CN112694982B - Candida utilis and application thereof - Google Patents
Candida utilis and application thereof Download PDFInfo
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- CN112694982B CN112694982B CN202110171558.6A CN202110171558A CN112694982B CN 112694982 B CN112694982 B CN 112694982B CN 202110171558 A CN202110171558 A CN 202110171558A CN 112694982 B CN112694982 B CN 112694982B
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- candida utilis
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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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- C—CHEMISTRY; METALLURGY
- 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
- C02F2101/20—Heavy metals or heavy metal compounds
Abstract
The invention discloses Candida utilis and application thereof, relates to the technical field of microorganism application, and provides Candida utilis (Candida utilis) KUZ 511 with the preservation date of 2020, 10 months and 28 days, the preservation unit is China center for type culture Collection, and the preservation number is CCTCC M2020649. The invention also provides application of the Candida utilis (Candida utilis) KUZ 511 in adsorption of radioactive uranium ions. The invention has the beneficial effects that: the Candida utilis (Candida utilis) KUZ 511 has a good adsorption effect on radioactive uranium ions, the maximum uranium resistance of the Candida utilis (Candida utilis) KUZ reaches 1200mg/L, the maximum enrichment amount of uranium is 456.75mg/g, and the maximum enrichment amount and the uranium resistance of the Candida utilis in the invention are far higher than those of uranium in the prior art.
Description
Technical Field
The invention relates to the field of microbial remediation, and particularly relates to candida utilis and application thereof in uranium adsorption.
Background
Nuclear energy is an important energy source to replace fossil energy as an economic and clean energy source that is widely developed. Uranium (1) 235 U) is the main nuclear energy raw material and has large demand. However, the mining of excess uranium ore results in leakage of uranium ions into the natural environment. Leakage of harmful uranium into the environment can lead to many diseases, such as cancer and leukemia, due to acute toxicity of uranium and severe internal irradiation. The long half-life and trace concentrations of radioactivity u (vi) can migrate through the biological chain and be bio-enriched, ultimately threatening human health. Thus, a material is selected that is suitable and effective for uranium removalIs important.
As a new environment-friendly, efficient and economical sewage treatment and recycling technology, biotechnology has been increasingly paid attention to by people. The nuclear waste has complex components, wide related range, multiple influencing factors and different properties, and a plurality of researchers find that the effect is good when the microorganisms are used as the biological enrichment agent to remove uranium from the solution. Biological enrichment is one of the most attractive alternatives and has significant advantages over traditional methods. Meanwhile, it is environment-friendly and does not cause any secondary environmental pollution. The mechanism of interaction between microorganisms and radionuclides such as uranium has become an interesting research project for many researchers.
At present, the main methods for removing U (VI) from wastewater are as follows: ion exchange, electrochemical techniques, chemical precipitation and membrane filtration. Compared with the conventional removal method, the biological enrichment is more effective and more environment-friendly, and only a small amount of by-products are generated. Many researchers use various microorganisms, such as algae and fungi, for removing u (vi) from aqueous solutions. Although the methods can realize the adsorption purification of the U (VI) ions, the methods have the defects of high cost, secondary pollution and incapability of achieving the recycling of both the adsorbent and the U (VI).
In recent years, the removal of heavy metal ions from dilute solution by using biological materials is increasingly researched, and a lot of important progresses are obtained, the removal and recovery of U (VI) by using the biological technology is superior to the traditional treatment method in the aspects of investment, operation management, process and the like, and the characteristics make the environmental microbial wastewater treatment technology go forward in the direction of no toxicity, no harm and no secondary pollution.
The author is doctor's academic paper of Liu Lei's "mechanism and biological effect research of uranium enrichment by microorganisms" and records that U is biologically enriched by Candida utilis, but the maximum biological enrichment capacity is only 41.15mg/g, and the industrial application requirements cannot be met.
Disclosure of Invention
The invention aims to provide candida utilis capable of improving the biological enrichment of uranium.
The invention solves the technical problems through the following technical means:
in a first aspect, the invention provides a Candida utilis (Candida utilis) KUZ 511, wherein the Candida utilis is preserved in 28 days 10 months 2020, the preservation unit is China center for type culture Collection, and the preservation number is CCTCC NO: M2020649.
In a second aspect, the invention provides the use of Candida utilis (Candida utilis) KUZ 511 for adsorbing radioactive uranium ions, in particular radioactive uranium ions in sewage.
In a third aspect, the invention provides a method for removing radioactive uranium ions in sewage, which mainly comprises the following steps:
s1, inoculating the Candida utilis KUZ 51 of the first aspect into a liquid culture medium for culture, and centrifuging to obtain thalli;
s2, adjusting the pH value of the sewage containing radioactive uranium ions, adding the bacterial cells obtained in the step S1 into the sewage, and stirring and adsorbing the radioactive uranium ions.
