CN105039164B - A kind of microalgae immobilized cultivation method - Google Patents
A kind of microalgae immobilized cultivation method Download PDFInfo
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- CN105039164B CN105039164B CN201510503798.6A CN201510503798A CN105039164B CN 105039164 B CN105039164 B CN 105039164B CN 201510503798 A CN201510503798 A CN 201510503798A CN 105039164 B CN105039164 B CN 105039164B
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- polyvinyl alcohol
- absorbent cotton
- softening
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Abstract
The present invention discloses a kind of microalgae immobilized cultivation method, is related to field of microbial culture technology, the problem of to prevent hosqt media selection not at that time, and cannot achieve the industrialized production of microalgae using Immobilized culture method culture microalgae.The microalgae immobilized cultivation method includes: to be inoculated with microalgae cell on the polyvinyl alcohol absorbent cotton of softening using the polyvinyl alcohol absorbent cotton of softening as hosqt media to carry out microdisk electrode；The quality for the fountain solution being adsorbed in polyvinyl alcohol absorbent cotton is 3 times -6 times of polyvinyl alcohol absorbent cotton quality；The dry weight for the microalgae cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is 5g-100g；When carrying out microdisk electrode, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs the culture medium of 300mL-2000mL per minute.Microalgae immobilized cultivation method provided by the invention is used for microculture field.
Description
Technical field
The present invention relates to field of microbial culture technology more particularly to a kind of microalgae immobilized cultivation methods.
Background technique
Microalgae (Microalgae) is a kind of autophyte widely distributed in land, ocean, it can using solar energy,
Carbon dioxide and water pass through photosynthesis Synthetic Oil, starch, carbohydrate and a variety of high added value bioactive substances；
Moreover, because the micro algae growth period is short, biomass accumulation ability is higher than terrestrial plant, and therefore, microalgae is that one kind potentially can be again
The raw materials for production of the raw energy.
In order to improve micro algae biomass accumulation ability, promote the optical energy utilization efficiency in microalgae mass culture, people are to micro-
The cultural method of algae is explored, and two kinds of microalgae culture methods of Maitland culture and Immobilized culture method are proposed；Wherein, fixed
Changing cultivation is the surface layer that microalgae is fixed on to hosqt media, and it is wet to be able to maintain cell colony by supplement during the cultivation process
Culture medium to maintain cell high-efficient to grow；And currently, when using Immobilized culture method culture microalgae, common hosqt media one
As be high molecular material, these high molecular materials can be the natural fibers such as the already existing linen-cotton of nature, or people
Geosynthetics, such as polytetrafluoroethylene (PTFE), acrylic are to chemical fibres such as the nonwettable material of water or polyester, nylon.Wherein,
Natural fiber is easy the pollution by exogenous biological (such as mould) and corrodes；Although chemical fibre can in synthetic material
The defect for avoiding natural fiber from corroding vulnerable to exogenous biological；But water is not infiltrated；Therefore, using artificial synthesized material
When carrying out the Immobilized culture of microalgae as hosqt media, culture medium cannot be uniformly distributed on hosqt media, be unable to fully
Meet the needs of microalgae is to culture solution in Immobilized culture method, leads to frustule growth restriction；And the weatherability of chemical fibre is poor,
It is especially easy to decompose under conditions of extraneous illumination, service life is limited.Therefore, Immobilized culture method culture microalgae is being used
When, the selection of hosqt media is very important；It is the industrialized production that cannot achieve microalgae if hosqt media selection is improper
's.
Summary of the invention
The purpose of the present invention is to provide a kind of microalgae immobilized cultivation methods, to prevent hosqt media from selecting not at that time,
The problem of industrialized production of microalgae cannot achieve using Immobilized culture method culture microalgae.
To achieve the goals above, the invention provides the following technical scheme:
A kind of microalgae immobilized cultivation method, using the polyvinyl alcohol absorbent cotton of softening as hosqt media, to the poly- of softening
It is inoculated with microalgae cell on vinyl alcohol absorbent cotton, then carries out microdisk electrode, obtains cultured microalgae；Wherein,
The polyvinyl alcohol absorbent cotton of softening is by polyvinyl alcohol absorbent cotton and the fountain solution being adsorbed in polyvinyl alcohol absorbent cotton
Composition；The quality for the fountain solution being adsorbed in polyvinyl alcohol absorbent cotton is 3 times -6 times of polyvinyl alcohol absorbent cotton quality；Every square
The dry weight for the microalgae cell that the polyvinyl alcohol absorbent cotton surface of rice softening is inoculated with is 5g-100g；When carrying out microdisk electrode, often put down
The polyvinyl alcohol absorbent cotton of square rice softening absorbs the culture medium of 300mL-2000mL per minute.
Preferably, the polyvinyl alcohol absorbent cotton of the softening is obtained using following methods: polyvinyl alcohol absorbent cotton is soaked
It steeps in fountain solution, is adsorbed onto fountain solution on polyvinyl alcohol absorbent cotton, the polyvinyl alcohol absorbent cotton softened；Or,
By on fountain solution spray to polyvinyl alcohol absorbent cotton, so that polyvinyl alcohol absorbent cotton is absorbed fountain solution, softened
Polyvinyl alcohol absorbent cotton.
Preferably, the mass area ratio of the polyvinyl alcohol absorbent cotton is 100g/m2-1000g/m2。
Preferably, the temperature of the immersion fluid is greater than 0 and is less than or equal to 80 DEG C.
Preferably, the immersion fluid is water or culture medium.
Preferably, the microalgae cell being inoculated on the polyvinyl alcohol absorbent cotton of the softening is covered on the polyvinyl alcohol of softening
On absorbent cotton, and the microalgae cell being inoculated on the polyvinyl alcohol absorbent cotton of the softening covers the polyvinyl alcohol water suction of softening
The 100% of continuous surface area.
Preferably, the dry weight for the microalgae cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is
15g-32g。
Preferably, the microalgae cell is spherical microalgae cell, the polyvinyl alcohol absorbent cotton surface of every square metre of softening
The dry weight for the microalgae cell being inoculated with is 5g-15g；Or,
The microalgae cell is filamentous microalgae cell, and the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with
Microalgae cell dry weight be 10g-32g；Or,
The microalgae cell is shuttle-type microalgae cell or microalgae many cells disjunctor；The polyvinyl alcohol of every square metre of softening is inhaled
The dry weight for the microalgae cell that water silk floss surface is inoculated with is 15g-20g.
Preferably, when carrying out the microdisk electrode, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs per minute
The culture medium of 600mL-1200mL.
Preferably, it is greater than 300 μm of olm in intensity of illumination-2·s-1Visible light under or humidity less than 20% in the environment of
When carrying out microdisk electrode, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs per minute is greater than 1000mL, is less than or equal to
The culture medium of 2000mL；Or,
In less than 300 μm olm of intensity of illumination-2·s-1Visible light under or humidity be greater than 20% in the environment of carry out it is micro-
When algae is cultivated, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs per minute is more than or equal to 300mL, the culture less than 1000mL
Base；Or,
It is equal to 300 μm of olm in intensity of illumination-2·s-1Visible light under or humidity be equal to 20% in the environment of carry out it is micro-
When algae is cultivated, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs the culture medium of 1000mL per minute.
Compared with prior art, the beneficial effect of microalgae immobilized cultivation method provided by the invention is:
For the present invention using the polyvinyl alcohol absorbent cotton that softens as hosqt media, the polyvinyl alcohol absorbent cotton of softening is by poly- second
Enol absorbent cotton and the fountain solution being adsorbed in polyvinyl alcohol absorbent cotton form；And polyvinyl alcohol absorbent cotton surface be distributed with it is several
Micropore, these micropores can make polyvinyl alcohol absorbent cotton have good water suction and the capacity of water supply；And polyvinyl alcohol absorbent cotton is good
Good water absorbing capacity can guarantee that polyvinyl alcohol absorbent cotton uniformly adsorbs culture medium in the case where abundant wetting；Polyvinyl alcohol
The good capacity of water supply of absorbent cotton enables microalgae adequately to provide culture medium when immobilizing culture for microalgae cell, with
Maintain the growth of microalgae cell.In addition, the production cost of polyvinyl alcohol absorbent cotton of the present invention is low, it is not easy mouldy, tool
There are good toughness and wearability, therefore, when the present invention is using the polyvinyl alcohol absorbent cotton that softens as hosqt media, service life
It is higher, it can be used for industrializing Immobilized culture microalgae.
