Pseudomonas stutzeri C1-1 and application thereof

Abstract

The invention provides a pseudomonas stutzeri (Stutzeria stutzeri) C1-1 and an application of the pseudomonas stutzeri (Stutzeria stutzeri) C1-1. The preservation number of the pseudomonas stutzeri is CGMCC (China General Microbiological Culture Collection Center) NO.32082, the pseudomonas stutzeri is preserved in the China General Microbiological Culture Collection Center, the preservation date is September 26th, and the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The embodiment of the invention also provides an application of the Stutzeria stutzeri (Stutzeria stutzeri) C1-1 in short-cut denitrification of the sewage. According to the present invention, the existing nitrogen source and the existing carbon source in the sewage can be utilized to convert NO3 <->-N into NO2 <->-N, the good NO2 <->-N accumulation effect is provided, and the nitrite converted from the nitrate can be stably accumulated during the reaction process.

Description

Technical Field

[0001] The present application relates to the technical field of sewage treatment, and in particular to a Pseudomonas stutzeri (Stutzerimonas stutzeri) C1-1 and its application. Background Art

[0002] With the proposal of the "dual carbon" goal, greenhouse gas emissions in sewage treatment have received widespread attention. The important direct emission source in the biological denitrification unit of sewage treatment is the intermediate product N2O gas in the denitrification stage. The short-range denitrification (PD) process can use denitrifying bacteria to convert NO3--N into NO2--N by controlling the amount of carbon source added, that is, to control the product of complete denitrification to the stage where only NO2--N is generated, thereby reducing the emission of the back-end product N2O gas.

[0003] Finding short-range denitrifying functional bacteria that can stably accumulate NO2--N is of great value for sewage treatment. Summary of the invention

[0004] In view of this, the purpose of the present application is to provide a Pseudomonas stutzeri C1-1 and its application.

[0005] The present application provides Pseudomonas stutzeri C1-1, with a deposit number of CGMCCNO.32082, deposited in the General Microbiology Center of China National Committee for the Preservation of Microorganisms, with a deposit date of September 26, and a deposit address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0006] The present application also provides an application of the aforementioned Pseudomonas stutzeri C1-1 in short-range denitrification of sewage.

[0007] In some embodiments, the sewage is municipal sewage or industrial sewage.

[0008] In some of the embodiments, the carbon-nitrogen ratio of the sewage is adjusted to 8-12.

[0009] The present application also provides a Stutzeri C1-1 inoculum solution, which is prepared using the Stutzeri C1-1 as described above.

[0010] The present application also provides a method for preparing the aforementioned Stutzeri C1-1 inoculum, comprising:

[0011] The Pseudomonas stutzeri C1-1 with the deposit number of CGMCC NO.32082 was inoculated into the denitrification medium and cultured for 2.9-3.1 days to obtain the Pseudomonas stutzeri C1-1 bacterial liquid; the culture temperature was 28-32°C;

[0012] The Pseudomonas stutzeri C1-1 bacterial solution is centrifuged, the supernatant is removed, and a buffer having the same volume as the supernatant is mixed with the obtained precipitate to obtain the Pseudomonas stutzeri C1-1 inoculum solution.

[0013] The embodiments of the present application also provide a method for short-range denitrification of wastewater, using the Stutzerimonas stutzeri C1-1 inoculum as described above, or the Stutzerimonas stutzeri C1-1 inoculum prepared by the method for preparing the Stutzerimonas stutzeri C1-1 inoculum as described above.

[0014] In some embodiments, including:

[0015] The inoculation solution of Pseudomonas stutzeri C1-1 is inoculated into the denitrification medium at a concentration of 0.8-1.2% by volume; and the culture is carried out anaerobically at 28-32° C. for 35-37 hours.

[0016] In some embodiments, the carbon-nitrogen ratio of the denitrification medium is greater than or equal to 4.

[0017] In some embodiments, the carbon-nitrogen ratio of the denitrification medium is 8-12 BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the present application or related technologies, the drawings required for use in the embodiments or related technical descriptions are briefly introduced below. Obviously, the drawings described below are only embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0019] Figure 1 Schematic diagram of the growth of Pseudomonas stutzeri C1-1 under anaerobic conditions.

