Anaerobic treatment method of heparin sodium wastewater

Abstract

The invention discloses an anaerobic treatment method for heparin sodium wastewater, and belongs to the field of water pollution prevention and control. The invention provides an arthrobacter glass fly A1 which is preserved in the China Center for Type Culture Collection on May 23, 2024, and the preservation number of the arthrobacter glass fly A1 is CCTCC (China Center for Type Culture Collection) NO: M20241041. The arthrobacter sp. Can tolerate the salinity of 8.0%, and can still effectively secrete protease under the stress of high salt. The strain can provide an effective strain resource for strengthening anaerobic biological treatment of high-salt and high-protein wastewater such as heparin sodium wastewater, so that the treatment cost is remarkably reduced, and the strain has important application value.

Description

Technical Field

[0001] The invention belongs to the field of water pollution prevention and control, and specifically relates to an anaerobic treatment method for sodium heparin wastewater. Background Art

[0002] High-salinity organic wastewater mainly comes from chemical, pharmaceutical, food and pesticide related industries. In addition to organic pollutants, this type of wastewater also contains high concentrations of soluble inorganic ions (Cl - , SO4 2- , Na + , Ca 2+ ). At present, the treatment methods for high-salinity wastewater mainly include biological method, membrane separation method and high-efficiency evaporation method. The biological method relies on microorganisms to remove pollutants and is mainly divided into anaerobic biological treatment and aerobic biological treatment.

[0003] High-salt and high-protein wastewater mainly comes from food industries such as meat pickling and casing production, as well as pharmaceutical industries such as heparin sodium production. For example, heparin sodium production wastewater is a typical high-salt and high-protein wastewater, with a salinity of usually above 3.6% and a high COD content (mainly protein-derived organic matter). Anaerobic biological methods are more suitable for the treatment of high-concentration organic wastewater, but how to improve the treatment capacity in a high-salt environment is the key.

[0004] In order to improve the anaerobic treatment efficiency of high-salt and high-protein wastewater such as sodium heparin wastewater, based on the concept of bioaugmentation, constructing a highly salt-tolerant protein-degrading bacterial community is one of the feasible solutions to improve the treatment efficiency. Bioaugmentation technology does not require changes to the treatment process, has low cost of use, and shows strong application potential in wastewater treatment and pollutant biodegradation. Summary of the invention

[0005] Purpose of the invention: This patent provides an anaerobic treatment method for sodium heparin wastewater, specifically using a highly salt-tolerant protease-producing Arthrobacterium glabratum, which can tolerate 8.0% salinity and can still effectively secrete protease under high salt stress. The bacteria can effectively strengthen the anaerobic biological treatment of high-salt and high-protein wastewater such as sodium heparin wastewater, thereby significantly reducing the treatment cost and having important application value.

[0006] To achieve the above object, the technical solution of the present invention is:

[0007] The first object of the present invention is to provide an anaerobic treatment method for sodium heparin wastewater, the method comprising: using highly salt-tolerant and protease-producing Arthrobacterprotophormiae A1 to strengthen the anaerobic treatment of sodium heparin wastewater, the highly salt-tolerant and protease-producing Arthrobacterprotophormiae A1 was deposited in the China Center for Type Culture Collection on May 23, 2024, with an address in Wuhan, China, and a preservation number of CCTCC NO: M 20241041.

[0008] Optionally, in some embodiments of the present invention, the 16SrDNA sequence of the original glass fly Arthrobacter A1 is shown as SEQ ID NO.1.

[0009] Optionally, in some embodiments of the present invention, the method specifically includes constructing a sodium heparin wastewater-degrading bacterial community for anaerobic treatment of sodium heparin wastewater, and fermenting for 10-15 days at an inoculation amount of 5%-15%, pH 6.5-7.5 and 30-38°C, wherein the sodium heparin wastewater-degrading bacterial community includes salt-tolerant methanogenic sludge and Protothecobacterium glabratum A1, and the salt-tolerant methanogenic sludge is methanogenic sludge with a salinity tolerance of 3.0%-3.6%.

[0010] Optionally, in some embodiments of the present invention, in the sodium heparin wastewater anaerobic degradation bacterial community, the volume ratio of the salt-tolerant methanogenic activated sludge to the original glass fly Arthrobacter A1 activation liquid is 5:1-15:1, preferably 10:1.

[0011] Optionally, in some embodiments of the present invention, the salt-tolerant methanogenic sludge is methanogenic sludge having a salinity tolerance of 3.0%-3.2%.

[0012] Optionally, in some embodiments of the present invention, the method for cultivating salt-tolerant methanogenic sludge includes using anaerobic methanogenic sludge as the initial inoculum and peptone as the sole carbon source and nitrogen source, and taming salt-tolerant methanogenic sludge capable of tolerating a salinity of more than 3.0% by gradiently increasing salt stress pressure.

[0013] Optionally, in some embodiments of the present invention, the Arthrobacterium vitae produces protease under anaerobic conditions and has a salt tolerance of 4.0% or more.