In order to guarantee that the efficiency of industrial Immobilized culture microalgae, the present invention also make polyvinyl alcohol absorbent cotton adsorb sole mass
3 times -6 times of fountain solution, the polyvinyl alcohol absorbent cotton of obtained softening can not only guarantee microalgae cell in the polyethylene of softening
The success rate being inoculated on alcohol absorbent cotton, additionally it is possible to prevent the fountain solution contained in the polyvinyl alcohol absorbent cotton due to softening excessive,
The loss of microalgae cell on the caused polyvinyl alcohol absorbent cotton for being seeded in softening.
On this basis, the present invention is also by the inoculum density for the microalgae cell being inoculated with to the polyvinyl alcohol surface of softening
It is defined, the dry weight 5g-100g for the microalgae cell for being inoculated with the polyvinyl alcohol absorbent cotton surface of every square metre of softening,
This is prevented while guaranteeing the utilization rate of polyvinyl alcohol absorbent cotton of softening because of inoculated more microalgae cells, makes microalgae
The problem of cell falls off from the polyvinyl alcohol absorbent cotton of softening；So polyvinyl alcohol absorbent cotton surface of the present invention to softening
The inoculum density for the microalgae cell being inoculated with, which is defined, can make what microalgae cell during the cultivation process can be stable to be attached to
On the polyvinyl alcohol absorbent cotton of softening.
In addition, the present invention during microdisk electrode, by controlling the supply amount of culture medium, makes the poly- of every square metre of softening
Vinyl alcohol absorbent cotton absorbs the culture medium of 300mL-2000mL per minute, makes the water-holding capacity of culture medium Yu polyvinyl alcohol absorbent cotton
Match, to ensure that the microalgae cell being seeded on the polyvinyl alcohol absorbent cotton of softening there can be sufficient culture medium to supply
It gives, and the microalgae cell being seeded on the polyvinyl alcohol absorbent cotton of softening will not be caused to be detached from because culture medium supply amount is excessive
The polyvinyl alcohol absorbent cotton of softening.
Therefore, the present invention is by adsorbing the quality of fountain solution to polyvinyl alcohol absorbent cotton, absorbing water to the polyvinyl alcohol of softening
The restriction of the supply amount of culture medium during the inoculum density and microdisk electrode of the microalgae cell that continuous surface is inoculated with reduces micro-
The energy consumption and cost of algae culture, improve the efficiency of Immobilized culture method culture microalgae, meet the needs of microalgae industry culture.
Specific embodiment
The microalgae immobilized cultivation method that embodiment provides in order to further illustrate the present invention, does below with reference to the present invention
Detailed description.
A kind of microalgae immobilized cultivation method provided by the invention is that the polyvinyl alcohol absorbent cotton that will soften is situated between as substrate
Matter is inoculated with microalgae cell on the polyvinyl alcohol absorbent cotton of softening, then carries out microdisk electrode, obtains cultured cell；Its
In,
The polyvinyl alcohol absorbent cotton of softening is by polyvinyl alcohol absorbent cotton and the wetting being adsorbed in polyvinyl alcohol absorbent cotton
Liquid composition；The quality for the fountain solution being adsorbed in polyvinyl alcohol absorbent cotton is 3 times -6 times of polyvinyl alcohol absorbent cotton quality；Often put down
The dry weight for the microalgae cell that the polyvinyl alcohol absorbent cotton surface of square rice softening is inoculated with is 5g-100g；When microdisk electrode, every square
The polyvinyl alcohol absorbent cotton of rice softening absorbs the culture medium of 300mL-2000mL per minute.
In above-mentioned microalgae immobilized cultivation method, the present invention is soft using the polyvinyl alcohol absorbent cotton that softens as hosqt media
The polyvinyl alcohol absorbent cotton of change is made of polyvinyl alcohol absorbent cotton and the fountain solution being adsorbed in polyvinyl alcohol absorbent cotton；And
Several micropores are distributed in polyvinyl alcohol absorbent cotton surface, these micropores can make polyvinyl alcohol absorbent cotton have it is good water suction and
The capacity of water supply；And the good water absorbing capacity of polyvinyl alcohol absorbent cotton can guarantee polyvinyl alcohol absorbent cotton in abundant the case where soaking
Lower uniform absorption culture medium；The good capacity of water supply of polyvinyl alcohol absorbent cotton enables microalgae to fill when immobilizing culture
Dividing is that microalgae cell provides culture medium, to maintain the growth of microalgae cell.In addition, polyvinyl alcohol water suction of the present invention
Continuous production cost is low, is not easy mouldy, has good toughness and wearability, therefore, the present invention is inhaled with the polyvinyl alcohol softened
When water silk floss is as hosqt media, service life is higher, can be used for industrializing Immobilized culture microalgae.
In order to guarantee that the efficiency of industrial Immobilized culture microalgae, the present invention also make polyvinyl alcohol absorbent cotton adsorb sole mass
3 times -6 times of fountain solution, the polyvinyl alcohol absorbent cotton of obtained softening can not only guarantee microalgae cell in the polyethylene of softening
The success rate being inoculated on alcohol absorbent cotton, additionally it is possible to prevent the fountain solution contained in the polyvinyl alcohol absorbent cotton due to softening excessive,
The loss of microalgae cell on the caused polyvinyl alcohol absorbent cotton for being seeded in softening.
On this basis, the present invention is also by the inoculum density for the microalgae cell being inoculated with to the polyvinyl alcohol surface of softening
It is defined, the dry weight 5g-100g for the microalgae cell for being inoculated with the polyvinyl alcohol absorbent cotton surface of every square metre of softening,
This is prevented while guaranteeing the utilization rate of polyvinyl alcohol absorbent cotton of softening because of inoculated more microalgae cells, makes microalgae
The problem of cell falls off from the polyvinyl alcohol absorbent cotton of softening；So polyvinyl alcohol absorbent cotton surface of the present invention to softening
The inoculum density for the microalgae cell being inoculated with, which is defined, can make what microalgae cell during the cultivation process can be stable to be attached to
On the polyvinyl alcohol absorbent cotton of softening.
In addition, the present invention, during microdisk electrode, by controlling the supply amount of culture medium, the measurement index of supply amount is
The polyvinyl alcohol absorbent cotton of every square metre of softening absorbs the quality of culture medium per minute.During specific microdisk electrode, often put down
The polyvinyl alcohol absorbent cotton of square rice softening absorbs the culture medium of 300mL-2000mL per minute, and culture medium and polyvinyl alcohol is made to absorb water
Continuous water-holding capacity matches, to ensure that the microalgae cell being seeded on the polyvinyl alcohol absorbent cotton of softening can have sufficiently
Culture medium supply, and will not cause to be seeded in micro- on the polyvinyl alcohol absorbent cotton of softening because culture medium supply amount is excessive
Frustule is detached from the polyvinyl alcohol absorbent cotton of softening.
Therefore, the present invention is by adsorbing the quality of fountain solution to polyvinyl alcohol absorbent cotton, absorbing water to the polyvinyl alcohol of softening
The restriction of the supply amount of culture medium during the inoculum density and microdisk electrode of the microalgae cell that continuous surface is inoculated with reduces micro-
The energy consumption and cost of algae culture, improve the efficiency of Immobilized culture method culture microalgae, make, meet the need of microalgae industry culture
It wants.
It should be noted that culture medium is with continous way or clearance-type into culture vessel during carrying out microdisk electrode
It is passed through, so that humectation wet condition (the i.e. water storage of polyvinyl alcohol absorbent cotton always during microdisk electrode of polyvinyl alcohol absorbent cotton
Limit of power), this state when specific operation, is realized by controlling the supply amount of culture medium.
Below from the acquisition of polyvinyl alcohol absorbent cotton of softening, the inoculation of microalgae cell and the culture of microalgae cell three
Microalgae immobilized cultivation method provided by the invention is further elaborated in aspect.