[0020] Figure 2 The figure shows a comparison of NO3--N degradation by four species of Pseudomonas stutzeri in denitrification medium.

[0021] Figure 3 This is a comparison of the short-term denitrification effects of four species of Pseudomonas stutzeri in denitrification culture medium.

[0022] Figure 4 NO3 of Pseudomonas stutzeri C1-1 under different C / N conditions - -N degradation situation.

[0023] Figure 5 NO2 of Pseudomonas stutzeri C1-1 under different C / N conditions - -N accumulation situation.

[0024] Figure 6 NO2 of Pseudomonas stutzeri C1-1 at C / N=8 - -N cumulative stability verification.

[0025] Figure 7 NO3 of Pseudomonas stutzeri C1-4 under different C / N conditions - -N degradation situation.

[0026] Figure 8 NO2 of Pseudomonas stutzeri C1-4 under different C / N conditions - -N accumulation situation. DETAILED DESCRIPTION

[0027] In order to make the objectives, technical solutions and advantages of the present application more clearly understood, the present application is further described in detail below in combination with specific embodiments and with reference to the accompanying drawings.

[0028] It should be noted that, unless otherwise defined, the technical terms or scientific terms used in the embodiments of the present application should have the common meanings understood by persons with ordinary skills in the field to which the present application belongs.

[0029] Nitrifying, denitrifying and anaerobic ammonium oxidizing microorganisms are widely present in wetlands, supporting the operation of wetland nitrogen cycles. The artificial wetland sediments that treat aquaculture wastewater are rich in various types of denitrifying bacteria, and the strains generally tolerate high C / N wastewater environments and are naturally associated with other nitrogen cycle microorganisms. The purpose of this application is to isolate and screen a denitrifying bacterium from artificial wetland sediments, and to explore the key C / N conditions for its growth and stable accumulation of nitrite nitrogen, to support its application as a functional bacterial agent in the PD / A process.

[0030] The present application embodiment provides a short-range denitrifying bacteria: Pseudomonas stutzeri (Stutzerimonas stutzeri) C1-1. The deposit number is CGMCC NO.32082, deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration, the deposit date is September 26, 2024, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0031] The Pseudomonas stutzeri C1-1 has a good short-range denitrification effect and can use the existing nitrogen and carbon sources in sewage to convert NO3 - -N turns into NO2 - -N, and has good NO2 - -N accumulation effect, the ability to stably accumulate nitrite converted from nitrate during the reaction process. At the same time, it can reduce oxygen consumption by 50% during sewage treatment, reducing sewage treatment costs and greenhouse gas effects. It also has the advantages of being able to grow rapidly under anaerobic conditions, and is suitable for colonization in anaerobic ammonia oxidation sludge in small quantities for short-term denitrification. The short-term denitrification efficiency at the appropriate carbon source dosage in sewage can be as high as 91% or more, and can stably maintain the accumulation of nitrite nitrogen for more than 24 hours. It can be used for front-end treatment of sewage, and the product NO2 obtained after treating sewage - -N can provide substrate for other denitrification functional bacteria to make up for NO2 in urban sewage - -N concentration is low.

[0032] Based on the same inventive concept, the present application also provides an application of the above-mentioned Pseudomonas stutzeri C1-1 in short-range denitrification of sewage, wherein the sewage can be municipal sewage or industrial sewage.

[0033] In some of the embodiments, the carbon-nitrogen ratio of the sewage can be adjusted to be above 4 so that Pseudomonas stutzeri C1-1 can have better short-range denitrification ability.

[0034] In some of the embodiments, the carbon-nitrogen ratio of the sewage can be adjusted to 8-12 so that Pseudomonas stutzeri C1-1 can have better short-range denitrification ability.

[0035] Based on the same inventive concept, the present application embodiment also provides a Pseudomonas stutzeri (Stutzerimonas stutzeri) C1-1 inoculation solution, which is prepared using the Pseudomonas stutzeri (Stutzerimonas stutzeri) C1-1 as described above.