[0014] Further optionally, in some embodiments of the present invention, the salt tolerance of the original glass fly Arthrobacter A1 is up to 8.0%.

[0015] Further optionally, in some embodiments of the present invention, the salinity tolerance of the original glass fly Arthrobacter A1 is 4.0%-8.0%.

[0016] Optionally, in some embodiments of the present invention, the enzyme activity of the protease reaches above 25 U / mL.

[0017] Further optionally, in some embodiments of the present invention, when the salt tolerance of the original glass fly Arthrobacter A1 is 4.0%-6.0%, the enzyme activity of the protease reaches more than 35U / mL.

[0018] Further optionally, in some embodiments of the present invention, when the salt tolerance of the original glass fly Arthrobacter A1 is 6.0%-8.0%, the enzyme activity of the protease reaches more than 25U / mL.

[0019] Optionally, in some embodiments of the present invention, the salinity of the sodium heparin wastewater reaches above 4.0%.

[0020] Optionally, in some embodiments of the present invention, the protein degradation rate of the sodium heparin wastewater is above 70%, and the COD removal rate is above 50%.

[0021] Further optionally, in some embodiments of the present invention, the protein degradation rate of the sodium heparin wastewater is above 80%, and the COD removal rate is above 60%.

[0022] Further optionally, in some embodiments of the present invention, the protein degradation rate of the sodium heparin wastewater is above 90%, and the COD removal rate is above 70%.

[0023] Optionally, in some embodiments of the present invention, the methane production reaches more than 150 mL / g-COD.

[0024] The second object of the present invention is to provide an anaerobic degradation bacterial community for sodium heparin wastewater, comprising salt-tolerant methanogenic sludge and any of the above-mentioned highly salt-tolerant protease-producing Arthrobacterium glabratum.

[0025] Optionally, in some embodiments of the present invention, the volume ratio of the salt-tolerant methanogenic sludge bacteria liquid to the activated liquid of Arthrobacter glabraensis is 5:1-15:1, preferably 10:1.

[0026] Beneficial effects: The anaerobic biological treatment of high-salt organic wastewater such as sodium heparin has always been one of the difficult problems in the field of environmental protection. Compared with the prior art, the method disclosed in the present invention can construct a protein-degrading bacterial community that can tolerate a salinity of more than 4.5%. When treating actual sodium heparin wastewater with a salinity of 4.3%, the COD removal rate and protein degradation rate reached 76.3% and 92.5%, respectively. Therefore, the technical method described in this patent can provide an effective bacterial strain resource for strengthening the anaerobic treatment of high-salt and high-protein wastewater such as sodium heparin wastewater, thereby significantly reducing the treatment cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 : The process flow of strain screening and salt-tolerant degradation bacterial community construction of the present invention.

[0028] Figure 2 : Changes in COD removal efficiency during adaptive evolution.

[0029] Figure 3 : Changes in protein degradation rates during adaptive evolution.

[0030] Figure 4 : Colony morphology of strain A1.

[0031] Figure 5 : Salt tolerance of the strain.

[0032] Figure 6 :Effect of salinity on the growth of Arthrobacterium truncatum A1.

[0033] Figure 7 :Effect of salinity on protease secretion from Arthrobacterium glabratum A1.

[0034] Figure 8 :Effect of salinity on protein degradation by Arthrobacterium protothecoides A1.

[0035] Fig. 9 :Construction of anaerobic degradation bacterial community for sodium heparin wastewater.

[0036] Fig.10 :Application effect of anaerobic degradation bacteria in sodium heparin wastewater. DETAILED DESCRIPTION

[0037] On the one hand, the present invention discloses an anaerobic treatment method for sodium heparin wastewater, which belongs to the field of water pollution prevention and control. Figure 1 The process flow of strain screening and salt-tolerant degradation bacterial community construction of the present invention is shown. The method adopts a "top-down" adaptive evolution strategy of anaerobic bacterial community combined with directional screening of highly salt-tolerant protease-producing bacteria to construct an anaerobic degradation bacterial community for sodium heparin wastewater. The main steps include:

[0038] (1) First, a "top-down" adaptive evolution strategy was adopted to domesticate salt-tolerant methanogenic sludge that could tolerate salinity exceeding 3.0% by gradiently increasing salt stress;

[0039] (2) To address the problem of protein hydrolysis inhibition under high salt stress, the salt-tolerant methanogenic sludge was used as the bacterial strain source to screen out highly salt-tolerant protease-producing strains;

[0040] (3) The selected highly salt-tolerant protease-producing strains were compounded with salt-tolerant methane-producing sludge to further construct a highly salt-tolerant protein-degrading bacterial community (tolerant to salinity above 4.5%), thereby effectively improving the anaerobic treatment effect of sodium heparin wastewater.

[0041] As a preferred improvement of the present invention, the salt tolerance adaptive evolution of the anaerobic bacteria is carried out in a continuous flow fully mixed anaerobic reactor, the initial inoculum is anaerobic methanogenic sludge, the inoculum amount is 25-35g-TS / L, peptone is used as the only carbon source and nitrogen source, and the influent concentration is 6.0-10.0g / L.

