CN103981083B - The closed mixotrophic cultivation method of a kind of micro-algae - Google Patents

The closed mixotrophic cultivation method of a kind of micro-algae Download PDF

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CN103981083B
CN103981083B CN201410254108.3A CN201410254108A CN103981083B CN 103981083 B CN103981083 B CN 103981083B CN 201410254108 A CN201410254108 A CN 201410254108A CN 103981083 B CN103981083 B CN 103981083B
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algae
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photoreactor
feeder
charging opening
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CN103981083A (en
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刘晃
吴凡
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Fishery Machinery and Instrument Research Institute of CAFS
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Abstract

The present invention relates to microdisk electrode technology, be specifically related to the closed mixotrophic cultivation method of a kind of micro-algae and culture systems thereof, substratum, sterilized water and micro-algae algae kind being added in feeder, being then delivered to constant speed flowing in photoreactor and cultivating 7-10 days, does is intensity of illumination 40-100 μm of ol? m -2s -1, culture temperature is 25-37 DEG C, and flow velocity is 0.2-0.6m/s; Algae liquid in photoreactor flow in results separator, the algae liquid of the 70%-90% in results separator is collected mouth as fresh algae liquid from algae liquid and is discharged, the algae liquid of residue 10%-30% passes through reflux pump as micro-algae algae kind, be back in feeder from algae kind charging opening, continue constant speed flowing cultivation after supplemental medium and sterilized water simultaneously, constantly be cycled to repeat above-mentioned culturing step, realize the closed cultured continuously of micro-algae.Cultural method of the present invention achieves the continuous prodution of micro-algae, improves the productive rate of micro-algae, reduces production cost.

