CN104178446A - Application of acetylcholine and analogs thereof in promoting microalgae growth and microalgae grease accumulation - Google Patents
Application of acetylcholine and analogs thereof in promoting microalgae growth and microalgae grease accumulation Download PDFInfo
- Publication number
- CN104178446A CN104178446A CN201410421791.5A CN201410421791A CN104178446A CN 104178446 A CN104178446 A CN 104178446A CN 201410421791 A CN201410421791 A CN 201410421791A CN 104178446 A CN104178446 A CN 104178446A
- Authority
- CN
- China
- Prior art keywords
- vagusstoff
- algae
- micro
- analogue
- growth
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000009825 accumulation Methods 0.000 title claims abstract description 42
- 239000004519 grease Substances 0.000 title claims abstract description 30
- 230000001737 promoting effect Effects 0.000 title claims abstract description 7
- 230000012010 growth Effects 0.000 title abstract description 35
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 title abstract description 11
- 229960004373 acetylcholine Drugs 0.000 title abstract description 7
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims abstract description 64
- GGRLKHMFMUXIOG-UHFFFAOYSA-M 2-acetyloxyethyl(trimethyl)azanium;hydroxide Chemical compound [OH-].CC(=O)OCC[N+](C)(C)C GGRLKHMFMUXIOG-UHFFFAOYSA-M 0.000 claims description 142
- 230000005791 algae growth Effects 0.000 claims description 53
- 241000195493 Cryptophyta Species 0.000 claims description 41
- 229960001231 choline Drugs 0.000 claims description 16
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 claims description 13
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 claims description 10
- RZZPDXZPRHQOCG-OJAKKHQRSA-M CDP-choline(1-) Chemical compound O[C@@H]1[C@H](O)[C@@H](COP([O-])(=O)OP([O-])(=O)OCC[N+](C)(C)C)O[C@H]1N1C(=O)N=C(N)C=C1 RZZPDXZPRHQOCG-OJAKKHQRSA-M 0.000 claims description 10
- 235000019743 Choline chloride Nutrition 0.000 claims description 10
- 229960003178 choline chloride Drugs 0.000 claims description 10
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 claims description 10
- 229960001284 citicoline Drugs 0.000 claims description 10
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 claims description 10
- KIZQNNOULOCVDM-UHFFFAOYSA-M 2-hydroxyethyl(trimethyl)azanium;hydroxide Chemical compound [OH-].C[N+](C)(C)CCO KIZQNNOULOCVDM-UHFFFAOYSA-M 0.000 claims description 8
- 240000002900 Arthrospira platensis Species 0.000 claims description 7
- 235000016425 Arthrospira platensis Nutrition 0.000 claims description 7
- 241000195585 Chlamydomonas Species 0.000 claims description 7
- 241000168525 Haematococcus Species 0.000 claims description 7
- 241001300629 Nannochloropsis oceanica Species 0.000 claims description 7
- 235000009754 Vitis X bourquina Nutrition 0.000 claims description 7
- 235000012333 Vitis X labruscana Nutrition 0.000 claims description 7
- 235000014787 Vitis vinifera Nutrition 0.000 claims description 7
- 238000005286 illumination Methods 0.000 claims description 7
- 229940082787 spirulina Drugs 0.000 claims description 7
- 150000003248 quinolines Chemical group 0.000 claims description 3
- 241000219095 Vitis Species 0.000 claims 3
- 239000002028 Biomass Substances 0.000 abstract description 39
- 238000000034 method Methods 0.000 abstract description 15
- 239000002858 neurotransmitter agent Substances 0.000 abstract description 6
- 230000008569 process Effects 0.000 abstract description 4
- 230000035508 accumulation Effects 0.000 description 38
- 239000007788 liquid Substances 0.000 description 13
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 12
- 239000000047 product Substances 0.000 description 11
- 230000005526 G1 to G0 transition Effects 0.000 description 9
- 240000009108 Chlorella vulgaris Species 0.000 description 6
- 238000005070 sampling Methods 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 5
- 240000006365 Vitis vinifera Species 0.000 description 4
- 150000003839 salts Chemical group 0.000 description 4
- 241000894007 species Species 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 239000003375 plant hormone Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000195645 Auxenochlorella protothecoides Species 0.000 description 2
- 241000195654 Chlorella sorokiniana Species 0.000 description 2
- 235000007089 Chlorella vulgaris Nutrition 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 241000195646 Parachlorella kessleri Species 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000005984 hydrogenation reaction Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 230000001360 synchronised effect Effects 0.000 description 2
- 241000760893 Deasonia multinucleata Species 0.000 description 1
- 241000305506 Desmodesmus Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241001518671 Multiformis Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000002551 biofuel Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000004807 desolvation Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- DCXXMTOCNZCJGO-UHFFFAOYSA-N tristearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC DCXXMTOCNZCJGO-UHFFFAOYSA-N 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for adding neurotransmitter acetylcholine or acetylcholine analogs in a microalgae growth process to enhance microalgae biomass quantity and grease content, belonging to the field of microalgae biology. Different concentrations of neurotransmitter acetylcholine or acetylcholine analogs in different microalgae growth periods are added to analyze the influence of different concentrations of acetylcholine or acetylcholine analogs added in different periods on the Chlorella biomass quantity and grease content. The invention provides the addition of the neurotransmitter acetylcholine or acetylcholine analogs in the microalgae growth process for the first time, and provides a new idea for synchronously promoting microalgae growth and grease accumulation by using a neurotransmitter.