Further, the method also comprises a recycling step S3, wherein the recycling step S3 is to collect the mushroom dregs in the tail liquid enriched in the step S2, add a desorbent aqueous solution into the mushroom dregs, desorb, separate and recycle the mushroom dregs, and obtain the candida utilis and radioactive uranium ions.
Preferably, the liquid culture medium in the step S1 is a YEDP culture medium, the culture temperature is 20-32 ℃, the culture time is 20-28h, the culture mode is shaking culture, and the rotation speed adopted by the shaking culture is 100-250 r/min.
Preferably, in the step S2, the pH value of the radioactive uranium ion-containing sewage is adjusted to 4.0-7.0, the temperature of stirring and adsorption is 20-32 ℃, the adsorption time is 30-60min, and the stirring rotation speed is 50-150 r/min.
Preferably, the mass ratio of the bacteria to the sewage in the step S2 is 1:200-1000, and the concentration of radioactive uranium ions in the sewage is 2-500 mg/L.
Preferably, the step S3 is implemented by desorbing at the speed of 100-150r/min for 10-30min, the aqueous desorbent solution is an ammonium carbonate aqueous solution with the mass concentration of 5%, and the mass ratio of the bacterial residues to the aqueous desorbent solution is 2: 1.
The invention has the beneficial effects that:
1. the Candida utilis (Candida utilis) KUZ 511 provided by the invention has high surface protein quality, has a good adsorption effect on radioactive uranium ions, has the maximum uranium resistance of 1200mg/L, has the maximum enrichment of 456.75mg/g, is far higher than the maximum enrichment of uranium and the maximum uranium resistance of Candida utilis in the prior art, widens the concentration range of radioactive uranium ions in biological sewage treatment, and protects the environment.
2. The method for removing radioactive uranium ions in sewage provided by the invention has a simple process, can effectively separate the radioactive uranium ions in the sewage, can recycle Candida utilis (Candida utilis) KUZ 511 serving as an adsorbent, can discharge the residual water after the radioactive uranium ions are detected to be in accordance with the sewage discharge standard, can recycle the residual water, and has good social benefit and economic benefit.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Test materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The specific techniques or conditions not specified in the examples can be performed according to the techniques or conditions described in the literature in the field or according to the product specification.
Wherein the concentration of U (VI) in the solution is determined using inductively coupled plasma atomic emission spectrometry (ICP-AES) (ICP 6300, Thermo Fisher Scientific, USA).
Wherein the maximum adsorption capacity of the candida utilis for enriching uranium is calculated through a Langmuir model
q e -the amount of u (vi) adsorbed by the biosorbent, mg/g; c e -u (vi) equilibrium concentration, mg/L; q m -maximum removal, mg/g; b-Langmuir constant, L/mg.
Wherein the YEDP (Yeast Extract Peptone Dextrose-Medium) culture Medium comprises the following components: 1L of distilled water contains 10g of yeast powder, 20g of peptone and 20g of glucose, wherein the solid medium contains 20g of agar.
Example 1 screening, purification and identification of highly resistant Candida utilis (Candida utilis) KUZ 511
(1) Soil near the uranium radionuclide uranium ore in northern mountain of Gansu is taken as a sample, the collected sample is put into a self-sealing bag and brought back to a laboratory, 1 g of soil with radionuclide uranium is taken and dissolved in a 100 ml conical flask, then 1 ml of soil solution is taken and added into 9 ml of deionized water for gradual dilution, and soil suspensions with different dilutions are formed. And (3) coating the diluted soil solution on YEDP culture media with different uranium concentrations, culturing at 28 ℃ until microbial spots grow, and then continuously separating and purifying the microorganisms.
(2) The strain is cultured in enrichment mode, namely the strain is inoculated to YEDP culture media with different concentrations of U (VI), the strain is cultured at constant temperature of 28 ℃ until stains appear, the strain shows better colony growth condition under the stress condition of low concentration of U (VI), when the concentration of U (VI) is more than 200mg/L U (VI), the biomass of the strain rapidly decreases along with the increase of the concentration of U (VI), finally the strain resisting 1200mg/L U (VI) is separated out, and the strain is repeatedly purified and stored in a refrigerator at 4 ℃ for later use.
The morphological and physiochemical characteristics of the strain are as follows:
colony color: milky white, aerobic: aerobic, bacterial colony size: (3.5 to 4.5) μm × (7 to 13) μm, growth temperature: 28 degrees, optimum pH: 4-7, reproduction mode: and (4) sprouting.
