CN107502550A - A kind of method of frustule broken wall treatment - Google Patents
A kind of method of frustule broken wall treatment Download PDFInfo
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Abstract
The invention belongs to technical field of microalga biology.A kind of method of frustule broken wall, comprises the steps successively：(1) microalgae cell is placed in the culture medium of suitable microalgae cell growth, the metal ion added in its culture medium, in microalgae cell in growth course, part metals ion can pass through physisorption and adsorb in microalgae cell wall surface；Part metals ion can be by reacting to be adsorbed onto cell wall with the charged particles of microalgae cell wall surface；Remaining part metals ion can be adsorbed onto on cell membrane via the transport protein of microalgae cell wall surface；(2) the microalgae cell culture of metal ion will have been adsorbed until harvest, microalgae cell solid is separated with medium liquid；(3) the frustule liquid of harvest is subjected to broken wall treatment with normal pressure microwave, obtains the microalgae cell by broken wall treatment.The method of the frustule broken wall of the present invention can improve the microwave frequency measurment treatment effect of frustule.
Description
Technical field
The invention belongs to technical field of microalga biology, and in particular to a kind of method for improving frustule broken wall treatment efficiency.
Background technology
Microalgae is could to distinguish the small algae group of its form under the microscope, it has been found that has tens of thousands of kinds.They constitute generation
Maximum primary producer colony, accounts for more than the 32% of global gross photosynthesis in boundary.Microalge aboundresources, containing higher
The composition such as protein, grease and soluble polysaccharide, without leaf, stem, root, do not produce lignocellulose hard-to-degrade biomass,
Ground is not striven with traditional agriculture, does not destroy the ecosystem because of harvest.Because the quantum that microalgae conversion solar can be biomass energy is imitated
Rate is up to 2-10% (and the quantum efficiency of terrestrial plant is both less than 1%), and micro algae growth speed is exceedingly fast (daily biomass
Increment can reach 1-3 times), therefore can produce substantial amounts of high-quality biological matter (algae oil, algae using microalgae photosynthetic efficiency height
Albumen and polysaccharides) come preparing biological diesel oil, polyunsaturated fatty acid and feed addictive there is outstanding advantage.
The growth that there is microalgae certain molecular mechanism can be told from water body to itself simultaneously is beneficial or useless
Metal ion.Accepted using the researchers in the advantage times whole world of the metal ion in microalgae adsorbed water body.It is primarily due to
The growth rate of microalgae cell quickly, so the speed of microalgae adsorbing metal ions is quickly, can save time and energy；Microalgae
The CO in environment can be absorbed in cell growth process2To carry out photosynthesis production biomass, product is pure green, to life
State environmental beneficial, and it is harmless to user.And microalgae production annual can be carried out, and be operated and simple and convenient, can
Recycle, cost is low, has the faster speed of growth, higher photosynthetic efficiency, bigger ratio compared with higher plant
Surface area, the adsorption capacity that absorption accounts for own biological quality 10% is at best able to, high selectivity, secondary pollution will not be produced,
Batch is produced and can used in continuous production, can be used for the higher or relatively low condition of underwater gold category amount.
Microalgae cell can most metal ion in adsorbed water body, such as K, Mg, Ca, Fe, Sr, Co, Mn, Ni, V,
The metal ions such as Zn, As, Cd, Mo, Pb, Se and Al.And the microalgae species that can be used for adsorbing metal ions is also very various, such as
Chlorella (Chlorella sp.), small ring algae (Cyclotella sp.), grid algae (Scenedesmus sp.), spirulina
(Spirulina sp.), split an algae (Stichococcus sp.), Mao Zhizao (Stigeoclonium sp.), brown algae
(Phaeodactylum sp.) etc..Different algae kinds is different to the adsorption capacity of same metal ion species, either microalgae
The species and condition of culture of cell, the species of metal ion and concentration, and the state of frustule, all to frustule adsorbing metal
The ability of ion has a great impact.The frustule for having adsorbed metal ion produces diversified new old generation by photosynthesis
Product, such as algae oil, algae albumen and polysaccharides etc. are thanked, can be utilized by subsequent production high value-added product.
