CN104371929A - Nannochloropsis sp. mutant strain and applications thereof - Google Patents
Nannochloropsis sp. mutant strain and applications thereof Download PDFInfo
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- CN104371929A CN104371929A CN201410456584.3A CN201410456584A CN104371929A CN 104371929 A CN104371929 A CN 104371929A CN 201410456584 A CN201410456584 A CN 201410456584A CN 104371929 A CN104371929 A CN 104371929A
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- nannochloropsis oceanica
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- 241000509521 Nannochloropsis sp. Species 0.000 title claims abstract description 10
- 241001300629 Nannochloropsis oceanica Species 0.000 claims description 45
- 241000195493 Cryptophyta Species 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 16
- 239000002253 acid Substances 0.000 claims description 10
- 238000005286 illumination Methods 0.000 claims description 10
- 150000002632 lipids Chemical class 0.000 claims description 10
- 239000002917 insecticide Substances 0.000 claims description 8
- 239000013535 sea water Substances 0.000 claims description 8
- 235000008935 nutritious Nutrition 0.000 claims description 7
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 6
- 239000000460 chlorine Substances 0.000 claims description 6
- 229910052801 chlorine Inorganic materials 0.000 claims description 6
- 244000005700 microbiome Species 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 3
- 230000007613 environmental effect Effects 0.000 abstract description 5
- 239000000575 pesticide Substances 0.000 abstract description 2
- 239000004519 grease Substances 0.000 abstract 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 9
- 229930195729 fatty acid Natural products 0.000 description 9
- 239000000194 fatty acid Substances 0.000 description 9
- 150000004665 fatty acids Chemical class 0.000 description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 8
- 239000007789 gas Substances 0.000 description 8
- 239000002551 biofuel Substances 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 5
- 235000011089 carbon dioxide Nutrition 0.000 description 5
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 230000001105 regulatory effect Effects 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 4
- GHVNFZFCNZKVNT-UHFFFAOYSA-N decanoic acid Chemical compound CCCCCCCCCC(O)=O GHVNFZFCNZKVNT-UHFFFAOYSA-N 0.000 description 4
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 4
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 4
- 239000002994 raw material Substances 0.000 description 3
- ADHNUPOJJCKWRT-JLXBFWJWSA-N (2e,4e)-octadeca-2,4-dienoic acid Chemical compound CCCCCCCCCCCCC\C=C\C=C\C(O)=O ADHNUPOJJCKWRT-JLXBFWJWSA-N 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- 239000005632 Capric acid (CAS 334-48-5) Substances 0.000 description 2
- 239000005639 Lauric acid Substances 0.000 description 2
- 235000021355 Stearic acid Nutrition 0.000 description 2
- HXWJFEZDFPRLBG-UHFFFAOYSA-N Timnodonic acid Natural products CCCC=CC=CCC=CCC=CCC=CCCCC(O)=O HXWJFEZDFPRLBG-UHFFFAOYSA-N 0.000 description 2
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 2
- 235000021342 arachidonic acid Nutrition 0.000 description 2
- 229940114079 arachidonic acid Drugs 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000012159 carrier gas Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- VZCCETWTMQHEPK-UHFFFAOYSA-N gamma-Linolensaeure Natural products CCCCCC=CCC=CCC=CCCCCC(O)=O VZCCETWTMQHEPK-UHFFFAOYSA-N 0.000 description 2
- VZCCETWTMQHEPK-QNEBEIHSSA-N gamma-linolenic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCC(O)=O VZCCETWTMQHEPK-QNEBEIHSSA-N 0.000 description 2
- 235000020664 gamma-linolenic acid Nutrition 0.000 description 2
- 229960002733 gamolenic acid Drugs 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- BUTPBERGMJVRBM-UHFFFAOYSA-N methanol;methylsulfinylmethane Chemical compound OC.CS(C)=O BUTPBERGMJVRBM-UHFFFAOYSA-N 0.000 description 2
- 231100000219 mutagenic Toxicity 0.000 description 2
- 230000003505 mutagenic effect Effects 0.000 description 2
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- SECPZKHBENQXJG-FPLPWBNLSA-N palmitoleic acid Chemical compound CCCCCC\C=C/CCCCCCCC(O)=O SECPZKHBENQXJG-FPLPWBNLSA-N 0.000 description 2
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000004576 sand Substances 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- TUNFSRHWOTWDNC-HKGQFRNVSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCC[14C](O)=O TUNFSRHWOTWDNC-HKGQFRNVSA-N 0.000 description 2
- LKOVPWSSZFDYPG-WUKNDPDISA-N trans-octadec-2-enoic acid Chemical compound CCCCCCCCCCCCCCC\C=C\C(O)=O LKOVPWSSZFDYPG-WUKNDPDISA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N EtOH Substances CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 238000003723 Smelting Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000010411 cooking Methods 0.000 description 1
- 239000000498 cooling water Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000002803 fossil fuel Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000008236 heating water Substances 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000009318 large scale farming Methods 0.000 description 1
- 230000004130 lipolysis Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 230000026267 regulation of growth Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/89—Algae ; Processes using algae
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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
- C12N1/125—Unicellular algae isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K97/00—Accessories for angling
- A01K97/04—Containers for bait; Preparation of bait
- A01K97/045—Preparation of bait; Ingredients
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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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Abstract
The present invention relates to a Nannochloropsis sp. mutant strain and applications thereof, wherein the taxonomy name of the Nannochloropsis sp. mutant strain is Nannochloropsis sp. The Nannochloropsis sp. mutant strain of the present invention has advantages of strong high temperature resistance, strong pesticide tolerance, high grease content, stable performance, and strong environmental adaptability.
