CN110699350B - Immobilized microbial inoculum and preparation method and application thereof - Google Patents

Immobilized microbial inoculum and preparation method and application thereof Download PDF

Info

Publication number
CN110699350B
CN110699350B CN201910718561.8A CN201910718561A CN110699350B CN 110699350 B CN110699350 B CN 110699350B CN 201910718561 A CN201910718561 A CN 201910718561A CN 110699350 B CN110699350 B CN 110699350B
Authority
CN
China
Prior art keywords
immobilized
acinetobacter
microbial inoculum
immobilized microbial
bacterial suspension
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.)
Active
Application number
CN201910718561.8A
Other languages
Chinese (zh)
Other versions
CN110699350A (en
Inventor
陈惠明
蒋自胜
翟德勤
余艳鸽
郑茂盛
商放泽
俞静雯
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.)
China Power Construction Ecological Environment Group Co ltd
Original Assignee
China Power Construction Ecological Environment Group Co ltd
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 China Power Construction Ecological Environment Group Co ltd filed Critical China Power Construction Ecological Environment Group Co ltd
Priority to CN201910718561.8A priority Critical patent/CN110699350B/en
Publication of CN110699350A publication Critical patent/CN110699350A/en
Application granted granted Critical
Publication of CN110699350B publication Critical patent/CN110699350B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/10Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
    • C12N11/12Cellulose or derivatives thereof
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/10Inorganic compounds
    • C02F2101/16Nitrogen compounds, e.g. ammonia

Abstract

The invention relates to the technical field of microbial inoculum immobilization, and particularly discloses an immobilized microbial inoculum, and a preparation method and application thereof. The immobilized microbial inoculum comprises acinetobacter and an immobilized carrier, wherein the immobilized carrier is rice bran. The preparation method of the immobilized microbial inoculum at least comprises the following steps: culturing the acinetobacter to obtain a bacterial suspension; and sterilizing the bran coat, uniformly mixing the bran coat and the bacterial suspension under the aseptic condition, and drying to obtain the immobilized microbial inoculum. According to the invention, the immobilized microbial inoculum is prepared, and the acinetobacter is loaded into the immobilized carrier, so that the physiological activity and stability of the microorganism are not affected, and an aerobic and anoxic microenvironment is provided for the microorganism, thereby being beneficial to the growth of the microbial inoculum and the performance of denitrification and denitrification of the microbial inoculum.

