CN106754390B - The albuminiferous chlorella of one plant height and its cultural method and application - Google Patents
The albuminiferous chlorella of one plant height and its cultural method and application Download PDFInfo
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Abstract
The invention discloses a chlorellas, it is named as chlorella (Chlorella sorokiniana) BL-ch1, it is preserved in China typical culture collection center on November 8th, 2016, address: wuchang, wuhan Luo Jia Shan, deposit number are as follows: CCTCC M 2016626.The cultural method of algae strain are as follows: algae strain is seeded on culture medium, inoculum concentration 5-15%, liquid amount 400-800mL/L, initial pH is 6.0-8.0,28 DEG C -33 DEG C of cultivation temperature.The chlorella that the present invention screens, protein content is high, and high to the degradation rate of total nitrogen, total phosphorus, ammonia nitrogen, can be used for the multiple fields such as protein production, feed preparation, sewage treatment.
Description
Technical field
The present invention relates to the albuminiferous chlorella of a plant height and its cultural method and applications, belong to algae bio technology neck
Domain.
Background technique
Microalgae is that one kind can be carried out photoautotrophic microorganism, primary producer is in the ecosystem, in entire object
Huge, high production efficiency is acted in matter circulation, development prospect is tempting.A large amount of nitrogen phosphorus members can be absorbed and utilized in microalgae during the growth process
Element, therefore denitrogenation dephosphorizing deep purifying sewage is carried out using the post-processing unit culture microalgae of sewage treatment plant, early in 1957
Year Oswald etc. is it has been suggested that be used as a kind of biosystem for replacing the activated sludge in sewage treatment for microalgae.Hereafter
The oxidation pond technology established on artificial mode water body helotisn self-cleaning basis is widely used and develops.Separately
Outside, microalgae also has the characteristics that growth is fast, harvest time is short, high rich in grease, photosynthetic utilization efficiency, can be used as food and biology
One of main source of the energy also has been to be concerned by more and more people.
Currently, in relation to using microalgae administer waste water and as the research of food and bioenergy it is also relatively fewer, mostly
It is in the laboratory lab scale stage of Strain selection, and from the point of view of having been reported, the generally existing condition of culture of algae strain utilized
It is required that it is high, protein yield is low, frustule wall is relatively thick that albumen is led to problems such as to be difficult to be utilized by organism, and be difficult to realize wide
General application.Therefore, excellent algae is screened to have great importance for solving the above problems.
Summary of the invention
For the above-mentioned prior art, the object of the present invention is to provide the albuminiferous chlorella of a plant height and its cultural method and
Using.
To achieve the above object, the present invention adopts the following technical solutions:
The first aspect of the present invention provides a chlorella, is named as chlorella (Chlorella sorokiniana)
BL-ch1 is preserved in China typical culture collection center on November 8th, 2016, address: wuchang, wuhan Luo Jia Shan,
Deposit number are as follows: CCTCC NO:M 2016626.
Algae strain is isolated from the excrement pond and waste discharge irrigation canals and ditches on Taian Shandong Ningyang pig farm, and algae strain is unicellular, spherical shape,
Cell wall is thin, and 3-6 μm of cell dia.Cell green, chromatoplast cup-shaped account for cell major part, have a pyrenoids.
The second aspect of the present invention provides the cultural method of the albuminiferous chlorella of a plant height, step are as follows: is inoculated with algae strain
To culture medium, inoculum concentration is 5-15% (preferably 10%), and liquid amount is 400-800mL/L (preferably 600mL/L), initially
PH be 6.0-8.0 (preferably 7.0), 28 DEG C -33 DEG C of cultivation temperature.
In above-mentioned cultural method, the composition of the culture medium are as follows: in No. 10 culture mediums of Zhu Shi add 1 ‰ glucose,
0.1 ‰ dipotassium hydrogen phosphates, 0.1 ‰ urea and 0.1 ‰ glycine.
The third aspect of the present invention provides application of the above-mentioned chlorella in sewage treatment.