Description
Technical field
The invention belongs to Microalgae biotechnology field, be specifically related to the application in promoting micro algae growth and microalgae grease accumulation of a kind of vagusstoff and analogue thereof.
Background technology
Micro-algae abundant species, can cultivate in a large number and not occupy cultivated land; Growth cycle is short, about 3-5 days of doubling time, and yield per unit is tens times and even hundreds of times of grain; Photosynthetic efficiency is high, can effectively utilize sun power, inorganics is converted into the nutritive ingredients such as protein, polysaccharide, polyunsaturated fatty acid, can be used for food, medicine and energy aspect; Fat content is high, reaches the 20%-70% of dry weight, far above land plant, is the promising biological raw material of a kind of very tool.Utilize at present the micro-algae of cultivation to accumulate grease with production biofuel, become and utilized the most popular research field of solar energy development renewable resources, there is powerful market potential and outstanding social value.
Compare with high oil crops, the biochemical composition of micro-algae is cultivated, and salt forms and the impact of envrionment conditions change is larger, as nitrogen lacks the especially accumulation of triacylglycerol of grease that easily stimulates micro-algae, temperature reduces the accumulation of the micro-algae polyunsaturated fatty acid of meeting promotion etc., therefore need to further investigate the optimum produce oil culture condition of micro-algae.
Cultivate the research that salt forms and envrionment conditions accumulates impact to microalgae grease more, and that the Chemical Regulation of plant hormone isoreactivity molecule affects the research of microalgae grease accumulation is less, the result of study of the impact of higher plant being grown from plant hormone, can find out that plant hormone has very strong regulating effect to the growth of higher plant, therefore study its impact on micro algae growth, for the amount of biomass and the oil and fat accumulation speed that improve micro-algae, have great importance.
It is lower that micro-algae autotrophy system and heterotrophism system are compared Biomass accumulation, and the measure that therefore can significantly promote growth to improve Biomass accumulation has low-cost advantage micro-algae autotrophy system to setting up plays an important role.In prior art, in micro-algae autotrophy system, generally adopt two-step approach to produce grease, under the optimal conditions of first stage, reach Fast Growth to form under biomass and stress conditions subsequently with accumulation grease.Under Fast Growth, oil and fat accumulation is few, and under stress conditions, Biomass accumulation is suppressed, integrates, and final grease yield is on the low side, has reduced the practical value that micro-algae autotrophy system is produced grease.When therefore, invention can make micro-algae Fast Growth, the single stage method method of synchronous accumulation grease is produced grease technique functions to keying action to setting up efficient micro-algae autotrophy system cheaply.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of method of utilizing vagusstoff and analogue thereof to promote micro algae growth and the synchronous accumulation of grease.
For solving above technical problem, the present invention adopts following technical scheme:
Vagusstoff and analogue thereof promote at the same time in the accumulation of micro algae growth and microalgae grease be applied in protection scope of the present invention within.
Wherein, described micro-algae is any one in chlorella, spirulina, chlamydomonas, Nannochloropsis oceanica, haematococcus pulvialis, the new green alga of rich oil, multinuclear green alga (Deasonia multinucleata), multiform grid algae (Desmodesmus multiformis), palm net algae (Palmodictyon varium) and grape algae.
Wherein, in the substratum of micro-algae, add the analogue of vagusstoff or vagusstoff, the adjustment period that the time of interpolation being micro algae growth or logarithmic phase.Preferably at micro-algae OD
680value be 0～0.5 o'clock (adjustment period) add vagusstoff or vagusstoff analogue, most preferably at micro-algae OD
680value be 0 o'clock (adjustment period) add vagusstoff or vagusstoff analogue; Or preferably at micro-algae OD
680value is that 1.2～1.6 o'clock (logarithmic phase) adds vagusstoff or vagusstoff analogue, most preferably at micro-algae OD
680value is that 1.5 o'clock (logarithmic phase) adds vagusstoff or vagusstoff analogue.
Wherein, vagusstoff micro algae growth adjustment period addition be 0.1～5 μ g/L, be preferably 0.5～5 μ g/L, most preferably 5 μ g/L.