Description

Immobilized microbial inoculum and preparation method and application thereof
Technical Field
The invention relates to the technical field of microbial inoculum immobilization, in particular to an immobilized microbial inoculum and a preparation method and application thereof.
Background
The rapid development of industry and agriculture and the production activities of human beings bring serious pollution to the water environment. The exceeding of nitrogen is the most common pollution problem in water body pollution, which can cause eutrophication of water body, cause mass propagation of algae, reduce dissolved oxygen in the water body and cause black and odorous water body, thereby not only endangering the water safety of human beings, but also seriously influencing the ecological balance of the water body; in addition, excessive nitrogen discharge can cause the nitrogen balance to be destroyed, thereby causing the water body to lose the self-cleaning function. Therefore, the sewage rich in nitrogen can be discharged into the natural environment after the sewage is subjected to denitrification treatment and reaches the standard.
Biological denitrification is a denitrification method widely applied at present and has the advantages of good treatment effect, stable and reliable treatment process, convenient operation and management and the like. The traditional biological denitrification is completed by utilizing the aerobic autotrophic nitrification of nitrifying bacteria and the anaerobic heterotrophic denitrification of denitrifying bacteria, and the biological denitrification needs to provide two reactors with oxygen and oxygen respectively to carry out nitrification and denitrification respectively so as to achieve the aim of denitrification. Therefore, the reactor occupies a large space, the operation cost is high, and the process is complicated.
Immobilization is the physical or chemical means to confine free microorganisms or enzymes in a specific spatial region for more widespread and effective use. In the sewage with flow dynamics, because the flowing of the water body, the selection of the immobilized carrier material directly influences the physiological activity and stability of the microorganism, even the loss of the microorganism, the selection of the immobilized carrier which can not only ensure enough microorganism amount in unit volume, but also protect the microorganism to be in a high activity state becomes a hotspot of research.
Disclosure of Invention
Aiming at the problems of high cost, complex process, reduction of strain activity by an immobilization process and the like of the existing biological denitrification, the invention provides an immobilized microbial inoculum.
And a method for producing the immobilized microbial agent.
And the application of the immobilized microbial inoculum in the treatment of nitrogen-containing sewage.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
an immobilized bacteria agent comprises acinetobacter and an immobilized carrier, wherein the acinetobacter is preserved in the China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC NO. 16620; the immobilized carrier is rice bran.
The immobilized microbial inoculum provided by the invention has the following advantages:
the invention preserves the acinetobacter in the form of immobilized bacteria agent, loads the acinetobacter into the bran coat, solves the problem of difficult preservation of the acinetobacter, ensures that the physiological activity and stability of the microorganism are not affected, provides an aerobic and anoxic microenvironment for the microorganism, is beneficial to the growth of the bacteria agent and the performance of denitrification and denitrification of the bacteria agent, is also beneficial to keeping high viable bacteria concentration and improving the rate of degrading nitrogen-containing sewage when in use, and has the advantages of convenient application, separation and repeated use.
The invention takes the bran coat with small mass as the immobilized carrier, and is easily influenced by wind when being scattered to the water body, so that the immobilized microbial inoculum can be uniformly dispersed into the sewage, floats on the water surface, and is not easy to sink into the water to cause secondary pollution to the water body; meanwhile, the bran coat has obvious porous characteristic, large specific surface area and strong adsorption capacity, and after the immobilized microbial agent taking the bran coat as a carrier is added into sewage, the immobilized microbial agent can promote nitrogen-containing pollutants to migrate to the bran coat carrier, so that the bran coat can simultaneously enrich acinetobacter and the pollutants, and the contact between microorganisms and the pollutants is increased; the bran coat can also provide an independent microenvironment for the acinetobacter, so that the direct invasion of external adverse factors to the acinetobacter is shielded, and the removal efficiency of the acinetobacter to nitrogen-containing pollutants in a polluted environment is improved; and the bran coat as an organic matter can provide nutrient substances for the propagation of the acinetobacter and provide a nutritional basis for keeping high viable bacteria concentration of the acinetobacter, so that the bran coat immobilized microbial inoculum can realize the integration of enrichment and degradation of pollutants, and is beneficial to promoting the degradation of nitrogen-containing pollutants in sewage.
