CN110157620A - A kind of cultural method improving purple ball algae synthesis phycoerythrin content - Google Patents
A kind of cultural method improving purple ball algae synthesis phycoerythrin content Download PDFInfo
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- CN110157620A CN110157620A CN201910313268.3A CN201910313268A CN110157620A CN 110157620 A CN110157620 A CN 110157620A CN 201910313268 A CN201910313268 A CN 201910313268A CN 110157620 A CN110157620 A CN 110157620A
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Abstract
The invention discloses a kind of cultural method for improving purple ball algae synthesis phycoerythrin content, the purpose of this step is to induce purple ball frustule more to synthesize phycoerythrin by changing environmental condition, reducing temperature and reducing intensity of illumination.The purple ball algae of this method culture, phycoerythrin content are 109.2~205.3mg/L, and biomass is 5.3~7.7g/L, have important value in fields such as medicine, food.
Description
Technical field
The invention belongs to Marine Microbial Biotechnology: A Review fields, and in particular to a kind of to improve purple ball algae synthesis phycoerythrin content
Cultural method.
Background technique
Phycoerythrin is a kind of food grade colorant that water solubility is fabulous, is primarily present in the outermost end in phycobilisome, as
The main light filling antenna of purple ball algae frustule participates in photosynthesis, mainly by the color base and apoprotein of open chain tetrapyrrol(e) structure
It is formed by thioether bond covalent bond, phycoerythrin is because it is with good fluorescence intensity, anti-oxidant, removing free radical, high color
The conditions such as degree have very extensive purposes in industries such as food, cosmetics, drugs, also, phycoerythrin is alternatively arranged as fluorescence mark
Note object is applied in fields such as molecular biology, clinical medicine, for example, it can on optical dynamic treatment of tumor, because its safety,
Natural characteristic is as a kind of good photosensitizer great potential.
It is a kind of more original that purple ball algae Porphyridium purpureum is that Naegeli had found for the first time in 1849
Unique unicellular red algae.It belongs to Rhodophyta (Rhodophyta), former Rhodophyceae (Protoflorideophyceae), purple ball
Cutleriales (Porphyridiales), Zi Qiu algae section (Porphyridiaceae), Porphyridium (Porphyridium).Purple ball algae
Have the characteristics that biomass yield is high, high added value product is abundant in numerous microalgae strains.And purple ball algae is distributed in sea
In the soil of water, fresh water, salt water and humidity, there is stronger salt-resistance, purple ball frustule can synthesize a variety of lifes in growth course
Active substances, before the industries such as medicine, cosmetics, food, health care product, weaving, printing and dyeing, metallurgical petroleum have wide application
Scape.Purple ball frustule can synthesize various bioactivators during the growth process, include: (phycobniliprotein is divided into phycobniliprotein
Phycoerythrin PE, phycocyanin PC, apcA-PC, wherein phycoerythrin content is most, account about 84%), purple ball algae
Polysaccharide (predominantly extracellular sulfuric acid lipopolysaccharides), polyunsaturated fatty acid (predominantly arachidonic acid ARA and eicosapentaenoic acid
EPA).Wherein, purple ball algae polysaccharides have been successfully applied in food, drug, clinical medicine etc., insatiable hunger because of its oxidation resistance
With fatty acid ARA, EPA are also relatively different degrees of applies on the energy, health care product and drug, market relative maturity, and
The market of phycoerythrin etc. is relatively thin, and the businessman of the products such as autonomous sale phycoerythrin domestic at present only has Shanghai Lang Ya
Biotechnology Co., Ltd one, other agents are completely dependent on import, thus its price is very expensive.
Main source one of of the purple ball algae as phycoerythrin, the biomass of intracorporal phycoerythrin content with itself
There is vital determinant to the extraction of phycoerythrin.In addition to extraction purification phycoerythrin in existing research, how to increase
Add raw material sources ever more important, however, because production capacity limits, the method for cultivating purple ball algae usually pass through additional nutrient with
And the single culture means such as low temperature, light conditions are maintained to achieve the purpose that improve phycoerythrin content, this certain journey of mode
The content of phycoerythrin, but the problem that the relatively low phycoerythrin content of generally existing biomass content is not high enough can be improved on degree.
Summary of the invention
It is an object of the invention in place of overcome the deficiencies in the prior art, provide a kind of purple ball algae synthesis algae red egg of raising
The cultural method of Bai Hanliang solves the problems in above-mentioned background technique.
The technical solution adopted by the present invention to solve the technical problems is: providing a kind of purple ball algae synthesis algae red egg of raising
The cultural method of Bai Hanliang is cultivated purple ball algae seed liquor using Discrete control temperature and intensity of illumination, including is walked as follows
It is rapid:
(1) control temperature is cultivated 1~3 day at 145~185 μm of ol/m2s in 23~27 DEG C, intensity of illumination；Make purple ball algae
Cell adapted growing environment rapidly enters growth logarithmic phase.