Description
Technical field
The present invention relates to microorganism field, particularly relate to a strain and intend Nannochloropsis oceanica mutant strain ENN11-5 and application thereof.
Background technology
In recent years, Global Oil resource is increasingly exhausted, and energy shortage problem is day by day serious, and the problem of environmental pollution, ecological damage etc. is simultaneously day by day serious, and developing green, clean biofuel replace traditional fossil fuel, have become international research focus.In numerous raw materials of biofuel, micro-algae has that photosynthetic efficiency is high, growth cycle is short, oleaginousness is high, environmental compatibility is strong, be easy to the features such as genetic improvement, is considered to the biomass resource of the most potential petroleum replacing.Microalgae biomass can not only produce the multiple biofuel such as biofuel, high-quality aviation spirit through biological smelting, and can also develop the products such as platform chemicals, high-protein food or feed, can be widely used in industrial or agricultural and field of traffic.
Intending Nannochloropsis oceanica (Nannochloropsis sp.) is a kind of important micro-algae of marine products economy, its fast growth, strong stress resistance, and lipid acid composition is simple, and being rich in EPA, is the very potential algae kind of one.The optimum growth temperature intending Nannochloropsis oceanica is 25 DEG C ~ 30 DEG C, when temperature is more than 35 DEG C, grows suppressed, frustule will be caused dead gradually if be in high temperature for a long time.China's most area scorching temperature in summer, intends Nannochloropsis oceanica and is difficult to carry out large-scale farming in summer.In addition, in the outdoor large scale culturing of micro-algae, the pollution such as protozoon, bacterium is easily grown, the growth of severe contamination algae strain.Although use of insecticide can effectively suppress to pollute, algae strain can be injured, cause algae cell activity to decline and the underproduction, frustule when insecticide concentration is too high, can be caused dead.If by the algae strain of mutagenic obtained pesticide resistant and high temperature, the underproduction problem because use of insecticide and high temperature bring will be reduced, stable to plan Nannochloropsis oceanica, high yielding culture is significant.
Summary of the invention
For the shortcoming of prior art, a strain is the object of the present invention is to provide to intend Nannochloropsis oceanica mutant strain and application thereof.
For reaching this goal of the invention, the present invention by the following technical solutions:
The invention provides a strain and intend Nannochloropsis oceanica mutant strain ENN11-5, for Nannochloropsis sp., be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, postcode: 100101, preservation date is on June 18th, 2014, and deposit number is CGMCC No.9321.
The plan Nannochloropsis oceanica mutant strain ENN11-5 that the present invention is separated has sterilant and high temperature tolerance ability is strong, and fat content is high, stable performance, the advantage that environmental compatibility is strong.
Present invention also offers the cultural method intending Nannochloropsis oceanica mutant strain ENN11-5, described method comprises: be seeded in sea water medium by plan Nannochloropsis oceanica algae strain ENN11-5 green cell, wherein intensity of illumination is 50-500 μm of ol/m2.s, and pH value is 7-9, and temperature is 25-42 DEG C.
Present invention also offers and intend the application of Nannochloropsis oceanica mutant strain ENN11-5 in the cultivation of plan Nannochloropsis oceanica high temperature.
