CN103436481A - Preparation method and application of brewer yeast spore serving as novel adsorbing agent - Google Patents
Preparation method and application of brewer yeast spore serving as novel adsorbing agent Download PDFInfo
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- CN103436481A CN103436481A CN2013104209971A CN201310420997A CN103436481A CN 103436481 A CN103436481 A CN 103436481A CN 2013104209971 A CN2013104209971 A CN 2013104209971A CN 201310420997 A CN201310420997 A CN 201310420997A CN 103436481 A CN103436481 A CN 103436481A
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Abstract
The invention discloses a preparation method and an application of a brewer yeast spore. The preparation method of the brewer yeast spore comprises the steps of cultivating a brewer yeast single colony and preparing the brewer yeast spore. The prepared brewer yeast spore can be used for absorbing metal ions or fat-like substances, shows stronger adsorption capacity and has a broad application prospect. According to the preparation method of the brewer yeast spore, massive yeast cells can be obtained by cultivating a deltadit1 strain, then wall breaking and purifying are carried out to obtain the brewer yeast spore target product, and corresponding application can be carried out without acquiring expensive chitosan; the preparation method of the brewer yeast spore is easy to operate and environment-friendly.
Description
Technical field
The present invention relates to Microbial resources and Environmental Biotechnology field, specifically relate to a kind of preparation method and application thereof of take the new adsorbent that the yeast saccharomyces cerevisiae spore is raw material.
Background technology
Chitosan is obtained through deacetylation by chitin, its chemical name is Chitosan (1-4)-2-amino-beta--D-Glucose, its the most significant characteristic is its adsorptive power, the amino in its molecule and with it adjacent hydroxyl and many metal ions (as Hg
2+, Ni
2+, Cu
2+, Pb
2+, Ca
2+, Ag
+deng) all can form stable inner complex, therefore can be used for administering heavy metal wastewater thereby, purified tap water and separating metal ion etc. in hydrometallurgy.In addition, chitosan can be adsorbed dyestuff, protein, amino, nucleic acid, enzyme, hydracid element etc. by complexing and ion exchange, for the processing of waste water from dyestuff, dyeing waste water, food industrial wastewater, thus environment purification, to protect mankind health.At present the existing chitosan that utilizes adsorbs some metal ions in sewage (as Cu
2+, Zn
2+, Hg
2+deng) report, thereby alleviate water, pollute.Chitosan, except the application aspect environmental protection, also has very large purposes in other respects, such as controlling the synthetic of cholesterol, and Antibacterial activity, Control and prevention hypertension and as fields such as healthcare products, foodstuff additive.
At present, the method that domestic industry is produced chitosan is mainly that to take shrimp, crab shell be raw material, utilize the steps such as sour decalcification, alkali deproteinated, potassium permanganate decolouring to make product, but due to raw material shrimp shell or crab shell not the quality of easily collecting and raw material can not guarantee to stablize, also there is corrupt possibility in time length, and strong acid or highly basic processing can cause the pollution of environment.And, along with the increasing of people's product consumption and the reinforcement of environmental consciousness, be badly in need of excavating the chitosan production method made new advances.
Summary of the invention
The purpose of this part is to summarize some aspects of embodiments of the invention and briefly introduces some preferred embodiments.May do a little simplification or omit to avoid the making purpose of this part, specification digest and denomination of invention fuzzy in this part and the application's specification digest and denomination of invention, and this simplification or omit can not be for limiting the scope of the invention.
Problem in view of existing in the preparation method of above-mentioned and/or existing yeast saccharomyces cerevisiae spore and application, proposed the present invention.
Therefore, the purpose of this invention is to provide a kind of preparation method of yeast saccharomyces cerevisiae spore, to reach the purpose of the yeast saccharomyces cerevisiae spore for preparing specific end use.
