CN112226388B - Novel enzyme-producing species of propionibacteriaceae and application thereof - Google Patents
Novel enzyme-producing species of propionibacteriaceae and application thereof Download PDFInfo
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- 241000894007 species Species 0.000 title abstract description 14
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- 230000001580 bacterial effect Effects 0.000 claims abstract description 51
- 102000013142 Amylases Human genes 0.000 claims abstract description 16
- 108010065511 Amylases Proteins 0.000 claims abstract description 16
- 235000019418 amylase Nutrition 0.000 claims abstract description 16
- 239000004382 Amylase Substances 0.000 claims abstract description 15
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- 235000019270 ammonium chloride Nutrition 0.000 description 1
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- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
- B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
- C12N9/20—Triglyceride splitting, e.g. by means of lipase
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- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
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Abstract
The invention discloses a new enzyme-producing species of propionibacteriaceae and application thereof. The new zymogenic species of the propionibacteriaceae has the strain name of 99B, is preserved in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms at 8-11.2020, and has the preservation number of CGMCC No. 20523. The strain can produce amylase and lipase, can synchronously carry out heterotrophic nitrification-aerobic denitrification, is a functional new species of propionibacteriaceae, and has wide application prospects in the aspects of kitchen waste treatment, bacterial manure preparation, sewage treatment and the like.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a new enzyme-producing species of propionibacteriaceae and application thereof.
Background
Propionibacteriaceae (Propionibacteriaceae) belongs to the bacterial domain (Bacteria) -actinomycetales (actinobacteriia) -actinomycetes (actinomycetes) -Propionibacteriales (Propionibacteriales). The family is established by E.A. Delweichi in 1957, and currently comprises 23 effective description genera, wherein 10 genera including Auratiococcus, Brooklawnia, Granulococcus, Micropruina, Nigerium, Propionicella, Propioniclava, Propioniferax, Propionibacterium, Raineella and the like have only 1 effective description species, and 5 genera including Aesturtiimicrobium, Brevibacter, Desertihabines, Marininecoccus, Propionimonas and the like have only 2 effective description species. Indicating that the intergeneric diversity of this family is relatively high.
Amylases, which are a generic term for enzymes that hydrolyze starch and glycogen, are important carbohydrate hydrolyzing enzymes; the lipase can catalyze reactions such as lipolysis, ester exchange, ester synthesis and the like, and both are important industrial enzyme preparation varieties and are widely applied to industries such as environmental protection, feed, daily chemicals and the like. The ammonia nitrogen can consume a large amount of dissolved oxygen in the water body, so that the water body is anoxic, and meanwhile, the water body eutrophication can be caused. The traditional nitrifying bacteria are autotrophic bacteria, the required growth environment is strict, the growth speed is slow, the mass culture is difficult, and the application is limited. Heterotrophic nitrifying bacteria can directly utilize organic matter nutrition in the water body to grow and degrade ammonia nitrogen, have strong adaptability and high growth speed, and can remove COD and ammonia nitrogen at the same time. Traditionally, denitrification is carried out under anaerobic condition, while some heterotrophic nitrifiers can carry out aerobic denitrification, which has great significance for removing nitrogen pollutants.
Disclosure of Invention
The invention aims to provide a new species of enzyme production of propionibacteriaceae and application thereof.
In a first aspect, the invention provides a strain of Oleericoccum amyloliquefaciem 99B of the family Propionibacterium.
The invention protects a classified name of Olericoccum amyloliquefaciens 99B of propionibacteriaceae, which is stored in the common microorganism center of China Committee for culture Collection of microorganisms (CGMCC for short, the address: No. 3 of West Lu No. 1 of the North Jing city Kogyo of the morning district, the postal code 100101 of the institute of microbiology of China academy of sciences), the storage number is CGMCC No.20523, on 8-11-2020.
In a second aspect, the present invention protects a novel use of Oleericoccum amyloliquefaciens of Propionibacterium family or a bacterial suspension thereof or a culture solution thereof or a fermentation product thereof or a microbial agent containing the same.
The invention protects application of Oleericoccum amyloliquefaciens of propionibacteriaceae or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in producing or preparing amylase.