Wherein, vagusstoff is 0.1～5 μ g/L at the addition of the logarithmic phase of micro algae growth, is preferably 0.25～5 μ g/L, most preferably 5 μ g/L.
Wherein, vagusstoff analogue micro algae growth adjustment period addition be 50～200 μ g/L, be preferably 150～200 μ g/L, most preferably 200 μ g/L.
Wherein, vagusstoff analogue is 50～200 μ g/L at the addition of the logarithmic phase of micro algae growth, is preferably 150～200 μ g/L, most preferably 200 μ g/L.
Wherein, the analogue of described vagusstoff is choline, choline chloride 60, bursine, citicoline or phosphatidylcholine, preferably choline.
Wherein, micro-algae culture temperature is 25～30 ℃, preferably 30 ℃; Cultivation intensity of illumination is 2000～4000lux, is preferably 3000lux.
Vagusstoff and analogue thereof in promoting microalgae grease accumulation be applied in protection scope of the present invention within.
Wherein, described micro-algae is any one in chlorella, spirulina, chlamydomonas, Nannochloropsis oceanica, haematococcus pulvialis, the new green alga of rich oil, multinuclear green alga, multiform grid algae, palm net algae and grape algae.
Wherein, in the substratum of micro-algae, add the analogue of vagusstoff or vagusstoff, the adjustment period that the time of interpolation being micro algae growth or logarithmic phase.Preferably at micro-algae OD
680value be 0～0.5 o'clock (adjustment period) add vagusstoff or vagusstoff analogue, most preferably at micro-algae OD
680value be 0 o'clock (adjustment period) add vagusstoff or vagusstoff analogue; Or preferably at micro-algae OD
680value is that 1.2～1.6 o'clock (logarithmic phase) adds vagusstoff or vagusstoff analogue, most preferably at micro-algae OD
680value is that 1.5 o'clock (logarithmic phase) adds vagusstoff or vagusstoff analogue.
Wherein, vagusstoff micro algae growth adjustment period addition be 0.1～5 μ g/L, be preferably 0.5～5 μ g/L, most preferably 5 μ g/L.
Wherein, vagusstoff is 0.1～5 μ g/L at the addition of the logarithmic phase of micro algae growth, is preferably 0.25～5 μ g/L, most preferably 5 μ g/L.
Wherein, vagusstoff analogue micro algae growth adjustment period addition be 50～200 μ g/L, be preferably 150～200 μ g/L, most preferably 200 μ g/L.
Wherein, vagusstoff analogue is 50～200 μ g/L at the addition of the logarithmic phase of micro algae growth, is preferably 150～200 μ g/L, most preferably 200 μ g/L.
Wherein, described vagusstoff analogue is choline, choline chloride 60, bursine, citicoline or phosphatidylcholine, preferably choline.
Wherein, micro-algae culture temperature is 25～30 ℃, preferably 30 ℃; Cultivation intensity of illumination is 2000～4000lux, is preferably 3000lux.
Application in vagusstoff and analogue promotion micro algae growth thereof is also within protection scope of the present invention.
Wherein, described micro-algae is any one in chlorella, spirulina, chlamydomonas, Nannochloropsis oceanica, haematococcus pulvialis, the new green alga of rich oil, multinuclear green alga, multiform grid algae, palm net algae and grape algae.
Wherein, in the substratum of micro-algae, add the analogue of vagusstoff or vagusstoff, the adjustment period that the time of interpolation being micro algae growth or logarithmic phase.Preferably at micro-algae OD
680value be 0～0.5 o'clock (adjustment period) add vagusstoff or vagusstoff analogue, most preferably at micro-algae OD
680value be 0 o'clock (adjustment period) add vagusstoff or vagusstoff analogue; Or preferably at micro-algae OD
680value is that 1.2～1.6 o'clock (logarithmic phase) adds vagusstoff or vagusstoff analogue, most preferably at micro-algae OD
680value is that 1.5 o'clock (logarithmic phase) adds vagusstoff or vagusstoff analogue.
Wherein, vagusstoff micro algae growth adjustment period addition be 0.1～10 μ g/L, be preferably 0.5～10 μ g/L, preferably 5 μ g/L.
Wherein, vagusstoff is 0.1～10 μ g/L at the addition of the logarithmic phase of micro algae growth, is preferably 0.25～10 μ g/L, most preferably 10 μ g/L.
Wherein, vagusstoff analogue micro algae growth adjustment period addition be 50～200 μ g/L, be preferably 150～200 μ g/L, most preferably 200 μ g/L.
Wherein, vagusstoff analogue is 50～200 μ g/L at the addition of the logarithmic phase of micro algae growth, is preferably 150～200 μ g/L, most preferably 200 μ g/L.
