CN106867909A - A kind of cultural method of very thin Euglena - Google Patents
A kind of cultural method of very thin Euglena Download PDFInfo
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Abstract
The invention discloses a kind of cultural method of very thin Euglena, it comprises the following steps：Heterotrophic culture is carried out to very thin Euglena, in exponential phase of growth mid-term, the very thin Euglena in blowing part is supplemented with part nutrients and water, continues to cultivate.The beneficial effects of the invention are as follows can quickly, high yield turn out utilization rate very thin Euglena high, greatly reduce the production cost of very thin Euglena.
Description
Technical field
The present invention relates to biological technical field, particularly a kind of method suitable for being cultivated very thin Euglena.
Background technology
Very thin Euglena belongs to Euglenophyta Euglena, and cell is generally spindle, and minority is cylinder, and acellular wall, table matter is soft,
Shape is variable, and 50 μm of diameter, is one of life earliest on the earth, and before appearing in more than 5 hundred million years, gene is not changed in all the time, is
A kind of efficient photosynthetic plant, with photoautotrophy growth and breeding, while chemoheterotrophy can be carried out again.Very thin Euglena is due to having
Rich in abundant nutriment, and with the training mode that can carry out chemoheterotrophy culture have it is advantageous excellent
Gesture, therefore be a kind of algae kind for having nutritive value and industrialization potential concurrently.
The main training method of very thin Euglena has autotrophy, raises together with and heterotrophism.Autotrophy is used current pilot scale culture more, but certainly
Support and there is the adverse effects such as culture density is low, growth cycle is long, low production efficiency, floor space big, harvesting difficulty.And use honest and clean
Under valency, suitable and widely used organic carbon source glucose are for the condition of culture of sole carbon source, the Heterotrophic culture of very thin Euglena is not
Influenceed by the natural environment and climate condition such as illumination, temperature, growth rate is fast, and culture density is high, and production process is easily controlled, and yield can
Greatly improve.
Current existing Heterotrophic culture mode is mainly batch culture, stream plus batch culture and the training of batch (-type) fed-batch
Support.Wherein batch culture is the training method for most generally using.But lag phase is more long during batch culture has incubation, culture
The aging growth of frustule slows down after reaching stationary phase, and culture latter stage concentration of glucose wasting problem higher, makes training
Support yield reduction.
The content of the invention
It is an object of the invention to provide a kind of cultivation cycle is short, the culture of high yield, utilization of carbon source rate very thin Euglena high
Method.
The technical scheme is that,
A kind of cultural method of very thin Euglena, its step includes：Heterotrophic culture is carried out to very thin Euglena, it is interim in exponential growth
Phase, the very thin Euglena in blowing part is supplemented with part nutrients and water, continues to cultivate.
In middle exponential growth, carry out that " the very thin Euglena of blowing, the selection amount of very thin Euglena is 1/2 daily（Account for culture liquid
Long-pending 50%）, it is supplemented with nutrients and water, the selection amount of nutrients and water is 1/2（Account for the 50% of nutrient solution volume）, continue to train
Support " operation once, until cultivation cycle terminates.
The composition of the Heterotrophic culture liquid includes：Glucose 20g/L, dusty yeast 5g/L, K2HPO30.3g/L, KH2PO3
0.2g/L, MgSO4·7H2O 0.1g/L, TE liquid microelement 1mL/L, remaining is water, and wherein TE liquid microelements composition includes
H3BO3 2.81g/L、MnCl 2·4H2O 1.82g/L、ZnSO4·7H2O 0.23g/L、Na2MoO4·2H2O 0.34g/L、
CuSO4·5H2O 0.05g/L and Co (NO3) 2·6H2O 0.06g/L。
The Heterotrophic culture liquid pH value is 3.0 ~ 6.0.
The composition of the extra-nutrition thing is included per addition glucose 4-5g, dusty yeast 4-5g/ in 1000mL nutrient solutions
L, K2HPO30.05g, KH2PO30.01g, MgSO4·7H2O 0.02g, TE liquid microelements 0.5mL.
The inoculum concentration of very thin Euglena is the volume accounting of nutrient solution 10% during the Heterotrophic culture.
25-28 DEG C of cultivation temperature during the Heterotrophic culture, blowing air amount 1.0L/min adds 0.05% organosilicon
Defoamer.
The exponential phase of growth mid-term is the 3rd day to the 4th day from inoculated and cultured.
Preferably, each blowing 1/2（Account for the 1/2 of total volume of culture）Very thin Euglena.
The beneficial effects of the invention are as follows can quickly, high yield turn out utilization rate very thin Euglena high, greatly reduce
The production cost of very thin Euglena.
Brief description of the drawings
Fig. 1 is ventilate autotrophy and heterotrophic growth curve synoptic diagram of the invention,
Fig. 2 is logarithmic phase feed supplement growth curve schematic diagram of the invention.