The Acinetobacter provided by the invention is an aerobic denitrifying bacterium, the Acinetobacter is obtained by enrichment screening through a biological culture means, the Acinetobacter has the characteristic of playing a role in denitrification under aerobic conditions, and the aerobic denitrifying bacterium strain is adopted to replace the conventional anaerobic denitrifying bacterium, so that the nitrification and the denitrification can be carried out in one reaction when the biological denitrification treatment of sewage is carried out, thereby simplifying the sewage treatment process, reducing the capital construction operation cost and improving the biological denitrification effect, and the Acinetobacter has good application prospect in the aspect of nitrogen-containing sewage treatment.
Preferably, the nucleotide sequence of 16S rDNA of Acinetobacter has a sequence as shown in SEQ ID No. 1.
The 16S rDNA gene sequence of the acinetobacter provided by the invention is shown as follows:
SEQ ID NO.1:
GGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATTCTGATCCGCGATTACTAGCGATTCCGACTTCATGGAGTCGAGTTGCAGACTCCAATCCGGACTACGATCGGCTTTTTGAGATTAGCATCCTATCGCTAGGTAGCAACCCTTTGTACCGACCATTGTAGCACGTGTGTAGCCCTGGCCGTAAGGGCCATGATGACTTGACGTCGTCCCCGCCTTCCTCCAGTTTGTCACTGGCAGTATCCTTAAAGTTCCCGACATTACTCGCTGGCAAATAAGGAAAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAGCCATGCAGCACCTGTATGTAAGTTCCCGAAGGCACCAATCCATCTCTGGAAAGTTCTTACTATGTCAAGGCCAGGTAAGGTTCTTCGCGTTGCATCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCATTTGAGTTTTAGTCTTGCGACCGTACTCCCCAGGCGGTCTACTTATCGCGTTAGCTGCGCCACTAAAGCCTCAAAGGCCCCAACGGCTAGTAGACATCGTTTACGGCATGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCATGCTTTCGCACCTCAGCGTCAGTGTTAGGCCAGATGGCTGCCTTCGCCATCGGTATTCCTCCAGATCTCTACGCATTTCACCGCTACACCTGGAATTCTACCATCCTCTCCCACACTCTAGCTAACCAGTATCGAATGCAATTCCCAAGTTAAGCTCGGGGATTTCACATTTGACTTAATTAGCCGCCTACGCGCGCTTTACGCCCAGTAAATCCGATTAACGCTTGCACCCTCTGTATTACCGCGGCTGCTGGCACAGAGTTAGCCGGTGCTTATTCTGCGAGTAACGTCCACTATCTCTAGGTATTAACTAAAGTAGCCTCCTCCTCGCTTAAAGTGCTTTACAACCATAAGGCCTTCTTCACACACGCGGCATGGCTGGATCAGGCTTGCGCCCATTGTCCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCGGATCATCCTCTCAGACCCGCTACAGATCGTCGCCTTGGTAGGCCTTTACCCCACCAACTAGCTAATCCGACTTAGGCTCATCTATTAGCGCAAGGTCCGAAGATCCCCTGCTTTCTCCCGTAGGACGTATGCGGTATTAGCATTCCTTTCGAAATGTTGTCCCCCACTAATAGGCAGATTCCTAA。
the length of the sequence is 1282bp, and the accession number of the gene sequence in GenBank is MK 163532. The gene sequence of the strain is uploaded to GenBank for comparison, and the comparison result shows that: the gene sequence of the strain has the highest similarity with the sequence of Acinetobacter. Therefore, the strain is judged to belong to Acinetobacter (Acinetobacter) by combining morphological characteristics of the bacteria, and is named as Acinetobacter sp.a 3.
Specifically, the strain Acinetobacter sp.A3 of the invention is deposited in China general microbiological culture Collection center (CGMCC) at 24/10/2018, and the address is as follows: the preservation number of the Xilu-Shi is CGMCC NO. 16620.
Preferably, the immobilized microbial agent is used for loading the thallus in the bacteria suspension of the acinetobacter onto the immobilized carrier by adopting an adsorption method.
The application adopts the adsorption method to directly utilize the acinetobacter to carry out loading, removes the complicated extraction step of the acinetobacter, and has convenient manufacture and low cost.
Preferably, the mass ratio of the immobilization carrier to the bacteria suspension of acinetobacter is 4.5-5.5: 1.5-2.5.
Preferably, the proportion of the immobilized carrier and the bacteria suspension is selected, so that the quantity of the acinetobacter in the immobilized bacteria agent is moderate, and if the quantity of the acinetobacter is too small, the degradation rate is influenced; if the number of the acinetobacter is too large, the immobilized carrier is supersaturated for adsorbing the microorganisms, the microorganisms are desorbed to enter a water body in the process of degrading pollutants, however, the physiological activity of the microorganisms in the water body is low, and the stability is poor, so that the proportion of the immobilized carrier and the bacterial suspension is preferably controlled.
Preferably, the water absorption of the bran coat is 35-45%.