Experiment proves that 94.31% is reached as high as using the degradation rate of chlorella of the invention to total nitrogen in sewage, to total
The degradation rate of phosphorus reaches as high as 90.07%, reaches as high as 99.21% to the degradation rate of ammonia nitrogen.To the excellent of sewage treatment.
The fourth aspect of the present invention provides application of the above-mentioned chlorella in protein production.Concrete application method are as follows: will be small
Ball algae is cultivated by above-mentioned cultural method, obtains algae solution；Algae solution is centrifuged, is concentrated, it is dry to get the algae powder containing albumen.
Application of the above-mentioned chlorella in preparation aquatic livestock open-mouthed bait and/or fish meal substitute is also guarantor of the invention
Protect range.
Beneficial effects of the present invention:
(1) chlorella that present invention screening obtains, protein content can be up to 71% or more, moreover, in high yield albumen
While, chlorella of the invention is high to total nitrogen, total phosphorus, the degradation rate of ammonia nitrogen in sewage, can be used for protein production, feed system
The multiple fields such as standby, sewage treatment.
(2) chlorella protein content is relatively high, can be used as an important sources of single cell protein, still, mostly
The miniature green alga of number all has one layer of tough and tensile cell wall compared with thick fiber element, to affect organism to the benefit of its protein
With digestibility also decreases.And the chlorella that the present invention screens, cell wall is thin, and the content of protein is high, non-
Often be conducive to absorption and utilization of the organism to chlorella protein.
Detailed description of the invention
Fig. 1: the cellular morphology microphoto of algae strain of the present invention；
Fig. 2: maximum brief (Maximum Parsimony) tree based on the building of 18S rDNA fragment sequence；(frame internal labeling
Be sample to be tested sequence)；
Fig. 3: the experimental rig figure of application of the chlorella in degradation waste water.
Specific embodiment
Instrument involved in following embodiments, reagent, material etc. are unless otherwise noted existing in the prior art
Conventional instrument, reagent, material etc., can be obtained by regular commercial sources.Experimental method involved in following embodiments, inspection
Survey method etc. is unless otherwise noted existing routine experiment method, detection method etc. in the prior art.
Embodiment 1: separation, screening and the identification of algae strain
The acquisition of 1 sample
Chlorella separates sample collecting location: the excrement pond and waste on Taian Shandong Ningyang pig farm discharge irrigation canals and ditches.
Separation, the screening of 2 algaes strain
2.1 test materials: the present embodiment uses culture medium for No. 10 culture mediums of Zhu Shi, adds 1% in solid medium
The agar of (mass percent) does not add agar in fluid nutrient medium.Culture medium constituent is shown in Table 1.
Table No. 10 culture medium constituents of 1 Zhu Shi
2.2 test method
2.2.1 the sample of acquisition is placed in 30 DEG C of illumination boxs and is cultivated 5-7 days, after to appear turn green, with Zhu Shi 10
Number fluid nutrient medium carries out 10,50,100,500 and 1000 times of gradient dilutions, and 100 μ L of sample after dilution is taken to be uniformly coated on Zhu Shi
On No. 10 solid mediums, it is placed in 30 DEG C of illumination boxs and cultivates；Picking list when observation has single algae to be born long in incubation
One algae falls access equipped in the test tube of No. 10 fluid nutrient mediums of Zhu Shi；Algae solution microscopy is taken after turning green, purebred algae strain is chosen and carries out
Expand numerous.
2.2.2 learn from else's experience diluted waste water sample 300mL, and loaded in 1L conical flask, 121 DEG C of high pressure sterilization 30min are accessed
Algae solution 100mL, in 30 DEG C, 180r/min continuously cultivates 5d, primary every sampling for 24 hours, measures chlorophyll, total phosphorus, total nitrogen, ammonia
The indexs such as nitrogen, nitrite.
2.3 index determining
Measure chlorophyll, total phosphorus, total nitrogen, ammonia nitrogen, nitrite and the protein content in purebred algae strain culture.