Preferably, described high temperature is 38-42 DEG C.
Present invention also offers and intend the application of Nannochloropsis oceanica mutant strain ENN11-5 when intending use of insecticide in Nannochloropsis oceanica cultivation.
Preferably, the effective chlorine density of described sterilant is 25ppm.
Present invention also offers and intend the application of Nannochloropsis oceanica mutant strain ENN11-5 in lipid acid is produced.
Present invention also offers and intend the application of Nannochloropsis oceanica mutant strain ENN11-5 in nutritious prod, feed or bait are produced.
Compared with prior art, the present invention has following beneficial effect:
The present invention be separated the plan Nannochloropsis oceanica mutant strain obtained there is sterilant and high temperature tolerance ability is strong, fat content is high, stable performance, advantage that environmental compatibility is strong.
Accompanying drawing explanation
Fig. 1 is the growth curve chart of cellar culture plan Nannochloropsis oceanica of the present invention ENN11-5 in 3cm column reactor.
Fig. 2 is the growth curve chart of cellar culture plan Nannochloropsis oceanica of the present invention ENN11-5 in 5cm column reactor.
Fig. 3 is the growth curve chart that Insecticide cultivates plan Nannochloropsis oceanica ENN11-5 of the present invention in 3cm column reactor.
Fig. 4 is the temperature changing curve diagram cultivating plan Nannochloropsis oceanica ENN11-5 of the present invention at 5cm pillar high temperature pressure.
Fig. 5 is the growth curve chart cultivating plan Nannochloropsis oceanica ENN11-5 of the present invention at 5cm pillar high temperature pressure.
Embodiment
For better the present invention being described, be convenient to understand technical scheme of the present invention, typical but non-limiting embodiment of the present invention is as follows:
In the typical embodiment of one of the present invention, the plan Nannochloropsis oceanica mutant strain ENN11-5 that the present invention is separated, for Nannochloropsis sp., be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, postcode: 100101, preservation date is on June 18th, 2014, and deposit number is CGMCC No.9321.
The plan Nannochloropsis oceanica mutant strain ENN11-5 that the present invention is separated has sterilant and high temperature tolerance ability is strong, fat content is high, stable performance, advantage that environmental compatibility is strong.
Plan Nannochloropsis oceanica ENN11-5 provided by the invention can grow at 25 DEG C, 26 DEG C, 27 DEG C, 28 DEG C, 29 DEG C, 30 DEG C, 31 DEG C, 32 DEG C, 33 DEG C, 34 DEG C, 35 DEG C, 36 DEG C, 37 DEG C, 38 DEG C, 39 DEG C, 40 DEG C, 41 DEG C or 42 DEG C.
Plan Nannochloropsis oceanica ENN11-5 provided by the invention can grow under the available chlorine pressure of 5ppm, 10ppm, 15ppm, 20ppm or 25ppm.
The cultural method of plan Nannochloropsis oceanica mutant strain ENN11-5 of the present invention, comprise plan Nannochloropsis oceanica algae strain ENN11-5 green cell to be seeded in sea water medium and cultivate, wherein intensity of illumination is 50 μm of ol/m
2.s-500 μm ol/m
2.s, pH value is 7-9, and temperature is 25 DEG C-42 DEG C.
In the cultural method of plan Nannochloropsis oceanica ENN11-5 of the present invention, intensity of illumination can be 50 μm of ol/m
2.s, 100 μm of ol/m
2.s, 150 μm of ol/m
2.s, 200 μm of ol/m
2.s, 250 μm of ol/m
2.s, 300 μm of ol/m
2.s, 350 μm of ol/m
2.s, 400 μm of ol/m
2.s, 450 μm of ol/m
2.s or 500 μm of ol/m
2.s; PH value can be 7,7.2,7.5,7.8,8,8.1,8.5,8.8 or 9; Temperature can be 25 DEG C, 26 DEG C, 27 DEG C, 28 DEG C, 29 DEG C, 30 DEG C, 31 DEG C, 32 DEG C, 33 DEG C, 34 DEG C, 35 DEG C, 36 DEG C, 37 DEG C, 38 DEG C, 39 DEG C, 40 DEG C, 41 DEG C or 42 DEG C.
Plan Nannochloropsis oceanica ENN11-5 of the present invention can be applied in and intend in the cultivation of Nannochloropsis oceanica high temperature, and wherein, described high temperature can be 38 DEG C, 39 DEG C, 40 DEG C, 41 DEG C or 42 DEG C.