One, the acquisition of polyvinyl alcohol absorbent cotton:
The polyvinyl alcohol absorbent cotton that the present invention softens is obtained using one of following two method:
The first: polyvinyl alcohol absorbent cotton is dipped into fountain solution, is adsorbed onto fountain solution on polyvinyl alcohol absorbent cotton,
The polyvinyl alcohol absorbent cotton softened；
Second: by fountain solution spray to polyvinyl alcohol absorbent cotton, so that polyvinyl alcohol absorbent cotton is absorbed fountain solution, obtain
The polyvinyl alcohol absorbent cotton of softening.
It should be noted that the polyvinyl alcohol absorbent cotton of above-mentioned softening can also be obtained using other methods, it is not limited only to
Above two method；And fountain solution can be water or the culture medium of other suitable algae sound fields.Moreover, the temperature of fountain solution of the present invention
Degree is greater than 0 and is less than or equal to 80 DEG C, while this temperature range can guarantee that fountain solution is liquid, keeps polyvinyl alcohol absorbent cotton steady
Fixed presence, will not aging and decompose.
In addition, the surface shape of above-mentioned polyvinyl alcohol absorbent cotton is arbitrary structures, such as planar structure or curved-surface structure；Especially
When the surface for being polyvinyl alcohol absorbent cotton is curved-surface structure, the actual surface area of polyvinyl alcohol absorbent cotton is than common plane structure
When polyvinyl alcohol absorbent cotton actual surface area it is big；And the actual surface area of polyvinyl alcohol absorbent cotton is big, then opposite softening
Polyvinyl alcohol absorbent cotton surface area it is big, to improve microalgae cell in the reality on the surface of the polyvinyl alcohol absorbent cotton of softening
Border inoculation efficiency.
Moreover, the quality of the above-mentioned fountain solution being adsorbed in polyvinyl alcohol absorbent cotton is the 3 of polyvinyl alcohol absorbent cotton quality
It is very important for -6 times again, when the quality for the fountain solution being adsorbed in polyvinyl alcohol absorbent cotton is polyvinyl alcohol absorbent cotton quality
When less than 3 times, polyvinyl alcohol absorbent cotton still has certain hardness, is unfavorable for being inoculated with, and absorbs water when being adsorbed on polyvinyl alcohol
When the quality of fountain solution in silk floss is that polyvinyl alcohol absorbent cotton quality is greater than 6 times, microalgae cell can be because of the polyvinyl alcohol softened
It is lost in absorbent cotton containing more fountain solution.
In addition, present invention also defines polyvinyl alcohol in order to enable polyvinyl alcohol absorbent cotton adequately to absorb fountain solution
The thickness range of absorbent cotton is 1mm-10mm, and the range of weight per unit area is 100g/m2-1000g/m2；Preferably, polyethylene
The thickness range of alcohol absorbent cotton is 2-4mm, and weight per unit area range is 150g/m2-350g/m2.It is excellent in polyvinyl alcohol absorbent cotton
Under the thickness range of choosing and the range of weight per unit area, microalgae cell can be inoculated into the poly- second after softening to greatest extent
Enol absorbent cotton surface, improves the rate of vaccination of microalgae cell.
Two, the inoculation of microalgae cell:
When the present invention is inoculated with microalgae cell on the polyvinyl alcohol absorbent cotton of softening, used microalgae cell includes grid
Algae, haematococcus pluvialis, chlorella, micro- quasi- ball algae, triangle brown fat algae, Du Shi algae, chrysophyceae, Huang Sizao or spirulina, but not only in
This；And inoculation method also includes immersion method, spray process, suction method or semar technique, but not only in this, and specific inoculating facility can
Adaptability selection is carried out according to selected microalgae product cell kind and inoculation method, this will not be repeated here.
In addition, keeping microalgae thin while in order to guarantee that microalgae cell can make full use of the polyvinyl alcohol absorbent cotton of softening
Born of the same parents can be stable the polyvinyl alcohol absorbent cotton surface for being adsorbed on softening, be inoculated with microalgae on the polyvinyl alcohol absorbent cotton to softening
When cell, inoculum density of the microalgae cell on the polyvinyl alcohol absorbent cotton of softening is defined complete to meet microalgae cell
The surface of the polyvinyl alcohol absorbent cotton of softening is covered, the polyvinyl alcohol absorbent cotton surface that inoculum density is softened with every square metre is connect
The dry weight of the microalgae cell of kind is measured；And when the surface of polyvinyl alcohol absorbent cotton of softening is completely covered in microalgae cell, inoculation
Microalgae cell on to the polyvinyl alcohol absorbent cotton of softening covers the 100% of the polyvinyl alcohol absorbent cotton surface area of softening.This
When, the dry weight for the microalgae cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is 5g-100g.If every square
When the dry weight of microalgae cell that the polyvinyl alcohol absorbent cotton surface of rice softening is inoculated with is less than 5g, when inoculation, microalgae cell cannot
Make full use of the space on the polyvinyl alcohol absorbent cotton surface of softening；And if the polyvinyl alcohol absorbent cotton surface of every square metre of softening
When the dry weight for the microalgae cell being inoculated with is greater than 100g, when inoculation, microalgae cell is easy the polyvinyl alcohol absorbent cotton table from softening
Emaciated face is fallen.
And in order to guarantee the utilization rate of the polyvinyl alcohol absorbent cotton softened, the inoculum density of preferred microalgae cell are as follows: every
The dry weight of microalgae cell that the polyvinyl alcohol absorbent cotton surface of square metre softening is inoculated with is 15g-32g.Meanwhile it is thin according to microalgae
The inoculum density of the type of born of the same parents, different types of microalgae cell is also different.Several specific microalgae cell types are given below
When, inoculum density of the microalgae cell in the polyvinyl alcohol absorbent cotton of softening:
The first: microalgae cell is spherical microalgae cell: thin including microalgaes such as haematococcus pluvialis, chlorella or micro- quasi- ball algaes
Born of the same parents, but it is not limited only to this；Such microalgae cell dense arrangement easy to form；Therefore, the polyvinyl alcohol of every square metre of softening is inhaled
The dry weight for the microalgae cell that water silk floss surface is inoculated with is 5g-15g；
Second: microalgae cell is filamentous microalgae cell, including the microalgae cells such as Huang Sizao or spirulina, but is not only limited
In this；Such microalgae cell loose arrangement easy to form；Therefore, the polyvinyl alcohol absorbent cotton surface of every square metre of softening is connect
The dry weight of the microalgae cell of kind is 10g-32g；
The third: microalgae cell is shuttle-type microalgae cell or microalgae many cells disjunctor, such as shuttle-type microalgae cell includes grid
Algae or triangle brown fat algae, but it is not limited only to this；The microalgae that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is thin
The dry weight of born of the same parents is 15g-20g.
Three, the culture of microalgae cell:
The present invention needs to adjust the supply amount of culture medium when carrying out microalgae cell culture, it is made to absorb water with polyvinyl alcohol
Continuous water-holding capacity matches, and absorbs 300mL- per minute particular by the polyvinyl alcohol absorbent cotton of every square metre of softening of control
The culture medium of 2000mL is realized.Preferably, the polyvinyl alcohol absorbent cotton that can further control every square metre of softening is per minute
Absorb the culture medium of 600mL-1200mL.
Moreover, the supply amount of culture medium also considers illumination condition locating for hosqt media when carrying out microalgae cell culture
Or when damp condition, it is also necessary to the supply amount of defined medium, so that the culture of microalgae cell carries out at optimum conditions.
Specifically, being greater than 300 μm of olm in intensity of illumination-2·s-1Visible light under or humidity be less than or equal to 20% ring
When carrying out microdisk electrode under border, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs per minute is greater than 1000mL, is less than or equal to
The culture medium of 2000mL.This is because intensity of illumination is greater than 300 μm of olm-2·s-1Visible light under, intensity of illumination is stronger, and
Humidity be less than or equal to 20% in the environment of when, humidity is lower, and needing to supply more culture mediums in this case could remain micro-
The culture of frustule；Moreover, intensity of illumination is bigger or humidity is smaller, the amount of the culture medium of supply is bigger.
It is less than or equal to 300 μm of olm in intensity of illumination-2·s-1Visible light under or humidity be greater than 20% in the environment of into
When row microdisk electrode, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs per minute is more than or equal to 300mL, less than 1000mL's
Culture medium.This is because intensity of illumination is less than or equal to 300 μm of olm-2·s-1Visible light under, if intensity of illumination compared in addition institute
Locating environment can be more dark, and humidity be greater than 20% in the environment of when, humidity is larger, needs to reduce culture medium in this case
Supply amount is just enough to ensure that the water consumption and nutritive salt supply of microalgae cell growth, realizes energy consumption during microalgae Immobilized culture
Reduction；Moreover, intensity of illumination is smaller or humidity is bigger, the amount of the culture medium of supply is fewer.