The measuring method of These parameters is as follows:
(1) total phosphorus: ammonium molybdate spectrophotometric method；
(2) total nitrogen: potassium persulfate oxidation method；
(3) ammonia nitrogen: reagent colorimetric method；
(4) nitrite: alpha-naphthylamine colorimetric method；
(5) protein content: Kjeldahl's method；
(6) chlorophyll: heat ethanol methods.
2.4 test result
2.4.1 algae strain isolate and purify
By isolating and purifying, 7 plants of single algae strains, number 1-1,2-1,3-1,4-1,5-1,6-1,7-1 are obtained.In optics
Under 100 times of microscope, the cell of 7 plants of algaes is spherical, Dan Sheng, and can see and contain more apparent chromatoplast into the cell,
Has the morphological feature of Chlorophyta, this 7 plants of algaes of Preliminary Identification are green alga.
2.4.2 the screening of algae strain
(1) measurement of total nitrogen content and its degradation rate
It the results are shown in Table 2.
2 total nitrogen content of table (mg/L) and degradation rate measurement
As can be seen from Table 2: in addition to 5-1,7-1, each algae strain total nitrogen degradation rate is up to 75% or more.
(2) measurement of total phosphorus content and its degradation rate
It the results are shown in Table 3.
3 total phosphorus content of table (mg/L) and degradation rate measurement
As can be seen from Table 3: in addition to 4-1, each algae strain total phosphorus degradation rate is up to 78% or more.
(3) measurement of ammonia-nitrogen content and its degradation rate
It the results are shown in Table 4.
4 ammonia-nitrogen content of table (mg/L) and degradation rate measurement
Each algae strain ammonia nitrogen degradation rate is higher as can be seen from Table 4.
(4) content of nitrite measures
It the results are shown in Table 5.
5 content of nitrite of table measures (mg/L)
Each algae strain does not have obvious effect to nitrite as can be seen from Table 5.
(5) measuring chlorophyll content
It the results are shown in Table 6.
6 measuring chlorophyll content of table (μ g/L)
Each algae strain chlorophyll content persistently increases as can be seen from Table 6, illustrates each algae strain well-grown.
(6) protein content and biomass estimation
It the results are shown in Table 7.
7 protein content of table and biomass
6-1 algae strain protein content is up to 67.9% as can be seen from Table 7.
The algae strain 6-1 protein content highest it can be seen from the above results, and the degradation rate of total nitrogen, total phosphorus, ammonia nitrogen is reachable
75% or more.To obtain a plant height lay eggs it is white, and to the algae of good waste water treatment effect strain, be named as BL-ch1.
The identification of 3 algaes strain
3.1 morphologic observation
The Main Biological of algae strain 6-1 are as follows: algae strain be it is unicellular, spherical, cell wall is thin, 3-6 μm of cell dia.Carefully
Born of the same parents' green, chromatoplast cup-shaped account for cell major part.With a pyrenoids.Microphoto such as Fig. 1 institute of algae strain cellular morphology
Show.
3.2 Molecular Identification
3.2.1 method
3.2.1.1 algal gene group is extracted using CTAB (Cetyltrimethylammonium bromide) method, and
With reference to " Molecular systematics ", the specific method is as follows:
10mL algae solution is taken, 8000rpm is centrifuged 10min, and 100 μ L TE are added, and vibrates suspension cell, adds 800 μ L cracking
Liquid and 0.8 μ L beta -mercaptoethanol, sufficiently oscillation mixes rapidly, is placed in 65 DEG C of warm bath 30min, overturns for several times back and forth therebetween, then plus
Enter 700 μ L chloroforms, mix well, 12000rpm is centrifuged 5min, and supernatant is transferred in new centrifuge tube after centrifugation, and 700 μ are added
L combination buffer is sufficiently mixed uniformly, is then added in centrifugal adsorbing column, places 1min, and 12000rpm is centrifuged 1min,
Fall the waste liquid in collecting pipe, add 700 μ L washing buffers, 12000rpm is centrifuged 1min, washes repeatedly 3 times, last time
12000rpm be centrifuged 2min, sufficiently removing washing buffer, take out centrifugal adsorbing column, be inserted in a clean 1.5mL from
Heart pipe, and 50 μ L elution buffers are added, after placing 1min, it is centrifuged 1min in 12000rpm, centrifugal adsorbing column is taken out, takes 10 μ L
Electrophoresis.
