CN105969664A - Method for promoting oil accumulation of microalgae by adding high-concentration organic wastewater into natural seawater - Google Patents
Method for promoting oil accumulation of microalgae by adding high-concentration organic wastewater into natural seawater Download PDFInfo
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- CN105969664A CN105969664A CN201610322867.8A CN201610322867A CN105969664A CN 105969664 A CN105969664 A CN 105969664A CN 201610322867 A CN201610322867 A CN 201610322867A CN 105969664 A CN105969664 A CN 105969664A
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- organic wastewater
- water
- concentrated organic
- high concentrated
- natural sea
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/12—Unicellular algae; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
- C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
Abstract
The invention discloses a method for promoting oil accumulation of microalgae by adding high-concentration organic wastewater into natural seawater, belonging to the technical field of microalga biology. The method comprises the following steps: by using natural seawater with 1-10% of high-concentration organic wastewater as a culture medium of the experimental group, carrying out culture under continuous illumination conditions until the microalgae stop growing, and carrying out centrifugal separation to collect the algae. The result indicates that the oil yield of the microalgae cultured in the natural seawater with high-concentration organic wastewater is obviously enhanced, so the method lowers the culture cost of microalgae and is worthy of popularization and use.
Description
Technical field
The present invention relates to technical field of microalga biology, particularly relate to one and add highly concentrated in natural sea-water
The method that degree organic wastewater promotes microalgae grease accumulation.
Background technology
Along with developing rapidly of economy and society, the mankind are to Fossil fuels such as coal, oil, natural gass
Demand is more and more higher.But the large-scale use of Fossil fuel seriously exacerbates energy crisis and the whole world
Climate change, therefore, seek a kind of new regenerative resource in the urgent need to.Microalgae is owing to having
Cell proliferation is fast, growth cycle is short, carbon sequestration capacity is strong, photosynthetic efficiency is high, environmental suitability by force, no
Take the features such as agriculture plantation and be widely used as producing the raw material of biodiesel.Yet with microalgae
The problem that toxigenic capacity is high constrains the large-scale commercial of microalgae oil-producing and produces.Therefore, one is sought
The micro-algae culture medium of low cost becomes focus and the focus of research.
Many researcheres have studied and utilize waste water to cultivate microalgae.Ting Cai et al. is in artificial sea
Water with the addition of the industrial wastewater anaerobism high concentrated organic wastewater of different proportion to cultivate cytoalgae, wherein
When the ratio of high concentrated organic wastewater is 3%, cytoalgae obtains maximum biomass yield (150.9
± 8.6mgL-1d-1) and oils and fats productivity (19.9 ± 1.2mgL-1d-1) (Cai et al., 2013).
Joo-Young Jung et al. cultivates scenedesmus obliquus with the natural sea-water of different proportion and BG11, wherein when
Maximum Biomass (1.1g/L) and fatty acid methyl ester yield is obtained when the ratio of natural sea-water is 10%
(9%) (Jung et al., 2015).But preparation artificial seawater and BG11 need to consume substantial amounts of fresh water
Resource.Comparatively speaking, sea water, rich reserves, cover the 71% of earth surface, containing being conducive to
The abundant mineral of marine organism growth, mainly Na+, Cl-, Mg2+, SO42-, Ca2+.Wherein
Sea water is famous with high salinity, this accumulation being conducive to stimulating microalgae grease, micro-hence with cultivation in sea water
Algae has the biggest development prospect.But it is again worthless for directly utilizing cultivation in sea water microalgae, because
Micro algae growth is except needs mineral, in addition it is also necessary to the nutrient substance that nitrogen, phosphorus etc. are abundant.And nitrogen in sea water,
The content of the nutrient substance such as phosphorus is at a fairly low, is insufficient for the needs of micro algae growth.High concentration organic waste
Containing nutrient substance such as abundant nitrogen, phosphorus inside water.If directly discharged, serious environment can be caused
Pollute.If high concentrated organic wastewater being added to as culture medium to cultivate microalgae in sea water, sea water
Can be as the derivant of microalgae grease accumulation, high concentrated organic wastewater can be as the battalion of micro algae growth
The source of supporting, thus reach the doulbe-sides' victory purpose of micro algae growth and oil and fat accumulation.And sea water rich reserves, become
This is relatively low, and high concentrated organic wastewater have also been obtained process simultaneously.So with the addition of high concentration organic waste
It is a kind of economic and environment-friendly effective method that the natural sea-water of water cultivates microalgae.