But due to the metabolism product of microalgae, portion is wrapped up by cell membrane in the cell, in the microalgae biomass system of utilization
During taking high-quality value-added product, how to break microalgae cell wall so that intracellular thing can discharge high efficiency, low cost
One of technical bottleneck to exterior space to turn into this industry development.
The content of the invention
The technical problem to be solved of the present invention is to provide a kind of method that can improve frustule broken wall treatment efficiency.
In order to solve the above technical problems, solution provided by the invention is：A kind of method of frustule broken wall, is wrapped successively
Include following step：(1) microalgae cell is placed in the culture medium of suitable microalgae cell growth, the metal added in its culture medium
Ion, in microalgae cell in growth course, part metals ion can pass through physisorption and adsorb in microalgae cell wall
Surface；Part metals ion can be by reacting to be adsorbed onto cell wall with the charged particles of microalgae cell wall surface；
Remaining part metals ion can be adsorbed onto on cell membrane via the transport protein of microalgae cell wall surface；(2) will absorption
The microalgae cell culture of metal ion separates microalgae cell solid with medium liquid until harvest, the frustule of acquisition
Moisture content is 60%-90%；(3) the frustule liquid of harvest is subjected to broken wall treatment, and microwave treatment power with normal pressure microwave
For 600-1000W, treatment temperature is 70-90 DEG C, obtains the microalgae cell by broken wall treatment.
Preferably, microalgae cell culture is until the 7-10 days time of harvest.
Preferably, microalgae cell solid is separated with medium liquid by the method for filtering or centrifuging.
Preferably, broken wall treatment processing time is 8-12min.
Preferably, microwave produces the double action of heat damage and mechanical damage for microalgae cell wall in step (3), by
In the presence of metal ion, under the electromagnetic field effect changed caused by microwave, electric field can be induced in metallic surface, is inducted
Electric field causes surface current, so as to cause Joule heat, exacerbates heat damage effect of the microwave to microalgae cell wall.
Preferably, the frequency of microwave is 2450MHZ or 915MHZ.
Preferably, metal ion is Cd2+、Hg2+、K、Mg、Ca、Fe、Sr、Co、Mn、Ni、V、Zn、As、Cd、Mo、Pb、
One or more combination in Se and Al；Microalgae cell is chlorella, small ring algae, grid algae, spirulina, splits an algae, be Mao Zhizao, brown
Refer to the one or more combination in algae.
Preferably, culture medium is one kind in SE culture mediums, Zarrouk culture mediums, SM culture mediums.
Compared with prior art, the beneficial effects of the invention are as follows：Due to frustule in growth course by physics or
Biological agent has adsorbed a certain amount of metal ion in frustule surface or inside, in the process of microwave treatment microalgae cell
In, in addition to heat damage of the microwave of routine for microalgae cell wall, mechanical damage, microwave magnetic field induces in metal surface
Electric field, induced electric field cause surface current, so as to cause Joule heat, exacerbate the fuel factor of microwave treatment, due to metal and micro-
The cooperative gain effect of ripple, frustule broken wall treatment efficiency is improved, therefore the microalgae cell for having adsorbed metal ion is passing through
After microwave treatment, cell dia reduces 12%-18%, and the fractal dimension of cell adds 9%-16%, i.e. this patent
The method of frustule broken wall can improve the microwave frequency measurment treatment effect of frustule.
Embodiment
Further detailed description is done to the present invention with reference to embodiment.