For solving the problems of the technologies described above, the invention provides following technical scheme: a kind of preparation method of yeast saccharomyces cerevisiae spore, comprise, the cultivation of yeast saccharomyces cerevisiae list bacterium colony, adopt the method for homologous recombination to knock out responsible coding Saccharomyces Cerevisiae in S K1 conidial cell wall outermost layer o,o-Dityrosine layer DIT1 gene, the yeast saccharomyces cerevisiae defect bacterial strain obtained is rule on YPAD solid medium flat board, and 28 ℃～32 ℃ cultivations grow yeast saccharomyces cerevisiae list bacterium colony; The preparation of yeast saccharomyces cerevisiae spore, picking yeast saccharomyces cerevisiae list bacterium colony is cultivated 22h～26h in the YPAD solid medium, then bacterium liquid is transferred in the YPAce substratum, collecting cell after cultivating, in the liquor kalii acetici that to forward concentration to be 1%～3% again, cultivate, the concentration that makes cell in liquor kalii acetici is 2 * 10
7/ ml～4 * 10
7/ ml, then centrifugal, by PBS damping fluid washing for the yeast saccharomyces cerevisiae thalline after centrifugal, process 0.5h～1.5h with a certain amount of lyticase under 36 ℃～38 ℃ after, carry out ultrasonic disruption, obtain the yeast saccharomyces cerevisiae spore of unbound state.
As a kind of preferred version of the preparation method of yeast saccharomyces cerevisiae spore of the present invention, wherein: also comprise, the yeast saccharomyces cerevisiae spore is carried out to purifying and lyophilize is processed.
A kind of preferred version as the preparation method of yeast saccharomyces cerevisiae spore of the present invention, wherein: described the yeast saccharomyces cerevisiae spore is carried out to purifying, comprise, preparation Percoll solution, by the cell of processing 0.5%TritonX-100 solution washing, to remove the impurity such as lyase wherein, then by cell suspension in 0.5%TritonX-100 solution, prepare the Percoll solution of different gradient 50%～80% different concns gradients; The purifying of yeast saccharomyces cerevisiae spore, according to joining successively from the high density to the lower concentration in centrifuge tube, finally add brewing yeast cell suspension by described Percoll solution, centrifugal, and supernatant liquid is removed, and obtains the yeast saccharomyces cerevisiae spore of purifying.
As a kind of preferred version of the preparation method of yeast saccharomyces cerevisiae spore of the present invention, wherein: the cultivation of described yeast saccharomyces cerevisiae list bacterium colony, comprise, design of primers is as follows according to yeast saccharomyces cerevisiae DIT1 gene order design primer:
Upstream primer P1
AATTTGTTAATATCCTAATTCGGTAAAGCTTTGTCGAGACATTAACAAAACGGATCCCCGGGTTAATTAA
Downstream primer P2
TGTTTAAGTAAAAGAACAAAAAGGTAGACCAATGTAGCGCTCTTACTTTAGAATTCGAGCTCGTTTAAAC；
Pcr amplification, utilize upstream primer P1 and downstream primer P2 to carry out pcr amplification to plasmid pFA6a-His3MX6, obtains the fragment that knocks out containing yeast saccharomyces cerevisiae DIT1 gene upstream and downstream sequence;
Knocking out of DIT1 gene, the his fragment that the method for transformation by Lithium Acetate/PEG obtains pcr amplification imports to respectively in the haploid cell 4B and 16D of Saccharomyces Cerevisiae in S K1, the row filter of going forward side by side;
The screening of defective bacterial strain, the conversion bacterial strain obtained after the knocking out of DIT1 gene is applied on SD-His defective flat board, after growing bacterium colony, the bacterium colony of picking certain number extracts genome, the performing PCR of going forward side by side checking, contrasted with the genome with wild type strain, if be proved to be successful, again monoploid is merged on the SD-Leu-Arg flat board, obtain the diploid bacterial strain of defective;
The checking primer is as follows:
Upstream primer: CATAAATTGTGCTCCTCCGC
Downstream primer: CATTGCAGTGTCTCGAAACC.
