CN115322908B - Salt-tolerant aureobasidium pullulans and application thereof in dephosphorization of water body - Google Patents
Salt-tolerant aureobasidium pullulans and application thereof in dephosphorization of water body Download PDFInfo
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- CN115322908B CN115322908B CN202210620840.2A CN202210620840A CN115322908B CN 115322908 B CN115322908 B CN 115322908B CN 202210620840 A CN202210620840 A CN 202210620840A CN 115322908 B CN115322908 B CN 115322908B
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- 241000223678 Aureobasidium pullulans Species 0.000 title claims abstract description 50
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 28
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims abstract description 66
- 229910052698 phosphorus Inorganic materials 0.000 claims abstract description 66
- 239000011574 phosphorus Substances 0.000 claims abstract description 66
- 239000010865 sewage Substances 0.000 claims abstract description 6
- 239000010842 industrial wastewater Substances 0.000 claims abstract description 4
- 238000004321 preservation Methods 0.000 claims description 8
- 229920001218 Pullulan Polymers 0.000 claims description 4
- 235000019423 pullulan Nutrition 0.000 claims description 4
- 238000000034 method Methods 0.000 abstract description 18
- 229920000388 Polyphosphate Polymers 0.000 abstract description 11
- 239000001205 polyphosphate Substances 0.000 abstract description 11
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- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 description 1
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- WBTCZEPSIIFINA-MSFWTACDSA-J dipotassium;antimony(3+);(2r,3r)-2,3-dioxidobutanedioate;trihydrate Chemical compound O.O.O.[K+].[K+].[Sb+3].[Sb+3].[O-]C(=O)[C@H]([O-])[C@@H]([O-])C([O-])=O.[O-]C(=O)[C@H]([O-])[C@@H]([O-])C([O-])=O WBTCZEPSIIFINA-MSFWTACDSA-J 0.000 description 1
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
- B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/347—Use of yeasts or fungi
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- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/105—Phosphorus compounds
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Abstract
The invention relates to the field of microorganisms, in particular to a salt-tolerant aureobasidium pullulans strain and application thereof in water dephosphorization. The Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 provided by the invention can remove phosphorus efficiently, and the Aureobasidium pullulans GC02-5-42 can secrete a large amount of EPS in the growth process, so that phosphorus elements can be enriched from the environment, and the phosphorus elements are stored in the form of polyphosphate, so that polyphosphate particles can be formed in cells or EPS, and further the phosphorus concentration in the environment is reduced, so that the effect of removing excessive phosphorus in the environment is achieved; the aureobasidium pullulans GC02-5-42 has excellent salinity tolerance capability, can grow normally under the salinity condition, can remove phosphorus or enrich phosphorus in a low salinity environment and also remove phosphorus or enrich phosphorus in a high salinity environment, and is suitable for removing excessive phosphorus in high-salt industrial wastewater or domestic sewage or ocean.
Description
Technical Field
The invention relates to the field of microorganisms, in particular to a salt-tolerant aureobasidium pullulans strain and application thereof in water dephosphorization.
Background
The common wastewater dephosphorization method of the sewage treatment plant at present mainly comprises a biological method, a chemical method and a crystallization method. The most widely applied and studied biological phosphorus removal method is EBPR (enhanced biological phosphorus removal method enhanced biological phosphorus removal), and phosphate in wastewater is removed mainly by utilizing the principles of aerobic phosphorus absorption and anaerobic phosphorus release of phosphorus accumulating bacteria (PAOs). In an anaerobic state, the polyphosphate bacteria decompose polyphosphate (Poly-P) in the body, release Adenosine Triphosphate (ATP), utilize the ATP released to absorb organic matters in the solution, store in the body in the form of Poly beta-hydroxybutyrate (PHB), and release phosphate, namely anaerobic phosphorus release; in the aerobic state, the polyphosphate accumulating bacteria take up phosphorus in the wastewater by utilizing energy released by decomposing the poly beta-hydroxybutyric acid (PHB) stored in the body, synthesize polyphosphate and store the polyphosphate in the body. Finally, in the aerobic state, the phosphorus accumulating bacteria which complete the process of excessive phosphorus absorption are discharged out of the system in the form of residual sludge in the discharge system, so as to achieve the aim of removing phosphorus in sewage.