The invention also discloses the application of the Olearicococcum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension or the culture solution or the fermentation product thereof or the microbial inoculum containing the same in the preparation of products for producing or preparing amylase.
The invention also protects the application of the Oleericoccum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension or the culture solution or the fermentation product thereof or the microbial inoculum containing the same in producing or preparing the lipase.
The invention also discloses the application of the Olearicococcum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension or the culture solution or the fermentation product thereof or the microbial inoculum containing the same in the preparation of products for producing or preparing lipase.
The invention also protects the application of the Olericiccocum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension or the culture solution or the fermentation product thereof or the microbial inoculum containing the Olericiccocum amyloliquefaciens in heterotrophic nitrification and/or aerobic denitrification.
The invention also discloses the application of the Olericiccocum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension or the culture solution or the fermentation product thereof or the microbial inoculum containing the Olericiccocumm amyloliquefaciens in the preparation of heterotrophic nitrification and/or aerobic denitrification products.
The heterotrophic nitrification refers to a process of oxidizing ammonia nitrogen into nitric acid nitrogen and nitrous acid nitrogen by heterotrophic microorganisms.
The aerobic denitrification refers to that the microorganisms reduce the nitric acid nitrogen and the nitrous acid nitrogen into nitrogen-containing gas (NO, N) under the aerobic condition2O、N2) The process of (1).
The invention also protects the application of the Olericiccocum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension or the culture solution or the fermentation product thereof or the microbial inoculum containing the Olericiccocum amyloliquefaciens in degrading ammonia nitrogen.
The invention also protects the application of the Olericiccocum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension or the culture solution or the fermentation product thereof or the microbial inoculum containing the Olericicococcum amyloliquefaciens in the preparation of products for degrading ammonia nitrogen.
The invention also protects the application of the Olericiccocum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension thereof or the culture solution thereof or the fermentation product thereof or the microbial inoculum containing the Olericicococcum amyloliquefaciens in the nitrogen pollutant removal or the kitchen waste treatment or the sewage treatment.
The invention also protects the application of the Olericiccocum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension thereof, the culture solution thereof, the fermentation product thereof or the microbial inoculum containing the Olericicococcum amyloliquefaciens in the preparation of products for removing nitrogen pollutants or treating kitchen waste or sewage.
In a third aspect, the invention features an amylase or lipase.
The amylase or lipase protected by the invention is obtained by fermenting and culturing Oleericoccum amyloliquefaciens of Propionibacterium family.
Further, the method for fermentation culture comprises the following steps: the propionibacteriaceae Oleericoccum amyloliquefaciens is inoculated into LB liquid medium for fermentation culture.
Further, the fermentation culture conditions are 30 ℃ and 150rpm shaking culture for 24 h.
The substrate upon which the amylase can act includes soluble starch.
The substrate for lipase action comprises a fat source (the fat source is formed by mixing a 2% polyvinyl alcohol solution and tributyrin according to a volume ratio of 1: 9).
In a fourth aspect, the invention provides a product, wherein the active ingredient is olearoccum amyloliquefaciens of propionibacteriaceae or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same;
the product has any one of the following functions 1) to 5):
1) producing or preparing an amylase;
2) producing or preparing a lipase;
3) heterotrophic nitrification and/or aerobic denitrification;
4) degrading ammonia nitrogen;
5) removing nitrogen pollutants, or treating kitchen waste or sewage.
In a fifth aspect, the invention protects any of the following a1) -a 4):
a1) a method for producing or preparing an amylase, comprising the step of subjecting an Oleericoccum amyloliquefacien of the family Propionibacterium to fermentation culture;
a2) a method for producing or preparing a lipase, comprising the step of subjecting Oleericoccum amyloliquefaciens of the family Propionibacterium to fermentation culture;
a3) a method for degrading ammonia nitrogen comprises the step of treating ammonia nitrogen with Oleericoccum amyloliquefaciens of Propionibacterium family or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same;
a4) a method for removing nitrogen pollutants or treating kitchen waste or sewage comprises the step of treating the nitrogen pollutants or the kitchen waste or the sewage by using Olearicococcum amyloliquefaciens of propionibacteriaceae or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same.
