CN103484372A - Culture method for increasing biomass and grease yield of microalgae - Google Patents
Culture method for increasing biomass and grease yield of microalgae Download PDFInfo
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- 239000004519 grease Substances 0.000 title claims abstract description 30
- 239000002028 Biomass Substances 0.000 title claims abstract description 17
- 238000012136 culture method Methods 0.000 title abstract 4
- 238000000034 method Methods 0.000 claims abstract description 21
- 241000224476 Nannochloropsis salina Species 0.000 claims abstract description 14
- 235000016709 nutrition Nutrition 0.000 claims abstract description 13
- 230000035764 nutrition Effects 0.000 claims abstract description 13
- 241001300629 Nannochloropsis oceanica Species 0.000 claims description 13
- 235000003784 poor nutrition Nutrition 0.000 claims description 11
- 239000011573 trace mineral Substances 0.000 claims description 11
- 235000013619 trace mineral Nutrition 0.000 claims description 11
- 230000012010 growth Effects 0.000 claims description 10
- 229910019142 PO4 Inorganic materials 0.000 claims description 6
- YOZNUFWCRFCGIH-BYFNXCQMSA-L hydroxocobalamin Chemical compound O[Co+]N([C@]1([H])[C@H](CC(N)=O)[C@]\2(CCC(=O)NC[C@H](C)OP(O)(=O)OC3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)C)C/2=C(C)\C([C@H](C/2(C)C)CCC(N)=O)=N\C\2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O YOZNUFWCRFCGIH-BYFNXCQMSA-L 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 6
- 239000010452 phosphate Substances 0.000 claims description 6
- 235000013343 vitamin Nutrition 0.000 claims description 6
- 239000011782 vitamin Substances 0.000 claims description 6
- 229930003231 vitamin Natural products 0.000 claims description 6
- 229940088594 vitamin Drugs 0.000 claims description 6
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 6
- 241000195493 Cryptophyta Species 0.000 claims description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 3
- 229930003756 Vitamin B7 Natural products 0.000 claims description 3
- 229960001103 hydroxocobalamin Drugs 0.000 claims description 3
- 235000004867 hydroxocobalamin Nutrition 0.000 claims description 3
- 239000011704 hydroxocobalamin Substances 0.000 claims description 3
- 238000005286 illumination Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 235000002639 sodium chloride Nutrition 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 239000011735 vitamin B7 Substances 0.000 claims description 3
- 235000011912 vitamin B7 Nutrition 0.000 claims description 3
- 230000005526 G1 to G0 transition Effects 0.000 claims description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 abstract description 6
- 229910052698 phosphorus Inorganic materials 0.000 abstract description 6
- 239000011574 phosphorus Substances 0.000 abstract description 6
- 238000012258 culturing Methods 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 abstract 1
- 239000002609 medium Substances 0.000 abstract 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 14
- 229910052757 nitrogen Inorganic materials 0.000 description 7
- 239000003921 oil Substances 0.000 description 4
- 239000002551 biofuel Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 230000005791 algae growth Effects 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- -1 iron ion Chemical class 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 230000011218 segmentation Effects 0.000 description 2
- 240000009108 Chlorella vulgaris Species 0.000 description 1
- 235000007089 Chlorella vulgaris Nutrition 0.000 description 1
- 241000264606 Tetradesmus dimorphus Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003245 coal Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 239000003345 natural gas Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a culture method for increasing the biomass and grease yield of microalgae in the technical field of algal biological culture. The culture method comprises the following steps: culturing the microalgae in a nutrition-rich culture medium until a logarithmic phase with a nutrition-rich and nutrition-poor two-stage culture method for Nannochloropsis salina at first, then concentrating and collecting the microalgae, and inoculating the microalgae to a phosphorus-free or microelement-free nutrition-poor medium with the microalgae for continuous culture. According to the method, additional nutrition does not need to be added, so that the cost is reduced, the biomass yield, the cell grease content and the final grease yield of the microalgae are greatly increased, and the shortcoming that only the cell grease content is increased but the final grease yield is still very low in a traditional nitrogen-free culture process is overcome; in the absence of phosphorus, the grease content of the microalgae reaches 46% and the biomass and grease yields are increased by 30% and 117% respectively; and in the absence of microelements, the grease content of the microalgae reaches 40% and the biomass and grease yields are increased by 65% and 133% respectively.
Description
Technical field
The present invention relates to the culture technique of algae bio, particularly a kind of cultural method that improves micro-algal biomass and grease yield.