It should be noted that being equal to 300 μm of olm in intensity of illumination-2·s-1Visible light under or humidity equal to 20%
When carrying out microdisk electrode under environment, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs the culture medium of 1000mL per minute.
In addition, when carrying out microalgae cell culture, in order to enable microalgae cell preferably to grow up, it is also necessary to thin to microalgae
The conditions such as illumination, humidity, temperature, carbon source, the pH value of environment locating for born of the same parents are adjusted.Wherein, environment locating for microalgae cell
Humidity, temperature, carbon source, pH value adjusting be that realization is adjusted by temperature, carbon source, the pH value to culture medium.And it cultivates
Carbon source is usually the CO by being passed through in culture medium in base2It is adjusted with the mixed gas of air, and the pH value in order to make culture medium
It maintains in the range of microalgae cell adapts to, when being passed through mixed gas, it is also necessary to consider CO2Adjust the pH value of culture medium
Amount used.In addition, the temperature and pH value range of environment locating for microalgae cell be by the microalgae cell of selected culture and
Fixed, it is not limited here.
It should be noted that it is -15 days 3 days that the present invention, which carries out each cultivation cycle when microdisk electrode, each cultivation cycle
Interior, culture medium is preferably passed through in a manner of clearance-type to culture vessel；When specifically being operated, it can be supplied into culture vessel
Then the culture medium of a period of time stops supply after its maximum water-holding capacity in the realization of polyvinyl alcohol absorbent cotton；Microalgae is thin
The culture medium that born of the same parents can make full use of polyvinyl alcohol absorbent cotton to save is grown, and is greatly consumed when microalgae cell is grown, followed by
It is continuous to think to supply culture medium in culture vessel, so as to be carried out herein after its maximum water-holding capacity of the realization of polyvinyl alcohol absorbent cotton
Culture medium supply, and so on, to achieve the purpose that clearance-type microdisk electrode；And this clearance-type microdisk electrode can be greatly
The energy consumption for reducing microdisk electrode, to realize the efficient utilization of medium nutrient content.
The present invention is described in further details below with reference to embodiment.
Embodiment one:
Firstly, selecting the polyvinyl alcohol absorbent cotton that shape is cuboid as hosqt media, length × width is 1m × 1m, thick
Degree is 2mm, and the surface of polyvinyl alcohol absorbent cotton is planar structure, weight per unit area 100g/m2；The poly- second that will be cut
It is 5 minutes so that it is dipped to soft that enol absorbent cotton, which is placed in 20 DEG C of water-soaking times, makes to be adsorbed in polyvinyl alcohol absorbent cotton
The quality of water is 6 times of polyvinyl alcohol absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, the polyvinyl alcohol absorbent cotton of softening is transferred in culture vessel as hosqt media, by the fresh and alive Huang of 8L
Silk algae seed liquor (cell density of yellow silk algae seed liquor is 4g/L), the table of polyvinyl alcohol absorbent cotton of the uniform spray in softening
Face, so that the dry weight for the yellow silk algae daughter cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is that 32g (connects
Kind density reaches 32g/m2), realize that yellow silk algae daughter cell substantially uniformity must be covered on the polyvinyl alcohol absorbent cotton surface of softening；
Then the polyvinyl alcohol absorbent cotton plane by the softening of the good yellow silk algae daughter cell of inoculation is disposed vertically (i.e. polyvinyl alcohol absorbent cotton
Length and thickness be formed by side and be in contact with the surface of culture vessel)；
Again, selection BG11 culture medium carries out microdisk electrode, and BG11 culture medium carries out supply from top to down by water pump
Onto the polyvinyl alcohol absorbent cotton of the softening of the good yellow silk algae daughter cell of inoculation, and BG11 culture medium supply amount is adjusted, so that often
The polyvinyl alcohol absorbent cotton BG11 culture medium supply amount per minute of square metre softening is 1200mL, careful to meet yellow silk algae
The culture medium demand of intracellular growth；Then, where the polyvinyl alcohol absorbent cotton for adjusting the softening of the good yellow silk algae daughter cell of inoculation
The environmental condition of cultivating system: artificial light source, the culture surface of light face Huang silk algae daughter cell, intensity of illumination are selected in illumination
It is set as 400 μm of olm-2·s-1, temperature control 20 DEG C, pH control be 7；Simultaneously by being passed through CO into BG11 culture medium2
With the mixed gas of air as carbon source, and CO2Volume ratio with air is 5:100；Continuous illumination culture in 24 hours is carried out, often
A cultivation cycle is set as 5 days, and incubation sustainable supply BG11 culture medium simultaneously maintains cell colony wet；Complete a culture
When the period, the supply that culture medium supplies is closed, Huang Sizao is harvested by the way of scraping.
The result shows that good yellow silk algae adhesion effect and growth result, culture week are maintained in each cultivation cycle substantially
18.2g/m can be achieved in phase2The yield of/d, the yellow silk frustule water content harvested is up to 85% or less.Persistently carry out 60
The culture experiment of batch, polyvinyl alcohol absorbent cotton surface is simultaneously not affected by apparent damage, without mouldy, and maintains preferable tough
Property, also maintain stable culture performance.Therefore, polyvinyl alcohol absorbent cotton can also be used in the microdisk electrode in next period,
Reduce energy consumption and production cost.
It should be noted that BG11 culture medium used in the present embodiment is existing standard medium, solvent is water, is matched
When processed, the NaNO of 1.5g is dissolved in 1L water3, 0.04g K2HPO4, 0.075g MgSO4·7H2O, the CaCl of 0.036g2·
2H2O, the FeCl of the citric acid of 6.0mg, 3.15mg3.6H2O, the two ethylenediamine hydrate tetraacethyl disodiums of 4.36mg, 0.02g
Na2CO3, 1mL microelement liquid storage；Wherein,
Microelement liquid storage is by water and H3BO3、MnCl2·4H2O、ZnSO4·7H2O、Na2MoO4·2H2O、CuSO4·
5H2O、Co(NO3)2·6H2O is formulated；And the H of 2.86g is dissolved in every liter of water3BO3, 1.81g MnCl2·4H2O、0.222g
ZnSO4·7H2O, the Na of 0.39g2MoO4·2H2O, the CuSO of 0.079g4·5H2O, the Co (NO of 49.4mg3)2·6H2O。
Embodiment two:
Firstly, select shape be cuboid polyvinyl alcohol absorbent cotton be used as hosqt media, length × width for 0.5m ×
0.5m, with a thickness of 3mm, the surface of polyvinyl alcohol absorbent cotton is planar structure, weight per unit area 350g/m2；It will cut
It is 10 minutes so that it is dipped to soft that good polyvinyl alcohol absorbent cotton, which is placed in 80 DEG C of water-soaking times, makes to be adsorbed on polyvinyl alcohol suction
The quality of water in water silk floss is 3 times of polyvinyl alcohol absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, the polyvinyl alcohol absorbent cotton of softening is transferred in culture vessel as hosqt media, by the fresh and alive rain of 2L
Raw haematococcus seed liquor (cell density of haematococcus pluvialis seed liquor is 4g/L), the uniform polyvinyl alcohol suction filtered in softening
The surface of water silk floss, so that the haematococcus pluvialis seed cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is dry
Weight is that (i.e. inoculum density reaches 12g/m to 12g2), realize that haematococcus pluvialis seed cell substantially uniformity must be covered on the poly- of softening
Vinyl alcohol absorbent cotton surface；Then the polyvinyl alcohol absorbent cotton plane that the softening of haematococcus pluvialis seed cell will be inoculated with is vertical
It places (i.e. the length and width of polyvinyl alcohol absorbent cotton is formed by side and is in contact with the surface of culture vessel)；
Again, the low nitrogen culture medium of selection BG11 carries out microdisk electrode, and BG11 is low, and nitrogen culture medium is uniformly sprayed in a manner of spraying
It drenches to the polyvinyl alcohol absorbent cotton surface for the softening for being inoculated with haematococcus pluvialis seed cell, and adjusts the low nitrogen culture medium of BG11 and supply
To amount, so that the polyvinyl alcohol absorbent cotton of the every square metre of softening low nitrogen culture medium supply amount of BG11 per minute is 600mL, with full
The culture medium demand of sufficient haematococcus pluvialis seed cell growth；Then, it adjusts and has been inoculated with the softening of haematococcus pluvialis seed cell
The environmental condition of cultivating system where polyvinyl alcohol absorbent cotton: lamp, light face are selected in outdoor glass shed, illumination
Microdisk electrode surface, intensity of illumination are set as 150 μm of olm-2·s-1, temperature control 35 DEG C, pH control be 8；Pass through simultaneously
CO is passed through into the low nitrogen culture medium of BG112With the mixed gas of air as carbon source, and CO2Volume ratio with air is 1.5:
100；Continuous illumination culture in 24 hours is carried out, each cultivation cycle is set as 12 days, the low nitrogen training of incubation sustainable supply BG11
It supports base and maintains cell colony wet；When completing a cultivation cycle, the supply that culture medium supplies is closed, is received by the way of scraping
Obtain haematococcus pluvialis.