(3) The strain selected in step (2) with greater resistance to U (VI) was cultured in liquid, the genome was extracted, PCR amplification was performed using yeast universal primers ITS1/ITS4 (5'-TCCGTA GGT GAA CCT GCG G-3'/5'-TCC TCC GCT TAT TGA TAT GC-3'), sequencing was performed by a biological company after electrophoretic analysis, the sequencing result was submitted to NCBI (national Center for Biotechnology information) database, homology comparison analysis was performed using BLAST program with ITS sequences existing in the database, and the strain was determined to be Candida utilis and named KUZ 511.
Example 2 Candida utilis (Candida utilis) KUZ 511 for adsorptive recovery of radioactive uranium ions
The method adopts Candida utilis (Candida utilis) KUZ 511 to absorb and recover radioactive uranium ions, and specifically comprises the following steps:
s1, inoculating the Candida utilis (Candida utilis) KUZ 511 screened in the example 1 into a YEDP liquid culture medium, carrying out shake culture at 28 ℃ and 100r/min for 24h, and then carrying out centrifugal separation to obtain thalli;
s2, adjusting the pH value of sewage containing radioactive uranium ions to 4.0 by hydrochloric acid, adding thalli, wherein the mass ratio of the thalli to the sewage is 1:400, stirring at 28 ℃ and 100r/min to adsorb the radioactive uranium ions, and the adsorption is carried out for 30min to reach saturation, wherein the concentration of the radioactive uranium ions in the sewage is 20 mg/L;
s3, centrifugally collecting the mushroom dregs in the enrichment tail liquid, adding an ammonium carbonate aqueous solution with the mass concentration of 5% into the mushroom dregs, enabling the mass ratio of the mushroom dregs to the ammonium carbonate aqueous solution to be 2:1, desorbing for 15min at the rotating speed of 100r/min, then centrifuging, taking the precipitate to obtain candida utilis, and taking the supernatant to obtain radioactive uranium ions. As a result, the recovery was found to be 89.76%.
Example 3 Candida utilis (Candida utilis) KUZ 511 for adsorptive recovery of radioactive uranium ions
The method for adsorbing and recovering radioactive uranium ions by adopting Candida utilis (Candida utilis) KUZ 511 comprises the following steps:
s1, inoculating the Candida utilis (Candida utilis) KUZ 511 in the embodiment 1 into a liquid culture medium, carrying out shaking culture at 30 ℃ and 150r/min for 20h, and then carrying out centrifugal separation to obtain thalli;
s2, adjusting the pH value of sewage containing radioactive uranium ions to 6.0 by hydrochloric acid, adding thalli, wherein the mass ratio of the thalli to the sewage is 1:500, stirring at 35 ℃ and 120r/min to adsorb the radioactive uranium ions, and the adsorption is carried out for 30min to reach saturation, wherein the concentration of the radioactive uranium ions in the sewage is 60 mg/L;
s3, centrifugally collecting the mushroom dregs in the enrichment tail liquid, adding an ammonium carbonate aqueous solution with the mass concentration of 5% into the mushroom dregs, enabling the mass ratio of the mushroom dregs to the ammonium carbonate aqueous solution to be 2:1, desorbing for 10min at the rotating speed of 150r/min, then centrifuging, taking the precipitate to obtain candida utilis, and taking the supernatant to obtain radioactive uranium ions. As a result, the recovery rate was determined to be 92.65%.
Example 4 Candida utilis (Candida utilis) KUZ 511 for adsorptive recovery of radioactive uranium ions
The method for adsorbing and recovering radioactive uranium ions by adopting Candida utilis (Candida utilis) KUZ 511 comprises the following steps:
s1, inoculating the Candida utilis (Candida utilis) KUZ 511 in the embodiment 1 into a liquid culture medium, carrying out shaking culture at 32 ℃ and 125r/min for 24 hours, and then carrying out centrifugal separation to obtain thalli;
s2, adjusting the pH value of sewage containing radioactive uranium ions to 7.0 by hydrochloric acid, adding thalli, wherein the mass ratio of the thalli to the sewage is 1:500, stirring at 30 ℃ and 120r/min to adsorb the radioactive uranium ions, and the adsorption is carried out for 40min to reach saturation, wherein the concentration of the radioactive uranium ions in the sewage is 100 mg/L;
s3, centrifugally collecting the mushroom dregs in the enrichment tail liquid, adding an ammonium carbonate aqueous solution with the mass concentration of 5% into the mushroom dregs, enabling the mass ratio of the mushroom dregs to the ammonium carbonate aqueous solution to be 2:1, desorbing for 10min at the rotating speed of 125r/min, then centrifuging, taking the precipitate to obtain candida utilis, and taking the supernatant to obtain radioactive uranium ions. As a result, the recovery rate was found to be 91.12%.
