CN112299562A - A method for promoting the degradation of carbon, nitrogen and phosphorus by microalgae using yeast secretions - Google Patents

A method for promoting the degradation of carbon, nitrogen and phosphorus by microalgae using yeast secretions Download PDF

Info

Publication number
CN112299562A
CN112299562A CN201910691228.2A CN201910691228A CN112299562A CN 112299562 A CN112299562 A CN 112299562A CN 201910691228 A CN201910691228 A CN 201910691228A CN 112299562 A CN112299562 A CN 112299562A
Authority
CN
China
Prior art keywords
microalgae
yeast
nitrogen
culture
phosphorus
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201910691228.2A
Other languages
Chinese (zh)
Inventor
刘晖
钟玉鸣
王伟
王琴
童英林
柳建良
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhongkai University of Agriculture and Engineering
Original Assignee
Zhongkai University of Agriculture and Engineering
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zhongkai University of Agriculture and Engineering filed Critical Zhongkai University of Agriculture and Engineering
Priority to CN201910691228.2A priority Critical patent/CN112299562A/en
Publication of CN112299562A publication Critical patent/CN112299562A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • 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
    • C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
    • C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; 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
    • C12N1/12—Unicellular algae; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; 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
    • C12N1/16—Yeasts; Culture media therefor
    • 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
    • C02F2101/105—Phosphorus compounds
    • 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
    • C02F2101/16—Nitrogen compounds, e.g. ammonia
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2209/00—Controlling or monitoring parameters in water treatment
    • C02F2209/08—Chemical Oxygen Demand [COD]; Biological Oxygen Demand [BOD]

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Mycology (AREA)
  • Botany (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Water Supply & Treatment (AREA)
  • Environmental & Geological Engineering (AREA)
  • Hydrology & Water Resources (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

本发明公开了一种利用酵母分泌物促进微藻降解碳、氮、磷的方法，其利用酵母的分泌物作为微藻的生长刺激因素，从而促进微藻降解碳、氮、磷的能力。本发明通过环保、绿色的方法促进微藻的蛋白合成与生长，可以更快速的增加微藻蛋白的合成量，提高其在养殖废水中对碳、氮、磷的处理效率，且操作方法简便易行。

Figure 201910691228

The invention discloses a method for promoting microalgae to degrade carbon, nitrogen and phosphorus by utilizing yeast secretions. The yeast secretions are used as growth stimulators of microalgae, thereby promoting the ability of microalgae to degrade carbon, nitrogen and phosphorus. The invention promotes the protein synthesis and growth of the microalgae through an environment-friendly and green method, can increase the synthesis amount of the microalgae protein more rapidly, improve the treatment efficiency of carbon, nitrogen and phosphorus in the breeding wastewater, and the operation method is simple and easy Row.