In any of the above applications, products or methods, the method of preparing the culture medium is specifically as follows: the fermentation culture is carried out by inoculating Oleericoccum amyloliquefaciens of Propionibacterium family in LB liquid medium. Further, the fermentation culture conditions are specifically as follows: shaking and culturing at 30 deg.C and 150rpm for 24 h.
The preparation method of the bacterial suspension comprises the following steps: inoculating Oleericoccum amyloliquefaciens of Propionibacterium family in LB liquid culture medium for fermentation culture to obtain culture solution; centrifuging the culture solution, and collecting thalli; and (4) resuspending the thalli by using sterile water to obtain a bacterial suspension. Further, the conditions of the fermentation culture are specifically 30 ℃ and 150rpm shaking culture for 24 h. The centrifugation condition is specifically centrifugation for 5min at 8000rpm and 4 ℃. And the step of resuspending the thalli by using sterile water is to resuspend the thalli by using equal volume of sterile water.
In any of the above uses or products or methods, the aforementioned propionibacteriaceae oleraceae olylloquefacium is propionibacteriaceae oleraceae olylloquefacium 99B CGMCC No. 20523.
The invention provides a new enzyme-producing species of propionibacteriaceae and application thereof, wherein the strain name of the new enzyme-producing species of propionibacteriaceae is 99B, the new enzyme-producing species of propionibacteriaceae is preserved in the China general microbiological culture Collection center (CGMCC) at 8-11.2020, and the preservation number is CGMCC No. 20523. Experiments prove that: the 99B CGMCC No.20523 strain can produce amylase and lipase, can synchronously perform heterotrophic nitrification-aerobic denitrification, is a new species of propionibacteriaceae, and has wide application prospect in the aspects of kitchen waste treatment, bacterial manure preparation, sewage treatment and the like.
Drawings
FIG. 1 is a 99B phylogenetic tree.
Deposit description
Latin name: oleericoccum amyloliquefaciens
The strain number is as follows: 99B
The preservation organization: china general microbiological culture Collection center
The preservation organization is abbreviated as: CGMCC (China general microbiological culture Collection center)
Address: xilu No. 1 Hospital No. 3 of Beijing market facing Yang district
The preservation date is as follows: year 2020, 8 and 11
Registration number of the preservation center: CGMCC No.20523
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The test methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
Examples 1, 99B isolation, identification and preservation of strains
Isolation of first, 99B Strain
99B strains were isolated from oil well production from north Heibei corridor at 6 months of 2020. The method comprises the following specific steps: diluting the produced liquid with sterile normal saline, coating the diluted produced liquid on an LB solid culture medium (the solvent is water, the solute and the concentration thereof are respectively 10g/L NaCl, 10g/L peptone, 5g/L yeast powder, 15g/L agar and pH is 7-8), standing and culturing at 30 ℃, picking out single bacterial colonies after the single bacterial colonies grow out, carrying out streak purification until pure bacterial strains are obtained, and naming the pure bacterial strains as 99B bacterial strains.
Identification of the second, 99B Strain
1. Morphological identification
The 99B strain forms a circular regular convex dark yellow colony on an LB solid culture medium (the solvent is water, the solute and the concentration thereof are respectively 10g/L NaCl, 10g/L peptone, 5g/L yeast powder and 15g/L agar, and the pH value is 7-8). The gram stain of the thallus is negative and spherical, and spores are produced.
2. Physiological and biochemical identification
The 99B strain can grow under the condition of 0-8 percent (mass fraction) of sodium chloride, wherein the strain grows optimally under the condition of 1-5 percent (mass fraction) of sodium chloride, and grows weakly under the condition of 8 percent (mass fraction) of sodium chloride.
3. Molecular identification
Inoculating the activated 99B strain into an LB liquid culture medium (the solvent is water, the solute and the concentration thereof are respectively 10g/L NaCl, 10g/L peptone, 5g/L yeast powder and pH 7-8), performing shaking culture at 30 ℃ and 150rpm for 24h, centrifuging 1mL of fresh culture solution at 4 ℃ and 8000rpm for 5min, collecting the thallus in a 2mL centrifuge tube, and extracting DNA by adopting a DNA extraction kit. After electrophoretic detection, PCR amplification is carried out by using a universal primer 8F/1492R. And (3) after the electrophoresis detection of the PCR product, determining a 16S rDNA gene sequence, wherein the specific sequence is shown as a sequence 1 in a sequence table.