Under the condition of the same mass, the carrier with higher water absorption rate can absorb more bacterial suspension; however, if the water absorption of the bran coat is too high, the microbial adsorption capacity is too high, the immobilized microbial agent can be desorbed when entering a water body, so that the waste of the acinetobacter is caused, the bran coat and the bacterial suspension are difficult to be uniformly mixed, and whether the distribution of the microbes in the bran coat is uniform or not is difficult to be ensured, so the proportion of the bran coat is optimized, and the microbial adsorption capacity of the immobilized carrier is moderate.
Preferably, the bacterial suspension is a bacterial suspension obtained by culturing the acinetobacter to a logarithmic growth phase, and the OD600 of the bacterial suspension is 0.60-0.63. And measuring the absorbance value of the bacterial suspension at the wavelength of 600nm by using an ultraviolet spectrophotometer, and recording as OD 600.
The growth speed of the microorganism in the logarithmic growth phase is the maximum, the number of cells is increased exponentially, and the selection of the bacterial suspension in the logarithmic growth phase can ensure that the prepared immobilized microbial inoculum has high growth speed and high biological activity.
And verifying the content of the acinetobacter in the bacterial suspension by using the OD600 value, and further controlling whether the bacterial suspension is in a logarithmic growth phase.
Furthermore, the invention also provides a preparation method of the immobilized microbial inoculum. The preparation method at least comprises the following steps:
step a, culturing the acinetobacter to obtain a bacterial suspension;
and b, sterilizing the bran coat, uniformly mixing the bran coat and the bacterial suspension under the aseptic condition, and drying to obtain the immobilized microbial inoculum.
Compared with the prior art, the preparation method of the immobilized microbial inoculum provided by the invention has the following advantages:
the invention loads the acinetobacter onto the bran coat by an adsorption method, has simple preparation method and mild conditions, is beneficial to industrial popularization, fixes the microorganism in the bran coat, and improves the biological activity and stability of the immobilized microbial inoculum by utilizing the characteristics of the bran coat.
Preferably, in step b, the conditions of the sterilization treatment are as follows: the sterilization temperature is 120-125 ℃, and the sterilization time is 25-35 min.
And the bran coat is sterilized, so that other bacteria are not introduced in the later process, the activity of the acinetobacter is further influenced, and even the effect of the microbial inoculum on treating nitrogen-containing sewage is influenced.
Preferably, in the step b, the drying temperature is 30-40 ℃.
The preferable drying temperature can remove the excessive water in the immobilized bacteria agent and does not influence the activity of the acinetobacter.
Preferably, in step b, the mixing condition is stirring at 100-300rpm for 8-12 min.
The preferred mixing conditions are such that the bacterial suspension and carrier are well mixed until the carrier material is wet, loose and non-caking, non-wall sticking.
Preferably, the invention also provides application of the immobilized microbial agent in the field of nitrogen-containing sewage treatment.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a graph showing denitrification performance of an immobilized fungicide provided in example 1 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The following examples are provided to better illustrate the embodiments of the present invention.
Example 1
The embodiment of the invention provides an immobilized microbial inoculum, which comprises acinetobacter and bran coat, wherein the acinetobacter is preserved in the China general microbiological culture Collection center (CGMCC), and the preservation number of the acinetobacter is CGMCC NO. 16620; the water absorption rate of the bran coat is 40%.
The mass ratio of the bran coat to the bacteria suspension of the acinetobacter is 5:2, and the acinetobacter in the bacteria suspension is in a logarithmic growth phase; the OD600 of the bacterial suspension was 0.613.
The preparation method of the immobilized microbial inoculum comprises the following steps:
step a, culturing the Acinetobacter suspension to a logarithmic growth phase;
and b, sterilizing the bran coat at 121 ℃ for 30min, stirring the bran coat and the bacterial suspension at a speed of 200rpm under an aseptic condition for 10min, uniformly mixing until the bran coat is wet, loose, free from caking and wall sticking, and drying at 35 ℃ to obtain the immobilized microbial inoculum.
Example 2
The embodiment of the invention provides an immobilized microbial inoculum, which comprises acinetobacter and bran coat, wherein the acinetobacter is preserved in the China general microbiological culture Collection center (CGMCC), and the preservation number of the acinetobacter is CGMCC NO. 16620; the water absorption of the bran coat is 45%.