Specific embodiment：
Embodiment is described in detail with reference to the figures above,
Embodiment 1
Ventilation autotrophy culture：
In the blake bottle of 1000mL, autotrophy culture of ventilating is carried out, condition of culture is：Very thin Euglena inoculum density is 0.33g/
L；The composition of Heterotrophic culture base includes：NaNO31.5g/L, K2HPO30.3g/L, KH2PO3 0.2g/L, MgSO4·7H2O 0.1g/
L, TE liquid microelement 1mL/L, remaining is water, and wherein TE solution formulas are H3BO3 2.81g/L、MnCl 2·4H2O
1.82g/L、ZnSO4·7H2O 0.23g/L、Na2MoO4·2H2O 0.34g/L、CuSO4·5H2O 0.05g/L and Co (NO3)2·6H2O 0.06g/L；28 DEG C of cultivation temperature, blowing air amount 1.0L/min adds 0.05% organic silicon defoamer.As a result
Display is as shown in Fig. 1.
Additional carbon glucose ventilation Heterotrophic culture
In the blake bottle of 1000mL, additional carbon glucose ventilation Heterotrophic culture is carried out, condition of culture is：Very thin Euglena connects
It is 0.33g/L to plant density；The composition of Heterotrophic culture base includes：Glucose 20g/L, NaNO31.5g/L, K2HPO30.3g/L,
KH2PO3 0.2g/L, MgSO4·7H2O 0.1g/L, TE liquid microelement 1mL/L, remaining is water, wherein TE solution formulas
It is H3BO32.81g/L、MnCl 2·4H2O 1.82g/L、ZnSO4·7H2O 0.23g/L、Na2MoO4·2H2O0.34g/L、
CuSO4·5H2O 0.05g/L and Co (NO3) 2·6H2O 0.06g/L；25 DEG C of cultivation temperature, blowing air 1.0L/min,
Add 0.05% organic silicon defoamer.Result is shown as shown in Fig. 1.
As shown in Figure 1, using the very thin Euglena of ventilation autotrophy culture, the very thin Euglena of ventilation autotrophy culture enters within the 8th day
Stationary phase, then culture terminates；Using the very thin Euglena of additional carbon glucose ventilation Heterotrophic culture, culture starts latter 2nd day
Very thin Euglena enters exponential phase of growth, and the very thin Euglena of ventilation Heterotrophic culture enters stationary phase within the 6th day, and then culture terminates.
Embodiment 2
Ventilation autotrophy culture：
In the blake bottle of 1000mL, autotrophy culture of ventilating is carried out, condition of culture is：Very thin Euglena inoculum density is 0.33g/
L；The composition of Heterotrophic culture base includes：NaNO31.5g/L, K2HPO30.3g/L, KH2PO3 0.2g/L, MgSO4·7H2O 0.1g/
L, TE liquid microelement 1mL/L, remaining is water, and wherein TE solution formulas are H3BO3 2.81g/L、MnCl 2·4H2O
1.82g/L、ZnSO4·7H2O 0.23g/L、Na2MoO4·2H2O 0.34g/L、CuSO4·5H2O 0.05g/L and Co (NO3)2·6H2O 0.06g/L；25 DEG C of cultivation temperature, blowing air amount 1.0L/min adds 0.05% organic silicon defoamer.As a result
Display is as shown in Fig. 1.
Additional carbon glucose ventilation Heterotrophic culture
In the blake bottle of 1000mL, additional carbon glucose ventilation Heterotrophic culture is carried out, condition of culture is：Very thin Euglena connects
It is 0.33g/L to plant density；The composition of Heterotrophic culture base includes：Glucose 20g/L, NaNO31.5g/L, K2HPO30.3g/L,
KH2PO3 0.2g/L, MgSO4·7H2O 0.1g/L, TE liquid microelement 1mL/L, remaining is water, wherein TE solution formulas
It is H3BO32.81g/L、MnCl 2·4H2O 1.82g/L、ZnSO4·7H2O 0.23g/L、Na2MoO4·2H2O0.34g/L、
CuSO4·5H2O 0.05g/L and Co (NO3) 2·6H2O 0.06g/L；28 DEG C of cultivation temperature, blowing air 1.0L/min,
Add 0.05% organic silicon defoamer.Result is shown as shown in Fig. 1.
As shown in Figure 1, using the very thin Euglena of ventilation autotrophy culture, the very thin Euglena of ventilation autotrophy culture enters within the 8th day
Stationary phase, then culture terminates；Using the very thin Euglena of additional carbon glucose ventilation Heterotrophic culture, culture starts latter 2nd day
Very thin Euglena enters exponential phase of growth, and the very thin Euglena of ventilation Heterotrophic culture enters stationary phase within the 6th day, and then culture terminates.