Summary of the invention
The invention provides a kind of high concentrated organic wastewater that adds in natural sea-water to cultivate microalgae
Method, is greatly facilitated the accumulation of microalgae grease, can well solve microdisk electrode cost high simultaneously
Problem.
For solving above-mentioned technical problem, the present invention provides technical scheme as follows:
A kind of method adding high concentrated organic wastewater promotion microalgae grease accumulation in natural sea-water, its
It is characterized as comprising the steps:
(1) little double-colored algae SDEC-6 is placed in BG11 culture medium carries out enrichment culture；
(2) preparation mixed culture medium, described mixed culture medium is to be by natural sea-water and ratio
The high concentrated organic wastewater composition of 1%-10%；
(3) the frustule seed liquor that step (1) prepares is inoculated into the mixing that step (2) prepares
In culture medium, postvaccinal biomass density is 0.06-0.07g/L；
(4) reaction unit that step (3) prepares is placed in phjytotron cultivation, condition of culture
For: temperature 25 ± 1 DEG C, 81 μm ol photons/m2/s continuous illuminations.
The method have the advantages that
(1) present invention adds high concentrated organic wastewater in natural sea-water and promotes microalgae grease accumulation
Method, hence it is evident that improve the oils and fats productivity of microalgae, can be as the preferable training method of one.
(2) present invention adds high concentrated organic wastewater in natural sea-water and promotes microalgae grease accumulation
Method, natural sea-water rich reserves, cost is relatively low, and microalgae obtains more satisfactory growth speed simultaneously
Rate, therefore reduces the cost of microdisk electrode.
(3) present invention adds high concentrated organic wastewater in natural sea-water and promotes microalgae grease accumulation
Method so that high concentrated organic wastewater have also been obtained process, environmental protection and economy simultaneously.
Accompanying drawing explanation
Fig. 1 is little double-colored algae SDEC-6 growth curve in different culture media.
Fig. 2 is that little double-colored algae SDEC-6 is in the oil and fat accumulation characteristic in different culture media.
Detailed description of the invention
The present invention is further elaborated by mode by the following examples.
1, algae strain, the microalgae kind choosing cultivation is little double-colored algae SDEC-6, and screening is from Ji Nanquan
Park, city, and it is stored in CHINESE FRESHWATER algae kind storehouse.
2, natural sea-water, is selected from the Huanghai Sea (Shandong Rizhao marine site), each nutrient in natural sea-water
The concentration of matter and mineral is respectively as follows: TN:10-50mg/L, NH3-N:1-10mg/L, TP:0.01-1
mg/L,pH:8-10,Na:10000-15000mg/L,Mg:1000-1500mg/L,Ca:800-1000
mg/L,K:200-500mg/L,Si:60-90mg/L。
3, high concentrated organic wastewater, is selected from Jinan ten side's environmental protection company, high concentrated organic wastewater
Initial water quality is: TN:2000-3000mg/L, NH3-N:2000-3000mg/L, TP:10-50
Mg/L, COD:5000-8000mg/L, pH:8-10.
