CN102550263A - Method for cultivating Botryococcus braunii - Google Patents
Method for cultivating Botryococcus braunii Download PDFInfo
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- CN102550263A CN102550263A CN2012100249246A CN201210024924A CN102550263A CN 102550263 A CN102550263 A CN 102550263A CN 2012100249246 A CN2012100249246 A CN 2012100249246A CN 201210024924 A CN201210024924 A CN 201210024924A CN 102550263 A CN102550263 A CN 102550263A
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Abstract
The invention relates to a method for cultivating Botryococcus braunii, which is technically characterized by including: using an inorganic carbon source, a nitrogen source and a phosphorus source additionally fed to adjust the concentration of nutrient salt in waste water; allowing the concentrations of the inorganic carbon source Na2CO3, the nitrogen source KNO3 and the phosphorus source KH2PO4 to be 5-15mg/L, 10-20mg/L and 1-2mg/L respectively; placing chlorella into culture solution at the temperature of 20 DEG C to 30 DEG C with lamination intensity 3000Lx-8000Lx and brightness darkness ratio 13h; and cultivating for 30 days with the brightness darkness ratio 11h, wherein the waste water can be domestic sewage, eutrophicating river water or lake water, and aquacultural water, and the waste water is used to cultivate high-concentration Botryococcus braunii rich in greases and hydrocarbons. By the method, the waste water is used for cultivating the botryococcus braunii with high material energy.
Description
Technical field:
The present invention relates to a kind of algae culture, the cultural method of particularly a kind of Blang's grape algae.
Background technology:
At present, global energy shortage and problem of environmental pollution impel renewable, the free of contamination new forms of energy of development and utilization.On the other hand, water pollution problems is also very outstanding, and the nutrition load of water body sharply increases and accumulation, makes the eutrophication process of water body accelerate, and water quality worsens rapidly, partial loss of function.Blang's grape algae (
Botryococcus braunii) can effectively utilize solar energy, through photosynthesis fixation of C O
2, inorganic matter is converted into high unsaturated alkane, grease equal energy source material.In addition, the nutritive element in the growth water body capable of using of Blang's grape algae reduces the nutrition load in the water body.If the production capacity of Blang's grape algae and the control of body eutrophication are combined, not only can improve the eutrophication degree of water body, and can produce bioenergy.
Summary of the invention:
The object of the invention provides a kind of algae kind and is easy to get, cultivates that water source is wide, the simple cost of cultural method is low and produce the cultural method of a kind of Blang's grape algae that material can be big.Technical scheme of the present invention is; It is characterized in that: inorganic carbon source, nitrogenous source, phosphorus source with adding are adjusted nutrient concentration in the sewage; Make the concentration of DIC, total nitrogen, total phosphorus be respectively 5 ~ 15mg/L, 10 ~ 20mg/L, 1 ~ 2mg/L, said inorganic carbon source, nitrogenous source, phosphorus source are respectively Na
2CO
3, KNO
3, KH
2PO
4Chlorella is put into above-mentioned culture fluid; 20 ℃ ~ 30 ℃ of temperature, intensity of illumination 3000Lx ~ 8000Lx, Light To Dark Ratio are that 13h:11h cultivated 30 days down; Said sewage is river, river, lake water and the aquaculture water of sanitary sewage and eutrophication, promptly cultivates the high density Blang's grape that becomes to be rich in grease and hydrocarbon with sewage.The present invention and prior art relatively have the algae kind and are easy to get, cultivate remarkable advantage wide with the water source, that the simple cost of cultural method is low and the generation material can be big.
Embodiment:
The medium that uses in following examples is as sewage stoste or the sewage culture fluid after optimizing.Sewage stoste is taken from river, river, lake water and the aquaculture pond water of sanitary sewage and eutrophication.
Blang's grape algae of using in following examples is available from Inst. of Hydrobiology, Chinese Academy of Sciences algae kind storehouse.
The biomass of the Blang's grape frustule that relates in following examples adopts the dry cell weight method, with the algae liquid of certain volume, filters the back and collects, and the adding distil water washing through freeze drying 72h, is weighed, and representes with the unit volume dry weight.
The total fat assay method reference literature that relates in following examples (Bligh E G, et al. Canadian Journal of Biochemistry and Physiology, 1959; Chen F, et al. Journal of Applied Physiology, 1991) in the method for Bligh etc. and Chen etc.; Take by weighing a certain amount of dry algae powder and suspend, add methyl alcohol/chloroform mixed liquor (2:1, V:V) concussion 2h with distilled water; Centrifugal, chloroform layer is transferred in the rotary evaporation bottle of weighing in advance, use Rotary Evaporators in the time of 60 ℃ with the chloroform evaporate to dryness; Weigh, the difference of quality is the TL amount before and after the rotary evaporation bottle, calculates total fat percentage composition with following formula.
Total fat percentage composition=(total fat weight/dry algae powder is heavy) * 100%.
