CN114292770B - Alcaligenes aquaticus, production method and application thereof - Google Patents
Alcaligenes aquaticus, production method and application thereof Download PDFInfo
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- CN114292770B CN114292770B CN202111456900.3A CN202111456900A CN114292770B CN 114292770 B CN114292770 B CN 114292770B CN 202111456900 A CN202111456900 A CN 202111456900A CN 114292770 B CN114292770 B CN 114292770B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
Abstract
The invention provides alcaligenes aquaticus, a production method and application thereof. An alcaligenes aquaticus, characterized in that the strain of alcaligenes aquaticus is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.21390. The invention selects the xylitol wastewater with low cost as a fermentation substrate of the alkali bacillus for water production, thereby reducing the pollution treatment investment of enterprises, and simultaneously reducing the production cost of the alkali bacillus for water production so as to realize the recycling utilization of waste.
Description
Technical Field
The invention belongs to the technical field of biology, and relates to alcaligenes aquaticus, a production method and application thereof.
Background
The cartap pesticide wastewater has the characteristics of high concentration of organic matters, deep chromaticity, large toxic effect, complex components, difficult degradation, poor biodegradability and the like. The method for degrading cartap pesticide wastewater by utilizing microorganisms has the advantages of low cost, no toxicity, no secondary pollution and the like, and is one of the most promising methods for removing pollution. At present, researches on cartap wastewater biological treatment technology are focused on aspects of treatment technology development, reactor treatment effect evaluation and the like, and researches on cartap characteristic pollution factor degradation bacteria are less.
The invention discloses a method for producing alcaligenes by using xylitol wastewater as a fermentation medium, wherein the alcaligenes produced by using the culture water is used for degrading cartap pesticide wastewater, and the adopted strain is the alcaligenes aquaticus which is screened from a pesticide wastewater system and stored in a national microorganism collection center.
In the practical application process of purifying COD and ammonia nitrogen in water by using the water producing alcaligenes, a large amount of thalli are needed, at present, the production of the water producing alcaligenes refers to the conventional biological fermentation mode, fermentation substrates mainly comprise bacterial culture media, such AS beef extract peptone culture media and LB culture media (the application number 202110317984.6 of the invention discloses using the LB culture media to carry out the water producing alcaligenes AS1 culture), the cost of raw materials of the culture media is calculated to be about 2000 yuan/ton according to market purchase price, and the cost of purifying the water by using the water producing alcaligenes is high due to high fermentation cost, so that the wide application of the culture media is limited.
Along with the continuous development and utilization of xylitol in the fields of foods and medicines, the xylitol market is pretty and the price is raised, so that new factories are built in various places, the old factories are expanded continuously, the yield is increased, the discharge amount of xylitol production wastewater is increased, and the municipal wastewater treatment load is increased. The applicant finds that the xylitol wastewater can be used for culturing the alkali bacillus in water production to replace beef extract peptone culture medium and LB culture medium, thereby changing waste into valuable.
Disclosure of Invention
The invention aims to provide alcaligenes aquaticus, a production method and application thereof.
An alcaligenes aquaticus, characterized in that the strain of alcaligenes aquaticus is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.21390.
The invention also provides a production method of the bacillus caldus, which is characterized in that sterilized xylitol wastewater with the pH of 6.0-9.0 is used as a culture medium for producing bacillus caldus.
Further, the production method comprises the following steps,
s1, preparing a culture medium: adjusting the pH value of xylitol wastewater to be between 6.0 and 9.0 by sodium hydroxide or sulfuric acid, sterilizing the xylitol wastewater in an autoclave at 115 ℃ for 30min, and taking the sterilized wastewater as a fermentation medium;
s2, preparing seed liquid: selecting 2-3 loops of a strain of the alcaligenes for producing water, inoculating the strain into a fermentation culture medium, and performing shaking culture for 24-48 hours at a rotating speed of 120rpm/min to obtain a bacterial liquid female parent after the culture is completed; inoculating a bacterial liquid female parent into a primary fermentation tank, wherein the inoculum size is 6-8%, and culturing for 12-24 hours at 25-37 ℃ by adopting a fermentation culture medium to obtain seed liquid;
s3, fermentation preparation of bacterial liquid: inoculating seed liquid into the secondary fermentation tank with inoculum size of 4-6%, culturing at 25-37deg.C for 18-48 hr with fermentation medium to obtain viable bacteria of Alkalibacterium in water production.
The invention also provides application of the bacillus caldus in wastewater treatment, which is characterized in that: the wastewater contains cartap pesticide.