Embodiment 3
Additional carbon glucose ventilation Heterotrophic culture, in exponential phase early stage blowing fluid infusion culture.
In the blake bottle of 1000mL, additional carbon glucose ventilation Heterotrophic culture is carried out, condition of culture is：Very thin Euglena
Inoculum density is 0.33g/L；The composition of Heterotrophic culture base includes：Glucose 20g/L, dusty yeast 5g/L, K2HPO30.3g/L,
KH2PO3 0.2g/L, MgSO47H2O 0.1g/L, TE liquid microelement 1mL/L, remaining is water, wherein TE solution formulas
It is H3BO32.81g/L、MnCl 2·4H2O 1.82g/L、ZnSO4·7H2O 0.23g/L、Na2MoO4·2H2O 0.34g/L、
CuSO4·5H2O 0.05g/L and Co (NO3) 2·6H2O 0.06g/L；25--28 DEG C of cultivation temperature, blowing air amount
1.0L/min, adds 0.05% organic silicon defoamer；Start within 1.5-2 days, the very thin Euglena of daily blowing 1/2, then mend
Equivalent nutrient solution is filled, continues to cultivate, wherein, the composition of the extra-nutrition thing includes：Glucose 4-5g, dusty yeast 5g/L,
K2HPO30.05g, KH2PO3 0.01g, MgSO4·7H2O 0.02g, TE liquid microelements 0.5mL.In incubation is continued,
The every 3 days antibiotics penicillin sodium salt 1mL of injection 300mg/L, every 8 days are a production cycle.Result shows as shown in Figure 2.
Additional carbon glucose ventilation Heterotrophic culture, in exponential phase mid-term blowing fluid infusion culture.
The experiment base of this experiment and " additional carbon glucose ventilate Heterotrophic culture, in exponential phase early stage blowing fluid infusion culture "
This is identical, and difference is that, at the 2.5th day ~ the 3.5th day, the very thin Euglena of daily blowing 1/2 is supplemented with equivalent nutrient solution,
Continue to cultivate.Result shows as shown in Figure 2.Additional carbon glucose ventilation Heterotrophic culture, in late exponential phase blowing fluid infusion training
Support.The basic phase of experiment of this experiment and " additional carbon glucose ventilate Heterotrophic culture, in exponential phase early stage blowing fluid infusion culture "
Together, difference is, starts within 3.5-4.5 days the, and the very thin Euglena of daily blowing 1/2 is supplemented with equivalent nutrient solution, continues
Culture.Result shows as shown in Figure 2.As shown in Figure 2, additional carbon glucose ventilation Heterotrophic culture, in exponential phase early stage blowing
Fluid infusion culture, the biomass stabilization of very thin Euglena is between 3.9-4.1g/L in nutrient solution；In exponential phase mid-term blowing fluid infusion training
Support, the biomass of very thin Euglena has the trend for growing steadily in nutrient solution, and biomass can be stablized in 10.2-12.0g/ substantially
L；In late exponential phase blowing fluid infusion culture, the biomass of very thin Euglena has larger amplitude in the previous day of sampling feed supplement in nutrient solution
The decline of degree, then there is the trend being gradually reduced, and biomass can be stablized between 8.0-8.3g/L substantially.
The cultural method of the very thin Euglena that the present invention is provided takes out part algae during Heterotrophic culture by silk blowing
Cell, then supplements the nutrient solution of certain volume, makes frustule continue to cultivate in Heterotrophic culture bottle.Using the method, significantly
Improve the culture yield of very thin Euglena, it is ensured that the vigor of frustule, while improve the utilization rate of carbon source, be conducive to scale
Change heterotrophism production；Additionally, being sampled by blowing, the nutritional ingredient in nutrient solution is remained, save production cost, it is to avoid waste
And environmental pollution, the easy optimal control of condition of culture.
Above-described embodiment is only intended to clearly illustrate example, and not to the restriction of implementation method.For institute
For the those of ordinary skill in category field, the change or change of other multi-forms can also be made on the basis of the above description
It is dynamic.There is no need and unable to be exhaustive to all of implementation method, and the obvious change or change thus extended out
Move within still in the protection domain of the invention.
Claims (9)
1. a kind of cultural method of very thin Euglena, it is characterised in that：Incubation step includes：Heterotrophic culture is carried out to very thin Euglena,
In exponential phase of growth mid-term, the very thin Euglena in blowing part is supplemented with part nutrients and water, continues to cultivate.
2. a kind of cultural method of very thin Euglena as claimed in claim 1, it is characterised in that：In middle exponential growth, daily
Carry out that " the very thin Euglena of blowing, the selection amount of very thin Euglena is account for nutrient solution volume 50%, is supplemented with nutrients and water, nutrients
With the selection amount of water be account for nutrient solution volume 50%, continue cultivate " operation once, until cultivation cycle terminates.