The result shows that haematococcus pluvialis cell growth is good in each cultivation cycle, 7.2g/ can be achieved in cultivation cycle
m2The yield of/d, the haematococcus pluvialis cell water content finally harvested is up to 80% hereinafter, the content astaxanthin of cell is reachable
2.4%.A cultivation cycle is completed, polyvinyl alcohol absorbent cotton can repeat the cultivation that next batch is carried out as hosqt media
Production persistently carries out the culture experiment of 30 batches, and polyvinyl alcohol absorbent cotton possesses stable culture performance, reduce energy consumption and
Production cost.
It should be noted that the solvent of the low nitrogen culture medium of BG11 used in the present embodiment is water, it is molten in 1L water when preparation
Solve the NaNO of 0.25g3, 0.04g K2HPO4, 0.075g MgSO4·7H2O, the CaCl of 0.036g2·2H2O, the lemon of 6.0mg
The FeCl of lemon acid, 3.15mg3.6H2O, the Na of the two ethylenediamine hydrate tetraacethyl disodiums of 4.36mg, 0.02g2CO3, 1mL it is micro
Elements Stock；Wherein,
Microelement liquid storage is by water and H3BO3、MnCl2·4H2O、ZnSO4·7H2O、Na2MoO4·2H2O、CuSO4·
5H2O、Co(NO3)2·6H2O is formulated；And the H of 2.86g is dissolved in every liter of water3BO3, 1.81g MnCl2·4H2O、0.222g
ZnSO4·7H2O, the Na of 0.39g2MoO4·2H2O, the CuSO of 0.079g4·5H2O, the Co (NO of 49.4mg3)2·6H2O。
Embodiment three:
Firstly, select shape be cuboid polyvinyl alcohol absorbent cotton be used as hosqt media, length × width for 2.0m ×
1.0m, with a thickness of 4mm, the surface of polyvinyl alcohol absorbent cotton is planar structure, weight per unit area 200g/m2；To cutting
The good low nitrogen culture medium of 50 DEG C of BG11 of polyvinyl alcohol absorbent cotton surface spraying, keeps the BG11 being adsorbed in polyvinyl alcohol absorbent cotton low
The quality of nitrogen culture medium is 5 times of polyvinyl alcohol absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, the polyvinyl alcohol absorbent cotton of softening is transferred in culture vessel as hosqt media, by 3L it is fresh and alive three
Angle brown fat algae seed liquor (cell density of the sub- liquid of triangle brown fat algae be 4g/L), uniform the polyvinyl alcohol for being applied to softening are inhaled
The surface of water silk floss, so that the dry weight for triangle brown fat algae that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is
(i.e. inoculum density reaches 20g/m to 20g2), realize that triangle brown fat algae daughter cell substantially uniformity must be covered on the polyethylene of softening
Alcohol absorbent cotton surface；Then the polyvinyl alcohol absorbent cotton plane for being inoculated with the softening of triangle brown fat algae daughter cell is disposed vertically
(i.e. the length and width of polyvinyl alcohol absorbent cotton is formed by side and is in contact with the surface of culture vessel)；
Again, the low nitrogen culture medium of selection BG11 carries out microdisk electrode, and BG11 is low, and nitrogen culture medium is uniformly sprayed in a manner of spraying
The polyvinyl alcohol absorbent cotton surface of the leaching extremely softening of inoculation triangle brown fat algae daughter cell, and adjust the low nitrogen culture medium supply of BG11
Amount, so that the polyvinyl alcohol absorbent cotton low nitrogen culture medium supply amount of BG11 per minute of every square metre of softening is 1000mL, with full
The culture medium demand of the sufficient careful intracellular growth of triangle brown fat algae；Then, it adjusts and has been inoculated with the softening of triangle brown fat algae daughter cell
The environmental condition of cultivating system where polyvinyl alcohol absorbent cotton: lamp, light face are selected in outdoor glass shed, illumination
Microdisk electrode surface, intensity of illumination are set as 300 μm of olm-2·s-1, temperature control 30 DEG C, pH control be 6；Pass through simultaneously
CO is passed through into the low nitrogen culture medium of BG112With the mixed gas of air as carbon source, and CO2Volume ratio with air is 8:100；
Clearance-type illumination cultivation is carried out, each cultivation cycle is set as 15 days, and incubation clearance-type supplies the low nitrogen culture medium of BG11 simultaneously
Maintain cell colony wet；When completing a cultivation cycle, the supply that culture medium supplies is closed, triangle is harvested by the way of scraping
Brown fat algae.
The result shows that in each cultivation cycle, 8.0g/ is can be achieved in cultivation cycle in triangle brown fat frustule well-grown
m2The yield of/d, the haematococcus pluvialis cell water content finally harvested is up to 80% hereinafter, completing a cultivation cycle, polyethylene
Alcohol absorbent cotton can repeat the breeding production that next batch is carried out as hosqt media, and the cultivation for persistently carrying out 32 batches is real
It tests, polyvinyl alcohol absorbent cotton possesses stable culture performance, reduces energy consumption and production cost.
Example IV:
Firstly, selecting the polyvinyl alcohol absorbent cotton that shape is cuboid as hosqt media, length × width is 0.5m × 1m,
With a thickness of 6mm, the surface of polyvinyl alcohol absorbent cotton is curved-surface structure, weight per unit area 150g/m2；It is poly- by what is cut
It is 2 minutes so that it is dipped to soft that vinyl alcohol absorbent cotton, which is placed in 40 DEG C of water-soaking times, makes to be adsorbed in polyvinyl alcohol absorbent cotton
The quality of water be 4 times of polyvinyl alcohol absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, by the polyvinyl alcohol absorbent cotton of softening immerse the fresh and alive chlorella seed liquor of 5L (chlorella seed liquor it is thin
Born of the same parents' density is 3.5g/L) so that the chlorella vulgaris daughter cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with
Dry weight is that (i.e. inoculum density reaches 5g/m to 5g2), realize that chlorella vulgaris daughter cell substantially uniformity must be covered on the polyethylene of softening
Alcohol absorbent cotton surface；Then the polyvinyl alcohol absorbent cotton plane for being inoculated with the softening of chlorella vulgaris daughter cell is disposed vertically (i.e.
The length and thickness of polyvinyl alcohol absorbent cotton are formed by side and are in contact with the surface of culture vessel)；
Again, selection BG11 culture medium carries out microdisk electrode, and BG11 culture medium carries out supply from top to down by water pump
Onto the polyvinyl alcohol absorbent cotton for the softening for being inoculated with chlorella vulgaris daughter cell, and BG11 culture medium supply amount is adjusted, so that often
The polyvinyl alcohol absorbent cotton BG11 culture medium supply amount per minute of square metre softening is 2000mL, careful to meet chlorella vulgaris
The culture medium demand of intracellular growth；Then, where the polyvinyl alcohol absorbent cotton for adjusting the softening for being inoculated with chlorella vulgaris daughter cell
The environmental condition of cultivating system: artificial light source, the culture surface of light face chlorella vulgaris daughter cell, intensity of illumination are selected in illumination
It is set as 800 μm of olm-2·s-1, temperature control 25 DEG C, pH control be 7；Simultaneously by being passed through CO into BG11 culture medium2
With the mixed gas of air as carbon source, and CO2Volume ratio with air is 5:100；Continuous illumination culture in 24 hours is carried out, often
A cultivation cycle is set as 7 days, and incubation sustainable supply BG11 culture medium simultaneously maintains cell colony wet；Complete a culture
When the period, the supply that culture medium supplies is closed, chlorella is harvested by the way of scraping.