3.2.1.2 sequence amplification and measurement
PCR amplification simultaneously measures core encoding ribosomal 18S rDNA fragment sequence.
50 μ L PCR reaction systems include: 10 × PCR buffer, 0.2mM dNTP, 2.5mM Mg2+, 1U Taq enzyme, mould
Plate DNA, forward and reverse primer, ddH2O.Response procedures are as follows: then 95 DEG C of denaturation 5min press 95 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 1min
Program 34 circulations, last 72 DEG C of extensions 10min.PCR product electrophoresis in 1.5% agarose gel.After PCR product is purified
Directly it is sequenced by Huada gene company.
3.2.1.3 data are analyzed
Using Blast function, obtained sample 18S rDNA fragment sequence is compared with GenBank database.
The sequence for downloading similar algae on GenBank, is compared gene order using Clustal X, is aided with SEAVIEW craft school
Just, morphological development tree is constructed using MEGA5, analyzes the systematic growth position of sample.
3.2.2 interpretation of result
3.2.2.1 sequence analysis
Through being sequenced, as shown in SEQ ID NO.1, clip size is sample 18S rDNA fragment sequence to be identified
1693bp.This sequence and data in NCBI Genbank database are carried out sequence homology to compare, find algae strain with
Chlorella sorokiniana affiliation is nearest, and sequence homology can reach 100%.
3.2.2.2 Phylogenetic Analysis
Phylogenetic tree (Fig. 2) based on the building of 18S rDNA fragment sequence shows (the i.e. 6-1 algae of sample to be tested Contig 1
Strain) it is close with Chlorella sorokiniana affiliation, it is located on same evolutionary branching.
According to microexamination and molecular sequences sequencing result comprehensive analysis, show that sample to be tested is a kind of chlorella
Chlorella sorokiniana is under the jurisdiction of Chlorophyta (Chlorophyta), Chlorophyceae (Chlorophyceae), Chlorococcum
Mesh (Chlorococcales), chlorella section (Chlorellaceae), Chlorella (Chlorella).
The sequence analysis of biological property analysis and 18s rDNA based on bacterial strain is as a result, algae strain 6-1 is accredited as bead
Algae is named as chlorella (Chlorella sorokiniana) BL-ch1, is preserved in Chinese Typical Representative on November 8th, 2016
Culture collection, address: wuchang, wuhan Luo Jia Shan, deposit number are as follows: CCTCC M 2016626.
Embodiment 2: the application in chlorella production albumen of the invention
The determination of algae powder amount needed for 1 determining the protein quantity
1.1 test material
Algae strain: the embodiment of the present invention 1 screens isolated chlorella (Chlorella sorokiniana) BL-ch1；
Culture medium: Zhu Shi 10, formula: sodium nitrate 0.1g, dipotassium hydrogen phosphate 0.01g, magnesium sulfate 0.25g, sodium carbonate
0.02g, sodium metasilicate 0.025g, iron chloride 0.008g, water 1L.
1.2 test method
Biomass detection --- weighing bottle method；
Protein content detection --- Kjeldahl's method.
1.3 test procedure
No. 10 culture mediums of Zhu Shi are prepared, BL-ch1 is into culture medium for inoculation algae strain, and 33 DEG C are cultivated 5 days, by algae solution 5000r/
Min is centrifuged 5min, then concentration gained algal gel is all moved to algal gel in weighing bottle, 70 DEG C dry with pure washing three times
Algae powder, weighs and calculates biomass, and algae powder surveys protein content.
1.4 test result
It the results are shown in Table 8.