Wherein, described vagusstoff analogue comprises choline, choline chloride 60, bursine, citicoline, phosphatidylcholine, preferably choline.
Wherein, micro-algae culture temperature is 25～30 ℃, preferably 30 ℃; Cultivation intensity of illumination is 2000～4000lux, is preferably 3000lux.
Adjustment period of the present invention, be defined as: when microorganism enters into a new environment, owing to will adapting to this site conditions original different from it, reproduction speed is very slow, maintains a lower level; Adjustment period preferred micro-algae OD
680value is in 0～0.5 scope.
Logarithmic phase of the present invention is defined as: microorganism has adapted to new environment, starts a large amount of Fast-propagations, and quantity rapid growth is logarithm or index curve shape, also claims exponential phase; The preferred micro-algae OD of logarithmic phase
680value is in 1.2～1.6 scopes.
Be defined as stationary phase of the present invention: microbe population has risen to certain degree, and because the consumption of nutrient etc. and the accumulation of harmful meta-bolites make the dramatic growth again of its quantity, increased numbers is substantially equal to death toll, in running balance period; Be preferably micro-algae OD stationary phase
680value is in 2.5～3.0 scopes.
Beneficial effect:
The present invention has following distinguishing feature and effect:
(1) the present invention proposes to add the analogue of vagusstoff or vagusstoff first in the process of growth of micro-algae, for using the analogue of neurotransmitter acetylcholine or vagusstoff to promote micro-algal biomass and oil and fat accumulation that new approaches are provided.
(2) by the analogue of supplement trace vagusstoff or vagusstoff, the accumulation of micro-algal biomass and the accumulation of grease can have obviously been promoted.
(3) in prior art, utilizing micro-algae to produce algae oil all will be divided into Biomass accumulation and coerce lower algae oil two stages of accumulation, micro algae growth is slow, accumulation grease efficiency is low, the present invention proposes to add to the different times of micro algae growth the analogue of vagusstoff or vagusstoff, add in nutrient solution and not only significantly promote the growth of micro-algae but also synchronously promote micro-algae efficiently to accumulate grease, realized with a kind of molecule and processed mode and the efficiency thereof of having improved micro algae growth and oil and fat accumulation.
Accompanying drawing explanation
Fig. 1 micro algae growth adjustment period, add the vagusstoff of different concns, the variation of chlorella amount of biomass.Wherein, X-coordinate is incubation time, the amount of biomass that ordinate zou is chlorella, and the absorbancy with frustule under 680nm wavelength represents,
represent not add vagusstoff,
the concentration that represents vagusstoff is 0.5 μ g/L,
the concentration that represents vagusstoff is 5 μ g/L,
the concentration that represents vagusstoff is 10 μ g/L.Data are the mean value of 3 parallel samplings.
Fig. 2 is at the logarithmic phase of micro algae growth, i.e. OD
680=1.5, add the vagusstoff of different concns, the variation of chlorella amount of biomass.Wherein, X-coordinate is incubation time, the amount of biomass that ordinate zou is chlorella, and the absorbancy with frustule under 680nm wavelength represents,
represent not add vagusstoff,
the concentration that represents vagusstoff is 0.5 μ g/L,
the concentration that represents vagusstoff is 5 μ g/L,
the concentration that represents vagusstoff is 10 μ g/L.Data are the mean value of 3 parallel samplings.
Fig. 3 chlorella not of the same race is added vagusstoff and is not added the comparison of vagusstoff dry weight.Micro algae growth adjustment period, add the vagusstoff of 0.5 μ g/L, the variation of different chlorella amount of biomass.Wherein, X-coordinate is different chlorellas, is that C.sorokiniana is Root Rot of Wheat chlorella successively, and C.kessleri is Kessler chlorella, and C.vulgaris is Chlorella vulgaris, and C.protothecoides is original chlorella; Ordinate zou is that the amount of biomass of chlorella changes, and data are the mean value of 3 parallel samplings.
Fig. 4 chlorella not of the same race is added vagusstoff and is not added the comparison of vagusstoff oil and fat accumulation.Micro algae growth adjustment period, add the vagusstoff of 0.5 μ g/L, the variation of oil and fat accumulation in chlorella.Wherein, X-coordinate is different chlorellas, is that C.sorokiniana is Root Rot of Wheat chlorella successively, and C.kessleri is Kessler chlorella, and C.vulgaris is Chlorella vulgaris, and C.protothecoides is original chlorella; Ordinate zou is that the oil and fat accumulation of chlorella changes, and data are the mean value of 3 parallel samplings.