Figure 201910691228

Description

Method for promoting microalgae to degrade carbon, nitrogen and phosphorus by using yeast secretion
Technical Field
The invention belongs to the technical field of wastewater reclamation and environmental protection, and particularly relates to a method for promoting microalgae to degrade carbon, nitrogen and phosphorus by using yeast secretion.
Background
At present, the water environment pollution problem in China is widely concerned, and the problems of ammonia nitrogen, total nitrogen, organic matters and novel toxic pollutants are endless. The total nitrogen emission control requirement is implemented in the new 'ten water entries' of the country for coastal land level and above cities, so that the requirement for water treatment is improved by a higher level. The microalgae wastewater recycling technology can effectively solve the problem.
The microalgae can efficiently remove pollutants such as COD, nitrogen, phosphorus and the like in the water body under the heterotrophic-autotrophic mixed condition and absorb greenhouse gases; meanwhile, the algae can be recovered to obtain resources and energy sources with high added values such as grease, protein and the like. Many studies have shown that: the treatment technology is suitable for treating various kinds of culture wastewater, and is a novel potential biological treatment technology for recycling wastewater phosphorus. In the last 50 th century, microalgae have been used in sewage and wastewater treatment projects, and the microalgae technology has not become the mainstream sewage treatment technology due to limitations of treatment efficiency, technology and the like. In recent years, with the emergence of crisis, a novel sustainable sewage treatment system becomes a trigger for the development of microalgae technology, and the microalgae technology becomes a hot spot of the next generation of sewage treatment research again. The microalgae recycling technology represented by the chlorella pyrenoidosa can well meet the practical requirement.
The method is derived from the degradation process of various nitrogen-containing pollutants for the ammonia nitrogen which is a pollutant accounting for more than 60 percent of the total nitrogen of the water body. The microalgae represented by the chlorella can efficiently convert nitrogen pollutants in a water body into in-vivo proteins. Under autotrophic culture, the protein content of the chlorella accounts for 42% -58% of the dry weight of cells, and data show that the protein amino acid composition in the chlorella is higher than the protein content standard which is issued by the World Health Organization (WHO) and the Food and Agriculture Organization (FAO) of the United nations and is applied to human nutrition, and the high protein content and the high growth rate of the chlorella enable the protein in the chlorella to be called as a substance which has the best substitute for edible protein; it also has higher emulsification function and is a high-quality protein source; the protein of the chlorella also has other nutritive values, and the contained protein and polypeptide have the potential of resisting oxidation, improving the immunity of human bodies, resisting tumors, viruses, fungi and the like. Therefore, if a method for increasing the content of proteins in microalgae represented by chlorella is obtained, it is advantageous to improve the economy of microalgae wastewater treatment, improve the treatment efficiency, and advance a technology for recycling wastewater.
Disclosure of Invention
The invention aims to: provides a method for promoting microalgae to degrade carbon, nitrogen and phosphorus in wastewater.
In order to achieve the above object, the present invention provides a method for promoting microalgae to degrade carbon, nitrogen and phosphorus by using yeast secretion, comprising the following steps:
s1, obtaining microalgae: purchasing microalgae or separating the microalgae from the culture wastewater, and culturing to obtain a microalgae culture;
s2, culturing yeast: selecting yeast, streaking on a sterilized YM solid culture medium plate, placing the plate in an illumination incubator, culturing for 7 days under the conditions of 30 ℃ and 2000Lux, transferring the plate onto an YM culture medium inclined plane, culturing for 5 days under the conditions of 30 ℃ and 2000Lux, transferring the plate into an M1 liquid culture medium for culturing for 2 days, and harvesting yeast culture after yeast growth is finished;
s3, obtaining yeast secretion: centrifuging or suction-filtering the yeast culture obtained in the step S2 to obtain yeast, then placing the obtained yeast in an M1 liquid culture medium with the volume of 1L, culturing for 2 days under the conditions of illumination all day and 25 ℃, measuring the removal rate of ammonia nitrogen and total nitrogen in the M1 culture medium, and finishing the culture when the removal rate reaches 70%; if the removal rate is lower than 70%, repeating the cultivation for 1-2 times under the conditions of illumination all day and 25 ℃, and then measuring the removal rate again until the removal rate reaches 70%; after the culture is finished, obtaining yeast secretion by centrifugation or suction filtration;
s4, strengthening microalgae: mixing and culturing the microalgae culture obtained in the step S1 and the yeast secretion obtained in the step S3 according to the volume ratio of 6: 1-1: 1 to obtain strengthened microalgae;
s5, putting the strengthened microalgae obtained in the S4 into culture wastewater, and culturing for 4-7 days to promote degradation of carbon, nitrogen and phosphorus;
wherein each liter of the M1 liquid culture medium comprises the following components: CH (CH)3COONa 3.68g，Na2HPO4 28.73mg， NH4Cl 57.27mg，CaCl2.2H2O17.2 mg, pH adjusted to 7.0 with NaOH.