8 algae silty amount of table and protein content
As can be seen from Table 8, chlorella BL-ch1 protein content is that 67% or so, 0.13g-0.24g algae powder measures bead
Algae BL-ch1 protein content no significant difference.
The optimization of 2 production albumen used mediums
2.1 test method
On the basis of No. 10 culture mediums of Zhu Shi, glucose, dipotassium hydrogen phosphate, urea, potassium nitrate and sweet are further added
Propylhomoserin optimizes the prescription of culture medium by orthogonal design, and the formula composition of orthogonal design is as shown in table 9.
9 orthogonal design formula composition of table
2.2 test result
The embodiment of the present invention 1 is screened into isolated chlorella (Chlorella sorokiniana) BL-ch1 inoculation
To the culture medium of different numbers, 33 DEG C are cultivated 5 days, and algae solution 5000r/min is centrifuged 5min, and gained algal gel is concentrated, and use is pure
Washing three times, then all moves to algal gel in weighing bottle, and 70 DEG C dry to obtain algae powder, weighs and calculates biomass, and algae powder is surveyed albumen and contained
Amount, the results are shown in Table 10.
The optimization of 10 culture medium prescription of table
As can be seen from Table 10, culture medium is preferably formulated as formula 9, formula composition are as follows: No. 10 culture mediums+1 ‰ of Zhu Shi
+ 0.1 ‰ glycine of+0.1 ‰ urea of ‰ dipotassium hydrogen phosphate of glucose+0.1.Algae strain is carried out using the culture medium prescription after optimization
Culture, is cultivated relative to No. 10 culture mediums of Zhu Shi, and biomass can be improved 47.54%, and protein content improves 2.98%.
The optimization of 3 high yield protein pellet algae condition of culture
3.1 test material
Algae strain: the embodiment of the present invention 1 screens isolated chlorella (Chlorella sorokiniana) BL-ch1；
Culture medium: culture medium prescription after optimization: sodium nitrate 0.1g, dipotassium hydrogen phosphate 0.11g, magnesium sulfate 0.25g, sodium carbonate
0.02g, sodium metasilicate 0.025g, iron chloride 0.008g, 1g glucose, 0.1g urea, 0.1g glycine water 1L.
3.2 test method
Chlorophyll detection --- ethanol extraction method；
Biomass detection --- weighing bottle method；
Protein content detection --- Kjeldahl's method.
The influence of different vaccination amount, liquid amount, initial pH, cultivation temperature to chlorella protein yield is investigated respectively.
3.3 test result
3.3.1 the influence (continuously cultivating 5d) of different vaccination amount, liquid amount to chlorophyll
11 different vaccination amount of table, influence of the liquid amount to chlorophyll (μ g/L)
As can be seen from Table 11, the inoculum concentration of algae strain is preferably 5%-10%, and liquid amount is advisable with 600mL/L.
3.3.2 the influence (continuously cultivating 5d) of different vaccination amount, liquid amount to biomass, protein content
12 different vaccination amount of table, influence of the liquid amount to biomass, protein content
As can be seen from Table 12, different vaccination amount and liquid amount do not make significant difference to algae strain biomass and protein content.
3.3.3 variation (continuously cultivate 6d) of the different initial pH values to pH value in growth course
Variation of the different initial pH values of table 13 to pH value in growth course
As can be seen from Table 13, pH value is at continuous ascendant trend in incubation, and chlorella BL-ch1 is in growth course
In its pH value up to 9 or more.
3.3.4 influence (continuously cultivate 6d) of the different initial pH values to algae strain chlorophyll
Influence of the different initial pH values of table 14 to algae strain chlorophyll (μ g/L)
As can be seen from Table 14, chlorella BL-ch1 chlorophyll is fallen after rising, and most suitable initial pH value is 6.00.
3.3.5 influence of the different initial pH values to biomass, protein content
Influence of the different initial pH values of table 15 to biomass, protein content
As can be seen from Table 15, different initial pH values influence chlorella BL-ch1 protein content not significant, with initial pH
Value is slightly excellent when being 7.