The mass ratio of the bran coat to the bacteria suspension of the acinetobacter is 4.5:2.5, and the acinetobacter in the bacteria suspension is in a logarithmic growth phase; the OD600 of the bacterial suspension was 0.63.
The preparation method of the immobilized microbial inoculum comprises the following steps:
step a, culturing the Acinetobacter suspension to a logarithmic growth phase;
and b, sterilizing the bran coat at 125 ℃ for 25min, stirring the bran coat and the bacterial suspension under an aseptic condition at the speed of 300rpm for 8min, uniformly mixing until the bran coat is wet, loose, free from caking and wall sticking, and drying at 40 ℃ to obtain the immobilized microbial inoculum.
Example 3
The embodiment of the invention provides an immobilized microbial inoculum, which comprises acinetobacter and bran coat, wherein the acinetobacter is preserved in the China general microbiological culture Collection center (CGMCC), and the preservation number of the acinetobacter is CGMCC NO. 16620; the water absorption of the bran coat is 35%.
The mass ratio of the bran coat to the bacteria suspension of the acinetobacter is 5.5:1.5, and the acinetobacter in the bacteria suspension is in a logarithmic growth phase; the OD600 of the bacterial suspension was 0.605.
The preparation method of the immobilized microbial inoculum comprises the following steps:
step a, culturing the Acinetobacter suspension to a logarithmic growth phase;
and b, sterilizing the bran coat at 120 ℃ for 35min, stirring the bran coat and the bacterial suspension under an aseptic condition at a speed of 100rpm for 12min, uniformly mixing until the bran coat is wet, loose, free from caking and wall sticking, and drying at 30 ℃ to obtain the immobilized microbial inoculum.
Example 4
The embodiment of the invention provides an immobilized microbial inoculum, which comprises acinetobacter and bran coat, wherein the acinetobacter is preserved in the China general microbiological culture Collection center (CGMCC), and the preservation number of the acinetobacter is CGMCC NO. 16620; the water absorption rate of the bran coat is 40%.
The mass ratio of the bran coat to the bacteria suspension of the acinetobacter is 5:2, and the acinetobacter in the bacteria suspension is in a logarithmic growth phase; the OD600 of the bacterial suspension was 0.153, i.e., the bacterial suspension of example 1 was diluted 1/4 times the original OD 600.
The preparation method of the immobilized microbial inoculum is also the same as that of the example 1 and is not repeated.
Example 5
The comparative example provides an immobilized microbial inoculum, which comprises acinetobacter and bran coat, wherein the acinetobacter is preserved in the China general microbiological culture Collection center (CGMCC), and the preservation number of the acinetobacter is CGMCC NO. 16620; the water absorption rate of the bran coat is 40%.
The mass ratio of the bran coat to the bacteria suspension of the acinetobacter is 5:2, and the OD600 of the bacteria suspension is 1.226, namely, the bacteria suspension in the embodiment 1 is concentrated to be 2 times of the original OD 600.
The preparation method of the immobilized microbial inoculum is also the same as that of the example 1 and is not repeated.
In order to better illustrate the technical solution of the present invention, further comparison is made below by means of a comparative example and an example of the present invention.
Comparative example 1
The comparative example provides an immobilized microbial inoculum, which comprises acinetobacter and zeolite, wherein the acinetobacter is preserved in the China general microbiological culture Collection center (CGMCC), and the preservation number of the acinetobacter is CGMCC NO. 16620; the water absorption of the zeolite was 40%.
The mass ratio of the zeolite to the acinetobacter bacterial suspension is 5:2, and the OD600 of the bacterial suspension is 2.453, i.e. the bacterial suspension in example 1 is concentrated to 4 times of the original OD 600.
The preparation method of the immobilized microbial inoculum is also the same as that of the example 1 and is not repeated.
Comparative example 2
The comparative example provides an immobilized microbial inoculum, which comprises acinetobacter and activated carbon, wherein the acinetobacter is preserved in the China general microbiological culture Collection center (CGMCC), and the preservation number of the acinetobacter is CGMCC NO. 16620; the water absorption of the activated carbon was 75%.
The mass ratio of the activated carbon to the bacteria suspension of acinetobacter is 5:2, and the OD600 of the bacteria suspension is 2.453, i.e. the bacteria suspension in example 1 is concentrated to 4 times of the original OD 600.
The preparation method of the immobilized microbial inoculum is also the same as that of the example 1 and is not repeated.