Description

The closed mixotrophic cultivation method of a kind of micro-algae
Technical field
The present invention relates to microdisk electrode technology, be specifically related to the closed mixotrophic cultivation method of a kind of micro-algae and culture systems thereof.
Background technology
Micro-algae (microalgae) is the unicellular algae existed with individual, chain or group form of a large class microcosmic, and size is not from several microns to hundreds of micron etc.Micro-algae is of a great variety, according to estimates nearly 200000-800000 kind, and wherein existing record has kind more than 35000.Micro-algae is divided into the micro-algae of autotrophy, heterotrophic microalgae and holds concurrently supports micro-algae.From the sixties in 20th century, since first Japan start to carry out the extensive both culturing microalgae of commercialization, people can realize micro-algae at multiple fields such as protective foods, medical material, beauty treatment, feeds and commercially produce so far.
The cultivation of micro-algae large-scale autotrophy is mainly adopted in two ways: open microalgae cultivation system and sealed microalgae cultivating system.Open cultivation system is exactly that outdoor utilizes natural sunlight to carry out both culturing microalgae, has the advantages such as expansion scale ratio is easier to, cost is lower.But open cultivation system is easily by the impact of external environment, as intensity of illumination, light application time, temperature and weather; Also the pollution of other algae kinds, bacterium and pathogenic microorganism is easily subject to.Sealed microalgae cultivating system refers to and utilizes substratum to carry out both culturing microalgae in airtight container.Sealed microalgae cultivating system can be divided into fermentor tank, culture bag, plate smooth biochemical reactor and cast light biochemical reactor; The cultivation of micro-algae large-scale heterotrophism utilizes the micro-algae of industrial fermentation process Heterotrophic culture, can save space, and improve output, avoid the pollution of assorted algae and bacterium, and culture condition easily controlling, is the development trend of micro-algae large-scale industrialized production.Part produce oil algae can make its training method transfer heterotrophism to by autotrophy by metabolic regulation, owing to not relying on illumination, does not also consume CO 2, common fermentor device can be utilized to cultivate, but strictly will keep sterile state in culturing process.
Micro-algae mixotrophic cultivation can utilize organism, carbon source, carries out photosynthesis simultaneously.In micro-algae mixotrophic cultivation process, photoautotrophy and chemoheterotrophy are synchronous and relatively independent processes.Illumination has impact to two pathways metabolisms, but influence degree is different.
Available Microalgae refer to those factorial praluction or application prospect, can by the kind of biotechnology mass propgation, in its cell, contained certain or some composition can be utilized by people.At present, Available Microalgae has a few class fronds such as spirulina, chlorella, Dunaliella salina, haematococcus pulvialis.Wherein, the fat content of chlorella is high, can be used as raw material prepared by the renewable energy resources.Chlorella can utilize at sun power and have function grow fast and accumulate grease.LiTingting etc. (2013) result of study shows: adopt different training method, carries out biomass and the lipid production experiment of chlorella.Holding concurrently under the condition of supporting, can obtain and compare independent light autotrophy or independent heterotrophism, better growth performance, chlorella lipid content also increases considerably.
At present, the cultivation of chlorella large-scale commercial is mainly carried out in bioreactor, and its floor space is large, the production cycle long, stand density is lower, in a disguised form adds production cost, limits extensive chlorella cultivation development.Finding the chlorella cultural method of a kind of high-density, low cost, is important to promotion chlorella aquaculture industry development pole.
Summary of the invention
The object of this invention is to provide the closed mixotrophic cultivation method of a kind of micro-algae, adopt closed cultured continuously mode, in culturing process, release the fresh algae liquid of part with constant speed and carry out processing purification, another part carries out backflow as algae kind and cultivates, supplementary corresponding substratum and sterilized water make it the closed cultured continuously realizing micro-algae simultaneously, improve the productive rate of micro-algae.
Another object of the present invention is to provide the closed mixotrophic cultivation system of a kind of micro-algae, and this system footprint area is few, structure is simple, cost is low.
In order to realize above technique effect, the present invention realizes as follows:
The closed mixotrophic cultivation system of a kind of micro-algae, is characterized in that: this culture systems is feeder, photoreactor, results separator and reflux pump are connected successively by pipeline;
The end face of described feeder is provided with substratum charging opening, sterilized water charging opening, initial algae kind charging opening and circulation algae kind charging opening, and the outlet of this feeder is connected with the import of photoreactor;
The outlet of described photoreactor is connected with the import of results separator;
Described results separator is provided with algae liquid and collects mouth and algae kind discharge outlet, and this algae kind discharge outlet is connected with described reflux pump, and the circulation algae kind charging opening refluxed on pump outlet and feeder communicates.Preferably, described photoreactor is duct type or flat photoreactor; Described results separator is centrifugal or air-flotation type separator.
The closed mixotrophic cultivation method of above-mentioned micro-algae, its step comprises:
A, substratum, sterilized water and initial micro-algae algae kind be added in feeder to be formed from substratum charging opening, sterilized water charging opening and initial algae kind charging opening respectively and cultivate mixed solution, then be delivered to constant speed flowing in photoreactor and cultivate 7-10 days, intensity of illumination is 40-100 μm of olm -2s -1, culture temperature is 25-37 DEG C, and the flow velocity of algae liquid in photoreactor is 0.2-0.6m/s.The concentration of described initial micro-algae algae kind is 50-100mg/L.Preferably, the flow velocity of algae liquid in photoreactor is 0.3m/s.