3.3.6 the influence (continuously cultivating 6d) of different temperatures and condition of culture to chlorophyll
The influence of 16 different temperatures of table and condition of culture to chlorophyll (μ g/L)
As can be seen from Table 16, growth tendency is best at 33 DEG C by chlorella BL-ch1.
3.3.7 the influence of different temperatures and condition of culture to biomass, protein content
The influence of 17 different temperatures of table and condition of culture to biomass, protein content
As can be seen from Table 17, chlorella BL-ch1 biomass and protein content highest at 28 DEG C of illumination.
It thereby determines that, high yield protein pellet algae BL-ch1 optimal condition of culture are as follows: inoculum concentration 10%, liquid amount 600mL/
L, initial pH value 7.0,28 DEG C -33 DEG C of temperature.
The measurement of 4 high yield protein pellet algae growth tendencies and protein content
18 chlorella BL-ch1 growth tendency of table and protein content
As can be seen from Table 18, high after pH value is first low, biomass is incremented by always, reaches 1.5660g/L, chlorophyll within the 8th day
Changes of contents trend is identical as biomass, and protein content gradually rises, and substantially remains in 70% or so after five days, reaches as high as
71%.
5 high yield protein pellet algae analysis of amino acids
5.1 test material
(algae solution is centrifuged chlorella BL-ch1 algae powder prepared by the present invention through 5000 turns/min, 5min, distills water washing three
It is secondary, 65 C overnights drying gained)
5.2 measuring method
Mailing to Shandong University carries out Contents of Amino Acids after sample preparation success, and instrument is Hitachi L-8900 type
High speed automatic amino acid analyzer.
The report of 5.3 amino acid analysis
Amino acid content in 19 chlorella BL-ch1 algae powder of table
Gained chlorella contains amino acid classes up to 17 kinds as the result is shown.
Amino acid content compares in 5.4 chlorella BL-ch1 algae powder and fish meal
Amino acid content compares in 20 chlorella BL-ch1 algae powder of table and fish meal
As can be seen from Table 20, compared with fish meal, the algae strain valine of BL-ch1, leucine are higher than fish meal, threonine, different
Leucine, phenylalanine are similar to fish meal, and methionine, histidine are lower than fish meal.
To sum up, the chlorella BL-ch1 that the present invention screens can carry out Heterotrophic culture, and albumen contains after formulation optimization
Amount reaches as high as 69% or more, and contains 17 kinds of amino acid, and two of them amino acid is higher than amino acid content in fish meal, three kinds of ammonia
Base acid content is substantially close with amino acid content in fish meal.Therefore, the chlorella powder that the present invention screens can be used as aquatic livestock
Open-mouthed bait and part substitution fish meal.
Embodiment 3: the application of chlorella of the invention in degradation waste water
1 experimental design
Experimental design is as shown in table 21.
The degradation waste water test design of 21 chlorella of table
The cultural method of algae solution: algae strain is the algae strain BL-ch1 that the embodiment of the present invention 1 is screened, and inoculum concentration is 10% (volume
Score), liquid amount 600mL/L, initial pH value 7.0,28 DEG C -33 DEG C of temperature.
This test considers that waste smell and sampling are convenient, uses finishing box with cover, it is poor to do illuminance with 3L triangular flask
Different comparison, as shown in Figure 3.
2 experimental results
The degradation (total nitrogen content mg/L) of 2.1 pairs of total nitrogens
As a result as shown in table 22.
Degradation results of the table 22 to total nitrogen
As can be seen from Table 22: processing 4,5,6, i.e., algae dirt than for the degradation rate of 1:1,4:1,8:1 to total nitrogen it is reachable
80% or more.
The degradation (total phosphorus content content mg/L) of 2.2 pairs of total phosphorus
As a result as shown in table 23.