Example 5 use of Candida utilis, Aspergillus niger and Mucor circinelloides for adsorption recovery of radioactive uranium ions
Comparative example 1
This comparative example differs from example 1 in that: candida utilis (Candida utilis) KUZ 511 is replaced by Candida utilis in the research on mechanism and biological effect of uranium enrichment by microorganisms in doctor' S academic paper, then the maximum uranium resistance of the Candida utilis is measured on YEDP culture media containing different concentrations of U (VI), and the maximum adsorption enrichment is obtained in operation steps S1 and S2.
Comparative example 2
This comparative example differs from example 1 in that: candida utilis (Candida utilis) KUZ 511 was replaced with Aspergillus Niger (Aspergillus Niger AS3.316, available from the institute of microbiology, China academy of sciences) and then the maximum uranium resistance was measured on YEDP medium containing different concentrations of U (VI), and the maximum adsorption enrichment was obtained in operation S1 and S2.
Comparative example 3
This comparative example differs from example 1 in that: candida utilis (Candida utilis) KUZ 511 was replaced with Mucor circinelloides (Mucor circinelloides AS3.2514, available from the institute of microbiology, China institute of sciences, culture Collection), and then the maximum uranium resistance was measured on YEDP media containing different concentrations of U (VI), and maximum adsorption enrichment was obtained in operations S1 and S2.
The maximum uranium resistance and the maximum adsorption enrichment of each of the strains of example 1 and comparative examples 1 to 3 were measured, and the results are shown in table 1.
Table 1 shows the results of the measurement of each strain
As can be seen from Table 1, the maximum uranium resistance and the maximum adsorption enrichment of other strains are significantly lower than that of Candida utilis (Candida utilis) KUZ 511 in the present invention.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (9)
1. Candida utilis (A. utilis:Candida utilis) KUZ 511, wherein the preservation number of the Candida utilis is CCTCC NO: M2020649.
2. Use of the candida utilis according to claim 1 for the adsorption of radioactive uranium ions.
3. Use of the candida utilis according to claim 1 for adsorbing radioactive uranium ions in sewage.
4. A method for removing radioactive uranium ions in sewage is characterized by comprising the following steps: s1, inoculating the Candida utilis of claim 1 into a liquid culture medium for culture, and centrifuging to obtain thalli; and S2, adjusting the pH value of the sewage containing radioactive uranium ions, adding the bacterial cells obtained in the step S1 into the sewage, and stirring and adsorbing the radioactive uranium ions.
5. The method of claim 4, further comprising a recycling step S3, wherein the recycling step S3 is to collect the mushroom dregs in the tail liquid enriched in the S2 step, and a desorbent aqueous solution is added into the mushroom dregs for desorption and separation recycling, so as to obtain the Candida utilis and the radioactive uranium ions.
6. The method of claim 5, wherein the aqueous desorbent solution is an aqueous ammonium carbonate solution, and the mass ratio of the mushroom residue to the aqueous desorbent solution is 2: 1.
7. The method according to claim 4 or 5, wherein the liquid medium in step S1 is a YEDP medium, and the pH of the radioactive uranium ion-containing wastewater is adjusted to 4.0 to 7.0 in step S2.
8. The method as set forth in claim 7, wherein the mass ratio of the bacteria to the sewage in the step S2 is 1: 200-1000.
9. The method of claim 8, wherein the concentration of radioactive uranium ions in the wastewater in step S2 is 2-500 mg/L.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013033456A2 (en) * | 2011-09-02 | 2013-03-07 | Board Of Trustees Of Michigan State University | Microbial nanowires and methods of making and using |
| WO2018144807A2 (en) * | 2017-02-02 | 2018-08-09 | Massachusetts Institute Of Technology | Engineered yeast as a method for bioremediation |
| CN110511880A (en) * | 2018-08-14 | 2019-11-29 | 西南科技大学 | A kind of resistance to uranium bacterium and its method for degradation treatment heavy metal accumulation plant |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013033456A2 (en) * | 2011-09-02 | 2013-03-07 | Board Of Trustees Of Michigan State University | Microbial nanowires and methods of making and using |
| WO2018144807A2 (en) * | 2017-02-02 | 2018-08-09 | Massachusetts Institute Of Technology | Engineered yeast as a method for bioremediation |
| CN110511880A (en) * | 2018-08-14 | 2019-11-29 | 西南科技大学 | A kind of resistance to uranium bacterium and its method for degradation treatment heavy metal accumulation plant |
Non-Patent Citations (3)
| Title |
|---|
| Bioaccumulation of uranium by Candida utilis: Investigated by water chemistry and biological effects;Lei Liu et al.;《Environmental Research》;20210102;第194卷;第1-9页 * |
| Effect of selenium on uranium biosorption;A. Nakajima;《Journal of Radioanalytical and Nuclear Chemistry》;20011231;第247卷(第2期);第347-350页 * |
| 微生物富集铀的机理及生物学效应研究;刘磊;《中国优秀博硕士学位论文全文数据库(博士)基础科学辑》;20190823(第8期);第1-3页 * |
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