The 99B strain was found to belong to the family propionibacteriaceae after sequence alignment using the NCBI database, but the similarity to existing strains was low. Phylogenetic trees were constructed using the software MEGA based on the 16S rDNA gene sequence, and the results are shown in fig. 1. The 99B strain had only 94.74% and 94.67% similarity to the closest strains Aescurginia kwangyangense and Aescurginia soli, respectively, of the developing tree, both of which were less than 95%.
As a result of comparison using the Ezbiochloud database, as shown in Table 1, the similarity of the strain Lutecoccus pertinenei most similar to other genera of Propionibacterium family was 95.22%, the similarity of the strain Microlucidus was 94.81%, the similarity of the strain Brooklawnia cerclae was 94.30%, the similarity of the strain Tessacccus flavescens was 94.16%, the similarity of the strain Naumannella huperziae was 93.57%, and the similarity of the strain Mariniuteicoccus flavus was 93.56%. Meanwhile, only 1 effective descriptor in 10 genera of the current 23 effective descriptors of the propionibacteriaceae and only 2 effective descriptors in 5 genera indicate that the intergeneric diversity of the propionibacteriaceae is relatively high. Therefore, in summary, it is shown that: the 99B strain is a newly discovered new genus strain belonging to the family propionibacteriaceae.
TABLE 1 alignment of the Ezbiocloud database
Deposit of the third, 99B Strain
The 99B strain is classified and named as Oleericoccum amyloliquefaciem, and is preserved in China general microbiological culture Collection center (CGMCC for short, the address: No. 3 of Beijing university Hokko No. 1 of North Chen West Lu, institute of microbiology, Chinese academy of sciences, postal code 100101) at 8-month and 11-month in 2020 with the preservation number of CGMCC No. 20523.
Example 2 detection of Amylase Activity of Strain 99B CGMCC No.20523
1. Adding 0.2 percent (mass fraction) of soluble starch and 1.5 percent (mass fraction) of agar into an LB liquid culture medium, and pouring a solid plate after sterilization to obtain a starch solid culture medium.
2. The activated strain 99B CGMCC No.20523 of example 1 was inoculated into LB liquid medium (10g/L sodium chloride, 10g/L peptone, 5g/L yeast powder, pH 7-8), and cultured at 30 ℃ and 150rpm for 24 hours with shaking to obtain 99B bacterial liquid. Meanwhile, the most similar Aescuariimiacross kwangyangensse strain and Aescuariimiacross soli strain to the developing tree of the strain 99B CGMCC No.20523 were used as controls.
3. And (3) inoculating 5 mu L of the fresh 99B bacterial liquid obtained in the step (2) on a starch solid culture medium, standing and culturing at 30 ℃ for 48h, and developing by using a Lu's classical liquid.
The results show that: an obvious transparent ring appears around the 99B CGMCC No.20523 bacterial colony, and the ratio of the transparent ring to the bacterial colony is 21/6-3.5, which shows that the 99B CGMCC No.20523 bacterial strain has better amylase activity. The most similar Aescurgirniicrobium kwangyangensse strain and Aescurgirniicrobium soli strain to the developing tree of the 99B CGMCC No.20523 strain have no amylase activity, further indicating that the 99B CGMCC No.20523 strain is a new strain belonging to the family Propionibacterium.
Example 3 detection of Lipase Activity of Strain of 99B CGMCC No.20523
1. Adding 2% (mass fraction) of fat source (prepared by mixing 2% of polyvinyl alcohol solution and tributyrin at a volume ratio of 1: 9) and 1.5% (mass fraction) of agar into LB liquid culture medium, sterilizing, and pouring into solid plate to obtain fat solid culture medium.
2. The activated strain 99B CGMCC No.20523 of example 1 was inoculated into LB liquid medium (10g/L sodium chloride, 10g/L peptone, 5g/L yeast powder, pH 7-8) and cultured with shaking at 30 ℃ and 150rpm for 24 hours to obtain a 99B bacterial solution.
3. And (3) inoculating 5 mu L of the fresh 99B bacterial liquid obtained in the step (2) on a fat solid culture medium, and standing and culturing for 48h at the temperature of 30 ℃.