Embodiment 1
A kind of method of frustule broken wall, comprises the steps successively：(1) chlorella is placed in 500ml SE culture mediums
It is interior, the Pb that mass concentration is 10ppm is added in the culture medium of chlorella growth2+, it is then 25 DEG C in environment temperature, illumination is strong
It is 12h illumination, 12 hours dark to spend for 5000lux, light application time, and it is thin to cultivate chlorella under conditions of throughput 50ml/min
Born of the same parents, Pb is adsorbed using chlorella cells growth course2+Metal, part Pb2+Ion can pass through physisorption and adsorb small
Ball frustule wall surface；Part Pb2+Ion can be by reacting to be adsorbed onto with the charged particles of chlorella cells wall surface
Cell wall；Remaining part Pb2+Ion can be adsorbed onto on cell membrane via the transport protein of chlorella cells wall surface；
(2) harvested after cultivating 7 days, chlorella cells are harvested using centrifugal method, chlorella cells solid is tried one's best with medium liquid
Separation, the moisture content for obtaining chlorella cells are 60%；(3) it is 10ml the chlorella cells liquid 5ml of harvest to be contained in into volume
Conical flask in, in processing procedure keep conical flask bottleneck be unlimited, the normal pressure microwave for being 2450MHZ with frequency enters
Row broken wall treatment, and microwave treatment power is 600W, treatment temperature is 70 DEG C, processing time 8min, obtained after processing through
Cross the microalgae cell of broken wall treatment.(4) the frustule sample after microwave treatment is by fixed, dehydration, critical point drying and plated film
Processing, most clearly image can be obtained under conditions of ensureing that frustule form is constant, be observed using ESEM (SEM) thin
Born of the same parents, obtain the metamorphosis image after frustule processing；SEM image is analyzed.The change of cell size uses MATLAB
Software carries out image procossing, obtains the change of microwave frequency measurment frustule size before and after the processing, and introduces the concept of fractal dimension,
To quantify the change of frustule form.Pb is not added with contrast culture medium2+And the unadsorbed Pb harvested2+Frustule and culture
Pb is with the addition of in base2+And what is harvested has adsorbed Pb2+Frustule, cell size and form after being handled by microwave frequency measurment
Change, finds adsorbed Pb2+Chlorella cells than unadsorbed Pb2+Chlorella cells its cell dias reduce 12%,
And the fractal dimension of cell adds 9%.Adsorb Pb2+Chlorella cells by microwave frequency measurment processing after destructiveness
More than unadsorbed Pb2+Chlorella cells.
Embodiment 2
A kind of method of frustule broken wall, comprises the steps successively：(1) Zarrouk that spirulina is placed in 500ml is trained
Support in base, the Cd that mass concentration is 5ppm is added in the culture medium of growth of spirulina platensis2+, it is then 28 DEG C in environment temperature, light
It is 8000lux according to intensity, light application time is 12h illumination, 12 hours dark, and spirulina is cultivated under conditions of throughput 50ml/min
Cell, Cd is adsorbed using spirulina cells growth course2+, part Cd2+Ion can pass through physisorption and adsorb in spiral
Frustule wall surface；Part Cd2+Ion can be adsorbed onto thin by being reacted with the charged particles of spirulina cells wall surface
Cell wall surface；Remaining part Cd2+Ion can be adsorbed onto on cell membrane via the transport protein of spirulina cells wall surface；
(2) harvested after cultivating 8 days；Spirulina cells are harvested using filter method, spirulina cells solid is tried one's best with medium liquid
Separation, the moisture content for obtaining spirulina cells are 80%；(3) it is 15ml the spirulina cells liquid 8ml of harvest to be contained in into volume
Conical flask in, in processing procedure keep conical flask bottleneck be unlimited, the normal pressure microwave for being 915MHZ with frequency enters
Row broken wall treatment, and microwave treatment power is 800W, treatment temperature is 80 DEG C, processing time 10min, obtained after processing through
Cross the microalgae cell of broken wall treatment；(4) the frustule sample after microwave treatment is by fixed, dehydration, critical point drying and plated film
Processing, most clearly image can be obtained under conditions of ensureing that frustule form is constant, be observed using ESEM (SEM) thin
Born of the same parents, obtain the metamorphosis image after frustule processing；SEM image is analyzed.The change of cell size uses MATLAB
Software carries out image procossing, obtains the change of microwave frequency measurment frustule size before and after the processing, and introduce fractal dimension [12]
Concept, to quantify the change of frustule form.Cd is not added with contrast culture medium2+And the unadsorbed Cd harvested2+Frustule with
Cd is with the addition of in culture medium2+And what is harvested has adsorbed Cd2+Frustule, cell size and shape after being handled by microwave frequency measurment
The change of state, find adsorbed Cd2+Spirulina cells than unadsorbed Cd2+Spirulina cells its cell dias reduce
15%, and the fractal dimension of cell adds 12%.Adsorb Cd2+Spirulina cells it is broken after microwave frequency measurment processing
Bad degree is more than unadsorbed Cd2+Spirulina cells.