As a preferred embodiment of the method of the present invention, in S1, the method for purchasing microalgae and the culturing comprises: adding microalgae into an M0 liquid culture medium, culturing for 2-3 days under the conditions of 25 ℃ and 5500-6000 Lux, and shaking for 2-3 times every day to obtain a microalgae culture;
wherein each liter of the M0 liquid culture medium comprises the following components: NaNO3 1500mg，K2HPO4 0.04mg， MgSO47H2O 75mg，CaCl22H2O36 mg, citric acid 6mg, EDTA 1mg, Na2CO3 20mg。
As a preferred technical solution of the method of the present invention, in S1, the method for separating microalgae from aquaculture wastewater and culturing comprises: taking 10-20L of aquaculture wastewater, rough filtering, centrifuging, removing suspended matters and large-scale plankton, then performing aeration and precipitation, removing the precipitate, performing suction filtration again, collecting the precipitate on a filter membrane, transferring the precipitate into an M0 liquid culture medium, culturing for 2-3 days under the condition of 25 ℃ 4000-5000 Lux full-day illumination, shaking for 2-3 times every day to obtain a primary culture, then inoculating the primary culture into an M0 liquid culture medium, culturing for 7 days under the conditions of 25 ℃ full-day illumination and 150rpm, measuring the removal rates of nitrate, total nitrogen and total phosphorus every day, replacing the M0 liquid culture medium when the removal rates respectively reach 50%, 40% and 40%, and obtaining the primary culture after the removal effects of the nitrate, the total nitrogen and the total phosphorus respectively reach 80%, 50% and 60%; diluting the primary culture, finding microalgae cells under a low magnification magnifier, sucking out, centrifuging at 5000 rpm, inoculating the obtained precipitate to M0 solid culture medium, and streaking for separation to obtain microalgae; the step of separating the microalgae from the culture wastewater can be repeated for 2-3 times, so that sufficient microalgae can be obtained. And then adding the microalgae into an M0 liquid culture medium, culturing for 2-3 days under the conditions of 25 ℃ and 5500-6000 Lux, and shaking for 2-3 times every day to obtain a microalgae culture.
Wherein each liter of the M0 liquid culture medium comprises the following components: NaNO3 1500mg，K2HPO4 0.04mg， MgSO47H2O 75mg，CaCl22H2O36 mg, citric acid 6mg, EDTA 1mg, Na2CO3 20mg；
The M0 solid culture medium comprises the following components per liter: NaNO3 1500mg，K2HPO4 0.04mg，MgSO47H2O 75mg/L，CaCl22H2O36 mg, citric acid 6mg, EDTA 1mg, Na2CO320mg, 2 wt% agar powder.
As a preferred embodiment of the method of the present invention, the purchased microalgae is Chlorella pyrenoidosa (Chlorella pyrenoidosa) or Chlorella pyrenoidosa.
As a preferable technical scheme of the method, in S3, the rotating speed of the centrifugation is not lower than 10000rpm, and the aperture of the suction filtration is not less than 0.22 μm.
Compared with the prior art, the invention has the following beneficial effects:
the invention promotes the synthesis and growth of protein of microalgae by an environment-friendly and green method, improves the degradation efficiency of microalgae on carbon, nitrogen and phosphorus in the culture wastewater, and has simple and easy operation method.
Drawings
FIG. 1 shows the result of the effect of yeast secretion in promoting the removal of COD from chlorella;
FIG. 2 shows the result of the effect of yeast secretion in promoting the removal of total nitrogen from chlorella;
FIG. 3 shows the result of the effect of yeast secretion in promoting the removal of total phosphorus from chlorella.
FIG. 4 shows that the secretion of test yeast promotes the average removal rate of COD, total nitrogen and total phosphorus in chlorella.
Detailed Description
In order to make the objects, technical solutions and advantageous technical effects of the present invention clearer, the present invention is further described in detail with reference to the following embodiments. It should be understood that the embodiments described in this specification are only for the purpose of illustrating the invention and are not to be construed as limiting the invention, and the parameters, proportions and the like of the embodiments may be suitably selected without materially affecting the results.
The culture media and the specific formulations used in the following examples are as follows:
m1 liquid medium per liter included: CH (CH)3COONa 3.68g，Na2HPO4 28.73mg，NH4Cl 57.27mg， CaCl2.2H2O17.2 mg, pH adjusted to 7.0 with NaOH.
M0 liquid medium per liter included: NaNO3 1500mg，K2HPO4 0.04mg，MgSO47H2O 75mg，CaCl22H2O36 mg, citric acid 6mg, EDTA 1mg, Na2CO3 20mg。
The M0 solid culture medium comprises the following components in each liter: NaNO3 1500mg，K2HPO4 0.04mg，MgSO47H2O 75mg/L，CaCl22H2O36 mg, citric acid 6mg, EDTA 1mg, Na2CO320mg, 2 wt% agar powder.
The remaining medium is a medium commonly used in the art.
The strain activation and culture method is a conventional method in the art, unless otherwise specified.
Example 1
1. Taking 30L of wastewater from a waterfowl breeding wastewater factory, removing suspended matters, large-scale plankton and oil stains in the wastewater after conventional isolation, then carrying out closed aeration for 30min, precipitating for 30min, and circulating for 2-3 times for later use.