The results show that: an obvious transparent ring appears around the 99B CGMCC No.20523 bacterial colony, and the ratio of the transparent ring to the bacterial colony is 22/7-3.14, which shows that the 99B CGMCC No.20523 bacterial strain has better lipase activity.
Example 4 detection of synchronous heterotrophic nitrification-aerobic denitrification function of 99B CGMCC No.20523 Strain
1. The activated strain 99B CGMCC No.20523 of example 1 was inoculated into LB liquid medium (10g/L sodium chloride, 10g/L peptone, 5g/L yeast powder, pH 7-8) and cultured with shaking at 30 ℃ and 150rpm for 24 hours to obtain a 99B bacterial solution.
2. And (3) centrifuging the fresh 99B bacterial liquid obtained in the step (1) for 5min at the temperature of 4 ℃ and at the rpm of 8000, collecting thalli, and suspending the thalli by using sterile water with the same volume to prepare 99B bacterial suspension.
3. Inoculating the 99B bacterial suspension obtained in the step 2 into an aerobic denitrification liquid culture medium (the solvent is water, the concentration of the solvent is 2g/L of glucose, 1.25g/L of sodium acetate, 0.25g/L of sodium chloride, 0.15g/L of magnesium sulfate heptahydrate, 0.35g/L of dipotassium phosphate trihydrate and 0.3g/L of sodium nitrate) according to 1% (v/v), and performing shaking culture at 30 ℃ and 150rpm for 7d by taking the culture medium without inoculated bacteria as a control group to obtain a inoculated bacteria culture solution.
The inoculation broth was tested using Griess reagent (purchased from Shanghai Richu Biotechnology Ltd.) and the results indicated that nitrite nitrogen was present in the inoculation broth, whereas nitrite nitrogen was not present in the control group, indicating that nitrate nitrogen was reduced to nitrite nitrogen by 99B CGMCC No. 20523. And (3) determining the total nitrogen concentration of the inoculated culture solution and the non-inoculated control by alkaline potassium persulfate digestion ultraviolet spectrophotometry according to the determination of the total nitrogen of the water quality of HJ 636-2012, and calculating the total nitrogen degradation rate according to the total nitrogen degradation rate { (the total nitrogen concentration of the non-inoculated control-the total nitrogen concentration of the inoculated culture solution)/the total nitrogen concentration of the non-inoculated control } × 100%.
The results show that: the total nitrogen degradation rate of the 99B CGMCC No.20523 strain is 44.12 percent. The 99B CGMCC No.20523 strain has aerobic denitrification function.
4. Inoculating the 99B bacterial suspension obtained in the step 2 into a heterotrophic nitrification liquid culture medium (the solvent is water, the concentration of the solvent is 2g/L of glucose, 1.25g/L of sodium acetate, 0.25g/L of sodium chloride, 0.15g/L of magnesium sulfate heptahydrate, 0.35g/L of dipotassium phosphate trihydrate and 0.18g/L of ammonium chloride respectively) according to 1% (v/v), and performing shaking culture at 30 ℃ and 150rpm for 7d by taking a culture medium without inoculated bacteria as a control to obtain a inoculated bacteria culture solution.
The inoculation medium was tested using Griess' reagent (purchased from shanghai ruichu biotechnology limited) and the results showed: nitrite nitrogen appears in the inoculation culture solution, but nitrite nitrogen does not appear in the control group, which indicates that ammonia nitrogen is oxidized into nitrite nitrogen by 99B CGMCC No. 20523. And (3) measuring the ammonia nitrogen concentrations of the inoculated culture solution and the non-inoculated control according to a standard HJ 535-2009 ammonia nitrogen measuring reagent spectrophotometry, and calculating the ammonia nitrogen degradation rate according to the ammonia nitrogen degradation rate { (non-inoculated control ammonia nitrogen concentration-inoculated culture solution ammonia nitrogen concentration)/non-inoculated control ammonia nitrogen concentration } × 100%. The results show that: the ammonia nitrogen degradation rate of the 99B CGMCC No.20523 strain is 59.14 percent. The 99B CGMCC No.20523 strain has heterotrophic nitrification function and can oxidize ammonia nitrogen into nitrite nitrogen and the like.