Further, the wastewater treatment comprises the following steps,
s1', anaerobic biochemical treatment, namely, after the pH value of the wastewater is regulated to 6-9, introducing the wastewater into an anaerobic biochemical device, wherein the anaerobic biochemical device is filled with activated carbon with the effective volume of 30% and the particle size of 1-6 meshes, and the culture conditions, namely, pH=6-9, and COD: n: p=200:5:1, do (dissolved oxygen) < 1.5mg/L, temperature=25-30 ℃, hydraulic retention time 24-48h;
s2', aerobic biochemical treatment, namely, adjusting the pH value of the effluent of the anaerobic biochemical device to 6-9, and then introducing the effluent into the aerobic biochemical device, wherein the aerobic biochemical device is filled with volcanic rock with the effective volume of 20% and activated carbon with the effective volume of 20% and the particle size of 30-80 meshes, and the culture conditions are as follows, wherein the pH value=6-9, and the COD: n: p=100:5:1, do (dissolved oxygen) 3-6mg/L, temperature=25-30 ℃, hydraulic retention time 24-48h;
the anaerobic biochemical device and the aerobic biochemical device are both inoculated with the living bacterial culture, and the number of living bacteria of the living bacterial culture reaches 1.5x10 9 CFU/ml, and the inoculation amount is 5 per mill.
The invention selects the xylitol wastewater with low cost as a fermentation substrate of the alkali bacillus for water production, thereby reducing the pollution treatment investment of enterprises, and simultaneously reducing the production cost of the alkali bacillus for water production so as to realize the recycling utilization of waste.
The alkaline bacillus aquaticus strain adopted by the invention is obtained by separating and screening cartap pesticide wastewater, and the biological preservation is described below.
Latin school name: alcaligenes aquatilis;
chinese translated name: producing alkali bacillus by water;
the preservation units are fully named: preserving in China general microbiological culture Collection center;
the preservation unit is abbreviated as: CGMCC;
deposit unit address: the institute of microorganisms of national academy of sciences of China, national institute of sciences, no. 1, no. 3, north Chen West Lu, the Korean region of Beijing;
preservation date: 12 months 17 days 2020;
preservation number: CGMCC No.21390.
Detailed Description
Example 1: production of water-producing alcaligenes agent
Preparation of a culture medium: adjusting pH=6.0 (pH 5.4, shandong Cheng sea environmental protection technology Co., ltd.) of xylitol wastewater with 30% sodium hydroxide solution, sterilizing at 115deg.C for 30min in an autoclave to eliminate the influence of mixed bacteria in wastewater on bulk fermentation, and using sterilized wastewater as fermentation medium;
seed liquid preparation: picking a water-producing alcaligenes strain 2 ring (the strain is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No. 21390), inoculating the strain in a sterilized fermentation medium, and performing shaking culture for 24 hours at the rotating speed of 120rpm/min to obtain a bacterial liquid female parent after the culture is completed; inoculating a bacterial liquid female parent into a primary fermentation tank, wherein the inoculum size is 6%, and culturing for 12 hours at 25 ℃ by introducing air to obtain seed liquid;
and (3) fermenting bacterial liquid: inoculating seed solution into a secondary fermentation tank, culturing at 25deg.C for 18 hr to obtain viable cell culture with bacterial density of 2X10 9 cfu/ml。
Example 2: production of water-producing alcaligenes agent
S1, preparing a culture medium: adjusting the pH=9.0 of the xylitol wastewater by using a sodium hydroxide solution with the mass concentration of 30% (the xylitol wastewater is the same as in example 1), sterilizing the xylitol wastewater in an autoclave at 115 ℃ for 30min so as to eliminate the influence of mixed bacteria in the wastewater on bulk fermentation, wherein the sterilized wastewater is used as a fermentation medium;
s2, preparing seed liquid: picking an alkali bacillus strain 3 ring (the strain is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No. 21390), inoculating the strain to a sterilized fermentation medium, and performing shaking culture for 48 hours at a rotating speed of 120rpm/min to obtain a bacterial liquid female parent after the culture is completed; inoculating a bacterial liquid female parent into a primary fermentation tank, wherein the inoculum size is 8%, and culturing for 24 hours at 37 ℃ by introducing air to obtain seed liquid;
s3, fermentation preparation of bacterial liquid: inoculating seed solution into a secondary fermentation tank, culturing at 37deg.C for 48 hr to obtain viable cell culture with bacterial density of 2X10 9 cfu/ml。
Example 3: degradation effect of water production alkaline bacillus agent on cartap wastewater
Cartap waste water: cod=33000 mg/L, salinity 13.11%, ph=9, ammonia nitrogen=3082 mg/L, wastewater source: hunan Hao Hua Biotechnology Co., ltd.
The cartap wastewater is diluted by a certain multiple for treatment, the wastewater treatment comprises the following steps of S1', anaerobic biochemical treatment, the wastewater is directly introduced into an anaerobic biochemical device, the anaerobic biochemical device is filled with activated carbon with the effective volume of 30% and the particle size of 1-6 meshes, and anaerobic culture parameters are as follows: ph=9, cod: n: p=200:5:1, do (dissolved oxygen) 1mg/mL, temperature=25-30 ℃, hydraulic retention time see table below;
s2', aerobic biochemical treatment, namely, adjusting the pH value of the water discharged from the anaerobic biochemical device to 6-9, and then introducing the water into the aerobic biochemical device, wherein the aerobic biochemical device is filled with volcanic rock with the effective volume of 20% and activated carbon with the effective volume of 20% and the particle size of 30-80 meshes, and the aerobic culture parameters are as follows: ph=9, cod: n: p=100:5:1, DO (dissolved oxygen) 4mg/mL, temperature 25-30deg.C, hydraulic retention time see table below;
the anaerobic biochemical device and the aerobic biochemical device are both inoculated with the living bacterial culture, and the living bacterial count of the living bacterial cultureUp to 1.5X10 9 CFU/ml, inoculum size 5%.