2. Obtaining and culturing microalgae: obtaining concentrated solution through suction filtration, and collecting precipitate on a filter membrane; washing the precipitate with sterile water for 1-2 times, transferring to an M0 liquid culture medium, culturing for 3 days at 25 ℃ under the condition of illumination of 4000-5000 Lux all day, and shaking for 3 times every day to obtain a primary culture; inoculating the primary culture in an M0 liquid culture medium, culturing under the conditions of illumination all day and 25 ℃ at 150rpm, measuring the removal efficiency of nitrate, total nitrogen and total phosphorus every day, and replacing the culture medium M0 after the removal efficiency of nitrate, total nitrogen and total phosphorus respectively reaches 50%, 40% and 40%, until the removal efficiency of nitrate, total nitrogen and total phosphorus respectively reaches 80%, 50% and 60%, so as to obtain a primary culture; diluting the primary culture, finding chlorella cells in a low magnification magnifier, sucking out, centrifuging at 5000 r/min, collecting precipitate, inoculating to M0 solid culture medium, and streaking, purifying and separating to obtain chlorella strain as wastewater. This step can be repeated 2-3 times. And then adding the wastewater chlorella strain into an M0 liquid culture medium, culturing for 2-3 days under the conditions of 25 ℃ and 5500-6000 Lux, and shaking for 2-3 times every day to obtain a chlorella culture.
3. The yeast is purchased from a standard strain bank by a purchasing method, and the strain is rhodotorula toruloides (Rhodosporidium toruloides). Yeast secretion was obtained as follows. Firstly, after the strain is activated and cultured, the yeast is harvested under the centrifugation condition of 10000 r/rpm, and the supernatant is removed. Then, the obtained yeast was placed in a 1L volume of M1 liquid medium, and cultured for 2 days under the conditions of light irradiation and 25 ℃ throughout the day, and the residual nitrogen in the supernatant M1 medium was measured, whereby the ammonia nitrogen removal rate was 79.5%, the total nitrogen removal rate was 75.1%, and the yeast culture was completed when the 70% standard was reached. After the culture is finished, the filtrate obtained by 0.22um suction filtration is the yeast secretion which needs to be used within 12-24 hours.
4. And (4) treating the wastewater. Mixing the obtained yeast secretion and chlorella culture according to the ratio of 1: 3 inoculating into a 5L sequencing batch reactor, and inoculating the treated aquaculture wastewater, wherein the pollutant concentrations are respectively as follows: 45-50mg/L of total nitrogen, 30-35mg/L of ammonia nitrogen, 36-40mg/L of total phosphorus, 800-1000mg/L of COD and 6-7 of pH. In the process of treating wastewater, only adding chlorella group, yeast secretion and chlorella control group,
after 7-day sequencing batch operation, the removal efficiency of COD, ammonia nitrogen, total nitrogen, phosphate and total phosphorus was compared, and the results are shown in FIGS. 1 to 3.
Example 2
1. Taking 20L of wastewater from a pig raising wastewater factory, removing suspended matters, large plankton and oil stains in the wastewater after conventional isolation, performing closed aeration for 25min, precipitating for 30min, and circulating for 2-3 times for later use.
2. Obtaining and culturing microalgae: the chlorella is chlorella pyrenoidosa purchased from a standard strain bank, and is transferred to an M0 liquid culture medium after being activated, the chlorella pyrenoidosa is cultured for 7 days under the conditions of illumination of the whole day, 25 ℃ and 150rpm, the removal efficiency of nitrate, total nitrogen and total phosphorus is measured every day, the culture medium M0 is replaced after the removal efficiency of the nitrate, the total nitrogen and the total phosphorus respectively reaches 50 percent, 40 percent and 40 percent, and a primary culture is obtained until the removal efficiency of the nitrate, the total nitrogen and the total phosphorus respectively reaches 80 percent, 50 percent and 60 percent; diluting the primary culture, finding chlorella cells in a low magnification magnifier, sucking out, centrifuging at 5000 r/min, collecting precipitate, inoculating to M0 solid culture medium, and streaking, purifying and separating to obtain chlorella strain as wastewater. This step can be repeated 2-3 times. And then adding the wastewater chlorella strain into an M0 liquid culture medium, culturing for 2-3 days under the conditions of 25 ℃ and 5500-6000 Lux, and shaking for 2-3 times every day to obtain a chlorella culture.
3. Yeast was Rhodotorula glutinis, a commercially available strain of Rhodotorula glutinis, obtained from a stock of standard strains, and was processed in the same manner as in example 1.
4. And (4) treating the wastewater. The obtained chlorella and yeast secretion are respectively put into the culture wastewater according to the proportion of 3:1 and 6:1 for use. The pollutant concentration of the aquaculture wastewater is respectively as follows: 35-40mg/L of total nitrogen, 30-35mg/L of ammonia nitrogen, 40mg/L of total phosphorus, 1200mg/L of COD (chemical oxygen demand) and 6-7 of pH. (the treatment effect is related to the purchased microalgae, the experiment is repeated for 5 times, the COD removal rate is 3:1, the TN and TP removal rates of the groups are the highest, the average removal rate is 79.8 percent and 86.3 percent respectively, the COD removal rates are close to each other, the 6:1 group is 86.8 percent, and the 3:1 group is 83.6 percent.
Appropriate changes and modifications to the embodiments described above will become apparent to those skilled in the art from the disclosure and teachings of the foregoing description. Therefore, the present invention is not limited to the specific embodiments disclosed and described above, and some modifications and variations of the present invention should fall within the scope of the claims of the present invention. Furthermore, although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