And meanwhile, further measuring the total nitrogen concentration of the inoculated culture solution and the non-inoculated culture control, and calculating the total nitrogen degradation rate according to the total nitrogen degradation rate { (total nitrogen concentration of non-inoculated culture control-total nitrogen concentration of inoculated culture solution)/total nitrogen concentration of non-inoculated culture control } × 100%. The results show that: the total nitrogen degradation rate of the 99B CGMCC No.20523 strain is 36.73 percent.
In conclusion, the 99B CGMCC No.20523 strain is heterotrophic nitrification-aerobic denitrification strain, can perform heterotrophic nitrification and denitrification in the presence of oxygen, and has synchronous heterotrophic nitrification-aerobic denitrification function.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the technical principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> micron Erhua Biotech (Beijing) Ltd
<120> novel enzyme-producing species of propionibacteriaceae and uses thereof
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1369
<212> DNA
<213> Artificial Sequence
<400> 1
tgcagtcgaa cggtaaggcc cttcggggta cacgagtggc gaacgggtga gtaacacgtg 60
agtaatgtgc ccttgactct gggataacag ttggaaacag ctgctaatac cggatatgac 120
ctgttcctgc atgggggtgg gtggaaagct ccggcggtca tggatcaact cgcggcctat 180
cagcttgttg gtgaggtagt ggctcaccaa ggcttcgacg ggtagccggc ctgagagggc 240
gaccggccac actgggactg agatacggcc cagactccta cgggaggcag cagtggggaa 300
tattgcacaa tgggcgcaag cctgatgcag caacgccgcg tgcgggatga cggccttcgg 360
gttgtaaacc gctttcagca gggacgaagc gcaagtgacg gtacctgcag aagaagcacc 420
ggccaactac gtgccagcag ccgcggtgat acgtagggtg cgagcgttgt ccggaattat 480
tgggcgtaaa gagcttgtag gcggttgatt gcgtcagaag tgaaatctca gggcttaacc 540
ctgagtctgc ttctgatacg ggtcgactag agggaagtag gggagaacgg aattcctggt 600
ggagcggtgg aatgcgcaga tatcaggagg aacaccggtg gcgaaggcgg ttctctggac 660
ttttcctgac gctgagaagc gaaagcgtgg ggagcaaaca ggcttagata ccctggtagt 720
ccacgccgta aacggtgggt actaggtgtg ggggacttcc acgttctctg tgccgtagct 780
aacgcattaa gtaccccgcc tggggagtac ggccgcaagg ctaaaactca aaggaattga 840
cggggccccg cacaagcggc ggagcatgcg gattaattcg atgcaacgcg aagaacctta 900
cctgggtttg acatgtaccg gaaacatctg gagacaggtg cccctttttg gtcggtacac 960
aggtggtgca tggctgtcgt cagctcgtgt cgtgagatgt tgggttaagt cccgcaacga 1020
gcgcaaccct cgtcctatgt tgccagcggg taatgccggg gactcatggg agaccgccgg 1080
ggccaactcg gaggaaggtg gggatgaggt caagtcatca tgccccttat gtccagggct 1140
tcacgcatgc tacaatggct ggtacaaagg gctgcgatgc cgcaaggttg agcgaatccc 1200
aaaaagccag tctcagttcg gattggggtc tgcaactcga ccccatgaag tcggagtcgc 1260
tagtaatcgc agatcagcaa cgctgcggtg aatacgttcc cggggcttgt acacaccgcc 1320
cgtcaagtca tgaaagtcgg taacacccga agccggtggc ccaaccctt 1369
Claims (14)
1. A strain of Oleericoccum amyloliquefacien 99B of Propionibacterium family has a preservation number of CGMCC No. 20523.
2. Use of Oleericoccum amyloliquefaciens of Propionibacterium family 99B CGMCC No.20523 or a bacterial suspension thereof or a bacterial agent containing the same in the production or preparation of amylase.
3. Use of Oleericoccum amyloliquefaciens of Propionibacterium family 99B CGMCC No.20523 or a bacterial suspension thereof or a microbial inoculum containing the same in producing or preparing lipase.