Comparison on the impact of chlorella amount of biomass when Fig. 5 vagusstoff analogue adds the adjustment period of chlorella.Micro algae growth adjustment period, add respectively the vagusstoff analogue of 50 μ g/L, 150 μ g/L and 200 μ g/L, the variation of different chlorella amount of biomass.Wherein, X-coordinate is blank and different vagusstoff analogue, is that CHOL is choline successively, and CH is choline chloride 60, and PC is bursine, and INN is citicoline, and CHCL is phosphatidylcholine; Ordinate zou is that the amount of biomass of chlorella changes, and data are the mean value of 3 parallel samplings.
Comparison on the impact of chlorella oil and fat accumulation when Fig. 6 vagusstoff analogue adds the adjustment period of chlorella.Micro algae growth adjustment period, add respectively the vagusstoff analogue of 50 μ g/L, 150 μ g/L and 200 μ g/L, the variation of different chlorella oil and fat accumulations.Wherein, X-coordinate is blank and different vagusstoff analogue, is that CHOL is choline successively, and CH is choline chloride 60, and PC is bursine, and INN is citicoline, and CHCL is phosphatidylcholine; Ordinate zou is the variation of chlorella oil and fat accumulation, and data are the mean value of 3 parallel samplings.
Embodiment
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand, the described content of embodiment is only for the present invention is described, and should also can not limit the present invention described in detail in claims.
In following examples, micro-algae is cultivated in accordance with the following methods: fresh microalgae cell is inoculated in the 250ml triangular flask that the basic Tap substratum that 100ml is fresh is housed according to 5% volume fraction, standing cultivation, and add vagusstoff at the different times of micro algae growth, therebetween, shake triangular flask every day 3～5 times, the microalgae cell of gathering when microalgae cell is bred the stationary phase to maximum density, algae liquid centrifugal concentrating, with deionized water wash, centrifugal, remove microalgae cell and cultivate salt, be prepared into without the wet algae of cultivating salt, as the use of oil and grease extracting.
Basic Tap substratum (every liter) formula used in following examples is as follows: 400.00mg NH
4cl, 50.00mgCaCl
22H
2o, 100.00mg MgSO
47H
2o, 98.80mgNa
2hPO
4, 61.73mg KH
2pO
4, 50.00mgNa
2eDTA2H
2o, 22.00mg ZnSO
47H
2o, 11.40mg H
3bO
3, 5.10mg MnCl
24H
2o, 5.00mgFeSO
47H
2o, 1.60mg CoCl
26H
2o, 1.16mg CuSO
45H
2o, 1.10mg (NH
4)
6mo
7o
244H
2o, 2420.00mg Trisma Base, 1ml Glacial Acetic.
In following examples, the grease of micro-algae obtains by the following method: in the micro-algae dry product of 1g of gained, add 5ml concentrated hydrochloric acid (mass concentration 36%-38%), mix, in 70 ℃ of water-baths, place 20min, add 5ml dehydrated alcohol, standingly in system, add 5mL ether again after cooling, vibration 1min, the centrifugal 2min of 4000rpm, obtain upper strata ether phase and lower sediment, upper strata ether is shifted out mutually, to the ether that adds 5mL in lower sediment, vibration 1min, the centrifugal 2min of 4000rpm, again upper strata ether is shifted out mutually, merge all upper stratas ether phase, concentrating under reduced pressure desolvation, obtain algae oil.Microalgae cell fat content is expressed as mgg
-1dw, i.e. microalgae grease content (%)=grease weight/micro-algae dry product weight * 100.
In following examples, the weight of micro-algae dry product refers to for putting forward micro-algae powder quality of oil, absorbancy by microalgae cell under 680nm can judge the upgrowth situation of micro-algae, treat that micro algae growth is to stationary phase, frustule is shaken up, get the centrifugal 3min of 10ml algae liquid 8000rpm, remove nutrient solution, by obtained algae mud dry 12h at 70 ℃, obtain micro-algae dry product, obtain micro-algae dry product quality in every ml algae liquid, be expressed as mg/ml, for carrying the micro-algae dry product of micro-algae dry product quality (the mg)=every ml algae liquid quality * of oil, for carrying micro-algae algae liquid of oil, amass.
Embodiment 1:
In order to investigate the impact of vagusstoff on chlorella growth and oil and fat accumulation, the defined medium of our configuration is in basic Tap substratum, to add the vagusstoff of different concns, and the concentration of controlling vagusstoff in substratum is 0 μ g/L, 0.125 μ g/L, 0.25 μ g/L, 0.5 μ g/L, 1 μ g/L, 5 μ g/L, 10 μ g/L.
Above-mentioned, with the inoculum size of volume fraction 5%, inoculate fresh chlorella respectively in respectively organizing nutrient solution, at 25 ℃, in the constant temperature illumination box of 3000lux, be cultured to after stationary phase, obtain algae liquid.In culturing process, every 24h, get algae liquid, under 680nm, measure the absorbancy of algae liquid, obtain the growth curve of chlorella.