Claims (5)

1.一种利用酵母分泌物促进微藻降解碳、氮、磷的方法，其特征在于，包括如下步骤：1. a method utilizing yeast secretion to promote microalgae degradation of carbon, nitrogen, phosphorus, is characterized in that, comprises the steps: S1、获取微藻：购买微藻或从养殖废水中分离得到微藻，经培养，得到微藻培养物；S1. Obtaining microalgae: purchasing microalgae or separating and obtaining microalgae from aquaculture wastewater, and culturing to obtain a microalgae culture; S2、培养酵母：挑取酵母菌在已灭菌的YM固体培养基平板上划线，置于光照培养箱中，在30度、2000Lux条件下培养7天，再转接到YM培养基斜面上，在30度、2000Lux条件下培养5天后，转接入M1液体培养基培养2天，待酵母生长完毕后，收获酵母培养物；S2. Yeast cultivation: pick yeast and streak on the sterilized YM solid medium plate, place it in a light incubator, cultivate it for 7 days under the conditions of 30 degrees and 2000Lux, and then transfer it to the YM medium slant. , after culturing for 5 days under the conditions of 30 degrees and 2000Lux, transfer to M1 liquid medium for 2 days, and harvest the yeast culture after the yeast growth is completed; S3、获得酵母分泌物：将S2所述酵母培养物离心或抽滤后收获酵母，然后将所述收获酵母置于1L体积的M1液体培养基，经过全天光照、25度条件下培养2天后，测定M1培养基中氨氮与总氮的去除率，当所述去除率达到70％，则培养完毕；如去除率低于70％，则重复全天光照、25度条件下培养1～2次，然后再次测定所述去除率直至达到70％；培养完毕后，通过离心或抽滤，得到酵母分泌物；S3, obtaining yeast secretion: the yeast culture described in S2 is centrifuged or suction filtered to harvest the yeast, and then the harvested yeast is placed in a 1L volume of M1 liquid medium, and after culturing for 2 days under the conditions of all-day light and 25 degrees , measure the removal rate of ammonia nitrogen and total nitrogen in M1 medium, when the removal rate reaches 70%, the cultivation is completed; if the removal rate is lower than 70%, repeat the cultivation under the conditions of all-day light and 25 degrees for 1 to 2 times , and then measure the removal rate again until it reaches 70%; after culturing, obtain yeast secretion by centrifugation or suction filtration; S4、强化微藻：将S1所得微藻培养物和S3所得酵母分泌物按照6:1～1:1的体积比例混合，在M1液体培养基中，25度、5500～6000Lux条件下培养5天，得到强化后的微藻；S4. Strengthening microalgae: Mix the microalgae culture obtained from S1 and the yeast secretion obtained from S3 according to the volume ratio of 6:1 to 1:1, and cultivate in M1 liquid medium at 25 degrees and 5500 to 6000Lux for 5 days , to obtain enhanced microalgae; S5、将S4所得强化后的微藻投入养殖废水中，培养4～7天，促进碳、氮、磷的降解；S5, put the enhanced microalgae obtained in S4 into aquaculture wastewater, and cultivate for 4 to 7 days to promote the degradation of carbon, nitrogen and phosphorus; 其中，每升所述M1液体培养基包括如下组分：CH3COONa 3.68g，Na2HPO4 28.73mg，NH4Cl57.27mg，CaCl2.2H2O 17.2mg，用NaOH调整pH至7.0。Wherein, each liter of the M1 liquid medium includes the following components: CH 3 COONa 3.68 g, Na 2 HPO 4 28.73 mg, NH 4 Cl 57.27 mg, CaCl 2 .2H 2 O 17.2 mg, and the pH was adjusted to 7.0 with NaOH. 2.根据权利要求1所述的利用酵母分泌物促进微藻降解碳、氮、磷的方法，其特征在于，S1中，所述购买微藻和所述培养的方法是：将微藻加入M0液体培养基中，在25度、5500～6000Lux条件下培养2～3天，每天摇动2～3次，获得微藻培养物；2. the method that utilizes yeast secretion according to claim 1 to promote microalgae to degrade carbon, nitrogen, phosphorus, is characterized in that, in S1, the method for described buying microalgae and described cultivating is: adding microalgae to MO. In the liquid medium, culture at 25 degrees and 5500-6000Lux for 2-3 days, shaking 2-3 times a day to obtain a microalgae culture; 其中，每升所述M0液体培养基包括如下组分：NaNO3 1500mg，K2HPO4 0.04mg，MgSO47H2O75mg，CaCl22H2O 36mg，柠檬酸6mg，EDTA 1mg，Na2CO3 20mg。Wherein, each liter of the MO liquid medium includes the following components: NaNO 3 1500 mg, K 2 HPO 4 0.04 mg, MgSO 4 7H 2 O 75 mg, CaCl 2 2H 2 O 36 mg, citric acid 6 mg, EDTA 1 mg, Na 2 CO 3 20mg. 3.