Background technology
The continuous consumption of tradition fossil oil and the mankind, to the continuous growth of energy demand, have caused day by day serious energy dilemma.There are some researches show, counting coal, oil and natural gas in the world will exhaust greatly from 2005 about 107 years, 35 years and 37 years.Seek a kind of environmental protection, renewable energy source becomes the large problem that countries in the world are needed solution badly.Utilize micro-algae production biofuel that a solution with great potential is provided.The contrast of micro-algae and other biological, because of its fast growth, floor space is little, training method is simple, and the part algae contains very high lubricant component and is paid close attention to more and more widely.Utilize micro-algae production biofuel to be considered to current unique a kind of bioenergy that can replace the conventional traffic transport fuel.Therefore, in order to improve grease yield and to reduce costs, current normal culture condition and culture process need to further be optimized and be improved.
Micro algae growth speed under the normal growth condition compares comparatively fast, but fat content is lower.At certain under special condition pressure, the grease that micro-algae often can enriched.Such as, the one piece of report (99 volumes, 11 phases, 4717-4722 page) on " Bioresource technology " periodical in 2008, find, the iron ion of high density can bring out the grease that micro-algae Chlorella vulgaris produces high-content.Yet this method need to increase the concentration of iron ion in nutrient solution, not only increased cost but also can cause the pollution of water body.It is current more common a kind of grease enriching method that nitrogen stress is cultivated, but under this condition, the speed of growth of micro-algae has been suppressed widely, causes final grease yield not improved significantly.Chinese patent " is supported the method that improves oil-producing microalgae biomass and oil and fat accumulation with rich two stage of nitrogen-nitrogen stress training strategy ", and (patent No. 201110192159.4) proposes to add 5.4g/L glucose in micro-algae Scenedesmus dimorphus nitrogen stress stage simultaneously with holding concurrently, and improved micro-algal biomass output and fat content (reaching 29.62%).However, this method is only suitable in the micro-algae of row growth of can holding concurrently, and has increased cost adding of high concentration glucose.Therefore, how to design or find a kind of cheaply, can improve fat content and again the very little cultural method of speed of growth impact is become to a current large problem of utilizing micro-algae production biofuel.
Summary of the invention
In order to solve the problem in background technology, the invention provides a kind of cultural method that improves micro-algal biomass and grease yield, by eutrophy and dystrophic second order segmentation, cultivate, improve microalgae cell fat content and total grease yield.
Technical scheme of the present invention: a kind of cultural method of intending Nannochloropsis oceanica N.salina biomass and grease yield that improves, carry out the cultivation of priority two-section type to intending Nannochloropsis oceanica N.salina, after first eutrophy cultivation, poor nutrition is cultivated.At first, by intending Nannochloropsis oceanica N.salina, be inoculated in the f/2 substratum that normally is rich in nutrition, initial pH is 8,24 ± 2 ℃ of temperature, illumination 79.29 μ mol m
-2s
-1, aerated culture is to logarithmic phase.Be rich in the f/2 substratum of nutrition, every liter contains: the artificial sea salt of 33.6g, 0.075gNaNO
3, 0.00565g NaH
2pO
42H
2o, 1ml trace element and 1ml VITAMIN.Wherein, every liter of trace element contains: 4.16g Na
2eDTA, 3.15g FeCl
36H
2o, 0.18g MnCl
24H
2o, 10mg CoCl
26H
2o, 10mgCuSO
45H
2o, 22mg ZnSO
47H
2o, 6mg Na
2moO
42H
2o.Every liter of VITAMIN contains: 100mg VITMAIN B1,0.5mg hydroxocobalamin(e) nd0.5mg vitamin H.Nannochloropsis oceanica N.salina to be intended is cultured to the logarithmic growth after date, by algae liquid centrifugal concentrating, then is inoculated in dystrophic f/2 substratum, continues to be cultured to the stationary phase of growing.In poor nutrition f/2 substratum, phosphate-containing, can still not contain trace element by phosphate-containing yet, and all the other components are identical with normal eutrophy f/2 substratum.
The present invention and prior art contrast have following beneficial effect: the method that the present invention adopts eutrophy and poor nutrition second order segmentation to cultivate, reduced cost without adding extra nutrition, and greatly promoted the yield of biomass of micro-algae, cell fat content and final grease yield.Overcome traditional nitrogen stress and cultivated and only increase the cell fat content, but final grease yield very low shortcoming still.Under scarce phosphorus condition, the fat content of micro-algae has reached 46%, and biomass and grease yield have improved respectively 30% and 117%.Lacking under micro-condition, the fat content of micro-algae has reached 40%, and biomass and grease yield have improved respectively 65% and 133%.
The accompanying drawing explanation
Fig. 1 is process flow diagram of the present invention.
Fig. 2 intends the growth curve of Nannochloropsis oceanica (N.salina) under the Different Nutrition condition in embodiments of the invention.