A kind of preferred version as the preparation method of yeast saccharomyces cerevisiae spore of the present invention, wherein: the cultivation of described yeast saccharomyces cerevisiae list bacterium colony, the YPAD solid medium be formulated as yeast extract 10g, peptone 20g, VITAMIN B4 30mg, agar 20g, constant volume is in 900mL distilled water, after 121 ℃ of sterilizing 15min, add 20% glucose of the independent sterilizing of 100mL.。
A kind of preferred version as the preparation method of yeast saccharomyces cerevisiae spore of the present invention, wherein: the preparation of described yeast saccharomyces cerevisiae spore, the YPAce substratum be formulated as yeast extract 10g, peptone 20g, VITAMIN B4 30mg, constant volume, in 900mL distilled water, after 121 ℃ of sterilizing 15min, is added 20% Potassium ethanoate of the independent sterilizing of 100mL.
Another object of the present invention is to provide a kind of metal ion adsorbent, and this sorbent material adopts the yeast saccharomyces cerevisiae spore prepared in the present invention.
For solving the problems of the technologies described above, the invention provides following technical scheme: a kind of yeast saccharomyces cerevisiae spore is as the application of metal ion adsorbent.
Application as yeast saccharomyces cerevisiae spore of the present invention as metal ion adsorbent, wherein: comprise, described yeast saccharomyces cerevisiae spore is carried out obtaining yeast saccharomyces cerevisiae spore powder after purifying and lyophilize processing; Described yeast saccharomyces cerevisiae spore powder is added in the solution that contains metal ion.
A further object of the present invention is to provide a kind of lipoid material sorbent material, and this sorbent material adopts the yeast saccharomyces cerevisiae spore prepared in the present invention.
For solving the problems of the technologies described above, the invention provides following technical scheme: a kind of yeast saccharomyces cerevisiae spore is as the application of lipoid material sorbent material.
Application as yeast saccharomyces cerevisiae spore of the present invention as lipoid material, wherein: comprise, described yeast saccharomyces cerevisiae spore is carried out obtaining yeast saccharomyces cerevisiae spore powder after purifying and lyophilize processing; Described yeast saccharomyces cerevisiae spore powder is added in lipoid material.
The present invention proposes a kind of preparation method and application of yeast saccharomyces cerevisiae spore, prepared yeast saccharomyces cerevisiae spore can be used for absorbing metal ion or lipoid material, and shows stronger adsorptive power, has wide practical use.The present invention directly obtains a large amount of yeast cell by cultivating △ dit1 bacterial strain, more in addition the broken wall purifying obtains the purpose product, just can do corresponding application, and without the expensive chitosan of acquisition, this method easy handling, environmental protection.
The accompanying drawing explanation
Fig. 1 is yeast saccharomyces cerevisiae spore adsorbing metal ions and the lipoid material principle schematic after removal conidial cell wall o,o-Dityrosine layer;
Fig. 2 utilizes described yeast saccharomyces cerevisiae spore to absorb Cu according to first embodiment of the invention
2+the effect schematic diagram;
Fig. 3 utilizes described yeast saccharomyces cerevisiae spore to absorb the effect schematic diagram of Bile Salts according to second embodiment of the invention.
Embodiment
For above-mentioned purpose of the present invention, feature and advantage can be become apparent more, below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in detail.
A lot of details have been set forth in the following description so that fully understand the present invention, but the present invention can also adopt other to be different from alternate manner described here and implement, those skilled in the art can be in the situation that do similar popularization without prejudice to intension of the present invention, so the present invention is not subject to the restriction of following public specific embodiment.
Secondly, alleged " embodiment " or " embodiment " refers to special characteristic, structure or the characteristic that can be contained at least one implementation of the present invention herein.Different local in this manual " in one embodiment " that occur not all refer to same embodiment, neither be independent or the embodiment mutually exclusive with other embodiment optionally.