Although the EBPR process has many advantages, it has some drawbacks that are difficult to overcome, for example, in order to achieve the ideal treatment effect, the system needs to strictly control the anaerobic-aerobic alternate condition, and has strict requirements on the oxygen concentration under the anaerobic-aerobic condition, and at the same time, external factors such as salinity, temperature, pH and carbon source type have great influence on the dephosphorization effect of the phosphorus accumulating bacteria. Thus, there is a great need for new, more stable microbial phosphorus removal pathways with greater environmental tolerance.
In recent years, many scholars have studied to find that the extracellular polymer (extracellular polymeric substance, EPS) contains a large amount of phosphorus and plays an important role in the process of absorbing/releasing phosphorus. Unlike the EBPR process which is most widely researched at present, the phosphorus removal technology based on the microbial extracellular polymer EPS has better environmental tolerance and stability, and the oxygen concentration of the environment does not need to be controlled in the treatment process.
In natural environment, some microorganisms secrete extracellular polymers (extracellular polymeric substance, EPS), which are composed of saccharides, proteins, nucleic acids, lipids, etc., and the basic functions of EPS include polymerizing microbial cells, surface adhesion, formation of flocs and biofilms, cell recognition, structural elements of biofilms, cell protection barriers, moisture retention to prevent cell dehydration, adsorption of exogenous organic substances, adsorption of inorganic ions, etc. The high-salt environment can inhibit the growth of microorganisms, so that the microorganisms cannot fully exert the treatment effect, and therefore, the traditional biological treatment method is not suitable for treating the high-salt environment. When the salinity in the environment is less than 2%, the phosphorus removal capability of the phosphorus accumulating bacteria is less influenced by the salinity, and when the biological phosphorus removal system is continuously impacted by the wastewater with the salinity of 2%, the phosphorus removal capability of the phosphorus accumulating bacteria is completely destroyed.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide the salt-tolerant aureobasidium pullulans and the application thereof in the dephosphorization of water bodies, wherein the aureobasidium pullulans has high-efficiency dephosphorization capability and excellent high-salt tolerance capability.
For this purpose, the invention provides the following technical scheme:
the invention provides Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42, which are preserved in China center for type culture collection, wherein the preservation number is CCTCC NO: m2022155, deposit address: chinese, university of Wuhan, mail code 430072, date of preservation: 2022, 25 d 2.
A microbial preparation comprising the aureobasidium pullulans or a treated product, an inactivated product and a metabolite thereof.
Optionally, the treatment comprises a culture, concentrate, dilution, dry or extract of the culture of Aureobasidium pullulans.
The use of said Aureobasidium pullulans or said microbial preparation for dephosphorization, phosphorus enrichment or polyphosphate preparation.
Optionally, the removing phosphorus includes removing phosphorus element in the environment or reducing phosphorus element concentration in the environment.
Optionally, the environment comprises a body of water or soil;
the water body comprises industrial wastewater, domestic sewage, rivers or oceans.
Optionally, the environment includes a high salinity environment and a low salinity environment; in the high salinity environment, the salinity is more than or equal to 2% and less than or equal to 5%; the salinity in the low salinity environment is less than 2 percent.
A method for identifying the Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 comprises identifying based on a sequence shown in SEQ ID NO. 1.
Alternatively, the nucleotide sequence of primer ITS4 is shown as SEQ ID NO. 2 and the nucleotide sequence of primer ITS5 is shown as SEQ ID NO. 3 based on primer ITS4 and primer ITS5 designed as shown as SEQ ID NO. 1.