Under plan Nannochloropsis oceanica ENN11-5 of the present invention can be applied in the environment intending use of insecticide in Nannochloropsis oceanica cultivation, the effective chlorine density of described sterilant is 25ppm.
Plan Nannochloropsis oceanica ENN11-5 of the present invention can be applied in the production of lipid acid, and described lipid acid is capric acid, lauric acid, myristic acid, palmitinic acid, Zoomeric acid, stearic acid, octadecenoic acid, octadecadienoic acid, gamma-linolenic acid, arachidonic acid and/or timnodonic acid.
Plan Nannochloropsis oceanica ENN11-5 of the present invention can be applied in the production of nutritious prod, feed or bait, and described nutritious prod, feed or bait comprise the nutritious prod of capric acid, lauric acid, myristic acid, palmitinic acid, Zoomeric acid, stearic acid, octadecenoic acid, octadecadienoic acid, gamma-linolenic acid, arachidonic acid and/or timnodonic acid, feed or bait.
Beneficial effect of the present invention is further illustrated below with reference to embodiment and specific embodiment.
the screening of specific embodiment 1ENN11-5 algae strain and acquisition
By the aseptic algae liquid of Nannochloropsis sp. through EMS mutagenic treatment, add clorox sterilant and oxyproline pressure treatment and primary dcreening operation, multiple sieve repeatedly, the final mutant strain ENN11-5 obtaining a kind of excellent property.This sudden change algae strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.9321.
specific embodiment 2ENN11-5 cellar culture is evaluated
1) 3cm column reactor is evaluated: be seeded in the sea water medium prepared by green for the ENN11-5 being in logarithmic phase swarm cell, make cell density reach OD
750between 0.8-1.2, with wild-type ENN11 for contrast.Culturing process intensity of illumination controls at 50-500umol/m
2.s, L:D=16:8 light dark period is cultivated.In incubation period, by passing into the carbonic acid gas of 1.5-2% and the mixed gas of air in nutrient solution, the pH value of substratum is regulated between 7-9.The column reactor that the reactor that cultivation uses is 30mm internal diameter, length 600mm.Cultivate 8 days, measure algae strain growth curve.
As shown in Figure 1, cultivate ENN11-5 growth under normal condition and be obviously better than wild strain ENN11, the speed of growth comparatively contrasts strain and improves 31.6%.
2) 5cm column reactor is evaluated: be seeded in the sea water medium prepared by green for the ENN11-5 being in logarithmic phase swarm cell, make cell density reach OD
750between 0.8-1.2, with wild-type ENN11 for contrast.Culturing process intensity of illumination controls at 50-500umol/m
2.s, L:D=16:8 light dark period is cultivated.In incubation period, by passing into the carbonic acid gas of 1.5-2% and the mixed gas of air in nutrient solution, the pH value of substratum is regulated between 7-9.The column reactor that the reactor that cultivation uses is 50mm internal diameter, length 600mm.Cultivate 7 days, measure algae strain growth curve.
As shown in Figure 2, cultivate ENN11-5 growth under normal condition and be obviously better than wild strain ENN11, the speed of growth comparatively contrasts strain and improves 24.0%.
specific embodiment 3ENN11-5 Insecticide is cultivated and is evaluated
Green for the ENN11-5 being in logarithmic phase swarm cell is seeded in the sea water medium prepared, makes cell density reach OD
750between 0.8-1.2, with wild-type ENN11 for contrast.Culturing process intensity of illumination controls at 50-500umol/m
2.s, L:D=16:8 light dark period is cultivated.In incubation period, by passing into the carbonic acid gas of 1.5-2% and the mixed gas of air in nutrient solution, the pH value of substratum is regulated between 7-9.The column reactor that the reactor that cultivation uses is 30mm internal diameter, length 600mm.Inoculation initially adds available chlorine 25ppm pressure, lucifuge process 8 hours, then illumination cultivation, cultivates 8 days, measures algae strain growth curve.