Embodiment 3
A kind of method of frustule broken wall, comprises the steps successively：(1) SM that haematococcus pluvialis are placed in 500ml is trained
Support in base, the Hg that mass concentration is 100ppb is added in the 500ml culture mediums of haematococcus pluvialis growing2+, then in environment temperature
Spend for 23 DEG C, intensity of illumination 6000lux, light application time 24h, haematococcus pluvialis are cultivated under conditions of throughput 50ml/min
Cell, Hg is adsorbed using haematococcus pluvialis cell growth process2+, part Hg2+Ion can pass through physisorption and adsorb
Haematococcus pluvialis cell wall surface；Part Hg2+Ion can be anti-by occurring with the charged particles of haematococcus pluvialis cell wall surface
Answer and be adsorbed onto cell wall；Remaining part Hg2+Ion can via the transport protein of haematococcus pluvialis cell wall surface and
It is adsorbed onto on cell membrane；(2) harvested after cultivating 10 days；Haematococcus pluvialis cell is harvested using centrifugal method, by haematococcus pluvialis
Cell solid separates as far as possible with medium liquid, and the moisture content for obtaining haematococcus pluvialis cell is 90%；The rain life of harvest is red
Ball frustule liquid 10ml is contained in the conical flask that volume is 20ml, and it is spacious that the bottleneck of conical flask is kept in processing procedure
Open, the normal pressure microwave for being 2450MHZ with frequency carries out broken wall treatment, and microwave treatment power is 1000W, treatment temperature 90
DEG C, processing time 12min, obtain the microalgae cell by broken wall treatment；(4) the frustule sample after microwave treatment is by solid
Fixed, dehydration, critical point drying and coating film treatment, it can obtain under conditions of ensureing that frustule form is constant and most clearly scheme
Picture, cell is observed using ESEM (SEM), obtain the metamorphosis image after frustule processing；SEM image is divided
Analysis.The change of cell size carries out image procossing using MATLAB softwares, obtains microwave frequency measurment frustule size before and after the processing
Change, and the concept of fractal dimension is introduced, to quantify the change of frustule form.Hg is not added with contrast culture medium2+And receive
The unadsorbed Hg obtained2+Frustule and culture medium in the addition of Hg2+And what is harvested has adsorbed Hg2+Frustule, by microwave
The change of cell size and form after broken wall treatment, finds adsorbed Hg2+Haematococcus pluvialis cell than unadsorbed Hg2+'s
Its cell dia of haematococcus pluvialis cell reduces 18%, and the fractal dimension of cell adds 16%.Adsorb Hg2+Rain life
Destructiveness of the haematococcus cell after microwave frequency measurment processing is more than unadsorbed Hg2+Haematococcus pluvialis cell.
The measurement on microwave frequency measurment efficiency is substantially weighed with grease extraction efficiency in the prior art, due to this specially
The addition of the metal ion of profit will cause the metabolic pathway of frustule oil-producing to change and cause cell itself oil-containing
Amount changes, so the addition that the change of grease extraction efficiency herein is probably metal ion influences frustule metabolic processes
Caused by, it is not that therefore this patent does not use grease extraction efficiency caused by the broken wall efficiency that metal ion influences microwave treatment
To weigh, by the change of microwave frequency measurment frustule size before and after the processing, and the concept of fractal dimension is introduced, it is thin to quantify algae
The change of born of the same parents' form, the fractal dimension of frustule is bigger, and the scrambling of frustule shape is bigger, i.e. the degree of crushing of frustule
Bigger, the efficiency of microwave frequency measurment processing is higher.