(2) control temperature is in 18~22 DEG C, intensity of illumination in 90~130 μm of ol/m2It is cultivated 7~9 days under s；Pass through change
Environmental condition reduces temperature and reduces intensity of illumination, purple ball frustule is induced more to synthesize phycoerythrin.Favors low temperature is in guarantor
Protein active is held, the synthesis that illumination is conducive to phycoerythrin is reduced.
(3) control temperature is 12~18 DEG C, intensity of illumination is in 35~75 μm of ol/m2It is cultivated 1~3 day under s, culture terminates.It is logical
It crosses and further changes environmental condition, further decrease temperature and reduce intensity of illumination, purple ball frustule is induced more to accumulate algae
Lactoferrin.Favors low temperature reduces the accumulation that illumination is conducive to phycoerythrin in keeping protein active.
In a preferred embodiment of the present invention, the purple ball algae includes purple ball algae Porphyridium purpureum
CoE1。
In a preferred embodiment of the present invention, the raw material algae solution is cultivated in artificial seawater culture medium.
In a preferred embodiment of the present invention, the preparation step of the raw material algae solution are as follows: will normally cultivate to growth logarithm
The purple ball algae algae solution of phase in the end of term is inoculated into artificial seawater culture medium, controls initial OD680Value is 0.4~0.6.
In a preferred embodiment of the present invention, the condition normally cultivated is 165 μm of 25 DEG C of temperature, intensity of illumination ol/
m2S, filtrated air ventilatory capacity 1L/min.
In a preferred embodiment of the present invention, growth logarithmic phase latter stage refers to the 8th day.Purple ball algae autotrophy culture was to 18 days biologies
To reach high value be about 8g/L~12g/L to amount, culture to 6~10 days Biomass yields reach high value be about 700~
900mg/L/d, i.e., 6-10 days are growth logarithmic phase latter stage.It points out to be inoculated into according to other experiments first in artificial seawater culture medium
The OD of eozoon amount680Value is optimum inoculation amount 0.4~0.6.
In a preferred embodiment of the present invention, the raw material algae solution is cultivated under the conditions of continuous light.
In a preferred embodiment of the present invention, using white fluorescence lamp as illumination source.
In a preferred embodiment of the present invention, control is segmented in illumination box, pillar bioreactor or open pond
Temperature and intensity of illumination processed cultivate purple ball algae seed liquor.
In a preferred embodiment of the present invention, the purple ball algae of end is cultivated, phycoerythrin content is 109.2~
205.3mg/L, biomass are 5.3~7.7g/L.
The technical program compared with the background art, it has the following advantages:
This method condition is simple, and environment is mild, is easy to control, and is not necessarily to additive, by changing environmental condition, reduces temperature
With reduction intensity of illumination, purple ball frustule is induced more to synthesize phycoerythrin.The purple ball algae of this method culture, phycoerythrin
Content is 109.2~205.3mg/L, and biomass is 5.3~7.7g/L, has important value in fields such as medicine, food.
Detailed description of the invention
Fig. 1 is the trend chart of biomass in purple ball algae incubation；
Fig. 2 is the trend chart of phycoerythrin content in purple ball algae incubation；
Wherein, each column histogram left side is comparative example 1, and the right side is embodiment 2.
Specific embodiment
Embodiment 1
The Discrete control temperature and intensity of illumination of the present embodiment improve the cultural method of purple ball algae synthesis phycoerythrin content,
It is cultivated using purple ball algae Porphyridium purpureum CoE1, comprising the following steps:
(0) normal culture: condition is 165 μm of 25 DEG C of temperature, intensity of illumination ol/m2S, filtrated air ventilatory capacity 1L/min,
The purple ball algae algae solution in normal culture to growth logarithmic phase latter stage is inoculated into artificial seawater culture medium, initial OD is controlled680Value is
0.4~0.6.
Wherein, artificial seawater culture medium prescription is as follows:
1 artificial seawater culture medium (ASW) of table
Table 1 ASW medium
1. trace meter salting liquid mother liquor (mg/L): ZnCl24.0, H3BO360.0 CoCl2·2H2O 4.0, CuCl2·
2H2O 4.0, MnCl2·4H2O 40.0, (NH4)6Mo7O24·4H2O 37.0。
2. chelated iron solution: FeCl3·4H2O 0.24g/100mL, Na2EDTA(pH7.6)0.05mol/L。
(1) control temperature is in 23 DEG C, intensity of illumination in 145 μm of ol/m2It is cultivated 1 day under s；
(2) control temperature is in 18 DEG C, intensity of illumination in 90 μm of ol/m2It is cultivated 7 days under s；
(3) control temperature is in 12 DEG C, intensity of illumination in 35 μm of ol/m2It is that culture terminates that 1 day is cultivated under s.Pass through segmentation
Temperature and intensity of illumination are controlled, the content that condition of culture improves purple ball algae synthesis phycoerythrin is changed.