Above-mentioned gained algae liquid is shaken up, get the centrifugal 3min of 10ml algae liquid 8000rpm, remove nutrient solution, by obtained algae mud dry 12h at 70 ℃, obtain micro-algae dry product, obtain micro-algae dry product quality in every ml algae liquid, be expressed as mg/ml, for carrying the micro-algae dry product of micro-algae dry product quality (the mg)=every ml algae liquid quality * of oil, for carrying micro-algae algae liquid of oil, amass.
The adjustment period of chlorella growth, add different concns vagusstoff on the impact of its amount of biomass and fat content in Table 1 and table 2 shown in.The adjustment period of chlorella growth, add different concns vagusstoff to see Fig. 1 to the impact of its amount of biomass.Result shows with respect to control group (concentration of vagusstoff is 0 μ g/L), chlorella growth adjustment period add vagusstoff can obviously improve amount of biomass and the fat content of chlorella, the raising degree of chlorella biomass and fat content and the concentration of vagusstoff have very large relation, when vagusstoff concentration is 5 μ g/L, it is the highest that the amount of biomass of chlorella and fat content all reach, and is respectively 1.26 times and 1.42 times of control group.
The adjustment period of table 1 chlorella growth, add the impact of different concns vagusstoff on its amount of biomass
The adjustment period of table 2 chlorella growth, add the impact of different concns vagusstoff on its fat content
Embodiment 2:
With the method similar to example 1, test, except Initial stage of culture does not add, vagusstoff, is cultured to logarithm initial stage of micro algae growth, i.e. OD
680value is 1.5, then to add concentration be outside the vagusstoff of 0 μ g/L, 0.125 μ g/L, 0.25 μ g/L, 0.5 μ g/L, 1 μ g/L, 5 μ g/L, 10 μ g/L, other parameter constants.
Chlorella growth logarithmic phase add different concns vagusstoff on the impact of its amount of biomass and fat content in Table 3 and table 4 shown in.Chlorella growth logarithmic phase adds different concns vagusstoff to see Fig. 2 to the impact of its amount of biomass.Result shows with respect to control group (concentration of vagusstoff is 0 μ g/L), in the logarithmic phase of chlorella growth, add vagusstoff can significantly improve amount of biomass and the fat content of chlorella, and the amplitude increasing higher than chlorella growth adjustment period add different concns vagusstoff.The raising degree of chlorella amount of biomass and fat content and the concentration of vagusstoff have very large relation, increase along with vagusstoff concentration, the amount of biomass of chlorella presents the trend increasing gradually, but the vagusstoff of high density but can cause the decline of chlorella fat content, consider the impact of vagusstoff on chlorella amount of biomass and fat content, select vagusstoff concentration 5 μ g/L, optimum concentration as chlorella growth and oil and fat accumulation, now the amount of biomass of chlorella and fat content are respectively 1.34 times and 1.45 times of control group.
Table 3 chlorella growth logarithmic phase adds the impact of different concns vagusstoff on its amount of biomass
Table 4 chlorella growth logarithmic phase adds the impact of different concns vagusstoff on its fat content
Chlorella not of the same race: growth adjustment period add 5 μ g/L, as shown in Figure 3, Figure 4, result shows that chlorella not of the same race is all significantly increased than the amount of biomass and the oil and fat accumulation that do not add vagusstoff after adding vagusstoff respectively to result.
Embodiment 3:
The method similar to example 1 tested, except Initial stage of culture does not add neurotransmitter acetylcholine, add concentration the stationary phase that is cultured to micro algae growth is again outside the vagusstoff of 0 μ g/L, 0.125 μ g/L, 0.25 μ g/L, 0.5 μ g/L, 1 μ g/L, 5 μ g/L, 10 μ g/L, other parameter constants.
Chlorella growth logarithmic phase add different concns vagusstoff on the impact of its amount of biomass and fat content in Table 5 and table 6 shown in.Result shows with respect to control group (concentration of vagusstoff is 0 μ g/L), in the stationary phase of chlorella growth, adds vagusstoff little on the raising impact of the amount of biomass of chlorella and fat content.
Table 5 chlorella growth adds the impact of different concns vagusstoff on its amount of biomass stationary phase
Table 6 chlorella growth adds the impact of different concns vagusstoff on its fat content stationary phase
Embodiment 4:
With the method similar to example 1, test, difference is that the object of experiment is changed to spirulina, chlamydomonas, Nannochloropsis oceanica, haematococcus pulvialis, the new green alga of rich oil, multinuclear green alga, multiform grid algae, palm net algae and grape algae, respectively in the logarithmic phase of above-mentioned algal grown, OD
680value is 1.5, to adding final concentration in its culture, is the vagusstoff of 5 μ g/L, other parameter constants.