The result shows that good chlorella adhesion effect and growth result are maintained in each cultivation cycle substantially, culture week
15.2g/m can be achieved in phase2The yield of/d, the chlorella cells water content harvested is up to 80% or less.Persistently carry out 70
The culture experiment of batch, polyvinyl alcohol absorbent cotton surface is simultaneously not affected by apparent damage, without mouldy, and maintains preferable tough
Property, also maintain stable culture performance.Therefore, polyvinyl alcohol absorbent cotton can also be used in the microdisk electrode in next period,
Reduce energy consumption and production cost.
Embodiment five:
Firstly, select shape be cuboid polyvinyl alcohol absorbent cotton be used as hosqt media, length × width for 0.5m ×
1.5m, with a thickness of 10mm, the surface of polyvinyl alcohol absorbent cotton is planar structure, weight per unit area 500g/m2；It will cut
It is 8 minutes so that it is dipped to soft that good polyvinyl alcohol absorbent cotton, which is placed in 60 DEG C of water-soaking times, makes to be adsorbed on polyvinyl alcohol suction
The quality of water in water silk floss is 4 times of polyvinyl alcohol absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, by the polyvinyl alcohol absorbent cotton of softening immerse the fresh and alive Du Shi algae seed liquor of 5L (Du Shi algae seed liquor it is thin
Born of the same parents' density is 5g/L) so that the Du Shi algae daughter cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is dry
Weight is that (i.e. inoculum density reaches 50g/m to 50g2), realize that Du Shi algae daughter cell substantially uniformity must be covered on the polyethylene of softening
Alcohol absorbent cotton surface；Then the polyvinyl alcohol absorbent cotton plane for being inoculated with the softening of Du Shi algae daughter cell is disposed vertically (i.e.
The length and thickness of polyvinyl alcohol absorbent cotton are formed by side and are in contact with the surface of culture vessel)；
Again, selection BG11 culture medium carries out microdisk electrode, and BG11 culture medium carries out supply from top to down by water pump
Onto the polyvinyl alcohol absorbent cotton for the softening for being inoculated with Du Shi algae daughter cell, and BG11 culture medium supply amount is adjusted, so that often
The polyvinyl alcohol absorbent cotton BG11 culture medium supply amount per minute of square metre softening is 1100mL, careful to meet Du Shi algae
The culture medium demand of intracellular growth；Then, where the polyvinyl alcohol absorbent cotton for adjusting the softening for being inoculated with Du Shi algae daughter cell
The environmental condition of cultivating system: artificial light source, the culture surface of light face Du Shi algae daughter cell, intensity of illumination are selected in illumination
It is set as 650 μm of olm-2·s-1, temperature control 35 DEG C, pH control be 7；Simultaneously by being passed through CO into BG11 culture medium2
With the mixed gas of air as carbon source, and CO2Volume ratio with air is 4:100；Continuous illumination culture in 24 hours is carried out, often
A cultivation cycle is set as 10 days, and incubation sustainable supply BG11 culture medium simultaneously maintains cell colony wet；Complete a training
When supporting the period, the supply that culture medium supplies is closed, Du Shi algae is harvested by the way of scraping.
The result shows that good Du Shi algae adhesion effect and growth result are maintained in each cultivation cycle substantially, culture week
19.2g/m can be achieved in phase2The yield of/d, the chlorella cells water content harvested is up to 79% or less.Persistently carry out 50
The culture experiment of batch, polyvinyl alcohol absorbent cotton surface is simultaneously not affected by apparent damage, without mouldy, and maintains preferable tough
Property, also maintain stable culture performance.Therefore, polyvinyl alcohol absorbent cotton can also be used in the microdisk electrode in next period,
Reduce energy consumption and production cost.
Embodiment six:
Firstly, select shape be cuboid polyvinyl alcohol absorbent cotton be used as hosqt media, length × width for 0.2m ×
0.2m, with a thickness of 6mm, the surface of polyvinyl alcohol absorbent cotton is planar structure, weight per unit area 1000g/m2；It will cut
It is 13 minutes so that it is dipped to soft that good polyvinyl alcohol absorbent cotton, which is placed in 45 DEG C of water-soaking times, makes to be adsorbed on polyvinyl alcohol suction
The quality of water in water silk floss is 6 times of polyvinyl alcohol absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, the polyvinyl alcohol absorbent cotton of softening is immersed into fresh and alive micro- quasi- ball algae seed liquor (micro- quasi- ball algae of 4.5L
The cell density of liquid is 3g/L) so that micro- quasi- ball algae that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with
The dry weight of cell is that (i.e. inoculum density reaches 15g/m to 15g2), it is soft to realize that micro- quasi- ball algae daughter cell substantially uniformity must be covered on
The polyvinyl alcohol absorbent cotton surface of change；Then the polyvinyl alcohol absorbent cotton plane of the softening of good micro- quasi- ball algae daughter cell will be inoculated with
It is disposed vertically (i.e. the length of polyvinyl alcohol absorbent cotton and thickness are formed by side and are in contact with the surface of culture vessel)；
Again, selection BG11 culture medium carries out microdisk electrode, and BG11 culture medium carries out supply from top to down by water pump
Onto the polyvinyl alcohol absorbent cotton for the softening for being inoculated with good micro- quasi- ball algae daughter cell, and BG11 culture medium supply amount is adjusted, so that
The polyvinyl alcohol absorbent cotton of every square metre of softening BG11 culture medium supply amount per minute is 800mL, to meet micro- quasi- ball algae
The culture medium demand of careful intracellular growth；Then, the polyvinyl alcohol absorbent cotton for being inoculated with the softening of good micro- quasi- ball algae daughter cell is adjusted
The environmental condition of the cultivating system at place: illumination selection artificial light source, the culture surface of the micro- quasi- ball algae daughter cell of light face,
Intensity of illumination is set as 200 μm of olm-2·s-1, temperature control 25 DEG C, pH control be 7；Simultaneously by BG11 culture medium
In be passed through CO2With the mixed gas of air as carbon source, and CO2Volume ratio with air is 6:100；Carry out 24 hours continuous light
According to culture, each cultivation cycle is set as 12 days, and incubation sustainable supply BG11 culture medium simultaneously maintains cell colony wet；It is complete
When at a cultivation cycle, the supply that culture medium supplies is closed, micro- quasi- ball algae is harvested by the way of scraping.
The result shows that maintaining good micro- quasi- ball algae adhesion effect and growth result in each cultivation cycle substantially, cultivate
18.2g/m can be achieved in period2The yield of/d, the chlorella cells water content harvested is up to 79% or less.Persistently carry out 50
The culture experiment of a batch, polyvinyl alcohol absorbent cotton surface is simultaneously not affected by apparent damage, without mouldy, and maintains preferable tough
Property, also maintain stable culture performance.Therefore, polyvinyl alcohol absorbent cotton can also be used in the microdisk electrode in next period,
Reduce energy consumption and production cost.
Embodiment seven:
Firstly, select shape be cuboid polyvinyl alcohol absorbent cotton be used as hosqt media, length × width for 0.5m ×
0.5m, with a thickness of 5mm, the surface of polyvinyl alcohol absorbent cotton is planar structure, weight per unit area 1000g/m2；It will cut
It is 10 minutes so that it is dipped to soft that good polyvinyl alcohol absorbent cotton, which is placed in 50 DEG C of water-soaking times, makes to be adsorbed on polyvinyl alcohol suction
The quality of water in water silk floss is 5 times of polyvinyl alcohol absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, the polyvinyl alcohol absorbent cotton of softening is transferred in culture vessel as hosqt media, by the fresh and alive spiral shell of 2L
It revolves algae seed liquor (cell density of spirulina seed liquor is 3g/L), it is uniform to filter in the polyvinyl alcohol absorbent cotton in softening
Surface so that the dry weight of spirulina seed cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated be 10g (i.e.