In order to better illustrate the characteristics of the immobilized bacteria agents provided in the examples of the present invention, the immobilized bacteria agents prepared in examples 1 to 5 and comparative examples 1 to 2 were subjected to performance tests.
Test example 1
150mL of the culture medium was added to 7 flasks, and sterilized at 120 ℃ for 30 min. Then, the immobilized bactericides prepared in examples 1 to 5 and comparative examples 1 to 2 were added into the conical flasks, the added amount of the bactericides was 0.5g by dry weight, the degradation effect on nitrate nitrogen was tested, the degradation efficiency is shown in the following table 1, and the degradation chart of example 1 is shown in fig. 1.
TABLE 1 degradation efficiency
Initial concentration of nitrate and nitrogen Time of denitrification Removal rate of nitro and nitrogen
Example 1 100mg/L 16h 100%
Example 2 100mg/L 16.1h 100%
Example 3 100mg/L 16.1h 100%
Example 4 100mg/L 18h 100%
Example 5 100mg/L 14h 100%
Comparative example 1 100mg/L 20h 35%
Comparative example 2 100mg/L 20h 5%
As can be seen from Table 1, the immobilized microbial inoculum provided by the embodiments 1-5 of the invention can completely degrade nitrate nitrogen with a concentration of 100mg/L within 18h, wherein the degradation rate of the embodiments 1-3 is better, the nitrate nitrogen can be completely degraded within about 16h, the immobilized microbial inoculum of the embodiments 5 is more excellent, and the nitrate nitrogen can be completely degraded within 14 h. The degradation effect of the immobilized bacteria agent prepared in the comparative examples 1-2 is poor, and the removal rate of nitrol in 20h is only 35% and 5%.
As can be seen from FIG. 1, the immobilized microbial inoculum prepared in example 1 completely degrades 100mg/L of nitrate nitrogen within 16h under aerobic conditions, and completely degrades accumulated nitrite nitrogen, thereby showing good denitrification effect. Unlike free bacteria, the immobilized bacteria prepared by the adsorption method can provide an aerobic and anoxic microenvironment for microorganisms, which is beneficial to the growth of the bacteria and the performance of denitrification and denitrification of the bacteria, and the immobilized bacteria prepared in examples 2 to 5 can completely degrade nitrate nitrogen and nitrite nitrogen within corresponding time.
It can be seen from the combination of examples 1 and 4 to 5 that the denitrification time for achieving a nitrate nitrogen removal rate of 100% is shortened as the concentration of acinetobacter in the immobilized microbial agent is increased.
In comparative examples 1-2, although the concentration of the bacterial suspension used was 4 times that of example 1, the removal rate of nitrol was much lower than that of example 1, and even lower than that of example 4, i.e., the degradation effect of the immobilized bacteria agent using the bacterial suspension concentration 4 times that of comparative examples 1-2 was much lower than that of example 5 using the bacterial suspension concentration 1/4 times.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
SEQUENCE LISTING
<110> Zhongji electric construction water environment governance technology Limited company
<120> immobilized microbial agent and preparation method and application thereof
<130> 2019
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1282
<212> DNA
<213> Artificial sequence
<400> 1
ggcggtgtgt acaaggcccg ggaacgtatt caccgcggca ttctgatccg cgattactag 60
cgattccgac ttcatggagt cgagttgcag actccaatcc ggactacgat cggctttttg 120
agattagcat cctatcgcta ggtagcaacc ctttgtaccg accattgtag cacgtgtgta 180
gccctggccg taagggccat gatgacttga cgtcgtcccc gccttcctcc agtttgtcac 240
tggcagtatc cttaaagttc ccgacattac tcgctggcaa ataaggaaaa gggttgcgct 300
cgttgcggga cttaacccaa catctcacga cacgagctga cgacagccat gcagcacctg 360
tatgtaagtt cccgaaggca ccaatccatc tctggaaagt tcttactatg tcaaggccag 420
gtaaggttct tcgcgttgca tcgaattaaa ccacatgctc caccgcttgt gcgggccccc 480
gtcaattcat ttgagtttta gtcttgcgac cgtactcccc aggcggtcta cttatcgcgt 540
tagctgcgcc actaaagcct caaaggcccc aacggctagt agacatcgtt tacggcatgg 600
actaccaggg tatctaatcc tgtttgctcc ccatgctttc gcacctcagc gtcagtgtta 660
ggccagatgg ctgccttcgc catcggtatt cctccagatc tctacgcatt tcaccgctac 720
acctggaatt ctaccatcct ctcccacact ctagctaacc agtatcgaat gcaattccca 780
agttaagctc ggggatttca catttgactt aattagccgc ctacgcgcgc tttacgccca 840
gtaaatccga ttaacgcttg caccctctgt attaccgcgg ctgctggcac agagttagcc 900
ggtgcttatt ctgcgagtaa cgtccactat ctctaggtat taactaaagt agcctcctcc 960
tcgcttaaag tgctttacaa ccataaggcc ttcttcacac acgcggcatg gctggatcag 1020
gcttgcgccc attgtccaat attccccact gctgcctccc gtaggagtct gggccgtgtc 1080
tcagtcccag tgtggcggat catcctctca gacccgctac agatcgtcgc cttggtaggc 1140
ctttacccca ccaactagct aatccgactt aggctcatct attagcgcaa ggtccgaaga 1200
tcccctgctt tctcccgtag gacgtatgcg gtattagcat tcctttcgaa atgttgtccc 1260
ccactaatag gcagattcct aa 1282