Experimental result, as shown in table 7～9, as can be seen from the table, is added the accumulation that vagusstoff can promote micro algae growth and microalgae grease in above-mentioned algae.
Table 7 vagusstoff promotes the accumulation of different algal species lipid acid
Table 8 vagusstoff promotes the accumulation of different algal species amount of biomass
Table 9 vagusstoff promotes the rate of increase of different algal species amount of biomass and oil and fat accumulation
Embodiment 5:
With the method similar to example 1, test, difference be chlorella growth adjustment period in different experimental group, add 50 μ g/L, 150 μ g/L, 200 μ g/L choline, choline chloride 60, hydrogenation choline, citicoline or phosphatidylcholine, result is as shown in Figure 5, Figure 6.Wherein CHOL represents choline, and CH represents hydrogenation choline, and PC represents phosphatidylcholine, and INN represents citicoline, and CHCL represents choline chloride 60.As can be seen from the figure, in chlorella, add choline precursor and analogue thereof, all can promote the accumulation of micro algae growth microalgae grease accumulation.
Claims (15)
1. vagusstoff and analogue thereof promote the application in micro algae growth and microalgae grease accumulation at the same time.
2. application according to claim 1, is characterized in that, described micro-algae is any one in chlorella, spirulina, chlamydomonas, Nannochloropsis oceanica, haematococcus pulvialis, the new green alga of rich oil, multinuclear green alga, multiform grid algae, palm net algae and grape algae.
3. application according to claim 1, is characterized in that, micro algae growth adjustment period, logarithmic phase adds vagusstoff or vagusstoff analogue in substratum; Vagusstoff is 0.1～5 μ g/L at the adjustment period addition of micro algae growth, and vagusstoff is 0.1～5 μ g/L at the logarithmic phase addition of micro algae growth; Vagusstoff analogue is 50～200 μ g/L at the adjustment period addition of micro algae growth, and vagusstoff analogue is 50～200 μ g/L at the logarithmic phase addition of micro algae growth.
4. according to the application described in claim 1 or 3, it is characterized in that, the analogue of described vagusstoff is choline, choline chloride 60, bursine, citicoline or phosphatidylcholine.
5. application according to claim 1, is characterized in that, micro-algae culture temperature is 25～30 ℃, and cultivation intensity of illumination is 2000～4000lux.
6. vagusstoff and analogue thereof the application in promoting microalgae grease accumulation.
7. application according to claim 6, is characterized in that, described micro-algae is any one in chlorella, spirulina, chlamydomonas, Nannochloropsis oceanica, haematococcus pulvialis, the new green alga of rich oil, multinuclear green alga, multiform grid algae, palm net algae and grape algae.
8. application according to claim 6, is characterized in that, micro algae growth adjustment period, logarithmic phase adds vagusstoff or vagusstoff analogue in substratum; Vagusstoff is 0.1～5 μ g/L at the adjustment period addition of micro algae growth, and vagusstoff is 0.1～5 μ g/L at the logarithmic phase addition of micro algae growth; Vagusstoff analogue is 50～200 μ g/L at the adjustment period addition of micro algae growth, and vagusstoff analogue is 50～200 μ g/L at the logarithmic phase addition of micro algae growth.
9. according to the application described in claim 6 or 8, it is characterized in that, described vagusstoff analogue is choline, choline chloride 60, bursine, citicoline or phosphatidylcholine.
10. application according to claim 6, is characterized in that, micro-algae culture temperature is 25～30 ℃, and cultivation intensity of illumination is 2000～4000lux.
The application in promoting micro algae growth of 11. vagusstoffs and analogue thereof.
12. application according to claim 11, is characterized in that, described micro-algae is that micro-algae is any one in chlorella, spirulina, chlamydomonas, Nannochloropsis oceanica, haematococcus pulvialis, the new green alga of rich oil, multinuclear green alga, multiform grid algae, palm net algae and grape algae.
13. application according to claim 11, is characterized in that, micro algae growth adjustment period, logarithmic phase adds vagusstoff or vagusstoff analogue in substratum; Vagusstoff is 0.1～10 μ g/L at the adjustment period addition of micro algae growth, and vagusstoff is 0.1～10 μ g/L at the logarithmic phase addition of micro algae growth; Vagusstoff analogue is 50～200 μ g/L at the adjustment period addition of micro algae growth, and vagusstoff analogue is 50～200 μ g/L at the logarithmic phase addition of micro algae growth.
14. according to the application described in claim 11 or 13, it is characterized in that, described vagusstoff analogue is choline, choline chloride 60, bursine, citicoline or phosphatidylcholine.