[0036] In some embodiments, the method for preparing the Stutzerimonas stutzeri C1-1 inoculum may include:

[0037] The Pseudomonas stutzeri C1-1 with the deposit number of CGMCC NO.32082 was inoculated into the denitrification medium and cultured for 2.9-3.1 days to obtain the Pseudomonas stutzeri C1-1 bacterial liquid; the culture temperature was 28-32°C;

[0038] The Pseudomonas stutzeri C1-1 bacterial solution is centrifuged, the supernatant is removed, and a buffer having the same volume as the supernatant is mixed with the obtained precipitate to obtain the Pseudomonas stutzeri C1-1 inoculum solution.

[0039] In some embodiments, the composition of the denitrification culture medium may include: 0.2g / LKNO3, 0.566g / L sodium citrate, 10.55g / L Na2HPO4·12H2O, 1.5g / LKH2PO4, 0.1g / L MgSO4·7H2O, and 2mL / L trace element solution (each liter of trace element solution may include 50.0g EDTA-Na2, 5.06g MnCl2·4H2O, 2.2g ZnSO4, 5.0gFeSO4·7H2O, 5.5g CaCl2, 1.61g CoCl2·6H2O, 1.57g CuSO4·5H2O). After culturing in this culture medium for about 3 days, a bacterial solution grown to a saturated bacterial concentration can be obtained.

[0040] Based on the same inventive concept, an embodiment of the present application also provides a method for short-range denitrification of sewage, using the Stutzerimonas stutzeri C1-1 inoculum as described above, or the Stutzerimonas stutzeri C1-1 inoculum prepared by the preparation method of the Stutzerimonas stutzeri C1-1 inoculum as described above.

[0041] In some embodiments, the method for short-range denitrification of sewage may include: inoculating the Pseudomonas stutzeri C1-1 inoculum in a denitrification medium at a concentration of 0.8-1.2% by volume; and anaerobic culturing at 28-32° C. for 35-37 hours.

[0042] In some embodiments, the carbon-nitrogen ratio of the denitrification medium is greater than or equal to 4, for example, the carbon-nitrogen ratio of the denitrification medium may be 8-12, so that Pseudomonas stutzeri C1-1 can have better short-range denitrification ability.

[0043] The technical solution of the present invention is further described below in conjunction with specific implementation methods.

[0044] The experimental methods in the following examples are all conventional methods unless otherwise specified.

[0045] The test materials used in the following examples, unless otherwise specified, were purchased from a conventional biochemical reagent store. The enrichment medium (pH = 7.0) contained the following reagents per liter: 0.2 g KNO3, 0.566 g sodium citrate, 5% (volume ratio) Weiss salt solution, which consisted of 25 g MgSO4·7H2O, 0.04 g MnSO4·H2O, 2.5 g NaCl, 0.05 g FeSO4·7H2O, and 6.5 g K2HPO4·3H2O. Denitrification medium (pH = 7.0-7.3), containing the following reagents per liter: 0.2 g KNO3, 0.566 g sodium citrate, 10.55 g Na2HPO4·12H2O, 1.5 g KH2PO4, 0.1 g MgSO4·7H2O, and 2 mL of trace element solution (including 50.0 g EDTA-Na2, 5.06 g MnCl2·4H2O, 2.2 g ZnSO4, 5.0 g FeSO4·7H2O, 5.5 g CaCl2, 1.61 g CoCl2·6H2O, 1.57 g CuSO4·5H2O per liter). Screening medium (pH = 7.0), each liter contains the following reagents: 1.0g KNO3, 8.5g sodium citrate, 1.0g KH2PO4, 1.0g asparagine, 1.0g MgSO4·7H2O, 0.2g CaCl2·6H2O, 0.05g FeCl3·6H2O and 1mL of bromothymol blue solution (0.1g bromothymol blue dissolved in 10mL anhydrous ethanol). Solid culture medium can be added with 2% agar in liquid culture medium and sterilized in a high pressure steam sterilizer at 121°C for 20min.

[0046] Example 1 Screening of Pseudomonas stutzeri

[0047] Screening method: Sediment samples (5-10 cm depth) were collected from a foxtail algae artificial wetland in Changsha County, Changsha City, Hunan Province. The following bacterial liquid transfer and inoculation operations were completed in a sterile operating table, and the anaerobic environment was created by filling argon in the anaerobic serum bottle.