The method of the middle Sawayama of the hydro carbons assay method reference literature that relates in following examples (Sawayama S, et al. Applied Microbiology and Biotechnology, 1992) etc.; Take by weighing the algae powder of certain mass, add the 15mL n-hexane and in mortar, fully grind, centrifugal then; Collect the n-hexane extract; Sediment is repeated said process more than 2 times, collect all n-hexane extracts, be transferred in the rotary evaporation bottle of weighing in advance; Place then on the Rotary Evaporators and under 30 ℃, evaporate n-hexane, weigh.The difference of quality is the total hydrocarbon quality before and after the rotary evaporation bottle, calculates the total hydrocarbon percentage composition with following formula.
Total hydrocarbon percentage composition=(total hydrocarbon weight/dry algae powder is heavy) * 100%.
Embodiment 1
In using the process of fish pond breeding wastewater, investigated the influence that temperature, intensity of illumination are grown to Blang's grape frustule as medium culture Blang grape algae.The growth conditions optimizing process is following: select 15 ℃, 20 ℃, 25 ℃, 30 ℃, 35 ℃ temperature and 1000Lx, 3000Lx, 5000Lx, 7000Lx, 9000Lx, the illumination condition of 11000Lx, carry out the combination of two kinds of conditions and cultivate.Cultivating end back collection frustule mensuration biomass.Optimization result shows: the Blang's grape algae algae in the sewage is 20 ℃ ~ 30 ℃ of temperature, well-grown under intensity of illumination 3000Lx ~ 8000Lx, and biomass reaches more than the 0.5g/L.Wherein temperature is 25 ℃, and light intensity is under the condition of 5000Lx, and it is best to grow, and its biomass reaches more than the 0.65g/L.
Embodiment 2
In the sanitary sewage of preliminary treatment, add the NaHCO of different quality
3, making the culture fluid that concentrations of inorganic carbon is 0 ~ 20mg/L respectively, Blang's grape algae inoculum concentration is 10%.At Light To Dark Ratio is that 13h:11h, temperature are that 25 ℃, intensity of illumination are to cultivate 30 days under the condition of 5000 Lx, wait to cultivate to finish the back suction filtration and collect frustule, freeze drying and prepare the algae powder after measure total lipid content and total hydrocarbon content.
The result shows: add concentrations of inorganic carbon in the sewage when being 5 ~ 15mg/L, Blang's grape algae algae well-grown also has high yield can effect, and biomass reaches as high as 0.6g/L, and total lipid content reaches as high as 17.42%, and total hydrocarbon content reaches as high as 34.50%;
Embodiment 3
In the lake water of eutrophication, add the KNO of different quality
3, making the culture fluid that nitrogen concentration is 2 ~ 20mg/L respectively, Blang's grape algae inoculum concentration is 10%.At Light To Dark Ratio is 13h:11h, and temperature is 25 ℃, cultivates 30 days under the condition of intensity of illumination 5000 Lx.Wait cultivate to finish the back and collect Blang's grape algae frustule with suction filtration, freeze drying and prepare the algae powder after measure total lipid content and total hydrocarbon content.
The result shows: when total nitrogen concentration was 10 ~ 20mg/L in the sewage, Blang's grape algae well-grown also had high yield ability effect, and biomass can reach 0.588g/L, and total lipid content reaches as high as 22.64%, and total hydrocarbon content reaches as high as 35.10%;
Embodiment 4
In the river of eutrophication, add the KH of different quality
2PO
4,Make the culture fluid that phosphorus concentration is 0.2 ~ 2mg/L respectively, Blang's grape algae inoculum concentration is 10%.At Light To Dark Ratio is 13h:11h, and temperature is 25 ℃, cultivates 30 days under the condition of intensity of illumination 5000 Lx.Wait cultivate to finish the back suction filtration and collect frustule, freeze drying and prepare the algae powder after measure total lipid content and total hydrocarbon content.
The result shows: when total phosphorus concentration was 1 ~ 2mg/L in the sewage, Blang's grape algae well-grown also had high yield ability effect, and biomass can reach 0.646g/L, and total lipid content reaches as high as 27.40%, and total hydrocarbon content reaches as high as 32.11%;
Embodiment 5
In the waste water of fish pond, add the NaHCO of different quality
3, KNO
3, KH
2PO
4, making concentrations of inorganic carbon is 5 ~ 15mg/L, when total nitrogen concentration was 10 ~ 20mg/L, total phosphorus concentration was the culture fluid of 1 ~ 2mg/L.Blang's grape algae inoculum concentration is 10%.At Light To Dark Ratio is 13h:11h, and temperature is 25 ℃, cultivates under the condition of intensity of illumination 5000 Lx 30 days, wait to cultivate to finish the back suction filtration and collect Blang's grape algae frustule, freeze drying and prepare the algae powder after measure total lipid content and total hydrocarbon content.
Optimization result shows: the external source concentrations of inorganic carbon is 10mg/L in the sewage, and total nitrogen concentration is 15mg/L, when total phosphorus concentration is 2mg/L; Blang's grape algae well-grown and have high yield can effect; Biomass can reach 0.635g/L, and total lipid content can reach 24.35%, and hydrocarbon content can reach 35.73%.