4. The application of the Oleericoccum amyloliquefaciens of the propionibacteriaceae or the bacterial suspension thereof or the microbial inoculum containing the same in heterotrophic nitrification and/or aerobic denitrification is disclosed, wherein the Oleericoccum amyloliquefaciens of the propionibacteriaceae is Oleericoccum amyloliquefaciens 99B CGMCC No. 20523.
5. Application of Oleericoccum amyloliquefaciens of Propionibacterium family or bacterial suspension thereof or microbial inoculum containing the same in preparation of heterotrophic nitrification and/or aerobic denitrification products, wherein the Oleericoccum amyloliquefaciens of Propionibacterium family is Oleericoccum amyloliquefaciens 99B CGMCC No. 20523.
6. Application of Oleericoccum amyloliquefaciens of Propionibacterium family or bacterial suspension thereof or microbial inoculum containing the same in degrading ammonia nitrogen, wherein the Oleericoccum amyloliquefaciens of Propionibacterium family is Oleericoccum amyloliquefaciens 99B CGMCC No.20523 of Propionibacterium family.
7. Application of Oleericoccum amyloliquefaciens of Propionibacterium family or bacterial suspension thereof or microbial inoculum containing the same in preparation of products for degrading ammonia nitrogen, wherein the Oleericoccum amyloliquefaciens of Propionibacterium family is Oleericoccum amyloliquefaciens 99B CGMCC No.20523 of Propionibacterium family.
8. Application of Oleericoccum amyloliquefaciens of Propionibacterium family or bacterial suspension thereof or microbial inoculum containing the same in nitrogen pollutant removal or kitchen waste treatment or sewage treatment, wherein the Oleericoccum amyloliquefaciens of Propionibacterium family is Oleericoccum amyloliquefaciens 99B CGMCC No.20523 of Propionibacterium family.
9. Application of Oleericoccum amyloliquefaciens of propionibacteriaceae or bacterial suspension thereof or microbial inoculum containing the same in preparation of products for removing nitrogen pollutants or treating kitchen waste or sewage, wherein the Oleericoccum amyloliquefaciens of propionibacteriaceae is Oleericoccum amyloliquefaciens 99B CGMCC No. 20523.
10. A microbial inoculum, the active ingredient of which is Oleericoccum amyloliquefaciens of Propionibacterium family or a bacterial suspension thereof, the microbial inoculum having any one of the following functions 1) to 5):
1) producing or preparing an amylase;
2) producing or preparing a lipase;
3) heterotrophic nitrification and/or aerobic denitrification;
4) degrading ammonia nitrogen;
5) removing nitrogen pollutants or treating kitchen waste or sewage;
the Propionibacterium family Oleaceae Oliericoccum amyloliquefaciens is Propionibacterium family Oleericoccum amyloliquefaciens 99B CGMCC No. 20523.
11. A method for producing or preparing an amylase, comprising the step of subjecting an Oleericoccum amyloliquefacien of the family Propionibacterium to fermentation culture, wherein the Oleericoccum amyloliquefacien of the family Propionibacterium is Oleericoccum amyloliquefacien 99B CGMCC No.20523 of the family Propionibacterium.
12. A method for producing or preparing a lipase, comprising the step of subjecting an Oleericoccum amyloliquefacien of the family Propionibacterium to fermentation culture, wherein the Oleericoccum amyloliquefacien of the family Propionibacterium is Oleericoccum amyloliquefacien 99B CGMCC No.20523 of the family Propionibacterium.
13. A method for degrading ammonia nitrogen comprises the step of treating ammonia nitrogen with Oleericoccum amyloliquefaciens of Propionibacterium family or bacterial suspension thereof or microbial inoculum containing the same, wherein the Oleericoccum amyloliquefaciens of Propionibacterium family is Oleericoccum amyloliquefaciens 99B CGMCC No.20523 of Propionibacterium family.
14. A method for removing nitrogen pollutants or treating kitchen waste or sewage comprises the step of treating the nitrogen pollutants or the kitchen waste or the sewage by using Oleericoccum amyloliquefaciens of Propionibacterium family or bacterial suspension thereof or microbial inoculum containing the same, wherein the Oleericoccum amyloliquefaciens of Propionibacterium family is Oleericoccum amyloliquefaciens 99B CGMCC No.20523 of Propionibacterium family.
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