Embodiment 1: the growth characteristics of little double-colored algae SDEC-6
1, frond incubation:
(1) little double-colored algae SDEC-6 is placed in BG11 culture medium carries out enrichment culture, BG11
Medium component is as follows: NaNO3 1.5g/L, K2HPO4 40mg/L, MgSO4 7H2O 75
mg/L,CaCl2·2H2O 36mg/L,Citric acid 6mg/L,Ferric ammonium citrate 6
mg/L,EDTANa2 1mg/L,Na2CO3 20mg/L,A5 solution 1mL/L.A5 solution:
H3BO3 2.86g/L,MnCl2·4H2O 1.86g/L,ZnSO4·7H2O 0.22g/L,Na2MoO4
·2H2O 0.39g/L,CuSO4·5H2O 0.08g/L,Co(NO3)2·6H2O 0.05g/L。
(2) the algae seed liquor that step (1) obtained dilute ten times after at 680nm its extinction
Degree is 0.172, and its dry weight is 0.06-0.07g/L.
(3) seed liquor obtained in step (2) being inoculated into high concentrated organic wastewater ratio is
In the mixed culture medium of 1%-10%, and with BG11, natural sea-water and high concentrated organic wastewater for cultivating
Base respectively as control experiment, in the controlled environment chamber in carry out constant temperature culture, in incubation, every day takes
Sample measures Biomass and carries out water analysis.
(4) treat that the frond in step (3) stops growing, algae solution is centrifuged results, outwells
Clear liquid, obtains algae mud.
(5) the algae mud in step (4) is carried out lyophilization, it is thus achieved that dry algae powder.
2, frond biomass estimation:
The every day same time takes 20mL algae solution centrifugal 10min layering, algae under the conditions of 5000r/min
Mud takes out and is placed in thermostatic drying chamber constant temperature drying at 60 DEG C and weighs mensuration biomass concentration (g/L).
Supernatant is used for analyzing concentration of nitrogen and phosphorus through 0.45 μm cellulose acetate ultrafiltration through membranes.The mensuration of total nitrogen
Using alkaline chitinase to clear up ultraviolet spectrophotometry (HJ636-2012), the mensuration of ammonia nitrogen uses
Berthelot spectrophotometry (HJ535-2009), total phosphorus yield uses ammonium molybdate spectrophotometric method
(GB11893-89)。
3, biomass estimation result:
(1) as it is shown in figure 1, at the simple medium and small double-colored algae SDEC-6 of high concentrated organic wastewater not
Can growth；In simple sea water the lowest due to nutrient substance, little double-colored algae SDEC-6 growth fraction is relatively
Slowly.In the natural sea-water that high concentrated organic wastewater adding proportion is 1%-10%, little double-colored algae
SDEC-6 all has growth.
(2) as shown in table 1, it is the natural sea-water of 1%-10% in high concentrated organic wastewater adding proportion
In, the Biomass of little double-colored algae SDEC-6 is apparently higher than the Biomass in BG11, natural sea-water；
Wherein in the natural sea-water that high concentrated organic wastewater adding proportion is 4%, little double-colored algae SDEC-6
Biomass can reach 0.38g/L, average organism matter productivity is 26.08mg/L/d, specific growth rate
For 0.18d-1。
4, interpretation of result:
Add the natural sea-water that high concentrated organic wastewater ratio is 1%-10% and can compare reason as one
The substitutive medium thought obtains biomass to cultivate little double-colored algae SDEC-6, this is because add at this
Concentration of nitrogen and phosphorus in proportion is relatively suitable for the growth of little double-colored algae SDEC-6, and in sea water
NaCl can increase the salinity in mixed culture medium thus stimulate algae metabolic activity accelerate algae
The growth of class, the high salinity in sea water can avoid the invasion of other microorganisms simultaneously.
The little double-colored algae SDEC-6 of table 1 final dry weight in different culture media, average organism matter productivity, ratio
Growth rate.