Utilize the total fat of Blang's grape algae that obtains, can process biodiesel, can be processed into gasoline, diesel oil use after contained total hydrocarbon materials extracts through ester exchange reaction.
List of references
[1]?Bligh?E?G,?Dyer?W?J.?A?rapid?method?of?total?lipid?extraction?and?purification[J].?Canadian?Journal?of?Biochemistry?and?Physiology,?1959,?37(8):?911-917
[2]?Chen?F,?John?M?R.?Effect?of?C/N?ratio?and?aeration?on?the?fatty?acid?composition?of?heterotrophic
?Chlorella?sorokiniana[J].?Journal?of?Applied?Physiology,?1991,?3:203-209
[3]?Sawayama?S,?Minowa?T,?Dotc?Y.?Growth?of?the?hydrocarbon?rich?microalga?
Botryococcus?braunii?in?secondarily?treated?sewage[J].?Applied?Microbiology?and?Biotechnology,?1992,?38(2):135-138
Claims (1)
1. the cultural method of Blang's grape algae; It is characterized in that: inorganic carbon source, nitrogenous source, phosphorus source with adding are adjusted nutrient concentration in the sewage; Make the concentration of DIC, total nitrogen, total phosphorus be respectively 5 ~ 15mg/L, 10 ~ 20mg/L, 1 ~ 2mg/L, said inorganic carbon source, nitrogenous source, phosphorus source are respectively Na
2CO
3, KNO
3, KH
2PO
4Chlorella is put into above-mentioned culture fluid; 20 ℃ ~ 30 ℃ of temperature, intensity of illumination 3000Lx ~ 8000Lx, Light To Dark Ratio are that 13h:11h cultivated 30 days down; Said sewage is river, river, lake water and the aquaculture water of sanitary sewage and eutrophication, promptly cultivates the high density Blang's grape algae that becomes to be rich in grease and hydrocarbon with sewage.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103571753A (en) * | 2012-07-30 | 2014-02-12 | 新奥科技发展有限公司 | Botryococcus braunii and application thereof |
| CN103571752A (en) * | 2012-07-30 | 2014-02-12 | 新奥科技发展有限公司 | Botryococcus braunii and application thereof |
| CN104496654A (en) * | 2014-12-03 | 2015-04-08 | 东莞市绿安奇生物工程有限公司 | Method for preparing and using low-heavy-metal-content chlorella cultivation raw material mother solution |
| CN115478019A (en) * | 2022-10-13 | 2022-12-16 | 暨南大学 | Culture method of hydrocarbon-producing botryococcus braunii SC-1 for producing squalene |
Citations (4)
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|---|---|---|---|---|
| CN101967450A (en) * | 2010-11-12 | 2011-02-09 | 河南师范大学 | Method for cultivating chlorella |
| CN101979497A (en) * | 2010-08-26 | 2011-02-23 | 北京芳能科技有限公司 | Culture method for efficiently inducing lipid accumulation in Botryococcus braunii |
| CN102174459A (en) * | 2011-03-10 | 2011-09-07 | 常州大学 | Method for promoting quick proliferation of Botryococcus braunii |
| CN102295983A (en) * | 2011-07-21 | 2011-12-28 | 上海辰浩生物技术有限公司 | Production method for comprehensive utilization of microalgae |
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2012
- 2012-02-06 CN CN2012100249246A patent/CN102550263A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101979497A (en) * | 2010-08-26 | 2011-02-23 | 北京芳能科技有限公司 | Culture method for efficiently inducing lipid accumulation in Botryococcus braunii |
| CN101967450A (en) * | 2010-11-12 | 2011-02-09 | 河南师范大学 | Method for cultivating chlorella |
| CN102174459A (en) * | 2011-03-10 | 2011-09-07 | 常州大学 | Method for promoting quick proliferation of Botryococcus braunii |
| CN102295983A (en) * | 2011-07-21 | 2011-12-28 | 上海辰浩生物技术有限公司 | Production method for comprehensive utilization of microalgae |
Non-Patent Citations (1)
| Title |
|---|
| 邓光，李夜光，胡鸿钧，齐雨藻，耿亚红，李中奎: "温度、光照和PH值对锥状斯氏藻和塔玛亚历山大藻光合作用的影响及光暗周期对其生长速率和生物量的影响", 《武汉植物学研究》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103571753A (en) * | 2012-07-30 | 2014-02-12 | 新奥科技发展有限公司 | Botryococcus braunii and application thereof |
| CN103571752A (en) * | 2012-07-30 | 2014-02-12 | 新奥科技发展有限公司 | Botryococcus braunii and application thereof |
| CN103571752B (en) * | 2012-07-30 | 2016-01-13 | 新奥科技发展有限公司 | Botryococcus braunii and application thereof |
| CN104496654A (en) * | 2014-12-03 | 2015-04-08 | 东莞市绿安奇生物工程有限公司 | Method for preparing and using low-heavy-metal-content chlorella cultivation raw material mother solution |
| CN115478019A (en) * | 2022-10-13 | 2022-12-16 | 暨南大学 | Culture method of hydrocarbon-producing botryococcus braunii SC-1 for producing squalene |
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Application publication date: 20120711 |