Fig. 3 intends the fat content of Nannochloropsis oceanica (N.salina) under the Different Nutrition condition in embodiments of the invention.
Fig. 4 intends the grease yield of Nannochloropsis oceanica (N.salina) under the Different Nutrition condition in embodiments of the invention.
Embodiment
Embodiment 1. the present embodiment improve the cultural method concrete steps following (process flow sheet is referring to Fig. 1) of intending Nannochloropsis oceanica (N.salina) biomass and grease yield:
1. the eutrophy of micro-algae is cultivated: will intend Nannochloropsis oceanica N.salina and be inoculated in the f/2 substratum that normally is rich in nutrition, initial pH is 8,24 ± 2 ℃ of temperature, illumination 79.29 μ mol m
-2s
-1, aerated culture is to logarithmic phase.Be rich in the f/2 substratum of nutrition, every liter contains: the artificial sea salt of 33.6g, 0.075g NaNO
3, 0.00565g NaH
2pO
42H
2o, 1ml trace element (T) and 1ml VITAMIN.Wherein, every liter of micro-T contains: 4.16g Na
2eDTA, 3.15g FeCl
36H
2o, 0.18g MnCl
24H
2o, 10mg CoCl
26H
2o, 10mg CuSO
45H
2o, 22mg ZnSO
47H
2o, 6mg Na
2moO
42H
2o.Every liter of VITAMIN contains: 100mg VITMAIN B1,0.5mg hydroxocobalamin(e) nd0.5mg vitamin H.
2. the collection of micro-algae is concentrated: Nannochloropsis oceanica N.salina to be intended is cultured to the logarithmic growth after date, by algae liquid middling speed centrifugal concentrating.
3. the poor nutrition of micro-algae is cultivated: the micro-algae after concentrating is inoculated in dystrophic f/2 substratum, continues to cultivate 3 days.In poor nutrition f/2 substratum, phosphate-containing, can still not contain micro-T by phosphate-containing yet, and all the other components are identical with normal eutrophy f/2 substratum.
When poor nutrition cultivation stage, cultivate and compare with original micro-algae, the micro algae growth Similar Broken Line of nitrogen stress, and the micro-algal biomass that lacks phosphorus and lack trace element all improves a lot, finally increased respectively 30% and 65%(Fig. 2).Aspect fat content, nitrogen stress, the micro-algae that lacks phosphorus and lack trace element reached respectively 38%, 46% and 40%(Fig. 3).Aspect final grease yield, cultivate and compare with original micro-algae, nitrogen stress, the micro-algae that lacks phosphorus and lack trace element improved respectively 42%, 117% and 133%(Fig. 4).Therefore, the grease yield that does not significantly improve micro-algae is cultivated in the poor nutrition of nitrogen stress.By contrast, the micro-algae that lacks phosphorus and scarce trace element has improved 117% and 133% than the grease yield of original micro-algae.
Claims (6)
1. one kind is improved the cultural method of intending Nannochloropsis oceanica N.salina biomass and grease yield, it is characterized in that micro-algae is carried out to the priority two-section type to be cultivated, i.e. first eutrophy poor nutrition cultivation after cultivating.
2. the method that micro-algae eutrophy of first stage is cultivated according to claim 1, is characterized in that being inoculated in the f/2 substratum that normally is rich in nutrition intending Nannochloropsis oceanica N.salina, and initial pH is 8,24 ± 2 ℃ of temperature, illumination 79.29 μ mol m
-2s
-1, aerated culture is to logarithmic phase.
3. the f/2 substratum that normally is rich in nutrition according to claim 2, is characterized in that every liter of f/2 substratum contains: the artificial sea salt of 33.6g, 0.075g NaNO
3, 0.00565g NaH
2pO
42H
2o, 1ml trace element and 1ml VITAMIN; Wherein, every liter of trace element contains: 4.16g Na
2eDTA, 3.15g FeCl
36H
2o, 0.18g MnCl
24H
2o, 10mgCoCl
26H
2o, 10mg CuSO
45H
2o, 22mg ZnSO
47H
2o, 6mg Na
2moO
42H
2o; Every liter of VITAMIN contains: 100mg VITMAIN B1,0.5mg hydroxocobalamin(e) nd0.5mg vitamin H.
4. the method that the poor nutrition of the micro-algae of subordinate phase is cultivated according to claim 1, it is characterized in that Nannochloropsis oceanica N.salina to be intended is cultured to the logarithmic growth after date, by algae liquid centrifugal concentrating, then be inoculated in dystrophic f/2 substratum, continue to be cultured to the stationary phase of growing.
5. poor nutrition f/2 substratum according to claim 4, is characterized in that not phosphate-containing, and all the other components are identical with normal eutrophy f/2 substratum.