As pointed in background technology in the present invention, the unicellular fungi of a large class of yeast general reference energy fermenting carbohydrate, most typical yeast is yeast saccharomyces cerevisiae (Saccharomyces cerevisiae).Yeast saccharomyces cerevisiae is under good nutrition and growth conditions, and growth rapidly, is carried out reproduction in the mode of the breeding of sprouting.But take acetate as unique or main carbon source, and be aided with (as only had the potassium acetate existence) under specified conditions such as lacking nitrogenous source, yeast saccharomyces cerevisiae will be bred in sporiparous mode, forms spore in born of the same parents.Cell now is called ascus.Ascus, after nature or artificial broken wall, can discharge thecaspore wherein.For thecaspore, its conidial cell wall forms by four layers, is followed successively by from inside to outside: seminose, beta-glucan, chitosan, o,o-Dityrosine.Because chitosan wherein has very large purposes industrial, so the present invention suppresses the generation of conidial cell wall o,o-Dityrosine layer by the means of genetic manipulation, then cracks ascus wall, thereby can make the chitosan layer of conidial cell wall come out fully; So, just can utilize chitosan to make corresponding purposes.The present invention has adopted and yeast saccharomyces cerevisiae is produced after the spore fragmentation to the mode of carrying out purifying has been used as novel sorbent material, after we experiment showed, that yeast saccharomyces cerevisiae produces the spore fragmentation, even purifying spore not directly adsorbs, also can reach certain effect.
The present invention prepares the yeast saccharomyces cerevisiae spore and comprises the following steps: adopt the method for homologous recombination to knock out responsible coding Saccharomyces Cerevisiae in S K1 conidial cell wall outermost layer o,o-Dityrosine layer DIT1 gene, the yeast saccharomyces cerevisiae defect bacterial strain obtained is rule on YPAD solid medium flat board, and 30 ℃ are cultured to and grow single bacterium colony; Wherein, the structure of yeast saccharomyces cerevisiae Δ dit1 mutant strain adopts following method to carry out:
1) design of primers: as follows according to yeast saccharomyces cerevisiae DIT1 gene order (GenBank) design primer:
Upstream primer
AATTTGTTAATATCCTAATTCGGTAAAGCTTTGTCGAGACATTAACAAAACGGATCCCCGGGTTAATTAA
Downstream primer
TGTTTAAGTAAAAGAACAAAAAGGTAGACCAATGTAGCGCTCTTACTTTAGAATTCGAGCTCGTTTAAAC
2) pcr amplification: utilize upstream primer P1 and downstream primer P2 to derive from Chinese Typical Representative culture collection center to plasmid pFA6a-His3MX6(, can buy acquisition) carry out pcr amplification, obtain the fragment that knocks out containing yeast saccharomyces cerevisiae DIT1 gene upstream and downstream sequence.
3) knocking out of DIT1 gene: the his fragment that the method for transformation by Lithium Acetate/PEG obtains pcr amplification imports to respectively in the haploid cell 4B and 16D of Saccharomyces Cerevisiae in S K1, the row filter of going forward side by side.
4) screening of defective bacterial strain: use SD-His defective flat board to be screened.By 3) the conversion bacterial strain that obtains is applied on SD-His defective flat board, and after growing bacterium colony, the bacterium colony of picking certain number extracts genome, and the performing PCR of going forward side by side checking, contrasted with the genome with wild type strain.If be proved to be successful, then monoploid is merged on the SD-Leu-Arg flat board, obtain the diploid bacterial strain of defective.
The checking primer is as follows:
Upstream primer: CATAAATTGTGCTCCTCCGC
Downstream primer: CATTGCAGTGTCTCGAAACC.