As shown in Figure 3, after adding available chlorine pressure, wild strain ENN11 is subject to obvious suppression, and stop growing the death that even turns yellow substantially, and ENN11-5 can recover rear continued growth, and resistance to sterilant ability is obviously better than contrasting strain.
specific embodiment 4ENN11-5 high temperature pressure is cultivated and is evaluated
Green for the ENN11-5 being in logarithmic phase swarm cell is seeded in the sea water medium prepared, makes cell density reach OD
750between 0.8-1.2, with wild-type ENN11 for contrast.Culturing process intensity of illumination controls at 50-500umol/m
2.s, L:D=16:8 light dark period is cultivated.In incubation period, by passing into the carbonic acid gas of 1.5-2% and the mixed gas of air in nutrient solution, the pH value of substratum is regulated between 7-9.The column reactor that the reactor that cultivation uses is 50mm internal diameter, length 600mm.Arrangement for heating or cooling water in receptacle for live fish temperature control, 3d starts pyroprocessing, and algae liquid temp reaches 38 DEG C, 4d to 6d, has every day 6h algae liquid temp all to reach 42 DEG C (temperature variation as shown in Figure 4).Cultivate 6 days, measure algae strain growth curve.
As shown in Figure 5, after pyroprocessing starts, wild-type ENN11 growth is slowed down, and start to decline at growth regulation 5 days biomasss, high temperature suppresses obviously.Compare wild strain, high temperature is compared low to ENN11-5 growth-inhibiting degree, and the speed of growth is totally higher than wild strain.After pyroprocessing, the speed of growth comparatively contrasts strain raising 15.7%.
the fat content of specific embodiment 5ENN11-5 algae strain
Green for the ENN11-5 being in logarithmic phase swarm cell is seeded in the column reactor of 30mm internal diameter, length 600mm, uses sea water medium to cultivate.Culturing process intensity of illumination controls at 50-500umol/m
2.s, L:D=16:8 light dark period is cultivated.In incubation period, by passing into the carbonic acid gas of 1.5-2% and the mixed gas of air in nutrient solution, the pH value of substratum is regulated between 7-9.Cultivation proceeds to the 8th day, is collected by algae liquid, obtains algae mud, algae mud is carried out vacuum lyophilization again by method that is centrifugal or natural subsidence.
After drying completes, measure its oil component content as following table 1, analytical procedure is as follows:
1) lipid acid extracts:
Getting the volume that 50mg or 100mg freeze-dried algae powder is placed on tool Telfnon bottle screw cap is in the phial of 15-20ml, place a little magnetic bar again, add 2-4ml 10%DMSO-Methanol solution, 40 DEG C of sand baths (beaker containing sand is placed on constant-temperature heating magnetic stirring apparatus) 5 minutes; Then stir extracting 30 minutes at 4 DEG C of lower magnetic forces, 3500 leave the heart, and transfer supernatant liquor is in another bottle.Remaining algae-residue adds the ether of 1:1 again, normal hexane 4-8ml 4 DEG C of lower magnetic forces stir extractings 1 hour, and 3500 leave the heart, and transfer supernatant liquor is in an above-mentioned bottle.Said process can be repeated until algae-residue bleaches.In above-mentioned merging extract, add pure water makes four (water, DMSO-Methanol, ether, normal hexane) volume ratio be 1:1:1:1, concussion phase-splitting, pipetting organic phase transfers in another phial, blow to one-tenth concentrated solution with nitrogen in stink cupboard, then transfer in 1.5ml plastic centrifuge tube weighed in advance, then dry up to constant weight with nitrogen.
2) fatty acid analysis:
After method is extracted above, with n-hexane dissolution, (chromatographic condition is carrier gas: nitrogen flow 1ml/min, hydrogen flowing quantity 30ml/min, air flow quantity 300ml/min to use Agilent 6820 gas chromatograph to carry out gas chromatographic analysis, injector temperature: 280 DEG C, detector temperature: 280 DEG C, detector type: FID, chromatographic column: DB-5 capillary chromatographic column (30m × 0.25mm, 0.25 μm), splitting ratio: 4:1.Analytical procedure: marker method (gas-chromatography nitrogen does carrier gas, is equivalent to the moving phase of liquid chromatography).
Do not have the accumulation of implementing measure induced lipolysis acid in the present embodiment, cultivate 8 days fatty acid content and reach 42.28%, EPA content reaches 3.4%.As taked corresponding measure, fatty acid content can be higher.