Claims (10)

1. An immobilized microbial inoculum, which is characterized in that: the culture medium comprises acinetobacter and an immobilized carrier, wherein the acinetobacter is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation number of the acinetobacter is CGMCC NO.16620, and the immobilized carrier is bran.
2. The immobilized inoculant according to claim 1, wherein: the nucleotide sequence of the 16S rDNA of the acinetobacter has a sequence shown as SEQ ID NO. 1.
3. The immobilized inoculant according to claim 1, wherein: the immobilized microbial agent adopts an adsorption method to load the thallus in the bacteria suspension of the acinetobacter onto the immobilized carrier.
4. The immobilized inoculant according to claim 3, wherein: the mass ratio of the immobilized carrier to the bacteria suspension of the acinetobacter is 4.5-5.5: 1.5-2.5.
5. The immobilized inoculant according to claim 3, wherein: the bacterial suspension is obtained by culturing the acinetobacter to logarithmic growth phase, and the OD600 of the bacterial suspension is 0.60-0.63.
6. A method for producing the immobilized fungicide according to any one of claims 1 to 5, characterized in that: at least comprises the following steps:
step a, culturing the acinetobacter to obtain a bacterial suspension;
and b, sterilizing the bran coat, uniformly mixing the bran coat and the bacterial suspension under the aseptic condition, and drying to obtain the immobilized microbial inoculum.
7. The method for preparing an immobilized microbial inoculum according to claim 6, wherein: in the step b, the conditions of the sterilization treatment are as follows: the sterilization temperature is 120-125 ℃, and the sterilization time is 25-35 min.
8. The method for preparing an immobilized microbial inoculum according to claim 6, wherein: in the step b, the drying temperature is 30-40 ℃.
9. The method for preparing an immobilized microbial inoculum according to claim 6, wherein: in the step b, the mixing condition is stirring for 8-12min under the condition of 100-300 rpm.
10. The use of the immobilized microbial agent of any one of claims 1 to 5 in the field of nitrogen-containing wastewater treatment.
CN201910718561.8A 2019-08-05 2019-08-05 Immobilized microbial inoculum and preparation method and application thereof Active CN110699350B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910718561.8A CN110699350B (en) 2019-08-05 2019-08-05 Immobilized microbial inoculum and preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910718561.8A CN110699350B (en) 2019-08-05 2019-08-05 Immobilized microbial inoculum and preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN110699350A CN110699350A (en) 2020-01-17
CN110699350B true CN110699350B (en) 2021-03-30