And 1L YPAD substratum be formulated as yeast extract 10g, peptone 20g, VITAMIN B4 30mg, constant volume (solid medium need add agar 20g) in 900mL distilled water, after 121 ℃ of sterilizing 15min, add 20% glucose of the independent sterilizing of 100mL.
Then the single bacterium colony obtained in step on picking, in the YPAD liquid nutrient medium, is cultivated 24h in 30 ℃ of shaking tables.Bacterium liquid is transferred in the YPAce substratum, and collecting cell after incubated overnight, forward in 2% Potassium ethanoate and cultivate, and the concentration that makes cell in Potassium ethanoate is 3 * 10
7/ ml, after cultivating 20h, after the centrifugal 10min of 5000rpm, PBS damping fluid washing for thalline is processed the 1h(625U/g cell with a certain amount of lyticase under 37 ℃), carry out ultrasonic disruption on ice: power 375watt, broken 1s, stop 2s, broken 10min, obtain the spore of unbound state and spore carried out to purifying and lyophilize.
Wherein, 1L YPACE liquid nutrient medium be formulated as yeast extract 10g, peptone 20g, VITAMIN B4 30mg, constant volume, in 900mL distilled water, after 121 ℃ of sterilizing 15min, is added 20% Potassium ethanoate of the independent sterilizing of 100mL.
In this embodiment, purifying comprises the steps: 0.5%Triton X-100 solution washing three times for the cell that first will process, to remove the impurity such as lyase wherein, then by cell suspension in 1mL0.5%Triton X-100 solution.Prepare the Percoll solution of different gradient 50%-80% different concns gradients:
(1) 8mL Percoll, 1mL0.5%Triton-X, 1mL2.5M sucrose;
(2) 7mL Percoll, 2mL0.5%Triton-X, 1mL2.5M sucrose;
(3) 6mL percoll, 3mL0.5%Triton-X, 1mL2.5M sucrose;
(4) 5mL Percoll, 4mL0.5%Triton-X, 1mL2.5M sucrose.
By above-mentioned Percoll solution 1mL according to join successively the centrifuge tube of 10mL from the high density to the lower concentration, finally add cell suspension, 15,000g, after 4 ℃ of centrifugal 1h, gradient solution will layering, and pure spore will be positioned at the bottom of centrifuge tube, supernatant liquid is removed to the lyophilize in freeze drier of the spore after purifying.
The first embodiment is as shown in Figure 2: the yeast saccharomyces cerevisiae spore is to Cu
2+absorption.
After △ dit1 bacterial strain produces the yeast saccharomyces cerevisiae spore, after preceding method purifying lyophilize, the spore powder that takes certainweight adds in centrifuge tube.The CuSO of preparation 1mmol/L
45H
2o (Cu
2+concentration is 64mg/L), to the CuSO that adds 1mL in centrifuge tube
45H
2o solution makes spore in suspended state, and after being placed in 30 ℃ of shaking tables reaction 5h, the centrifugal 10min of 15,000rpm, get Atomic Absorption Spectrometry Cu wherein for supernatant liquor
2+content also calculates absorbed Cu
2+content.
The second embodiment is as shown in Figure 3: the absorption of yeast saccharomyces cerevisiae spore to Bile Salts.
The cholyltaurine sodium solution of preparation 3mol/L, take above-mentioned spore powder, uses the cholyltaurine sodium solution suspension spore powder of 1mL, in 37 ℃ of shaking tables, reacts 2h.The centrifugal 10min of 15,000rpm, get supernatant liquid and measure wherein the cholyltaurine sodium content and calculate absorbed cholyltaurine sodium content with HPLC.Because Bile Salts is lipoid material, at this, be mainly in order to simulate in human body the absorption to lipid material, so carry out absorption reaction under 37 ℃.
Wherein, WT: wild-type yeast saccharomyces cerevisiae spore; Veg: yeast saccharomyces cerevisiae vegetative cell; △ dit1:DIT1 gene defection type yeast saccharomyces cerevisiae spore.