根据权利要求1所述的利用酵母分泌物促进微藻降解碳、氮、磷的方法，其特征在于，S1中，所述从养殖废水中分离得到微藻和所述培养的方法是：取10～20L养殖废水，经过粗滤、离心，去除悬浮物和大型浮游生物后闷曝、沉淀，去除沉淀再抽滤，收集滤膜上的沉淀物，转移到M0液体培养基中，在25度、4000～5000Lux全天光照条件下培养2～3天，每日摇动2～3次，获得初培物，然后接种于M0液体培养基中，在25度、全天光照、150转/min条件下培养7天，每天测定硝酸盐、总氮与总磷的去除率，当去除率分别达到50％、40％和40％后更换M0液体培养基，直至硝酸盐、总氮与总磷的去除效果分别达到80％、50％和60％后获得初步培养物；将初步培养物稀释，低倍放大镜下找到微藻细胞，吸出后5000转/min离心，所得沉淀物接种于M0固体培养基，经划线分离，得到微藻；然后将微藻加入M0液体培养基中，在25度、5500～6000Lux条件下培养2～3天，每天摇动2～3次，获得微藻培养物；3. the method that utilizes yeast secretion to promote microalgae to degrade carbon, nitrogen, phosphorus according to claim 1, is characterized in that, in S1, the described method that separates and obtains microalgae and described cultivation from culturing wastewater is: Take 10-20L of aquaculture wastewater, go through rough filtration and centrifugation, remove suspended solids and large plankton, then suffocate and precipitate, remove the sediment and then suction filter, collect the sediment on the filter membrane, transfer it to M0 liquid medium, and at 25 Cultivation for 2 to 3 days under 4000-5000Lux all-day light conditions, shaking 2-3 times a day to obtain primary culture, then inoculated in M0 liquid medium, at 25 degrees, all-day light, 150 rpm/min Cultivated for 7 days under the condition, the removal rate of nitrate, total nitrogen and total phosphorus was measured every day, when the removal rate reached 50%, 40% and 40%, respectively, the M0 liquid medium was replaced, until the nitrate, total nitrogen and total phosphorus were reduced. After the removal effect reaches 80%, 50% and 60% respectively, the preliminary culture is obtained; the preliminary culture is diluted, the microalgae cells are found under a low magnification magnifying glass, and centrifuged at 5000 rpm after suction, and the obtained precipitate is inoculated in M0 solid medium , and separated by streaking to obtain microalgae; then add the microalgae to the M0 liquid medium, cultivate at 25 degrees, 5500-6000Lux for 2-3 days, and shake 2-3 times a day to obtain a microalgae culture; 其中，每升所述M0液体培养基包括如下组分：NaNO3 1500mg，K2HPO4 0.04mg，MgSO47H2O75mg，CaCl22H2O 36mg，柠檬酸6mg，EDTA 1mg，Na2CO3 20mg；Wherein, each liter of the MO liquid medium includes the following components: NaNO 3 1500 mg, K 2 HPO 4 0.04 mg, MgSO 4 7H 2 O 75 mg, CaCl 2 2H 2 O 36 mg, citric acid 6 mg, EDTA 1 mg, Na 2 CO 3 20mg; 每升所述M0固体培养基包括如下组分：NaNO3 1500mg，K2HPO4 0.04mg，MgSO47H2O 75mg/L，CaCl22H2O 36mg，柠檬酸6mg，EDTA 1mg，Na2CO3 20mg，2wt％的琼脂粉。Each liter of the M0 solid medium includes the following components: NaNO 3 1500 mg, K 2 HPO 4 0.04 mg, MgSO 4 7H 2 O 75 mg/L, CaCl 2 2H 2 O 36 mg, citric acid 6 mg, EDTA 1 mg, Na 2 CO 3 20 mg, 2 wt% agar powder. 4.根据权利要求1所述的利用酵母分泌物促进微藻降解碳、氮、磷的方法，其特征在于，所述购买微藻是小球藻(Chlorella pyrenoidosa)或蛋白核小球藻。4. The method according to claim 1, wherein the purchased microalgae is Chlorella pyrenoidosa or Chlorella pyrenoidosa. 5.根据权利要求1所述的利用酵母分泌物促进微藻降解碳、氮、磷的方法，其特征在于，S3中，所述离心的转速不低于10000rpm，所述抽滤的孔径不小于0.22μm。5. the method that utilizes yeast secretion to promote microalgae to degrade carbon, nitrogen, phosphorus according to claim 1, is characterized in that, in S3, the rotating speed of described centrifugation is not less than 10000rpm, the pore size of described suction filtration is not less than 0.22μm.
CN201910691228.2A 2019-07-29 2019-07-29 A method for promoting the degradation of carbon, nitrogen and phosphorus by microalgae using yeast secretions Pending CN112299562A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910691228.2A CN112299562A (en) 2019-07-29 2019-07-29 A method for promoting the degradation of carbon, nitrogen and phosphorus by microalgae using yeast secretions