3. a kind of cultural method of very thin Euglena as claimed in claim 1, it is characterised in that：The composition of the Heterotrophic culture liquid
Including：Glucose 20g/L, dusty yeast 5g/L, K2HPO30.3g/L, KH2PO30.2g/L, MgSO4·7H2O 0.1g/L, TE
Liquid microelement 1mL/L, remaining is water, and wherein TE liquid microelements composition includes H3BO3 2.81g/L、MnCl 2·4H2O
1.82g/L、ZnSO4·7H2O 0.23g/L、Na2MoO4·2H2O 0.34g/L、CuSO4·5H2O 0.05g/L and Co
(NO3) 2·6H2O 0.06g/L。
4. a kind of cultural method of very thin Euglena as claimed in claim 1, it is characterised in that：The Heterotrophic culture liquid pH value exists
3.0~6.0。
5. a kind of cultural method of very thin Euglena as claimed in claim 1, it is characterised in that：The composition of the extra-nutrition thing
Including adding glucose 4-5g, dusty yeast 4-5g/L, K in every 1000mL nutrient solutions2HPO30.05g, KH2PO30.01g,
MgSO4·7H2O 0.02g, TE liquid microelements 0.5mL.
6. a kind of cultural method of very thin Euglena as claimed in claim 1, it is characterised in that：It is fine during the Heterotrophic culture
The inoculum concentration of thin Euglena is the volume accounting of nutrient solution 10%.
7. a kind of cultural method of very thin Euglena as claimed in claim 1, it is characterised in that：Trained during the Heterotrophic culture
25-28 DEG C of temperature is supported, blowing air amount 1.0L/min adds 0.05% organic silicon defoamer.
8. a kind of cultural method of very thin Euglena as claimed in claim 1, it is characterised in that：The exponential phase of growth mid-term is
From the 3rd day to the 4th day of inoculated and cultured.
9. a kind of cultural method of very thin Euglena as claimed in claim 1, it is characterised in that：The amount of each blowing is always trained to account for
Support the 1/2 of volume very thin Euglena.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107686813A (en) * | 2017-09-27 | 2018-02-13 | 青岛科海生物有限公司 | A kind of Euglena high-density cultivation method |
| CN108148763A (en) * | 2018-03-08 | 2018-06-12 | 青岛旭能生物工程有限责任公司 | A kind of method that the very thin Euglena yield of shake flask fermentation is improved by feeding method |
| CN108318307A (en) * | 2018-04-08 | 2018-07-24 | 南京大学 | Very thin Euglena measures the algae solution preparation method and application of aquatic particle bio-toxicity |
| CN108359608A (en) * | 2018-03-08 | 2018-08-03 | 青岛旭能生物工程有限责任公司 | A kind of method of the very thin Euglena of high density fermentation culture |
| CN109797104A (en) * | 2019-02-22 | 2019-05-24 | 中国科学院水生生物研究所 | A kind of chlorella Chlorella zofingiensis heterotrophism high-density cultivation method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101633894A (en) * | 2009-08-20 | 2010-01-27 | 北京芳能科技有限公司 | Culture medium of euglena gracilis and open type high-density culture method |
| CN104830691A (en) * | 2015-04-27 | 2015-08-12 | 长江大学 | Culture method of chlorella |
-
2017
- 2017-04-13 CN CN201710240311.9A patent/CN106867909A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101633894A (en) * | 2009-08-20 | 2010-01-27 | 北京芳能科技有限公司 | Culture medium of euglena gracilis and open type high-density culture method |
| CN104830691A (en) * | 2015-04-27 | 2015-08-12 | 长江大学 | Culture method of chlorella |
Non-Patent Citations (1)
| Title |
|---|
| 韩北忠: "《发酵工程》", 31 January 2013, 中国轻工业出版社 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107686813A (en) * | 2017-09-27 | 2018-02-13 | 青岛科海生物有限公司 | A kind of Euglena high-density cultivation method |
| CN108148763A (en) * | 2018-03-08 | 2018-06-12 | 青岛旭能生物工程有限责任公司 | A kind of method that the very thin Euglena yield of shake flask fermentation is improved by feeding method |
| CN108359608A (en) * | 2018-03-08 | 2018-08-03 | 青岛旭能生物工程有限责任公司 | A kind of method of the very thin Euglena of high density fermentation culture |
| CN108318307A (en) * | 2018-04-08 | 2018-07-24 | 南京大学 | Very thin Euglena measures the algae solution preparation method and application of aquatic particle bio-toxicity |
| CN109797104A (en) * | 2019-02-22 | 2019-05-24 | 中国科学院水生生物研究所 | A kind of chlorella Chlorella zofingiensis heterotrophism high-density cultivation method |
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Application publication date: 20170620 |