It should be noted that, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although with reference to preferred embodiment, the present invention is had been described in detail, those of ordinary skill in the art is to be understood that, can modify or be equal to replacement technical scheme of the present invention, and not breaking away from the spirit and scope of technical solution of the present invention, it all should be encompassed in the middle of claim scope of the present invention.
Claims (10)
1. the preparation method of a yeast saccharomyces cerevisiae spore is characterized in that: comprises,
The cultivation of yeast saccharomyces cerevisiae list bacterium colony, adopt the method for homologous recombination to knock out responsible coding Saccharomyces Cerevisiae in S K1 conidial cell wall outermost layer o,o-Dityrosine layer DIT1 gene, the yeast saccharomyces cerevisiae defect bacterial strain obtained is rule on YPAD solid medium flat board, and 28 ℃～32 ℃ cultivations grow yeast saccharomyces cerevisiae list bacterium colony;
The preparation of yeast saccharomyces cerevisiae spore, picking yeast saccharomyces cerevisiae list bacterium colony is cultivated 22h～26h in the YPAD solid medium, then bacterium liquid is transferred in the YPAce substratum, collecting cell after cultivating, in the liquor kalii acetici that to forward concentration to be 1%～3% again, cultivate, the concentration that makes cell in liquor kalii acetici is 2 * 10
7/ ml～4 * 10
7/ ml, then centrifugal, by PBS damping fluid washing for the yeast saccharomyces cerevisiae thalline after centrifugal, process 0.5h～1.5h with a certain amount of lyticase under 36 ℃～38 ℃ after, carry out ultrasonic disruption, obtain the yeast saccharomyces cerevisiae spore of unbound state.
2. the preparation method of yeast saccharomyces cerevisiae spore according to claim 1, is characterized in that: also comprise, the yeast saccharomyces cerevisiae spore is carried out to purifying and lyophilize processing.
3. the preparation method of yeast saccharomyces cerevisiae spore according to claim 2 is characterized in that: described the yeast saccharomyces cerevisiae spore carried out to purifying, comprises,
Preparation Percoll solution, by the 0.5%Triton X-100 solution washing for cell of processing, to remove the impurity such as lyase wherein, then by cell suspension in 0.5%Triton X-100 solution, prepare the Percoll solution of different gradient 50%～80% different concns gradients;
The purifying of yeast saccharomyces cerevisiae spore, according to joining successively from the high density to the lower concentration in centrifuge tube, finally add brewing yeast cell suspension by described Percoll solution, centrifugal, and supernatant liquid is removed, and obtains the yeast saccharomyces cerevisiae spore of purifying.
4. the preparation method of yeast saccharomyces cerevisiae spore according to claim 1 is characterized in that: the cultivation of described yeast saccharomyces cerevisiae list bacterium colony, comprise,
Design of primers is as follows according to yeast saccharomyces cerevisiae DIT1 gene order design primer:
Upstream primer P1
AATTTGTTAATATCCTAATTCGGTAAAGCTTTGTCGAGACATTAACAAAACGGATCCCCGGGTTAATTAA
Downstream primer P2
TGTTTAAGTAAAAGAACAAAAAGGTAGACCAATGTAGCGCTCTTACTTTAGAATTCGAGCTCGTTTAAAC；
Pcr amplification, utilize upstream primer P1 and downstream primer P2 to carry out pcr amplification to plasmid pFA6a-His3MX6, obtains the fragment that knocks out containing yeast saccharomyces cerevisiae DIT1 gene upstream and downstream sequence;
Knocking out of DIT1 gene, the his fragment that the method for transformation by Lithium Acetate/PEG obtains pcr amplification imports to respectively in the haploid cell 4B and 16D of Saccharomyces Cerevisiae in S K1, the row filter of going forward side by side;
The screening of defective bacterial strain, the conversion bacterial strain obtained after the knocking out of DIT1 gene is applied on SD-His defective flat board, after growing bacterium colony, the bacterium colony of picking certain number extracts genome, the performing PCR of going forward side by side checking, contrasted with the genome with wild type strain, if be proved to be successful, again monoploid is merged on the SD-Leu-Arg flat board, obtain the diploid bacterial strain of defective;
The checking primer is as follows:
Upstream primer: CATAAATTGTGCTCCTCCGC
Downstream primer: CATTGCAGTGTCTCGAAACC.