15. application according to claim 11, is characterized in that, micro-algae culture temperature is 25～30 ℃, and cultivation intensity of illumination is 2000～4000lux.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410421791.5A CN104178446B (en) | 2014-08-25 | 2014-08-25 | Application of acetylcholine and analogs thereof in promoting microalgae growth and microalgae grease accumulation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410421791.5A CN104178446B (en) | 2014-08-25 | 2014-08-25 | Application of acetylcholine and analogs thereof in promoting microalgae growth and microalgae grease accumulation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104178446A true CN104178446A (en) | 2014-12-03 |
| CN104178446B CN104178446B (en) | 2017-02-01 |
Family
ID=51959795
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410421791.5A Expired - Fee Related CN104178446B (en) | 2014-08-25 | 2014-08-25 | Application of acetylcholine and analogs thereof in promoting microalgae growth and microalgae grease accumulation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104178446B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112391293A (en) * | 2020-11-25 | 2021-02-23 | 宁波大学 | Method for preparing squalene by autotrophic culture of botryococcus through heterotrophic biomembrane adherence |
| CN112813015A (en) * | 2021-02-05 | 2021-05-18 | 深圳大学 | Promoter for increasing dry weight of euglena, euglena culture medium and application thereof |
-
2014
- 2014-08-25 CN CN201410421791.5A patent/CN104178446B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| ANNALIESE K. FRANZ ET AL.: "Phenotypic Screening with Oleaginous Microalgae Reveals Modulators of Lipid Productivity", 《ACS CHEM. BIOL.》, 22 March 2013 (2013-03-22) * |
| VICTORIA V. ROSHCHINA: "Evolutionary Considerations of Neurotransmitters in Microbial, Plant, and Animal Cells", 《MICROBIAL ENDOCRINOLOGY》, 31 December 2010 (2010-12-31) * |
| YOKO HORIUCHI ET AL.: "Evolutional study on acetylcholine expression", 《LIFE SCIENCES》, 31 December 2003 (2003-12-31) * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112391293A (en) * | 2020-11-25 | 2021-02-23 | 宁波大学 | Method for preparing squalene by autotrophic culture of botryococcus through heterotrophic biomembrane adherence |
| CN112813015A (en) * | 2021-02-05 | 2021-05-18 | 深圳大学 | Promoter for increasing dry weight of euglena, euglena culture medium and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104178446B (en) | 2017-02-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gim et al. | Comparison of biomass production and total lipid content of freshwater green microalgae cultivated under various culture conditions | |
| Feng et al. | Lipid accumulation and growth of Chlorella zofingiensis in flat plate photobioreactors outdoors | |
| Parmar et al. | Cyanobacteria and microalgae: a positive prospect for biofuels | |
| Yousuf | Fundamentals of microalgae cultivation | |
| US20090298159A1 (en) | Method for producing biodiesel from an alga | |
| KR101694711B1 (en) | Culturing method of microalgae for increasing lipid productivity | |
| JP2018520636A (en) | Omega-7 fatty acid composition, and method and application for culturing yellow-green algae to produce the composition | |
| CN102465098B (en) | Culture medium composition for culturing chlorella | |
| CN102268377A (en) | Method for improving lipid producing microalga biomass and lipid accumulation with two stage culture strategy of mixotrophic and nitrogen-rich-nitrogen-deficient culture | |
| CN109576315A (en) | A method of microalgae grease is produced using flue gas | |
| CN104152357A (en) | High-density culture method for improving chlorophyll and protein content of chlorella at same time | |
| CN105441524A (en) | Method for increasing yield of microalgae grease with saccharose as carbon source through co-culture | |
| Agwa et al. | Heterotrophic cultivation of Chlorella sp. using different waste extracts | |
| WO2015085631A1 (en) | Method for culturing botryococcus spp. with high yield | |
| CN110607238A (en) | Method for promoting grease accumulation of monoraphidium by combining nitrogen deficiency with strigolactone | |
| CN106480155B (en) | Method suitable for promoting haematococcus pluvialis to produce astaxanthin under high-temperature condition | |
| CN105713934B (en) | A method of producing microalgae grease | |
| CN104232559B (en) | The method of cultivating microalgae and the method for producing grease | |
| CN104178446A (en) | Application of acetylcholine and analogs thereof in promoting microalgae growth and microalgae grease accumulation | |
| CN107460217B (en) | Method for preparing microalgae grease through mixed culture | |
| CN103981106A (en) | High-yield DHA (Docosahexaenoic Acid) strain and application thereof | |
| CN106636264B (en) | Method for producing phycoerythrin and polyunsaturated fatty acid by utilizing haematococcus halophycus | |
| CN109929886A (en) | A method of utilizing walnut shell extract culture single needle algae generation diesel oil | |
| Bansal | Mixotrophic growth of Chlorella Sp. using Glycerol for the production of Biodiesel: A Review | |
| CN105713935A (en) | Method for producing lipid through mixed culture of microalgae |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170201 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |