after cell goes to the motionless stage, adjustment temperature is to 32 DEG C, and light intensity is to 600 μm ol/m2/ s, every 1 day with
The frequency of 80kHz and the power of 4000W/L are to culture fluid supersound process 8min；
(4), after cultivation terminates, collect cell, measure and extract the astaxanthin of intracellular.
Treatment effect is tested:
In incubation, the most ultrasonic group under astaxanthin productivity ratio the same terms improves 35%.
Embodiment 3
(1) in SE culture medium, the inoculum concentration with 3% accesses Chlorococcum；
(2) at 20 DEG C, 300 μm ol/m2Under/s, it is cultivated, incubation every 5 days, with the frequency of 20kHz and
The power of 6000W/L is to culture fluid supersound process 1min；
(3) after cell goes to the motionless stage, adjustment temperature is to 26 DEG C, and light intensity is to 1200 μm ol/m2/ s, every 4 days with
The frequency of 40kHz and the power of 10000W/L are to culture fluid supersound process 3min；
(4), after cultivation terminates, collect cell, measure and extract the astaxanthin of intracellular.
Treatment effect is tested:
In incubation, the most ultrasonic group under astaxanthin productivity ratio the same terms improves 29%.
Embodiment 4
(1) in A9 culture medium, the inoculum concentration with 15% accesses blood red Euglena；
(2) at 24 DEG C, 30 μm ol/m2Under/s, it is cultivated, incubation every 2 days, with the frequency of 30kHz and
The power of 1000W/L is to culture fluid supersound process 8min；
(3) after cell goes to the motionless stage, adjustment temperature is to 33 DEG C, and light intensity is to 200 μm ol/m2/ s, every 1 day with
The frequency of 60kHz and the power of 8000W/L are to culture fluid supersound process 12min；
(4), after cultivation terminates, collect cell, measure and extract the astaxanthin of intracellular.
Treatment effect is tested:
In incubation, the most ultrasonic group under astaxanthin productivity ratio the same terms improves 42%.
Embodiment 5
(1) in PHM-1 culture medium, the inoculum concentration with 10% accesses chlorella；
(2) at 26 DEG C, 200 μm ol/m2Under/s, it is cultivated, incubation every 3 days, with the frequency of 60kHz and
The power of 500W/L is to culture fluid supersound process 15min；
(3) after cell goes to the motionless stage, adjustment temperature is to 28 DEG C, and light intensity is to 400 μm ol/m2/ s, every 2 days with
The frequency of 70kHz and the power of 1000W/L are to culture fluid supersound process 20min；
(4), after cultivation terminates, collect cell, measure and extract the astaxanthin of intracellular.
Treatment effect is tested:
In incubation, the most ultrasonic group under astaxanthin productivity ratio the same terms improves 36%.
Embodiment 6
(1) in Z8 culture medium, the inoculum concentration with 20% accesses Haematocoocus Pluvialls；
(2) at 25 DEG C, 250 μm ol/m2Under/s, it is cultivated, incubation every 3 days, with the frequency of 40kHz and
The power of 4000W/L is to culture fluid supersound process 10min；
(3) after cell goes to the motionless stage, adjustment temperature is to 27 DEG C, and light intensity is to 1000 μm ol/m2/ s, every 3 days with
The frequency of 55kHz and the power of 3000W/L are to culture fluid supersound process 15min；
(4), after cultivation terminates, collect cell, measure and extract the astaxanthin of intracellular.
Treatment effect is tested:
In incubation, the most ultrasonic group under astaxanthin productivity ratio the same terms improves 33%.
Claims (6)
1. a supersonic induced promotion micro algae growth and the method for astaxanthin accumulation, it is characterised in that this technique includes following step
Rapid:
(1) in micro-algae culture medium, inoculation can accumulate the algae of astaxanthin；
(2) in 20～28 DEG C of temperature and 30～300 μm ol/m2Cultivate under/s light intensity, incubation be spaced 1～5 day/time,
Process with 20～60kHz frequencies and 500～6000W/L power ultrasonics；
(3) after cell goes to the motionless stage, adjust to 26～35 DEG C of temperature and 200～1200 μm ol/m2/ s light intensity, interval 1～
4 days/time, process with 40～80kHz frequencies and 1000～10000W/L power ultrasonics；
(4), after cultivation terminates, collect cell, measure and extract the astaxanthin of intracellular.
Method the most according to claim 1, it is characterised in that culture medium described in step (1) selects BBM culture medium, BG
11 culture medium, SE culture medium, A9 culture medium, PHM-1 culture medium, the one in Z8 culture medium；Inoculum concentration is 3%～20%；Produce
The algae of astaxanthin selects Haematocoocus Pluvialls, chlorella, Chlorococcum, the one in blood red Euglena.
Method the most according to claim 1, it is characterised in that in step (2), each sonication treatment time is 1～15min.
Method the most according to claim 3, it is characterised in that described in step (2), ultrasonic is 3～4 days/time, every super
Acoustical power is 1000～4000W/L, and each sonication treatment time is 3～20min.
Method the most according to claim 1, it is characterised in that in step (3), each sonication treatment time is 3～20min.
Method the most according to claim 5, it is characterised in that described in step (3), ultrasonic being spaced apart is 3～4 days/time；
Sonication treatment time is 5～15min every time.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022211302A1 (en) * | 2021-03-30 | 2022-10-06 | 고려대학교 산학협력단 | Method for enhancing biosynthesis of high value-added substance derived from microalgae by introducing ultrasonic stimulus |
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| CN104726445A (en) * | 2015-02-17 | 2015-06-24 | 中国计量学院 | Culture method for improving yield of botryococcus polysaccharides |
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- 2016-08-12 CN CN201610664400.1A patent/CN106086002B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104726445A (en) * | 2015-02-17 | 2015-06-24 | 中国计量学院 | Culture method for improving yield of botryococcus polysaccharides |
| CN104894102A (en) * | 2015-05-14 | 2015-09-09 | 邹宁 | Ultrasonic reinforcement culture method of microalgae |
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Non-Patent Citations (1)
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022211302A1 (en) * | 2021-03-30 | 2022-10-06 | 고려대학교 산학협력단 | Method for enhancing biosynthesis of high value-added substance derived from microalgae by introducing ultrasonic stimulus |
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