5. the preparation method of yeast saccharomyces cerevisiae spore according to claim 1, it is characterized in that: the cultivation of described yeast saccharomyces cerevisiae list bacterium colony, the YPAD solid medium be formulated as yeast extract 10g, peptone 20g, VITAMIN B4 30mg, agar 20g, constant volume is in 900mL distilled water, after 121 ℃ of sterilizing 15min, add 20% glucose of the independent sterilizing of 100mL.
6. the preparation method of yeast saccharomyces cerevisiae spore according to claim 1, it is characterized in that: the preparation of described yeast saccharomyces cerevisiae spore, the YPAce substratum be formulated as yeast extract 10g, peptone 20g, VITAMIN B4 30mg, constant volume, in 900mL distilled water, after 121 ℃ of sterilizing 15min, is added 20% Potassium ethanoate of the independent sterilizing of 100mL.
7. a yeast saccharomyces cerevisiae spore as described as claim 1～6 is as the application of metal ion adsorbent.
8. yeast saccharomyces cerevisiae spore according to claim 7, as the application of metal ion adsorbent, is characterized in that:
Described yeast saccharomyces cerevisiae spore is carried out obtaining yeast saccharomyces cerevisiae spore powder after purifying and lyophilize processing;
Described yeast saccharomyces cerevisiae spore powder is added in the solution that contains metal ion.
9. a yeast saccharomyces cerevisiae spore as described as claim 1～6 is as the application of lipoid material sorbent material.
10. yeast saccharomyces cerevisiae spore according to claim 9, as the application of lipoid material sorbent material, is characterized in that:
Described yeast saccharomyces cerevisiae spore is carried out obtaining yeast saccharomyces cerevisiae spore powder after purifying and lyophilize processing;
Described yeast saccharomyces cerevisiae spore powder is added in lipoid material.
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| CN112424234A (en) * | 2018-06-08 | 2021-02-26 | 麦基托科学公司 | Method for preparing chitosan |
| CN113862211A (en) * | 2021-09-13 | 2021-12-31 | 江南大学 | Application of chs3 saccharomyces cerevisiae spores and preparation method of beta-glucan microspheres |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107986454A (en) * | 2017-12-04 | 2018-05-04 | 湖南大学 | A kind of method using towel gourd load saccharomyces cerevisiae material degradation malachite green wastewater |
| CN112424234A (en) * | 2018-06-08 | 2021-02-26 | 麦基托科学公司 | Method for preparing chitosan |
| JP2021526810A (en) * | 2018-06-08 | 2021-10-11 | ミキト サイエンシズ インコーポレイテッドＭｙｋｉｔｏ Ｓｃｉｅｎｃｅｓ Ｉｎｃ． | How to make chitosan |
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| CN108658251A (en) * | 2018-07-03 | 2018-10-16 | 南京云实环境科技有限公司 | It is a kind of for the inorganic agent of textile waste, preparation method and its treatment process |
| CN108658251B (en) * | 2018-07-03 | 2021-11-16 | 南京乐透思环保科技有限公司 | Treating agent for textile wastewater, preparation method and treatment process thereof |
| CN113862211A (en) * | 2021-09-13 | 2021-12-31 | 江南大学 | Application of chs3 saccharomyces cerevisiae spores and preparation method of beta-glucan microspheres |
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Application publication date: 20131211 |