TABLE 2 Biochemical stage removal effect of Alkalobacterium Alkaloids from Water production on cartap wastewater
According to the experiment, in the application of cartap wastewater, the removal rate of COD and ammonia nitrogen reaches 88%, and the treatment effect is good.
Example 4 comparison of the Effect of different microbial Agents in the treatment of cartap wastewater
The experiment adopts the screened aerobic strengthening bacteria (model: bioPower300 PLUS) of national environmental protection technology Co., ltd.) of producing alcaligenes and Wuhan water to add 1 per mill of cartap wastewater in strain, and the same reaction conditions are as follows: experiments were performed at ph=7.5, COD concentration of feed water=4000 mg/L, ammonia nitrogen concentration=200 mg/L, temperature 35 ℃, dissolved oxygen=3.5, residence time=48 h, and experimental effects are shown in the following table.
TABLE 3 comparison of the treatment effects of different microbial agents
Through a comparison experiment, the alcaligenes aquaticus has a remarkable improvement on the removal effect of COD and ammonia nitrogen in cartap wastewater compared with aerobic strengthening bacteria (model: bioPower300 PLUS) of national environmental protection technology limited company of Wuhan water.
Example 5 production of Alkalibacterium by Water under different culture Medium Density conditions
The experiment sets a conventional beef extract culture medium and takes xylitol wastewater as a fermentation culture medium, and after culturing alcaligenes aquaticus for 48 hours in a shaking table, the bacterial density is observed to evaluate a fermentation product;
experimental conditions:
remarks: beef extract peptone medium: 10g of fish meal peptone, 3g of beef extract, 5g of sodium chloride and 1000ml of distilled water, wherein the pH=7-8;
xylitol fermentation medium: xylitol waste water 1000ml, ph=7.5.
Compared with beef extract peptone culture medium, the xylitol fermentation culture medium has obviously improved culture effect through a comparison experiment.
Claims (1)
1. A production method of alcaligenes aquaticus, the strain of which is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.21390, is characterized in that sterilized xylitol wastewater with pH of 6.0-9.0 is used as a culture medium of the alcaligenes aquaticus;
the production method comprises the following steps of S1, preparing a culture medium: adjusting the pH value of xylitol wastewater to be between 6.0 and 9.0 by sodium hydroxide or sulfuric acid, sterilizing the xylitol wastewater in an autoclave at 115 ℃ for 30min, and taking the sterilized wastewater as a fermentation medium;
s2, preparing seed liquid: selecting 2-3 loops of a strain of the alcaligenes for producing water, inoculating the strain into a fermentation culture medium, and performing shaking culture for 24-48 hours at a rotating speed of 120rpm/min to obtain a bacterial liquid female parent after the culture is completed; inoculating a bacterial liquid female parent into a primary fermentation tank, wherein the inoculum size is 6-8%, and culturing for 12-24 hours at 25-37 ℃ by adopting a fermentation culture medium to obtain seed liquid;
s3, fermentation preparation of bacterial liquid: inoculating seed liquid into the secondary fermentation tank with inoculum size of 4-6%, culturing at 25-37deg.C for 18-48 hr with fermentation medium to obtain viable bacteria of Alkalibacterium in water production.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110092485A (en) * | 2019-05-15 | 2019-08-06 | 江苏宜裕环保科技有限公司 | A kind of biological rejuvenation reactant and preparation method thereof for Industrial Wastewater Treatment |
| CN112408602A (en) * | 2020-11-27 | 2021-02-26 | 江苏蓝必盛化工环保股份有限公司 | Salt-tolerant denitrification technology in high-salt pesticide chemical wastewater |
| CN113215027A (en) * | 2021-03-25 | 2021-08-06 | 中国科学院微生物研究所 | Alcaligenes aquaticum AS1 and application thereof in sewage treatment |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110092485A (en) * | 2019-05-15 | 2019-08-06 | 江苏宜裕环保科技有限公司 | A kind of biological rejuvenation reactant and preparation method thereof for Industrial Wastewater Treatment |
| CN112408602A (en) * | 2020-11-27 | 2021-02-26 | 江苏蓝必盛化工环保股份有限公司 | Salt-tolerant denitrification technology in high-salt pesticide chemical wastewater |
| CN113215027A (en) * | 2021-03-25 | 2021-08-06 | 中国科学院微生物研究所 | Alcaligenes aquaticum AS1 and application thereof in sewage treatment |
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