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910691228.2A CN112299562A (en) 2019-07-29 2019-07-29 A method for promoting the degradation of carbon, nitrogen and phosphorus by microalgae using yeast secretions

Publications (1)

Publication Number Publication Date
CN112299562A true CN112299562A (en) 2021-02-02

Family

ID=74328973

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910691228.2A Pending CN112299562A (en) 2019-07-29 2019-07-29 A method for promoting the degradation of carbon, nitrogen and phosphorus by microalgae using yeast secretions

Country Status (1)

Country Link
CN (1) CN112299562A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114231454A (en) * 2021-12-21 2022-03-25 北京天诚众合科技发展有限公司 Bacteria-algae composition for degrading water body and directionally eliminating blue algae, production process and device
CN115558623A (en) * 2022-11-07 2023-01-03 宁夏农林科学院农业资源与环境研究所（宁夏土壤与植物营养重点实验室） Preparation method of microbial agent and organic fertilizer for treating cow breeding wastewater

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6443182A (en) * 1987-08-10 1989-02-15 Osaka Seiken Kk Mixed cultured product, production and method thereof
US20130078686A1 (en) * 2011-09-27 2013-03-28 Synthetic Genomics, Inc. Production and secretion of fatty acids and fatty acid derivatives
CN104212720A (en) * 2014-09-04 2014-12-17 山东大学 Method for producing microalgae biomass by using monosodium glutamate ion-exchanged mother liquid
CN105441524A (en) * 2016-01-27 2016-03-30 扬州大学 Method for increasing yield of microalgae grease with saccharose as carbon source through co-culture
CN106929422A (en) * 2017-03-15 2017-07-07 华南理工大学 A kind of method that chlorella and yeast co-culture purification yeast wastewater
CN108624506A (en) * 2018-05-16 2018-10-09 华南理工大学 The method of microalgae and yeast mixed culture purification biogas slurry coproduction microbial biomass
CN109536388A (en) * 2018-11-26 2019-03-29 江苏科技大学 A method of micro algae growth speed and protein content are improved using silk reeling wastewater