[0048] For every 10g of sediment sample, 100mL of 1× phosphate buffer solution (PBS) and a small amount of glass beads were added to a 250mL anaerobic serum bottle at 30°C, 150r / min constant temperature oscillation for 30min to fully release the microorganisms in the sediment. Then 10mL of the sediment suspension was transferred into 100mL of enrichment medium and oscillated at 30°C, 150r / min in an anaerobic serum bottle. During the enrichment stage, 10mL of the culture solution was aspirated and transferred to fresh enrichment medium every 2 days for a total of 28 days.

[0049] Take the enriched culture solution, apply gradient dilutions on the solid plate prepared with denitrification medium, and culture at a constant temperature of 30°C. Use an inoculation loop to separate the cultured colonies with different appearances and morphologies on the fresh denitrification solid medium, so as to obtain single colonies.

[0050] After three generations of purification, the microorganisms were inoculated on solid screening medium and cultured at 30° C. The strains that changed the color of the screening medium from green to blue were the rescreened strains.

[0051] Sequencing method: The four rescreened strains were numbered, and the genomic DNA of the isolated strains was extracted using the TlANamp Bacteria DNA Kit (TIANGEN). The 16S rDNA fragment of the strain DNA was amplified using universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGYTACCTTGT TACGACTT-3'). The amplified product was completed by Beijing Qingke Biotechnology Co., Ltd.

[0052] Sequencing results: The sequencing results are shown in SEQ 01. The sequencing results showed that the screened strains were Pseudomonas stutzeri (formerly known as Pseudomonas stutzeri, now changed to Stutzerimonas stutzeri), which were named Pseudomonas stutzeri C1-1, P2-1, C1-4 and G2-2 respectively.

[0053] Example 2 Preparation of Pseudomonas stutzeri Inoculum

[0054] Preparation method of inoculation solution: Pick and screen a single colony of Pseudomonas stutzeri on a solid culture medium, place it in a denitrification culture medium and culture it at 30°C, 150r / min constant temperature shaking for 3 days to obtain a bacterial solution that has grown to a saturated bacterial concentration. In order to avoid interference from residual nitrogen in the culture medium, the saturated bacterial solution is centrifuged at 4500rpm for 10min, the supernatant is discarded and resuspended with an equal amount of PBS buffer to obtain the inoculation solution.

[0055] Example 3 Screening of inoculation dose of inoculation solution

[0056] Experimental method: 100 mL of denitrification culture medium was inoculated with 10 mL and 1 mL of inoculum of Pseudomonas stutzeri C1-1 respectively. The culture medium was cultured at 30°C and 150 r / min with constant temperature shaking. Samples were taken every 4 hours to measure the OD600 of the bacterial solution to determine the growth progress of Pseudomonas stutzeri C1-1 at different time periods and different inoculation amounts.

[0057] Experimental results: Figure 1 shown.

[0058] Result analysis: According to Figure 1 , Pseudomonas stutzeri C1-1 enters the logarithmic growth phase at 8-12h and the stable phase at 12-16h. During the stable phase, OD600 can reach above 1.3. Compared with the initial 0.003-0.045, the biomass amplification is more than 100 times, which shows that the bacteria grows rapidly. When the inoculation amount is 10%, C1-1 grows faster than when the inoculation amount is 1%, and enters the logarithmic phase and the stable phase in 4h. During the stable phase of the two inoculation amounts, the bacterial solution concentrations are similar, and when the inoculation amount is 1%, OD600 preferentially rises to a peak value greater than 1.4. Therefore, when adding the bacterial agent, an inoculation amount of 1% can meet the conventional proliferation requirements, and the Pseudomonas stutzeri C1-1 strain is suitable for rapid production and amplification of bacterial agents.

[0059] Example 4 Denitrification Effects of Four Species of Pseudomonas stutzeri

[0060] Test method: According to the preparation method of the inoculum in Example 2, prepare the inoculum of the strains of Pseudomonas stutzeri C1-1, Pseudomonas stutzeri P2-1, Pseudomonas stutzeri C1-4 and Pseudomonas stutzeri G2-2 respectively. The four inoculums were cultured in denitrification medium respectively. The specific culture method is: add 1% inoculum to 100ml of denitrification medium, culture in a 250ml anaerobic serum bottle under hypoxia, 30°C, 150r / min conditions for 36h. Set up a control group without adding inoculum and culture under the same conditions. Take samples every 4h and filter through a 0.45μm filter to measure the NO2 - -N and NO3 - -N concentration.