Embodiment 2: the oil and fat accumulation characteristic of little double-colored algae SDEC-6
1, fat content measures:
Weigh the dry algae powder obtained in about 0.1g embodiment 1 in 50mL centrifuge tube, add 10mL
Chloroform/methanol (v/v=2:1) solution, processes 10min (frequency 20%) with Ultrasonic Cell Disruptor, then
4000r/min is centrifuged 10min, has been centrifuged solution and has divided biphase, supernatant is transferred to 60mL separatory
In funnel, whole extraction process is repeated twice.According to the volume of oils and fats extracting solution, add 0.9%
Sodium chloride solution (adding volume is the 1/5 of oils and fats extracting solution, about 4-5mL sodium chloride solution), fills
Divide and shake up 1min, and stand 15min.Measure the volume of low phase solution, and take 5mL low phase solution
In 10mL teat glass, dry up with nitrogen, glass tubing is placed in 60 DEG C of baking ovens and dries to constant weight
(about 30min).
2, fat content calculates:
In formula: LW is based on the fat content under dry weight, g/g
M1 algae dried bean noodles weight, g
The 10mL glass tubing dry weight of m2 band oils and fats, g
M0 10mL glass tubing dry weight, g
The volume of v low phase oils and fats, mL
3, fat content interpretation of result:
As in figure 2 it is shown, the little double-colored algae added in the natural sea-water of different proportion high concentrated organic wastewater
The fat content of SDEC-6 is obviously higher than the fat content in BG11 and natural sea-water, and this is
Owing to the salinity in sea water is conducive to stimulating the synthesis of oils and fats in frustule.Wherein high concentrated organic wastewater
Adding proportion when being 2% the fat content of little double-colored algae SDEC-6 reach the highest (50.13%), be
In BG11 1.72 times of fat content.When the adding proportion of high concentrated organic wastewater is 4% little double
The oils and fats productivity of color algae SDEC-6 reaches the highest (13.36mg/L/d), is oils and fats productivity in BG11
2.20 again.Therefore, from oil and fat accumulation data, with the addition of the natural sea-water of high concentrated organic wastewater
Little double-colored algae SDEC-6, the oils and fats obtained can be cultivated as the substitutive medium of a kind of low cost
It is significantly larger than the fat content in BG11 and natural sea-water.
Comparative example " method cultivating chroococcoid with sanitary sewage diluting high-concentration organic wastewater " (application
Numbers 201610034390.3) using high concentrated organic wastewater in is culture medium main body, by adding difference
The sanitary sewage of ratio cultivates chroococcoid, it is thus achieved that certain biomass yield and oil and fat accumulation, result
Show, when high concentrated organic wastewater is diluted 10 times, fat content (38.65%) and oils and fats productivity
(11.79mg/L/d) maximum is reached.The present invention uses natural sea-water to be culture medium main body, passes through
The high concentrated organic wastewater adding different proportion cultivates microalgae, adds high concentrated organic wastewater ratio and is
When 2%, fat content (50.13%) reaches maximum, and adding high concentrated organic wastewater ratio is 2%
Time, oils and fats productivity (13.36mg/L/d) reaches maximum.Drawing through comparing, the present invention uses
In natural sea-water add high concentrated organic wastewater promote microalgae grease accumulation method can be obvious
Improve fat content and the oils and fats productivity of frond.And natural sea-water storage capacity is very big, easily obtains,
Reducing toxigenic capacity, the large-scale commercial concurrently facilitating microalgae oil-producing produces.
The above is the preferred embodiment of the present invention, it is noted that general for the art
For logical technical staff, on the premise of without departing from principle of the present invention, it is also possible to make some changing
Entering and retouch, these improvements and modifications also should be regarded as protection scope of the present invention.
Claims (9)
1. in natural sea-water, add the method that high concentrated organic wastewater promotes microalgae grease accumulation,
It is characterized by comprise the steps:
(1) little double-colored algae SDEC-6 is placed in BG11 culture medium carries out enrichment culture；
(2) preparation mixed culture medium, described mixed culture medium is to be by natural sea-water and ratio
The high concentrated organic wastewater composition of 1%-10%；
(3) the frustule seed liquor that step (1) prepares is inoculated into the mixing that step (2) prepares
In culture medium, postvaccinal biomass density is 0.06-0.07g/L；
(4) reaction unit that step (3) prepares is placed in phjytotron cultivation, condition of culture
For: temperature 25 ± 1 DEG C, 81 μm ol photons/m2/ s continuous illumination.