Degradation results of the table 23 to total phosphorus
As can be seen from Table 23: the degradation of 4,5,6 pairs of total phosphorus of processing is up to 40% or more, wherein 5 pairs of total phosphorus of processing turn
Change up to 90%.
The degradation (ammonia-nitrogen content content mg/L) of 2.3 pairs of ammonia nitrogens
As a result as shown in table 24.
Degradation results of the table 24 to ammonia nitrogen
As can be seen from Table 24: processing 4,5,6 i.e. algae dirt than for 1:1,4:1,8:1 to the degradation rate of ammonia nitrogen up to 90% with
On.
The measurement (chlorophyll content μ g/L) of 2.4 chlorophyll
As a result as shown in Table 25.
The measurement result of 25 chlorophyll of table
As can be seen from Table 25: although slightly fluctuation can embody becoming for growth on 4,5,6 chlorophyll measurings of processing
Gesture illustrates algae strain well-grown in conjunction with culture color change.
Comprehensively consider result above, for chlorella BL-ch1 of the invention in phosphorus decomposing, drop nitrogen, total nitrogen initial value should be
400mg/L or so is more appropriate.
SEQUENCE LISTING
<110>Shandong Baolai-leelai Bio-engineering Co., Ltd.
The albuminiferous chlorella of<120>one plant heights and its cultural method and application
<130> 2016
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1693
<212> DNA
<213>algae strain 6-1
<400> 1
tgtctaagta taaactgctt tatactgtga aactgcgaat ggctcattaa atcagttata 60
gtttatttga tggtacctac tactcggata cccgtagtaa atctagagct aatacgtgcg 120
taaatcccga cttctggaag ggacgtattt attagataaa aggccgaccg ggctctgccc 180
gactcgcggt gaatcatgat aacttcacga atcgcatggc cttgcgccgg cgatgtttca 240
ttcaaatttc tgccctatca actttcgatg gtaggataga ggcctaccat ggtggtaacg 300
ggtgacggag gattagggtt cgattccgga gagggagcct gagaaacggc taccacatcc 360
aaggaaggca gcaggcgcgc aaattaccca atcctgacac agggaggtag tgacaataaa 420
taacaatact gggccttttc aggtctggta attggaatga gtacaatcta aaccccttaa 480
cgaggatcaa ttggagggca agtctggtgc cagcagccgc ggtaattcca gctccaatag 540
cgtatattta agttgctgca gttaaaaagc tcgtagttgg atttcgggtg gggcctgccg 600
gtccgccgtt tcggtgtgca ctggtagggc ccaccttgtt gccggggacg ggctcctggg 660
cttcactgtc cgggactcgg agtcggcgct gttactttga gtaaattaga gtgttcaaag 720
caggcctacg ctctgaatac attagcatgg aataacacga taggactctg gcctatcctg 780
ttggtctgta ggaccggagt aatgattaag agggacagtc gggggcattc gtatttcatt 840
gtcagaggtg aaattcttgg atttatgaaa gacgaactac tgcgaaagca tttgccaagg 900
atgttttcat taatcaagaa cgaaagttgg gggctcgaag acgattagat accgtcctag 960
tctcaaccat aaacgatgcc gactagggat cggcggatgt ttcttcgatg actccgccgg 1020
caccttatga gaaatcaaag tttttgggtt ccggggggag tatggtcgca aggctgaaac 1080
ttaaaggaat tgacggaagg gcaccaccag gcgtggagcc tgcggcttaa tttgactcaa 1140
cacgggaaaa cttaccaggt ccagacatag tgaggattga cagattgaga gctctttctt 1200
gattctatgg gtggtggtgc atggccgttc ttagttggtg ggttgccttg tcaggttgat 1260
tccggtaacg aacgagacct cagcctgcta aatagtcacg gttggttcgc cagccggcgg 1320
acttcttaga gggactattg gcgactagcc aatggaagca tgaggcaata acaggtctgt 1380
gatgccctta gatgttctgg gccgcacgcg cgctacactg atgcattcaa cgagcctagc 1440
cttggccgag aggcccgggt aatctttgaa actgcatcgt gatggggata gattattgca 1500
attattaatc ttcaacgagg aatgcctagt aagcgcaagt catcagcttg cgttgattac 1560
gtccctgccc tttgtacaca ccgcccgtcg ctcctaccga ttgggtgtgc tggtgaagtg 1620
ttcggattgg cgaccggggg cggtctccgc tctcggccgc cgagaagttc attaaaccct 1680
cccacctaga gga 1693
Claims (10)
1. a chlorella is named as chlorella (Chlorella sorokiniana) BL-ch1, on November 8th, 2016
It is preserved in China typical culture collection center, address: wuchang, wuhan Luo Jia Shan, deposit number are as follows: CCTCC M
2016626。
2. the cultural method of chlorella described in claim 1, which is characterized in that step are as follows: algae strain is seeded on culture medium,
Inoculum concentration is 5-15%, and liquid amount 400-800mL/L, initial pH are 6.0-8.0,28 DEG C -33 DEG C of cultivation temperature.