[0061] Experimental results: Figure 2 and Figure 3 shown.

[0062] Result analysis: referring to the changes in the control group, the natural decomposition of the substrate during the culture process can be ignored. Figure 2 It can be seen that Pseudomonas stutzeri strains P2-1, Pseudomonas stutzeri C1-4 and Pseudomonas stutzeri G2-2 can all achieve a 40% nitrate-nitrogen degradation rate within 24 hours of culture, but Pseudomonas stutzeri C1-1 can maintain a nitrate-nitrogen degradation rate of about 60% within 12 hours to 48 hours of culture, and has a more outstanding denitrification potential.

[0063] Depend on Figure 3 It can be seen that among the four species of Pseudomonas stutzeri, C1-1 of Pseudomonas stutzeri can achieve the most efficient nitrate degradation and nitrite accumulation effect, and the effect can be maintained for 48 hours. However, only trace amounts of NO2 were detected in C1-4 of Pseudomonas stutzeri. - In the culture medium of Pseudomonas stutzeri P2-1 and Pseudomonas stutzeri G2-2, almost no NO2 was detected. - -N increased significantly.

[0064] The purpose of the present invention is to isolate and screen a denitrifying bacteria from artificial wetland sediment, and explore the key C / N conditions for its growth and stable accumulation of nitrite nitrogen, to support its application as a functional bacterial agent in the PD / A process. Compared with Pseudomonas stutzeri, C1-4 is more suitable for forming a cooperative denitrification relationship with functional bacteria that utilize nitrite substrates in sewage treatment.

[0065] Example 5 Denitrification effects of four strains of Pseudomonas stutzeri.

[0066] Implementation method: On the basis of the denitrification medium, the carbon source and nitrogen source were adjusted, that is, Na2HPO4·12H2O 10.55 g / L, KH2PO4 1.5 g / L, MgSO4·7H2O 0.1 g / L, and 2 mL / L of trace element components were retained in the denitrification medium, and KNO3 and sodium citrate were added as nitrogen source and carbon source for the culture of Pseudomonas stutzeri C1-1 and Pseudomonas stutzeri C1-4 according to different experimental groups as shown in Table 1.

[0067] 1% inoculum (same as in Example 4) was added to 100 mL of denitrification culture medium with different C / N, and a blank control group with 1% PBS buffer was set up, and cultured for 36 hours under anoxic conditions at 30°C and 150 r / min. Samples were taken every 4 hours and filtered through a 0.45 μm filter to determine the NO2 - -N and NO3 - -N concentration.

[0068] Experimental results: Figure 4-Figure 8 As shown. Among them, Figure 4 NO3 of Pseudomonas stutzeri C1-1 under different C / N conditions - -N degradation situation. Figure 5 NO2 of Pseudomonas stutzeri C1-1 under different C / N conditions - -N accumulation situation. Figure 6 NO2 of Pseudomonas stutzeri C1-1 at C / N=8 - -N cumulative stability verification. Figure 7 NO3 of Pseudomonas stutzeri C1-4 under different C / N conditions - -N degradation situation. Figure 8 NO2 of Pseudomonas stutzeri C1-4 under different C / N conditions - -N accumulation situation.

[0069] Result analysis: Figure 4 and Figure 5 It can be seen that for Pseudomonas stutzeri C1-1, when C / N ≥ 4, NO3 - -N began to show obvious degradation. When C / N=8, 10, 12, NO3- The degradation rate of -N can reach more than 85% within 24 hours, among which when C / N=12, the degradation rate reaches 95.9%, which is the highest. In particular, when C / N=8, NO3 - -N showed a significant decrease of 70.8%, while when more carbon source was added, NO3 - A significant decrease in -N will be delayed to 16-20 hours. Figure 5 Medium NO2 - The change in -N concentration more clearly reflects that only in the high C / N environment of C / N=8, 10, and 12, Pseudomonas stutzeri C1-1 has NO2 - -N accumulation effect is manifested, and only when C / N=8, Pseudomonas stutzeri C1-1 can stably accumulate NO2 - -N exceeds 24h. In addition, under different carbon source addition conditions, Pseudomonas stutzeri C1-1 will express different denitrification pathways. Figure 6 It shows that when C / N=8, the C1-1 system can reach 31.72% NO2 in 12h. - -N accumulation rate. In the following 24h, NO2 - The -N accumulation rate can be maintained above 30% continuously.