Optionally, the method comprises the following steps:
and (3) taking DNA of the strain as a template, adopting primers ITS4 and ITS5 to carry out PCR amplification to obtain an ITS sequence, and carrying out sequence comparison on the obtained ITS sequence.
The technical scheme of the invention has the following advantages:
1. the Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 provided by the invention can effectively remove phosphorus, and the Aureobasidium pullulans GC02-5-42 can secrete a large amount of EPS in the growth process, so that phosphorus elements can be enriched from the environment, and the phosphorus can be stored in a polyphosphate form, so that polyphosphate particles can be formed in cells or EPS, and further the phosphorus concentration in the environment is reduced, so that the effect of removing excessive phosphorus in the environment is achieved;
furthermore, the aureobasidium pullulans GC02-5-42 has excellent salinity tolerance, can grow normally under the high salinity condition, can remove or enrich phosphorus in a low salinity environment, can remove or enrich phosphorus in a high salinity environment, and is suitable for removing excessive phosphorus in high-salt industrial wastewater or domestic sewage or ocean.
The Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 provided by the invention are preserved in China center for type culture collection, and the preservation number is CCTCC NO: m2022155, deposit address: chinese, university of Wuhan, mail code 430072, date of preservation: 2022, 25 d 2.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the characteristics of the fungus body form of Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 under a microscope in example 2 of the present invention;
FIG. 2 shows colony morphology characteristics of Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 in example 2 of the present invention during various growth periods; the left panel shows colony morphological features grown for 3 days, and the right panel shows colony morphological features grown for 7 days;
FIG. 3 shows the results of ITS-based tree construction of Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 in example 2 of the present invention;
FIG. 4 shows the growth of Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 in the phosphorus-rich medium with different salinity in example 3 of the present invention;
FIG. 5 shows the total phosphorus in the culture supernatant of Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 in example 4 according to the present invention as a function of the culture time;
FIG. 6 is a graph showing total phosphorus in the supernatant of Pichia pastoris culture as well as the change in culture time in example 4 of the present invention.
Detailed Description
The following examples are provided for a better understanding of the present invention and are not limited to the preferred embodiments described herein, but are not intended to limit the scope of the invention, any product which is the same or similar to the present invention, whether in light of the present teachings or in combination with other prior art features, falls within the scope of the present invention.
The specific experimental procedures or conditions are not noted in the examples and may be followed by the operations or conditions of conventional experimental procedures described in the literature in this field. The reagents or apparatus used were conventional reagent products commercially available without the manufacturer's knowledge.
The components of the medium referred to in the examples below are all commercially available products.
EXAMPLE 1 isolation and purification of strains
1. Preparing a separation medium: the 7 media were Malt Extract Agar (MEA), corn meal agar (CDA), corn Meal Agar (CMA), glucose agar (SDA), yeast maltose agar (YM), potato Dextrose Agar (PDA) and yeast peptone dextrose agar (YPD), respectively. The details are given in the following table.
TABLE 1 Total nutrient Components of fungal Medium
2. Dilution of the sediment samples: 200-210cm gravity column of ocean floor of Dongpacific GC02-5 station position by DY27 voyageSediment samples, which were diluted with sterile water (dilution volumes of 10, respectively -1 ，10 -2 ，10 -3 )。
3. Separating and purifying: the total nutrient medium in Table 1 above was diluted with sterile water (dilution volume ratio 1:5) to simulate the low nutrient conditions of a deep sea biosphere to yield an oligonutrient medium. The oligotrophic medium also contained 50. Mu.g/mL ampicillin and 50. Mu.g/mL streptomycin. 100. Mu.L of the diluted sediment samples were respectively spread on plates of 7 kinds of oligotrophic media and incubated at 22 ℃. After the oligotrophic medium to be coated with the sediment sample had grown out of colonies, colonies were picked and inoculated into the total nutrient medium (undiluted total nutrient medium in table 1). Until only one colony is present on one plate.