The high concentrated organic wastewater that adds in natural sea-water the most according to claim 1 promotes micro-
The method of algae oil and fat accumulation, it is characterised in that the described natural sea-water in step (2) is for by taking out
Filter off and removed wherein algae.
The high concentrated organic wastewater that adds in natural sea-water the most according to claim 2 promotes micro-
The method of algae oil and fat accumulation, it is characterised in that the sucking filtration filter membrane used by described natural sea-water is 0.45 μ
M cellulose acetate filter membrane.
The high concentrated organic wastewater that adds in natural sea-water the most according to claim 3 promotes micro-
The method of algae oil and fat accumulation, it is characterised in that each nutrient substance and mineral in described natural sea-water
Concentration is respectively as follows: TN:26.38 ± 0.04mg/L, and NH3-N:3.83 ± 0.76mg/L, TP:0.016 ± 0.00
mg/L,pH:9.37±0.02,Na:13920±142mg/L,Mg:1295±3mg/L,Ca:
988.8±13.6mg/L,K:346.6±7.2mg/L,Si:78.65±0.34mg/L。
The high concentrated organic wastewater that adds in natural sea-water the most according to claim 1 promotes micro-
The method of algae oil and fat accumulation, it is characterised in that the described high concentrated organic wastewater in step (2) is
By six layers of filtered through gauze centrifugal.
The high concentrated organic wastewater that adds in natural sea-water the most according to claim 5 promotes micro-
The method of algae oil and fat accumulation, it is characterised in that be to the speed that described high concentrated organic wastewater is centrifugal
4500-6500r/min, duration is 8-10min.
The high concentrated organic wastewater that adds in natural sea-water the most according to claim 6 promotes micro-
The method of algae oil and fat accumulation, it is characterised in that the initial water quality of described high concentrated organic wastewater is:
TN:2326.72 ± 20mg/L, NH3-N:2153.07 ± 25mg/L, TP:20.24 ± 0.1mg/L,
COD:6096.1 ± 65mg/L, pH:8.31 ± 0.1.
The high concentrated organic wastewater that adds in natural sea-water the most according to claim 1 promotes microalgae
The method of oil and fat accumulation, it is characterised in that the height in mixed culture medium described in described step (2)
The coefficient optimum proportion of concentration organic wastewater is 2%-6%.
The method adding high concentrated organic wastewater in natural sea-water the most according to claim 1,
It is characterized in that, the cultivation cycle of described step (4) is 12 days.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107460153A (en) * | 2017-09-12 | 2017-12-12 | 山东大学 | A kind of natural sea-water adds the method for gourmet powder fermenting mother liquor production spiral algal biomass |
| CN111548971A (en) * | 2020-06-02 | 2020-08-18 | 江苏海洋大学 | Preparation method of economic microalgae high-efficiency culture medium based on laver processing wastewater |
| CN115029248A (en) * | 2022-06-21 | 2022-09-09 | 昆明理工大学 | Method for improving microalgae lipid yield by utilizing recycled wastewater |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107460153A (en) * | 2017-09-12 | 2017-12-12 | 山东大学 | A kind of natural sea-water adds the method for gourmet powder fermenting mother liquor production spiral algal biomass |
| CN111548971A (en) * | 2020-06-02 | 2020-08-18 | 江苏海洋大学 | Preparation method of economic microalgae high-efficiency culture medium based on laver processing wastewater |
| CN115029248A (en) * | 2022-06-21 | 2022-09-09 | 昆明理工大学 | Method for improving microalgae lipid yield by utilizing recycled wastewater |
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