[0070] Depend on Figure 7 It can be seen that for Pseudomonas stutzeri C1-4, when C / N=12, the bacteria achieved the best degradation effect on nitrate nitrogen, degrading 33.2% of NO3 in 24 hours. - -N. is much lower than the 85% degradation efficiency achieved by Pseudomonas stutzeri C1-1 at 24h. Figure 8 , Pseudomonas stutzeri C1-4 only produces NO2 when C / N=8, 10 - -N accumulation effect is reflected in that the nitrite accumulation rate is only 13.0%, which is much lower than the nitrite accumulation rate of Pseudomonas stutzeri C1-1 when C / N=8.

[0071] Therefore, among the Pseudomonas stutzeri C1-1 and C1-4 with short-range denitrification function, the high C / N culture environment is more suitable for Pseudomonas stutzeri C1-1 to exert its denitrification function. Pseudomonas stutzeri C1-1 is a rare strain with good short-range denitrification effect. It can complete efficient short-range denitrification on the basis of saving carbon source cost. The bacteria can be used as functional bacteria in the biological denitrification process of sewage, has the potential to provide substrates for other functional bacteria, and can enhance the mutual cultivation of bacteria.

[0072] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present disclosure (including the claims) is limited to these examples. Based on the concept of the present disclosure, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of the different aspects of the embodiments of the present disclosure as described above, which are not provided in detail for the sake of simplicity.

[0073] While the disclosure has been described in conjunction with specific embodiments thereof, many alternatives, modifications and variations of these embodiments will be apparent to those skilled in the art in light of the foregoing description.

[0074] The embodiments of the present disclosure are intended to cover all such substitutions, modifications and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the embodiments of the present disclosure should be included in the scope of protection of the present disclosure.

Claims

1. A Pseudomonas stutzeri C1-1, characterized in that The deposit number is CGMCCNO.32082, and it is deposited in the General Microbiology Center of China Culture Collection Administration. The deposit date is September 26, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

2. Use of Pseudomonas stutzeri C1-1 as claimed in claim 1 in short-cut denitrification of sewage.

3. The use according to claim 2, characterized in that: The sewage is municipal sewage or industrial sewage.

4. The use according to claim 2, characterized in that: The carbon-nitrogen ratio of the sewage is adjusted to 8-12.

5. A Stutzeri C1-1 inoculum, prepared using the Stutzeri C1-1 of claim 1.

6. A method for preparing the inoculum of Pseudomonas stutzeri C1-1 as claimed in claim 5, characterized in that: include: The Pseudomonas stutzeri C1-1 with the deposit number of CGMCC NO.32082 was inoculated into the denitrification medium and cultured for 2.9-3.1 days to obtain the Pseudomonas stutzeri C1-1 bacterial liquid; the culture temperature was 28-32°C; The Pseudomonas stutzeri C1-1 bacterial solution is centrifuged, the supernatant is removed, and a buffer having the same volume as the supernatant is mixed with the obtained precipitate to obtain the Pseudomonas stutzeri C1-1 inoculum solution.

7. A method for short-cut denitrification of sewage, characterized in that: The Pseudomonas stutzeri (Stutzerimonas stutzeri) C1-1 inoculum liquid is prepared by the preparation method of the Pseudomonas stutzeri (Stutzerimonas stutzeri) C1-1 inoculum liquid according to claim 5 or the Pseudomonas stutzeri (Stutzerimonas stutzeri) C1-1 inoculum liquid according to claim 6.

8. The method for short-cut denitrification of sewage according to claim 7, characterized in that: include: The inoculation solution of Pseudomonas stutzeri C1-1 is inoculated into the denitrification medium at a concentration of 0.8-1.2% by volume; and the culture is carried out anaerobically at 28-32° C. for 35-37 hours.

9. The method for short-cut denitrification of sewage according to claim 8, characterized in that: The carbon-nitrogen ratio of the denitrification culture medium is greater than or equal to 4.

10. The method for short-cut denitrification of sewage according to claim 9, characterized in that: The carbon-nitrogen ratio of the denitrification culture medium is 8-12.

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