EXAMPLE 2 biological identification of fungi
The biological identification of fungi mainly comprises the following steps of:
the GC02-5-42 strain was obtained by isolation and purification in example 1. The morphological characteristics of the strain under a microscope are shown in figure 1, and colony morphological characteristics of the strain in different growth periods on a solid plate culture medium are shown in figure 2: the colony was grown in PDA medium for 7 days and had a diameter of 29mm, and initially had a yeast-like, viscous, reddish color, and under a microscope, a clear root-like colony with a node spore.
To further confirm the resulting GC02-5-42 strain, the purified fungus was identified using the ITS sequence (SEQ ID NO: 1). DNA of the obtained strain is extracted, primers ITS4 (5'-TCCGTAGGTGAACCTGCGG-3', SEQ ID NO: 2) and ITS5 (5'-TCCTCCGCTTATTGATAGC-3', SEQ ID NO: 3) are adopted and amplified by PCR technology to obtain ITS sequence (SEQ ID NO: 1), and the sequence is subjected to sequence alignment with the corresponding sequence in NCBI database, and the species relationship of the fungus is determined according to the standard that the similarity is more than or equal to 97%, preferably 98% to 100%, and the tree construction result based on the ITS is shown in FIG. 3. Combining the morphological detection result and the ITS identification, the GC02-5-42 strain obtained by the invention is discovered to be Aureobasidium pullulans (Aureobasidium pullulans). Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 are preserved in China center for type culture collection, and the preservation number is CCTCC NO: m2022155, deposit address: chinese, university of Wuhan, mail code 430072, date of preservation: 2022, 25 d 2.
EXAMPLE 3 growth of Aureobasidium pullulans in high salt Medium
Taking out Aureobasidium pullulans (Aureobasidium pullulans) GC02-5-42 strain preserved at-80deg.C, thawing on ice, streaking on PDA (total nutrient potato dextrose agar medium in Table 1 of example 1) plate for resuscitation, picking single colony after bacterial colony grows on the plate, inoculating into YPD liquid medium (total nutrient yeast peptone dextrose agar medium in Table 1 of example 1 without adding agar powder), and culturing to medium OD 600 When=0.6-1, aureobasidium pullulans GC02-5-42 seed solution was obtained.
Inoculating Aureobasidium pullulans GC02-5-42 seed solution into phosphorus-rich culture medium with different salinity according to 1% (v/v) inoculum size, culturing in constant temperature shaking table at 28deg.C and 120rpm, sampling every 24 hr, and detecting OD 600 To examine the growth of the strain in various media, the results are shown in FIG. 4, in which Brevibacterium pullulans GC02-5-42 can grow normally in a medium with a salinity of 5%, and the biomass and growth rate thereof are almost the same as those in a medium without salt.
The formulation of the phosphorus-rich medium is as follows: KNO of 2g/L 3 5g/L CH 3 COONa·3H 2 O, K of 0.05g/L 2 HPO 4 KH of 0.2g/L 2 PO 4 MgSO at 0.2g/L 4 ·7H 2 O,0.5g/L CaCl 2 The concentration of sodium chloride is 0g/L,30g/L,50g/L,100g/L or 150g/L. The salinity of the obtained phosphorus-rich culture medium is 0%,3%,5%,10% and 15%.
EXAMPLE 4 phosphorus-accumulating action of Aureobasidium pullulans
To control the phosphorus enrichment of Aureobasidium pullulans GC02-5-42, the following experiment was performed using another Pichia pastoris (preservation number CCTCC NO: M2020860, published by patent document CN 113373072A) as a control.