Family

ID=69193328

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910718561.8A Active CN110699350B (en) 2019-08-05 2019-08-05 Immobilized microbial inoculum and preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN110699350B (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6107067A (en) * 1998-07-06 2000-08-22 W.R. Grace & Co.-Conn. Porous, non-macroporous, inorganic oxide carrier body for immobilizing microorganisms for bioremediation
CN102747015A (en) * 2012-06-21 2012-10-24 北京理工大学 Denitrification acinetobacters and use thereof
CN104988089A (en) * 2015-06-13 2015-10-21 施瑞客(天津)生物技术有限公司 Probiotic bacterium preparation for chicken and preparing method of preparation

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6107067A (en) * 1998-07-06 2000-08-22 W.R. Grace & Co.-Conn. Porous, non-macroporous, inorganic oxide carrier body for immobilizing microorganisms for bioremediation
CN102747015A (en) * 2012-06-21 2012-10-24 北京理工大学 Denitrification acinetobacters and use thereof
CN104988089A (en) * 2015-06-13 2015-10-21 施瑞客(天津)生物技术有限公司 Probiotic bacterium preparation for chicken and preparing method of preparation

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Acinetobacter sp. strain A3 16S ribosomal RNA gene, partial sequence;Zheng,M;《NCBI GenBank》;20181119;第1页 *
Biodegradation of Ammonium by Suspended and Immobilized Cells of Acinetobacter sp. Y1;Lv, YK等;《ASIAN JOURNAL OF CHEMISTRY》;20131031;第25卷(第13期);第7243-7246页 *
Piggery wastewater treatment by Acinetobacter sp TX5 immobilized with spent mushroom substrate in a fixed-bed reactor;Yang, YL等;《SCIENCE OF THE TOTAL ENVIRONMENT》;20181210;第644卷;第1460-1468页 *
不动杆菌Acinetobacter sp.NG3固定化处理抗生素制药废水的研究;王俊峰等;《安徽农业科学》;20141212(第35期);第12633-12634页 *

Also Published As

Publication number Publication date
CN110699350A (en) 2020-01-17

Similar Documents

Publication Publication Date Title
Pai et al. Continuous degradation of phenol by Rhodococcus sp. immobilized on granular activated carbon and in calcium alginate
Yu et al. Enhanced nitrogen removal of low C/N wastewater in constructed wetlands with co-immobilizing solid carbon source and denitrifying bacteria
Nagadomi et al. Treatment of aquarium water by denitrifying photosynthetic bacteria using immobilized polyvinyl alcohol beads
CN102465103B (en) Aerobic denitrification methylobacterium phyllosphaerae and application thereof
CN102061276A (en) Pseudomonas sp. strain for biological denitrification under low temperature and application thereof
CN101812440A (en) Nitrifying bacteria vector for degrading ammonia nitrogen and preparation method thereof
CN101560486A (en) Achromobacter xylosoxidans strain for biological denitrificaion and application thereof
CN107487840B (en) Biological filter material for treating nitrate nitrogen in water
Zayed et al. Removal of organic pollutants and of nitrate from wastewater from the dairy industry by denitrification
CN108793433B (en) High-turbidity water quality emergency treatment technology
JP2014024002A (en) Organic waste liquid treatment method and organic waste liquid treatment apparatus
CN105217901A (en) A kind of method of wastewater biochemical removing total nitrogen
CN102465105B (en) Nitrous acid-type denitrification bacteria strain and application thereof
CN107473404B (en) Water purifying agent with self-formed block-shaped carbon carrier for fixing microorganisms and preparation method thereof
CN110699347B (en) Immobilized microbial inoculum and preparation method and application thereof
CN110699350B (en) Immobilized microbial inoculum and preparation method and application thereof
CN107488611A (en) A kind of efficient denitrification denitrifying bacterium and its application
CN102465106B (en) Bacterial strain used for short-cut denitrification for nitrogen removal and its application
CN100355675C (en) Method for treating phenol-contained waste water by anaerobic-aerobic coupled biological fluidizing bed
CN110697907A (en) Immobilized composite flora material and preparation method thereof
CN102978145B (en) Quinoline degrading bacteria QG6 with heterotrophic nitrification-aerobic denitrification function and phosphorous removal function and application thereof
JP2007117967A (en) Carrier for autotrophic ammonia oxidizing bacterium, ammonia nitrogen removing bacterium-deposited body and method for removing ammonia nitrogen
CN108975494B (en) Method for treating black smelly water by using graphene modified straw material
CN111470714B (en) Environment-friendly recycling method for aquaculture wastewater
CN109486697B (en) Bacillus licheniformis, preparation thereof and method for treating wastewater

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
GR01 Patent grant
GR01 Patent grant