Algae liquid in B, photoreactor flow in results separator, the algae liquid of the 70%-90% in results separator is collected mouth as fresh algae liquid from algae liquid and is discharged, the algae liquid of residue 10%-30% passes through reflux pump as micro-algae algae kind, be back in feeder from circulation algae kind charging opening, supplemental medium and sterilized water simultaneously, then flow to constant speed flowing in photoreactor and cultivates; Fresh algae liquid is collected after mouth discharges from algae liquid, and carries out dehydration through methods such as centrifugal, air supportings and gather in the crops.The micro-concentration of algae separated in described results separator is 3-4g/L, and total fat content is 35%-50%.
C, repeating step (A) and step (B).Constantly like this to be cycled to repeat, realize the closed cultured continuously of micro-algae, and the flow velocity of whole double sample culture systems to be 0.3m/s.
In feeder (1), add substratum makes the nutritive ingredient concentration of the cultivation mixed solution contained in feeder be 7-12g/L glucose, 1-3g/LKNO 3, 600-650mg/LNaH 2pO 4h 2o, 85-95mg/LNa 2hPO 42H 2o, 240-250mg/LMgSO 47H 2o, 9-10mg/LEDTA, 0.05-0.07mg/LH 3bO 3, 14-15mg/LCaCl 22H 2o, 6-8mg/LFeSO 47H 2o, 0.2-0.3mg/LZnSO 47H2O, 0.01-0.02mg/L (NH4) 6mo 7o 244H 2o, 0.1-0.2mg/LMnSO 4h 2o, 0.002-0.003mg/LCuSO 45H 2o.Preferably, the nutritive ingredient concentration of the cultivation mixed solution contained in feeder is 9g/L glucose, 1.7g/LKNO 3, 621mg/LNaH 2pO 4h 2o, 89mg/LNa 2hPO 42H 2o, 246.5mg/LMgSO 47H 2o, 9.3mg/LEDTA, 0.061mg/LH 3bO 3, 14.7mg/LCaCl 22H 2o, 6.95mg/LFeSO 47H 2o, 0.287mg/LZnSO 47H2O, 0.01235mg/L (NH4) 6mo 7o 244H 2o, 0.169mg/LMnSO 4h 2o, 0.00249mg/LCuSO 45H 2o.
Described micro-algae is chlorella.
The invention has the beneficial effects as follows:
1, the present invention adopts the closed mixotrophic cultivation method of micro-algae, micro-algae algae kind, substratum and sterilized water are invested in culture systems according to a certain percentage and carry out closed mixotrophic cultivation, and the cultured algae liquid of the 10-30% in culture systems is re-used as algae kind, like this be cycled to repeat the continuous prodution that training method achieves micro-algae, improve the productive rate of micro-algae.
2, the present invention cultivates the biomass of the micro-algae obtained and total fat content can reach 3-4g/L and 35%-50% respectively, the biomass of the micro-algae all obtained higher than common mixotrophic cultivation mode and total fat content.And the maximum biomass dry weight of micro-algae of the present invention's production can reach more than 1.8 times of Heterotrophic culture, light autotrophy cultivate more than 5.2 times, the yield of biomass that glucose consumption obtains can reach 0.82g/g, and under the same terms, Heterotrophic culture only has 0.34g/g.
3, the floor space of micro-algae closed mixotrophic cultivation system of the present invention's employing is few, structure is simple, cost is low, and the productive rate of micro-algae is high, can realize large-scale industrial production.
Accompanying drawing explanation
Fig. 1 is the structural representation of the closed mixotrophic cultivation system of micro-algae of the present invention.
Fig. 2 is the biomass of chlorella in embodiment 1 and the comparison diagram of total fat content and incubation time.
Wherein, 1 in Fig. 1 is feeder, and 2 is photoreactors, and 3 is results separators, 4 is reflux pumps, and 11 is substratum charging openings, and 12 is sterilized water charging openings, and 13 is initial algae kind charging openings, 14 is circulation algae kind charging openings, and 31 is that algae liquid collects mouth, and 32 is algae kind discharge outlets.
Embodiment
Below in conjunction with embodiment, the invention will be further described:
Embodiment 1
(1), the establishment of the closed mixotrophic cultivation system of micro-algae:
Feeder (1), photoreactor (2), results separator (3) and reflux pump (4) are connected to form the closed mixotrophic cultivation system of micro-algae successively by pipeline.Wherein, photoreactor (1) is duct type photoreactor; Results separator (3) are centrifugal separator.
The end face of described feeder (1) is provided with substratum charging opening (11), sterilized water charging opening (12) and initial algae kind charging opening (13) for supplementing.The outlet of feeder is connected with the import of photoreactor; The outlet of photoreactor (2) is connected with the import of results separator (3); Results separator is provided with algae liquid and collects mouth (31) and algae kind discharge outlet (32), in photoreactor, the 70-90% of cultured micro-algae algae liquid discharges from algae kind discharge outlet (32), and to dewater results through centrifugal or air supporting mode, the algae liquid of 10%-30% is discharged from algae kind discharge outlet (32), through reflux pump, squeeze in feeder from circulation algae kind charging opening (14).The concrete structure schematic diagram of the present embodiment as shown in Figure 1.
(2), the closed mixotrophic cultivation method of chlorella
(A), the configuration of substratum: configure substratum and carry out high-temperature sterilization process.
(B), the substratum configured, sterilized water and micro-algae algae kind are added in feeder (1) from substratum charging opening (11), sterilized water charging opening (12) and initial algae kind charging opening (13) respectively, the nutritive ingredient concentration of the cultivation mixed solution contained in feeder (1) is 9g/L glucose, 1.7g/LKNO 3, 621mg/LNaH 2pO 4h 2o, 89mg/LNa 2hPO 42H 2o, 246.5mg/LMgSO 47H 2o, 9.3mg/LEDTA, 0.061mg/LH 3bO 3, 14.7mg/LCaCl 22H 2o, 6.95mg/LFeSO 47H 2o, 0.287mg/LZnSO 47H2O, 0.01235mg/L (NH4) 6mo 7o 244H 2o, 0.169mg/LMnSO 4h 2o, 0.00249mg/LCuSO 45H 2o.The concentration of initial micro-algae algae kind is 50-100mg/L, and then micro-algae algae kind is delivered to constant speed flowing in photoreactor (2) and cultivates 7-10 days, intensity of illumination is 40-100 μm of olm -2s -1, culture temperature is 25-37 DEG C, and flow velocity is 0.3m/s.
(C) the algae liquid in photoreactor (2) flow in results separator (3), the algae liquid of the 70%-90% in results separator (3) is collected mouth (31) as fresh algae liquid from algae liquid and is discharged, residue 10%-30%, concentration be the algae liquid of 3-4g/L as micro-algae algae kind by reflux pump (4), be back in feeder (1) from circulation algae kind charging opening (14), supplemental medium and sterilized water simultaneously, then flow in photoreactor (2) and carries out constant speed flowing cultivation again.Subsequently cultured algae liquid is separated through results separator (3), fractional dose is that the algae liquid of 70%-90% collects mouth (31) discharge as fresh algae liquid from algae liquid, the concentration of residue 10%-30% is that the algae liquid of 3-4g/L is circulated to after in feeder (1) and cultivates, constantly recirculation, realizes the continuous prodution of chlorella.
From photoreactor, carry out sampling analysis, concrete outcome as shown in Figure 2, as can be seen from Figure, total fat content of chlorella constantly increases along with incubation time, can reach more than 45%, biomass then cultivate within the 3rd day, to reach maximum be 3.7g/L, next tend towards stability.