The concentration for the metal ion that this patent adds is advisable with not influenceing the growth of microalgae cell, due to different frustules pair
Different metals has different tolerance levels, therefore the addition concentration of metal ion is different, the concentration of different metal ions because
The adaptability of different algae kinds is different and different, is specifically advisable with not influenceing frustule growth, then in suitable temperature, light
According to normal culture microalgae cell under the conditions of intensity, light cycle period, ventilatory conditions etc..It is general using filter method harvest frustule
It is adapted to larger-size frustule, such as spirulina cells etc.；The less frustule of frustule size is harvested using centrifugal method,
Such as micro- plan ball frustule etc..It is above-mentioned to obtain the microalgae cell by broken wall treatment, by fixation, dehydration, critical point drying and
Coating film treatment, most clearly image can be obtained under conditions of ensureing that frustule form is constant, utilize ESEM (SEM) to see
Cell is examined, obtains the metamorphosis image after frustule processing.
Claims (8)
- A kind of 1. method of frustule broken wall, it is characterised in that comprise the steps successively：(1) microalgae cell is placed in the culture medium of suitable microalgae cell growth, the metal ion added in its culture medium, In growth course, part metals ion can pass through physisorption and adsorb in microalgae cell wall surface microalgae cell；Portion Divide metal ion can be by being reacted with the charged particles of microalgae cell wall surface to be adsorbed onto cell wall；Remaining portion Metal ion is divided to be adsorbed onto via the transport protein of microalgae cell wall surface on cell membrane；(2) the microalgae cell culture of metal ion will have been adsorbed until harvesting, microalgae cell solid separated with medium liquid, The moisture content of the frustule of acquisition is 60%-90%；(3) the frustule liquid of harvest is subjected to broken wall treatment with normal pressure microwave, and microwave treatment power is 600-1000W, place It is 70-90 DEG C to manage temperature, obtains the microalgae cell by broken wall treatment.
- 2. the method for a kind of frustule broken wall according to claim 1, it is characterised in that microalgae cell culture is until harvest 7-10 days time.
- A kind of 3. method of frustule broken wall according to claim 1, it is characterised in that the method by filtering or centrifuging Microalgae cell solid is separated with medium liquid.
- 4. the method for a kind of frustule broken wall according to claim 1, it is characterised in that broken wall treatment processing time is 8- 12min。
- 5. the method for a kind of frustule broken wall according to claim 1, it is characterised in that microwave is for microalgae in step (3) Cell membrane produces the double action of heat damage and mechanical damage, due to the presence of metal ion, in the electricity changed caused by microwave Under magnetic fields, electric field can be induced in metallic surface, induced electric field causes surface current, so as to cause Joule heat, aggravation Heat damage effect of the microwave to microalgae cell wall.
- A kind of 6. method of frustule broken wall according to claim 1, it is characterised in that the frequency of microwave be 2450MHZ or Person 915MHZ.
- 7. the method for a kind of frustule broken wall according to claim 1, it is characterised in that metal ion is Cd2+、Hg2+、K、 One or more combination in Mg, Ca, Fe, Sr, Co, Mn, Ni, V, Zn, As, Cd, Mo, Pb, Se and Al；Microalgae cell is bead Algae, small ring algae, grid algae, spirulina, split one or more combination in an algae, Mao Zhizao, brown algae.
- A kind of 8. method of frustule broken wall according to claim 1, it is characterised in that culture medium be SE culture mediums, One kind in Zarrouk culture mediums, SM culture mediums.
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| CN109187401A (en) * | 2018-09-12 | 2019-01-11 | 南京大学 | The method for measuring graphene subcellular proteomics in rice body |
| CN113355243A (en) * | 2021-06-04 | 2021-09-07 | 中国水产科学研究院黄海水产研究所 | Processing method for microwave-assisted extraction of microalgae active components |
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