Mixing at-80deg.CThe deposited Aureobasidium pullulans GC02-5-42 strain or Pichia pastoris is taken out, melted on ice, streaked and resuscitated on a PDA (the total nutrient potato dextrose agar medium in Table 1 of example 1) plate, after the colony grows out on the plate, single colony is selected and inoculated into YPD liquid medium (the total nutrient yeast peptone dextrose agar medium in Table 1 of example 1 is not added with agar powder) and cultured to the culture medium OD 600 When=0.6-1, aureobasidium pullulans GC02-5-42 seed solution or pichia pastoris seed solution was obtained.
Inoculating the seed solution into a phosphorus-rich culture medium according to an inoculum size of 1% (v/v), culturing in a constant temperature shaking table at 28 ℃ and 120rpm, sampling every 24 hours, centrifuging the sample 1200rpm for 10min, collecting the supernatant, and diluting 100 times by volume to obtain a sample to be tested.
The formula of the phosphorus-rich culture medium is as follows: KNO of 2g/L 3 5g/L CH 3 COONa·3H 2 O,0.05 g/L K 2 HPO 4 KH of 0.2g/L 2 PO 4 MgSO at 0.2g/L 4 ·7H 2 O。
The method for detecting the total phosphorus is measured according to the related detection method of the total phosphorus of the water body in GB/T12763.4-2007, and the method for detecting the total phosphorus by using potassium persulfate oxidative digestion and ascorbic acid reduction phosphomolybdic blue method is used.
The sample to be tested is oxidized by potassium persulfate at 110-120 ℃, the organic phosphorus compound is converted into inorganic phosphate, inorganic polymerized phosphorus is hydrolyzed into orthophosphate, the orthophosphate reacts with ammonium molybdate in an acidic medium to generate a phosphomolybdic yellow complex, the phosphomolybdic yellow complex is reduced into a phosphomolybdic blue complex by ascorbic acid in the presence of antimony potassium tartrate, and spectrophotometry is carried out at 882 nm. The total phosphorus concentration in water can be calculated as follows:
wherein:
c (TP-P) -the concentration of total phosphorus in a water sample is expressed in micromoles per cubic decimeter (mu mol/dm) 3 )；
-average absorbance of total phosphorus in the water sample;
A t -absorbance of turbidity in the water sample, which is 0 when turbidity correction is not required;
A b -blank absorbance;
a-standard working curve intercept;
b-standard work curve slope;
k-dilution factor.
Variation of total phosphorus in culture supernatants of Brevibacterium pullulans GC02-5-42 and Pichia guilliermondii over time, as shown in FIGS. 5 and 6, after 96 hours of cultivation, total phosphorus in culture supernatant of Brevibacterium pullulans GC02-5-42 was reduced from 120.32. Mu. Mol/L to 42.761. Mu. Mol/L, and total phosphorus in the environment was reduced by about 64.46%. While the control season also showed little change in total phosphorus in pichia pastoris.
In summary, the aureobasidium pullulans GC02-5-42 have good removal effect on total phosphorus in a water body in the process of removing excessive phosphorus elements in the water body, do not need to perform an anaerobic-aerobic stage, have higher tolerance on salinity compared with common PAOs, and have irreplaceable advantages in removing excessive phosphorus elements in high-salinity wastewater.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the invention.