After the present embodiment culture, phycoerythrin content is up to 109.2mg/L, and biomass content is up to 5.3g/L.
Embodiment 2
Embodiment 2 the difference from embodiment 1 is that:
(1) control temperature is in 25 DEG C, intensity of illumination in 165 μm of ol/m2It is cultivated 2 days under s；
(2) control temperature is in 20 DEG C, intensity of illumination in 110 μm of ol/m2It is cultivated 8 days under s；
(3) control temperature is in 15 DEG C, intensity of illumination in 55 μm of ol/m2It is that culture terminates that 2 days are cultivated under s.Pass through segmentation
Temperature and intensity of illumination are controlled, the content that condition of culture improves purple ball algae synthesis phycoerythrin is changed.
After the present embodiment culture, phycoerythrin content is up to 205.3mg/L, and biomass content is up to 8.4g/L.
Embodiment 3
Embodiment 3 the difference from embodiment 1 is that:
(1) control temperature is in 27 DEG C, intensity of illumination in 185 μm of ol/m2It is cultivated 3 days under s；
(2) control temperature is in 22 DEG C, intensity of illumination in 130 μm of ol/m2It is cultivated 9 days under s；
(3) control temperature is in 18 DEG C, intensity of illumination in 75 μm of ol/m2It is that culture terminates that 3 days are cultivated under s.Pass through segmentation
Temperature and intensity of illumination are controlled, the content that condition of culture improves purple ball algae synthesis phycoerythrin is changed.
After the present embodiment culture, phycoerythrin content is up to 189.2mg/L, and biomass content is up to 7.7g/L.
Comparative example 1
The present embodiment equally uses purple ball algae Porphyridium purpureum CoE1, carries out normal culture in 12 days and makees
For control experiment, regular culture conditions are 165 μm of 25 DEG C of temperature, intensity of illumination ol/m2S, filtrated air ventilatory capacity 1L/min.
Experimental result please refers to attached drawing 1~2, shows that this method can induce purple ball frustule compared to normal cultural method
More synthesis phycoerythrin.
The above is only the preferred embodiment of the present invention, the range implemented of the present invention that therefore, it cannot be limited according to, i.e., according to
Equivalent changes and modifications made by the invention patent range and description, should still be within the scope of the present invention.
Claims (10)
1. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content, which is characterized in that using Discrete control temperature and
Intensity of illumination cultivates purple ball algae seed liquor, includes the following steps:
(1) control temperature is in 23~27 DEG C, intensity of illumination in 145~185 μm of ol/m2It is cultivated 1~3 day under s；
(2) control temperature is in 18~22 DEG C, intensity of illumination in 90~130 μm of ol/m2It is cultivated 7~9 days under s；
(3) control temperature is in 12~18 DEG C, intensity of illumination in 35~75 μm of ol/m2It is cultivated 1~3 day under s, culture terminates.
2. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content according to claim 1, it is characterised in that:
The purple ball algae includes purple ball algae Porphyridium purpureum CoE1.
3. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content according to claim 1, it is characterised in that:
The raw material algae solution is cultivated in artificial seawater culture medium.
4. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content according to claim 1, which is characterized in that
The preparation step of the raw material algae solution are as follows: the purple ball algae algae solution in normal culture to growth logarithmic phase latter stage is inoculated into artificial seawater
In culture medium, initial OD is controlled680Value is 0.4~0.6.
5. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content according to claim 4, it is characterised in that:
The regular culture conditions are 165 μm of 25 DEG C of temperature, intensity of illumination ol/m2S, filtrated air ventilatory capacity 1L/min (normal culture
Condition is also control experiment condition).
6. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content according to claim 4, it is characterised in that:
Growth logarithmic phase latter stage refers to the 8th day.
7. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content according to claim 1, it is characterised in that:
The raw material algae solution is cultivated under the conditions of continuous light.
8. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content according to claim 1, it is characterised in that:
Using white fluorescence lamp as illumination source.
9. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content according to claim 1, it is characterised in that:
In illumination box, pillar bioreactor or open pond Discrete control temperature and intensity of illumination to purple ball algae seed liquor into
Row culture.
10. a kind of cultural method for improving purple ball algae synthesis phycoerythrin content according to claim 1, feature exist
In: the purple ball algae of end is cultivated, phycoerythrin content is 109.2~205.3mg/L, and biomass is 5.3~7.7g/L.
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| CN115197850A (en) * | 2022-06-22 | 2022-10-18 | 厦门大学 | Culture method for increasing yield of porphyridium polysaccharide |
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