3. cultural method as claimed in claim 2, which is characterized in that the composition of the culture medium are as follows: cultivated at Zhu Shi 10
1 ‰ glucose, 0.1 ‰ dipotassium hydrogen phosphates, 0.1 ‰ urea and 0.1 ‰ glycine are added in base.
4. cultural method as claimed in claim 2, which is characterized in that inoculum concentration 10%, liquid amount 600mL/L, initially
PH is 7.0.
5. application of the chlorella described in claim 1 in sewage treatment.
6. application as claimed in claim 5, which is characterized in that the sewage treatment includes total nitrogen, total phosphorus in degradation sewage
And ammonia nitrogen.
7. application of the chlorella described in claim 1 in protein production.
8. the use as claimed in claim 7, which is characterized in that application method are as follows: by chlorella described in claim 1 by power
Benefit require 2 described in cultural method cultivated, obtain algae solution；Algae solution is centrifuged, is concentrated, it is dry to get the algae containing albumen
Powder.
9. application of the chlorella described in claim 1 in preparation aquatic livestock open-mouthed bait and/or fish meal substitute.
10. a kind of albuminiferous method of life, which is characterized in that chlorella described in claim 1 is as described in claim 2
Cultural method carries out culture 4-6 days, obtains algae solution；Algae solution is centrifuged, is concentrated, it is dry.
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| CN107345208B (en) * | 2017-06-19 | 2020-05-05 | 南京师范大学 | Low-temperature growth green algae and application thereof in removing nitrogen and phosphorus in sewage |
| CN107541466B (en) * | 2017-08-02 | 2019-07-05 | 海南大学 | It is a kind of discharge aroma type smell the resistance to strong light of salt tolerant high temperature resistant chlorella and its application |
| CN107916226B (en) * | 2017-10-13 | 2020-11-27 | 浙江省农业科学院 | Screening of microalgae capable of efficiently treating livestock and poultry sewage in pig farm and realizing resource utilization |
| CN108753623A (en) * | 2018-06-12 | 2018-11-06 | 兰溪市沉默生物科技有限公司 | A kind of chlorella store method |
| JP2020174579A (en) * | 2019-04-18 | 2020-10-29 | 三菱重工機械システム株式会社 | Microalga culture method and microalga culture device |
| CN112538431B (en) * | 2020-12-16 | 2023-08-11 | 范秀娟 | Chlorella pyrenoidosa, preparation method of biological environment restoration liquid and application thereof |
| CN113481102B (en) * | 2021-08-19 | 2022-08-26 | 山西农业大学 | Chlorella sorokiniana strain as well as culture method and application thereof |
| CN113717854B (en) * | 2021-09-01 | 2023-05-02 | 东北农业大学 | Chlorella oleaginous ZM-5 and application thereof |
| CN114854596B (en) * | 2022-05-26 | 2023-04-14 | 珠海元育生物科技有限公司 | Chlorella protothecoides mutant strain with high protein yield and application thereof |
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