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6443182A (en) * 1987-08-10 1989-02-15 Osaka Seiken Kk Mixed cultured product, production and method thereof
US20130078686A1 (en) * 2011-09-27 2013-03-28 Synthetic Genomics, Inc. Production and secretion of fatty acids and fatty acid derivatives
CN104212720A (en) * 2014-09-04 2014-12-17 山东大学 Method for producing microalgae biomass by using monosodium glutamate ion-exchanged mother liquid
CN105441524A (en) * 2016-01-27 2016-03-30 扬州大学 Method for increasing yield of microalgae grease with saccharose as carbon source through co-culture
CN106929422A (en) * 2017-03-15 2017-07-07 华南理工大学 A kind of method that chlorella and yeast co-culture purification yeast wastewater
CN108624506A (en) * 2018-05-16 2018-10-09 华南理工大学 The method of microalgae and yeast mixed culture purification biogas slurry coproduction microbial biomass
CN109536388A (en) * 2018-11-26 2019-03-29 江苏科技大学 A method of micro algae growth speed and protein content are improved using silk reeling wastewater

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114231454A (en) * 2021-12-21 2022-03-25 北京天诚众合科技发展有限公司 Bacteria-algae composition for degrading water body and directionally eliminating blue algae, production process and device
CN115558623A (en) * 2022-11-07 2023-01-03 宁夏农林科学院农业资源与环境研究所（宁夏土壤与植物营养重点实验室） Preparation method of microbial agent and organic fertilizer for treating cow breeding wastewater
CN115558623B (en) * 2022-11-07 2023-10-17 宁夏农林科学院农业资源与环境研究所（宁夏土壤与植物营养重点实验室） Preparation method of microbial agent and organic fertilizer for treating cow breeding wastewater

Similar Documents

Publication Publication Date Title
Yunos et al. Harvesting of microalgae (Chlorella sp.) from aquaculture bioflocs using an environmental-friendly chitosan-based bio-coagulant
CN107055946A (en) A kind of method that utilization chlorella purifies pig farm fermentation waste water
CN108587915B (en) Chlorella W5 capable of removing heavy metals in water with high heavy metal content and application thereof
CN107129950A (en) A kind of active phycomycete community and preparation method for purifying domestic sewage
CN107082489B (en) Method for synchronously removing manganese and nitrate in underground water
CN114105307A (en) Method for removing Cr in wastewater by using living heterotrophic microalgae6+Method (2)
CN109081447B (en) A kind of Chlorella-Acinetobacter-Pseudomonas combined method for removing nitrogen and phosphorus from aquaculture wastewater
CN111977800A (en) Method for treating sewage by using immobilized chlorella and rhodotorula benthamii and application
CN108220165A (en) Method for removing carbon, nitrogen and phosphorus pollution of culture wastewater by using associated flora to strengthen microalgae
CN108546649B (en) Chlorella W2 capable of removing heavy metals in water with high heavy metal content and application thereof
CN117865335B (en) A method for treating high-ammonia nitrogen wastewater using mixed algae membrane
CN112299562A (en) A method for promoting the degradation of carbon, nitrogen and phosphorus by microalgae using yeast secretions
CN105600942B (en) A method of biological flocculation is formed using cyanobacterial bloom
CN109576159B (en) Chlorella W4 capable of removing heavy metals in water with high heavy metal content and application thereof
CN107828679B (en) A kind of Rhodococcus rhodochrous strain XHRR1 for purifying ammonia in aquaculture water and its application
CN109110927B (en) Method for removing nitrogen and phosphorus in culture wastewater by combining chlorella and acinetobacter
CN109576160B (en) A kind of Chlorella W3 capable of removing heavy metals in water with high heavy metal content and its application
CN109576161B (en) Chlorella W1 capable of removing heavy metals in water with high heavy metal content and application thereof
CN111690546B (en) Rhodosporidium toruloides ZDFY1801 and application thereof
CN102294227B (en) Preparation method and application method of chlorella seawater purifying biological adsorbent
CN109293156A (en) A joint control method of pond water quality and sediment
CN105695355B (en) A method for preparing bacteria and algae cells by two rounds of co-cultivation and flocculation
CN112028251A (en) Method for improving ammonia nitrogen removal effect of microalgae on livestock and poultry breeding wastewater
CN102399823A (en) Method for extracting bioflocculant
CN114044576B (en) A method for purifying piggery wastewater by attaching and culturing Scenedesmus

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20210202