The ENN11-5 8d of table 1 gas Chromatographic Determination collects total fat component of sample:
"/" represents and this component do not detected
Fatty acid component analysis shows, and its fat content is up to 42.28%, and wherein about 74.2% is C16-C18 lipid acid, 63.27% is C16 lipid acid, 10.95% is C18 lipid acid, and unsaturated fatty acids accounts for 45.15% of fatty acid total amount, can as the raw materials for production of biofuel.As can be seen here, algae strain ENN11-4 of the present invention can be used for the production of biofuel, bio-ethanol.
The unsaturated fatty acids of ENN11-4 accounts for 45.15% of fatty acid total amount, content of polyunsaturated fatty acid accounts for 16.0% of fatty acid total amount, wherein EPA content accounts for 8.055% of fatty acid total amount, and unsaturated fatty acid content enriches, and can be used for the production of polyunsaturated fatty acid.
specific embodiment 6 mutant strain ENN11-5 is used for the production of nutritious prod, feed, bait
Protein content in algae strain provided by the invention is 10 ~ 30%, and lipid content is 10 ~ 60%, and wherein the content of unsaturated fatty acids accounts for the 35%-60% of total fatty acid content, is the optimum feed stock source of cooking nutritious prod, feed, bait.
Applicant states, the present invention illustrates processing method of the present invention by above-described embodiment, but the present invention is not limited to above-mentioned processing step, does not namely mean that the present invention must rely on above-mentioned processing step and could implement.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of ancillary component, the concrete way choice etc. of raw material selected by the present invention, all drops within protection scope of the present invention and open scope.
Claims (8)
1. Nannochloropsis oceanica mutant strain ENN11-5 is intended in a strain, for Nannochloropsis sp., be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), deposit number is CGMCCNo.9321, and preservation date is on June 18th, 2014.
2. the as claimed in claim 1 cultural method intending Nannochloropsis oceanica mutant strain ENN11-5, is characterized in that, described method comprises and is seeded in sea water medium by plan Nannochloropsis oceanica algae strain ENN11-5 green cell, and wherein intensity of illumination is 50-500 μm of ol/m
2.s, pH value is 7-9, and temperature is 25-42 DEG C.
3. the Nannochloropsis oceanica mutant strain ENN11-5 that intends as claimed in claim 1 is intending the application in the cultivation of Nannochloropsis oceanica high temperature.
4. apply as claimed in claim 3, it is characterized in that, described high temperature is 38-42 DEG C.
5. the application of plan Nannochloropsis oceanica mutant strain ENN11-5 as claimed in claim 1 when intending use of insecticide in Nannochloropsis oceanica cultivation.
6. apply as claimed in claim 5, it is characterized in that, the effective chlorine density of described sterilant is 25ppm.
7. the application of plan Nannochloropsis oceanica mutant strain ENN11-5 as claimed in claim 1 in lipid acid is produced.
8. the application of plan Nannochloropsis oceanica mutant strain ENN11-5 as claimed in claim 1 in nutritious prod, feed or bait are produced.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP6218096B1 (en) * | 2016-12-31 | 2017-10-25 | 甲斐水産有限会社 | Nannochloropsis and its production method. |
| CN108977362A (en) * | 2017-06-05 | 2018-12-11 | 财团法人食品工业发展研究所 | The strain of Chlorococcum (CHLORELLA LEWINII) algae and its purposes |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100183744A1 (en) * | 2009-01-22 | 2010-07-22 | Aurora Biofuels, Inc. | Systems and methods for maintaining the dominance of nannochloropsis in an algae cultivation system |
| CN103468577A (en) * | 2013-09-22 | 2013-12-25 | 新奥科技发展有限公司 | Nannochloropsis sp mutant strain and application thereof |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100183744A1 (en) * | 2009-01-22 | 2010-07-22 | Aurora Biofuels, Inc. | Systems and methods for maintaining the dominance of nannochloropsis in an algae cultivation system |
| CN103468577A (en) * | 2013-09-22 | 2013-12-25 | 新奥科技发展有限公司 | Nannochloropsis sp mutant strain and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 万晶晶: "产油微藻的高密度培养工艺研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑 》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP6218096B1 (en) * | 2016-12-31 | 2017-10-25 | 甲斐水産有限会社 | Nannochloropsis and its production method. |
| JP2018108049A (en) * | 2016-12-31 | 2018-07-12 | 甲斐水産有限会社 | Nannochloropsis and production method thereof |
| CN108977362A (en) * | 2017-06-05 | 2018-12-11 | 财团法人食品工业发展研究所 | The strain of Chlorococcum (CHLORELLA LEWINII) algae and its purposes |
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