6. poor nutrition f/2 substratum according to claim 4, is characterized in that also can still not containing trace element by phosphate-containing, and all the other components are identical with normal eutrophy f/2 substratum.
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Cited By (8)
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| CN105132404A (en) * | 2015-10-12 | 2015-12-09 | 山东大学 | Method for fast accumulating grease through ultrasonic stimulation micro algae |
| CN105647825A (en) * | 2014-11-13 | 2016-06-08 | 中国科学院大连化学物理研究所 | Method for concurrently improving spirulina biomass and polysaccharide yield |
| CN105858894A (en) * | 2016-02-19 | 2016-08-17 | 南昌大学 | Method for nitrogen abundance transformation treatment of wastewater with high ammonia nitrogen |
| CN106635810A (en) * | 2016-12-06 | 2017-05-10 | 沈阳化工研究院有限公司 | Microalgae and method for culturing microalgae |
| CN107586720A (en) * | 2017-09-28 | 2018-01-16 | 新奥科技发展有限公司 | Microalgae culture method |
| CN110229755A (en) * | 2019-06-17 | 2019-09-13 | 昆明理工大学 | A method of promote bloom nitrogen stress to coerce lower microalgae grease using epiphysin and accumulates |
| CN113462578A (en) * | 2021-08-26 | 2021-10-01 | 海南绿藻世界生物科技有限公司 | Microalgae culture medium and culture method |
| CN116769847A (en) * | 2023-08-09 | 2023-09-19 | 德默特生物科技(珠海)有限公司 | Method for improving EPA content in pseudo-microalgae oil |
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Cited By (10)
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| CN105647825A (en) * | 2014-11-13 | 2016-06-08 | 中国科学院大连化学物理研究所 | Method for concurrently improving spirulina biomass and polysaccharide yield |
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| CN116769847A (en) * | 2023-08-09 | 2023-09-19 | 德默特生物科技(珠海)有限公司 | Method for improving EPA content in pseudo-microalgae oil |
| CN116769847B (en) * | 2023-08-09 | 2023-12-05 | 德默特生物科技(珠海)有限公司 | Method for improving EPA content in pseudo-microalgae oil |
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Effective date of registration: 20170302 Address after: 361000 Xiamen Fujian torch hi tech Zone Venture Park building 619C room Patentee after: Xiamen Blue Environmental Technology Co. Ltd. Patentee after: Shenzhen science and Technology Co., Ltd. Patentee after: Nanjing Abel of Engineering Technology Co. Ltd. Address before: 362331 Quanzhou Province, Nanan City, the town of new town, Hong Kong, the first new teacher Village Lane No. five, Lane 8 Patentee before: Chen Yimin |
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Address after: 361000 North 313A room, Chuang Chuang business park, torch high tech Zone, Xiamen, Fujian Co-patentee after: Shenzhen science and Technology Co., Ltd. Patentee after: Xiamen Bo Lu Biological Technology Co., Ltd. Co-patentee after: Nanjing Abel of Engineering Technology Co. Ltd. Address before: 361000 619C building, Chuang Chuang Chuang Chuang, Xiamen torch high tech Zone, Fujian Co-patentee before: Shenzhen science and Technology Co., Ltd. Patentee before: Xiamen Blue Environmental Technology Co. Ltd. Co-patentee before: Nanjing Abel of Engineering Technology Co. Ltd. |
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Address after: 361000 North 313A room, Chuang Chuang business park, torch high tech Zone, Xiamen, Fujian Co-patentee after: Shenzhen Siwen Technology Co., Ltd. Patentee after: Bolu (Xiamen) biological Co., Ltd Co-patentee after: Nanjing Abel of Engineering Technology Co. Ltd. Address before: 361000 North 313A room, Chuang Chuang business park, torch high tech Zone, Xiamen, Fujian Co-patentee before: Shenzhen Siwen Technology Co., Ltd. Patentee before: XIAMEN BLUE BIOLOGICAL TECHNOLOGY Co.,Ltd. Co-patentee before: Nanjing Abel of Engineering Technology Co. Ltd. |
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Effective date of registration: 20211019 Address after: 361000 North 313A room, Chuang Chuang business park, torch high tech Zone, Xiamen, Fujian Patentee after: Bolu (Xiamen) biological Co.,Ltd. Address before: 361000 North 313A room, Chuang Chuang business park, torch high tech Zone, Xiamen, Fujian Patentee before: Bolu (Xiamen) biological Co.,Ltd. Patentee before: Shenzhen Siwen Technology Co.,Ltd. Patentee before: Nanjing Abel of Engineering Technology Co.,Ltd. |
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