Claims (4)

1. the closed mixotrophic cultivation method of chlorella, is characterized in that: the culture systems that this culture method adopts is feeder (1), photoreactor (2), results separator (3) and reflux pump (4) are connected successively by pipeline;
The end face of described feeder (1) is provided with substratum charging opening (11), sterilized water charging opening (12), initial algae kind charging opening (13) and circulation algae kind charging opening (14), and the outlet of this feeder (1) is connected with the import of photoreactor (2);
The outlet of described photoreactor (2) is connected with the import of results separator (3);
Described results separator (3) is provided with algae liquid and collects mouth (31) and algae kind discharge outlet (32), this algae kind discharge outlet (32) is connected with described reflux pump (4), and circulation algae kind charging opening (14) on the outlet of reflux pump (4) and feeder (1) communicates;
Its culturing step comprises:
A, substratum, sterilized water and initial chlorella algae kind to be added in feeder (1) from substratum charging opening (11), sterilized water charging opening (12) and initial algae kind charging opening (13) respectively and to form cultivation mixed solution, the concentration of the initial chlorella algae kind in this cultivation mixed solution is 50-100mg/L; Then be delivered to constant speed flowing in photoreactor (2) and cultivate 7-10 days, intensity of illumination is 40-100 μm of olm -2s -1, culture temperature is 25-37 DEG C, and flow velocity is 0.2-0.6m/s;
Algae liquid in B, photoreactor (2) flow in results separator (3), the algae liquid of the 70%-90% in results separator (3) is collected mouth (31) as fresh algae liquid from algae liquid and is discharged, and to dewater results through centrifugal or air supporting mode, the algae liquid of residue 10%-30% passes through reflux pump (4) as chlorella algae kind, be back in feeder (1) from circulation algae kind charging opening (14), supplemental medium and sterilized water simultaneously, then flow to constant speed flowing in photoreactor (2) and cultivates;
C, repeating step (A) and step (B).
2. the closed mixotrophic cultivation method of chlorella according to claim 1, is characterized in that: described photoreactor (2) is duct type or flat photoreactor; Described results separator (3) is centrifugal or air-flotation type separator.
3. the closed mixotrophic cultivation method of chlorella according to claim 1, is characterized in that: the nutritive ingredient concentration of adding the cultivation mixed solution after substratum in described feeder (1) is 7-12g/L glucose, 1-3g/LKNO 3, 600-650mg/LNaH 2pO 4h 2o, 85-95mg/LNa 2hPO 42H 2o, 240-250mg/LMgSO 47H 2o, 9-10mg/LEDTA, 0.05-0.07mg/LH 3bO 3, 14-15mg/LCaCl 22H 2o, 6-8mg/LFeSO 47H 2o, 0.2-0.3mg/LZnSO 47H2O, 0.01-0.02mg/L (NH4) 6mo 7o 244H 2o, 0.1-0.2mg/LMnSO 4h 2o, 0.002-0.003mg/LCuSO 45H 2o.
4. the closed mixotrophic cultivation method of chlorella according to claim 1, is characterized in that: in described step (B), and the chlorella concentration separated from results separator is 3-4g/L, and total fat content is 35%-50%.
CN201410254108.3A 2014-06-09 2014-06-09 The closed mixotrophic cultivation method of a kind of micro-algae Expired - Fee Related CN103981083B (en)

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CN105154317A (en) * 2015-10-08 2015-12-16 扬州大学 Novel continuous microalgae culture reactor and using method thereof
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