Inoculum density reaches 10g/m2), realize that spirulina seed cell substantially uniformity must be covered on the polyvinyl alcohol absorbent cotton table of softening
Face；Then the polyvinyl alcohol absorbent cotton plane for being inoculated with the softening of spirulina seed cell is disposed vertically (i.e. polyvinyl alcohol suction
The length and width of water silk floss is formed by side and is in contact with the surface of culture vessel)；
Again, the low nitrogen culture medium of selection BG11 carries out microdisk electrode, and BG11 is low, and nitrogen culture medium is uniformly sprayed in a manner of spraying
It drenches to the polyvinyl alcohol absorbent cotton surface for the softening for being inoculated with spirulina seed cell, and adjusts the low nitrogen culture medium supply of BG11
Amount, so that the polyvinyl alcohol absorbent cotton low nitrogen culture medium supply amount of BG11 per minute of every square metre of softening is 300mL, to meet
The culture medium demand of spirulina seed cell growth；Then, the polyvinyl alcohol suction for being inoculated with the softening of spirulina seed cell is adjusted
The environmental condition of cultivating system where water is continuous: lamp, light face microdisk electrode table are selected in outdoor glass shed, illumination
Face, intensity of illumination are set as 20 μm of olm-2·s-1, temperature control 25 DEG C, pH control be 7；Simultaneously by the low nitrogen of BG11
CO is passed through in culture medium2With the mixed gas of air as carbon source, and CO2Volume ratio with air is 5:100；It carries out 24 hours
Continuous illumination culture, each cultivation cycle are set as 12 days, and the low nitrogen culture medium of incubation sustainable supply BG11 simultaneously maintains cell
Group is wet；When completing a cultivation cycle, the supply that culture medium supplies is closed, spirulina is harvested by the way of scraping.
The result shows that in each cultivation cycle, 8.0g/m is can be achieved in cultivation cycle in spirulina cells well-grown2/d
Yield, the spirulina cells water content finally harvested up to 75% hereinafter, complete a cultivation cycle, polyvinyl alcohol absorbent cotton
The breeding production for carrying out next batch as hosqt media can be repeated, the culture experiment of 40 batches, poly- second are persistently carried out
Enol absorbent cotton possesses stable culture performance, reduces energy consumption and production cost.
Embodiment eight:
Firstly, select shape be cuboid polyvinyl alcohol absorbent cotton be used as hosqt media, length × width for 0.2m ×
0.2m, with a thickness of 6mm, the surface of polyvinyl alcohol absorbent cotton is planar structure, weight per unit area 1000g/m2；It will cut
It is 13 minutes so that it is dipped to soft that good polyvinyl alcohol absorbent cotton, which is placed in 45 DEG C of water-soaking times, makes to be adsorbed on polyvinyl alcohol suction
The quality of water in water silk floss is 6 times of polyvinyl alcohol absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, by the polyvinyl alcohol absorbent cotton of softening immerse the fresh and alive spirulina seed liquor of 6L (spirulina seed liquor it is thin
Born of the same parents' density is 5g/L) so that the spirulina seed cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is dry
Weight is that (i.e. inoculum density reaches 23g/m to 23g2), realize that spirulina seed cell substantially uniformity must be covered on the polyethylene of softening
Alcohol absorbent cotton surface；Then the polyvinyl alcohol absorbent cotton plane for being inoculated with the softening of spirulina seed cell is disposed vertically (i.e.
The length and thickness of polyvinyl alcohol absorbent cotton are formed by side and are in contact with the surface of culture vessel)；
Again, selection BG11 culture medium carries out microdisk electrode, and BG11 culture medium carries out supply from top to down by water pump
Onto the polyvinyl alcohol absorbent cotton for the softening for being inoculated with spirulina seed cell, and BG11 culture medium supply amount is adjusted, so that often
The polyvinyl alcohol absorbent cotton BG11 culture medium supply amount per minute of square metre softening is 1600mL, careful to meet spirulina kind
The culture medium demand of intracellular growth；Then, where the polyvinyl alcohol absorbent cotton for adjusting the softening for being inoculated with spirulina seed cell
The environmental condition of cultivating system: artificial light source, the culture surface of light face spirulina seed cell, intensity of illumination are selected in illumination
It is set as 500 μm of olm-2·s-1, temperature control 25 DEG C, pH control be 7；Simultaneously by being passed through CO into BG11 culture medium2
With the mixed gas of air as carbon source, and CO2Volume ratio with air is 5:100；Continuous illumination culture in 24 hours is carried out, often
A cultivation cycle is set as 12 days, and incubation sustainable supply BG11 culture medium simultaneously maintains cell colony wet；Complete a training
When supporting the period, the supply that culture medium supplies is closed, spirulina is harvested by the way of scraping.
The result shows that good spirulina adhesion effect and growth result are maintained in each cultivation cycle substantially, culture week
15.2g/m can be achieved in phase2The yield of/d, the chlorella cells water content harvested is up to 79% or less.Persistently carry out 60
The culture experiment of batch, polyvinyl alcohol absorbent cotton surface is simultaneously not affected by apparent damage, without mouldy, and maintains preferable tough
Property, also maintain stable culture performance.Therefore, polyvinyl alcohol absorbent cotton can also be used in the microdisk electrode in next period,
Reduce energy consumption and production cost.
Embodiment nine:
Firstly, select shape be cuboid polyvinyl alcohol absorbent cotton be used as hosqt media, length × width for 2.0m ×
1.0m, with a thickness of 4mm, the surface of polyvinyl alcohol absorbent cotton is planar structure, weight per unit area 300g/m2；To cutting
Good 25 DEG C of water of polyvinyl alcohol absorbent cotton surface spraying, making the quality for the water being adsorbed in polyvinyl alcohol absorbent cotton is polyvinyl alcohol
3 times of absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, the polyvinyl alcohol absorbent cotton of softening is transferred in culture vessel as hosqt media, by the fresh and alive grid of 3L
Algae seed liquor (cell density of scenedesmus seed liquor be 2g/L), the surface of uniform the polyvinyl alcohol absorbent cotton for being applied to softening,
So that the dry weight for the scenedesmus seed that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is that (i.e. inoculum density reaches 15g
To 15g/m2), realize that scenedesmus seed cell substantially uniformity must be covered on the polyvinyl alcohol absorbent cotton surface of softening；It then will inoculation
The polyvinyl alcohol absorbent cotton plane of the softening of good scenedesmus seed cell is disposed vertically (the i.e. length and width of polyvinyl alcohol absorbent cotton
It is formed by side to be in contact with the surface of culture vessel)；
Again, the low nitrogen culture medium of selection BG11 carries out microdisk electrode, and BG11 is low, and nitrogen culture medium is uniformly sprayed in a manner of spraying
The polyvinyl alcohol absorbent cotton surface of the leaching extremely softening of inoculation scenedesmus seed cell, and the low nitrogen culture medium supply amount of BG11 is adjusted, make
Obtaining every square metre of polyvinyl alcohol absorbent cotton softened low nitrogen culture medium supply amount of BG11 per minute is 1600mL, to meet scenedesmus
The culture medium demand of seed cell growth；Then, it adjusts and has been inoculated with where the polyvinyl alcohol absorbent cotton of scenedesmus seed cell softening
Cultivating system environmental condition: outdoor glass shed, illumination select lamp, light face microdisk electrode surface, humidity
10%, it is 7 that temperature control is controlled in 30 DEG C, pH；Simultaneously by being passed through CO into the low nitrogen culture medium of BG112With the gaseous mixture of air
Body is as carbon source, and CO2Volume ratio with air is 5:100；Clearance-type illumination cultivation is carried out, each cultivation cycle is set as 15
It, the low nitrogen culture medium of incubation clearance-type supply BG11 simultaneously maintains cell colony wet；When completing a cultivation cycle, close
The supply that culture medium supplies, harvests scenedesmus by the way of scraping.
The result shows that in each cultivation cycle, 8.6g/m is can be achieved in cultivation cycle in scenedesmus cell well-grown2/ d's
Yield, up to 70% hereinafter, completing a cultivation cycle, polyvinyl alcohol absorbs water the haematococcus pluvialis cell water content finally harvested
Silk floss can repeat the breeding production that next batch is carried out as hosqt media, persistently carry out the culture experiment of 40 batches, gather
Vinyl alcohol absorbent cotton possesses stable culture performance, reduces energy consumption and production cost.