Sequence listing
<110> institute of natural resources third sea
<120> a salt-tolerant aureobasidium pullulans and application thereof in water dephosphorization
<160> 3
<170> SIPOSequenceListing 1.0
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tgcctgcgga aggatcatta aagagtaagg gtgctcagcg cccgacctcc aaccctttgt 60
tgttaaaact accttgttgc tttggcggga ccgctcggtc tcgagccgct ggggattcgt 120
cccaggcgag cgcccgccag agttaaacca aactcttgtt attaaaccgg tcgtctgagt 180
taaaattttg aataaatcaa aactttcaac aacggatctc ttggttctcg catcgatgaa 240
gaacgcagcg aaatgcgata agtaatgtga attgcagaat tcagtgaatc atcgaatctt 300
tgaacgcaca ttgcgcccct tggtattccg aggggcatgc ctgttcgagc gtcattacac 360
cactcaagct atgcttggta ttgggtgccg tccttagttg ggcgcgcctt aaagacctcg 420
gcgaggcctc accggcttta ggcgtagtag aatttattcg aacgtctgtc aaaggagagg 480
acttctgccg actgaaacct ttatttttct cggtc 515
<210> 2
<211> 19
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 2
tccgtaggtg aacctgcgg 19
<210> 3
<211> 19
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 3
tcctccgctt attgatagc 19
Claims (4)
1. The application of the aureobasidium pullulans in the dephosphorization of the water body is characterized in that the aureobasidium pullulans is prepared by the following steps ofAureobasidium pullulans) GC02-5-42, the preservation number is CCTCC NO: m2022155.
2. The use of claim 1, wherein the dephosphorization comprises removing phosphorus from the body of water or reducing the concentration of phosphorus in the body of water.
3. The use according to claim 2, characterized in that,
the water body comprises industrial wastewater, domestic sewage, rivers or oceans.
4. A use according to claim 2 or claim 3, wherein the body of water comprises a body of high salinity water and a body of low salinity water; in the high salinity water body, the salinity is more than or equal to 2% and less than or equal to 5%; the salinity of the low salinity water body is less than 2 percent.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5399495A (en) * | 1993-04-28 | 1995-03-21 | The Upjohn Company | Microbial degradation of chemical pollutants |
| CN106244471A (en) * | 2016-10-08 | 2016-12-21 | 南京工业大学 | The fungal bacterial strain of a kind of resistance to heavy metal Hg and application thereof |
| CN107058123A (en) * | 2017-03-10 | 2017-08-18 | 中国海洋大学 | A kind of Aureobasidium pullulans and its application |
-
2022
- 2022-06-01 CN CN202210620840.2A patent/CN115322908B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5399495A (en) * | 1993-04-28 | 1995-03-21 | The Upjohn Company | Microbial degradation of chemical pollutants |
| CN106244471A (en) * | 2016-10-08 | 2016-12-21 | 南京工业大学 | The fungal bacterial strain of a kind of resistance to heavy metal Hg and application thereof |
| CN107058123A (en) * | 2017-03-10 | 2017-08-18 | 中国海洋大学 | A kind of Aureobasidium pullulans and its application |
Non-Patent Citations (6)
| Title |
|---|
| "Efficient phosphorus removal by a novel halotolerant fungus Aureobasidium sp. MSP8 and the application potential in saline industrial wastewater treatment";Xiaoying Zeng 等;《Bioresource Technology》;第第334卷卷;摘要，第2、6、7页 * |
| "Fungalcommunityanalysisinthedeep-seasedimentsofthePacific Ocean assessedbycomparisonofITS,18Sand28SribosomalDNA regions";Wei Xu 等;《Deep-Sea ResearchI》;第109卷;全文 * |
| 以淀粉废水、麦芽根制备生物材料――短梗霉多糖;邵荣;许琦;刘珊珊;余晓红;汪志君;方维明;;食品科学(第08期);全文 * |
| 出芽短梗霉吸附水体中共存Cr(Ⅵ)、Cd(Ⅱ)重金属离子研究;张玉玲,张兰英,王显胜,王晓晖,高松;吉林大学学报(地球科学版)(第03期);全文 * |
| 张玉玲,张兰英,王显胜,王晓晖,高松.出芽短梗霉吸附水体中共存Cr(Ⅵ)、Cd(Ⅱ)重金属离子研究.吉林大学学报(地球科学版).2005,(第03期),全文. * |
| 邵荣 ; 许琦 ; 刘珊珊 ; 余晓红 ; 汪志君 ; 方维明 ; .以淀粉废水、麦芽根制备生物材料――短梗霉多糖.食品科学.2007,(第08期),全文. * |
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