Embodiment ten:
Firstly, select shape be cuboid polyvinyl alcohol absorbent cotton be used as hosqt media, length × width for 0.2m ×
0.2m, with a thickness of 6mm, the surface of polyvinyl alcohol absorbent cotton is planar structure, weight per unit area 450g/m2；It will cut
It is 7 minutes so that it is dipped to soft that good polyvinyl alcohol absorbent cotton, which is placed in 50 DEG C of water-soaking times, makes to be adsorbed on polyvinyl alcohol suction
The quality of water in water silk floss is 3.5 times of polyvinyl alcohol absorbent cotton quality, the polyvinyl alcohol absorbent cotton softened；
Then, by the polyvinyl alcohol absorbent cotton immersion 4L of softening fresh and alive scenedesmus seed liquor, (cell of scenedesmus seed liquor is close
Degree is 2.1g/L) so that the dry weight for the scenedesmus seed cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with is
(i.e. inoculum density reaches 18g/m to 18g2), realize that scenedesmus seed cell substantially uniformity must be covered on the polyvinyl alcohol water suction of softening
Continuous surface；Then the polyvinyl alcohol absorbent cotton plane for being inoculated with the softening of scenedesmus seed cell is disposed vertically (i.e. polyvinyl alcohol
The length and thickness of absorbent cotton are formed by side and are in contact with the surface of culture vessel)；
Again, selection BG11 culture medium carries out microdisk electrode, and BG11 culture medium carries out supply from top to down by water pump
Onto the polyvinyl alcohol absorbent cotton for the softening for being inoculated with scenedesmus seed cell, and BG11 culture medium supply amount is adjusted, so that often putting down
The BG11 culture medium supply amount that the polyvinyl alcohol absorbent cotton of square rice softening is per minute is 1600mL, raw to meet scenedesmus seed cell
Long culture medium demand；Then, the culture body where the polyvinyl alcohol absorbent cotton for the softening for being inoculated with scenedesmus seed cell is adjusted
The environmental condition of system: artificial light source, the culture surface of light face scenedesmus seed cell are selected in illumination, and intensity of illumination is set as
500μmol·m-2·s-1, temperature control 25 DEG C, pH control be 9；Simultaneously by being passed through CO into BG11 culture medium2And air
Mixed gas as carbon source, and CO2Volume ratio with air is 0.6:100；Carry out continuous illumination culture in 24 hours, Mei Gepei
Supporting cycle set is 10 days, and incubation sustainable supply BG11 culture medium simultaneously maintains cell colony wet；Complete a culture week
When the phase, the supply that culture medium supplies is closed, scenedesmus is harvested by the way of scraping.
The result shows that good scenedesmus adhesion effect and growth result, cultivation cycle are maintained in each cultivation cycle substantially
Interior achievable 16g/m2The yield of/d, the chlorella cells water content harvested is up to 79% or less.Persistently carry out 70 batches
Culture experiment, polyvinyl alcohol absorbent cotton surface is simultaneously not affected by apparent damage, without mouldy, and maintains preferable toughness, also
Maintain stable culture performance.Therefore, polyvinyl alcohol absorbent cotton can also be used in the microdisk electrode in next period, reduce
Energy consumption and production cost.
In the description of above embodiment, particular features, structures, materials, or characteristics can be at any one or more
It can be combined in any suitable manner in a embodiment or example.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (10)
1. a kind of microalgae immobilized cultivation method, which is characterized in that using the polyvinyl alcohol absorbent cotton of softening as hosqt media, to
It is inoculated with microalgae cell on the polyvinyl alcohol absorbent cotton of softening, then carries out microdisk electrode, obtains cultured microalgae；Wherein,
The polyvinyl alcohol absorbent cotton of softening is made of polyvinyl alcohol absorbent cotton and the fountain solution being adsorbed in polyvinyl alcohol absorbent cotton；
The quality for the fountain solution being adsorbed in polyvinyl alcohol absorbent cotton is 3 times -6 times of polyvinyl alcohol absorbent cotton quality；Every square metre soft
The dry weight for the microalgae cell that the polyvinyl alcohol absorbent cotton surface of change is inoculated with is 5g-100g；When carrying out microdisk electrode, every square metre
The polyvinyl alcohol absorbent cotton of softening absorbs the culture medium of 300mL-2000mL per minute.
2. microalgae immobilized cultivation method according to claim 1, which is characterized in that the polyvinyl alcohol of the softening absorbs water
Silk floss is obtained using following methods: polyvinyl alcohol absorbent cotton being dipped into fountain solution, fountain solution is made to be adsorbed onto polyvinyl alcohol suction
On water silk floss, the polyvinyl alcohol absorbent cotton that is softened；Or,
By on fountain solution spray to polyvinyl alcohol absorbent cotton, polyvinyl alcohol absorbent cotton is made to absorb fountain solution, the poly- second softened
Enol absorbent cotton.
3. microalgae immobilized cultivation method according to claim 1 or 2, which is characterized in that the polyvinyl alcohol absorbent cotton
Mass area ratio be 100g/m2-1000g/m2。
4. microalgae immobilized cultivation method according to claim 1, which is characterized in that the temperature of the immersion fluid is greater than 0
Less than or equal to 80 DEG C.
5. microalgae immobilized cultivation method according to claim 1 or 4, which is characterized in that the immersion fluid is water or training
Support base.
6. microalgae immobilized cultivation method according to claim 1, which is characterized in that be inoculated into the polyethylene of the softening
Microalgae cell on alcohol absorbent cotton is covered on the polyvinyl alcohol absorbent cotton of softening, and the polyvinyl alcohol for being inoculated into the softening is inhaled
The 100% of the polyvinyl alcohol absorbent cotton surface area of microalgae cell covering softening on water silk floss.
7. microalgae immobilized cultivation method according to claim 1 or 6, which is characterized in that every square metre of softening
The dry weight for the microalgae cell that polyvinyl alcohol absorbent cotton surface is inoculated with is 15g-32g.
8. microalgae immobilized cultivation method according to claim 1 or 6, which is characterized in that the microalgae cell is spherical
Microalgae cell, the dry weight for the microalgae cell that the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with are 5g-15g；Or,
The microalgae cell is filamentous microalgae cell, and the polyvinyl alcohol absorbent cotton surface of every square metre of softening is inoculated with micro-
The dry weight of frustule is 10g-32g；Or,
The microalgae cell is shuttle-type microalgae cell or microalgae many cells disjunctor；The polyvinyl alcohol absorbent cotton of every square metre of softening
The dry weight for the microalgae cell that surface is inoculated with is 15g-20g.
9. microalgae immobilized cultivation method according to claim 1, which is characterized in that when carrying out the microdisk electrode, often
The polyvinyl alcohol absorbent cotton of square metre softening absorbs the culture medium of 600mL-1200mL per minute.
10. microalgae immobilized cultivation method according to claim 1, which is characterized in that be greater than 300 μ in intensity of illumination
mol·m-2·s-1Visible light under or humidity less than 20% in the environment of carry out microdisk electrode when, every square metre softening poly- second
Enol absorbent cotton absorbs per minute is greater than 1000mL, the culture medium less than or equal to 2000mL；Or,
In less than 300 μm olm of intensity of illumination-2·s-1Visible light under or humidity be greater than 20% in the environment of carry out microdisk electrode
When, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs per minute is more than or equal to 300mL, the culture medium less than 1000mL；Or,
It is equal to 300 μm of olm in intensity of illumination-2·s-1Visible light under or humidity be equal to 20% in the environment of carry out microdisk electrode
When, the polyvinyl alcohol absorbent cotton of every square metre of softening absorbs the culture medium of 1000mL per minute.
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| CN103849615A (en) * | 2013-12-31 | 2014-06-11 | 浙江省海洋开发研究院 | Immobilized algal-bacterial symbiotic system for processing petroleum pollutants and application thereof |
| JP6038754B2 (en) * | 2013-09-24 | 2016-12-07 | 鹿島建設株式会社 | Seismic isolation method for existing structures |
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| JP6038754B2 (en) * | 2013-09-24 | 2016-12-07 | 鹿島建設株式会社 | Seismic isolation method for existing structures |
| CN103849615A (en) * | 2013-12-31 | 2014-06-11 | 浙江省海洋开发研究院 | Immobilized algal-bacterial symbiotic system for processing petroleum pollutants and application thereof |
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