[0042] As a preferred improvement of the present invention, the salt tolerance adaptive evolution conditions are: temperature 30-38°C, pH 6.5-7.5, hydraulic retention time 5-10 days. On the basis of COD removal rate > 70% in the steady state period, the salt stress pressure is gradually increased to obtain salt-tolerant methanogenic sludge that can tolerate salinity exceeding 3.0%.

[0043] As a preferred improvement of the present invention, highly salt-tolerant protease-producing bacteria are separated and screened from salt-tolerant methanogenic sludge. The separation and screening process includes: separation and purification in 4.0% salinity peptone solid culture medium, primary screening of protease production capacity at 4.0% salinity, and secondary screening of salt tolerance at 8.0% salinity.

[0044] On the other hand, according to the above screening method, the present invention provides a highly salt-tolerant, protease-producing protactin-producing Arthrobacterium glabratum A1, whose preservation number is CCTCC NO: M 20241041, the preservation time is May 23, 2024, and the preservation location is China Center for Type Culture Collection.

[0045] As a preferred improvement of the present invention, the 16S rDNA of the original Arthrobacter glabraeni has a nucleotide sequence as shown in SEQ ID NO.1.

[0046] As a preferred improvement of the present invention, the salt tolerance of the original Arthrobacter glabraensis under anaerobic conditions can reach more than 4.0%, even more than 6.0%, even up to 8.0%, and secrete protease.

[0047] As a preferred improvement of the present invention, the original glass fly Arthrobacter is isolated and screened from anaerobic methanogenic sludge that has undergone salt tolerance adaptive evolution. The adaptive evolution is carried out in a continuous flow fermentation mode, with peptone as the only carbon and nitrogen source, and the salt stress pressure gradient is increased to more than 3.0%.

[0048] As a preferred improvement of the present invention, the separation and screening process of the original glass fly Arthrobacter includes: dilution and separation culture in 4.0% salinity peptone solid culture medium, primary screening of protease production ability at 4.0% salinity and secondary screening of salt tolerance at 8.0% salinity.

[0049] The present invention also provides an application of the Arthrobacter viridis in anaerobic treatment of sodium heparin wastewater.

[0050] The application in the anaerobic treatment of sodium heparin wastewater includes: after the original glass fly Arthrobacter A1 is activated and cultured, it is compounded with salt-tolerant adaptively evolved methanogenic sludge (also referred to as salt-tolerant methanogenic sludge in the following embodiments) as an inoculum (volume ratio of 1:10); under the conditions of an inoculum amount of 10%, a pH of 7.0, and a temperature of 36.0°C, actual sodium heparin wastewater with a salinity of 4.3% is treated, and the COD removal rate and protein degradation rate reach 76.3% and 92.5%, respectively.

[0051] The present invention is further explained below with specific examples. However, it is easy for those skilled in the art to understand that the specific process conditions and results described in the examples are only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims.

[0052] Example:

[0053] The high salt-tolerant protease producing bacteria separation and fermentation medium involved in the following embodiments are as follows:

[0054] ①4.0% salinity peptone solid medium: 10.0 g / L tryptone, 20.0 g / L agar, 40 g / L sodium chloride, 0.2 g / L magnesium sulfate heptahydrate, 1.0 g / L potassium dihydrogen phosphate, 1.5 g / L potassium hydrogen phosphate, 0.25 g / L L-cysteine ​​hydrochloride monohydrate. In addition, 1.0 mL of vitamin solution and 1.0 mL of trace element solution are added to the unit volume (L) of the solution to adjust the pH to 7.0.

[0055] ②4.0% salinity clear zone medium: casein 16.0g / L, sucrose 30.0g / L, agar 20.0g / L, sodium chloride 40g / L, disodium hydrogen phosphate 1.0g / L, magnesium sulfate heptahydrate 0.5g / L, sodium nitrate 2.0g / L, ferrous sulfate 0.01g / L. Adjust the pH to 7.0.

[0056] ③4.0% salinity gelatin liquefaction medium: beef extract 3.0g / L, gelatin 200.0g / L, trypsin 5.0g / L, sodium chloride 40g / L, adjust the pH to 7.0.

[0057] ④LB medium: 10.0 g / L tryptone, 5.0 g / L yeast extract, 10.0 g / L sodium chloride, adjust the pH to 7.0.

[0058] ⑤ Salt-tolerant rescreening medium: 10 g / L tryptone, 80 g / L sodium chloride, 0.2 g / L magnesium sulfate heptahydrate, 1.0 g / L potassium dihydrogen phosphate, 1.5 g / L potassium hydrogen phosphate, 0.25 g / L L-cysteine ​​hydrochloride monohydrate. In addition, 1.0 mL vitamin solution and 1.0 mL trace element solution are added to the unit volume (L) of the solution to adjust the pH to 7.0.

[0059] ⑥Basic fermentation medium: 6.5 g / L tryptone, 0.2 g / L magnesium sulfate heptahydrate, 1.0 g / L potassium dihydrogen phosphate, 1.5 g / L potassium hydrogen phosphate, 0.25 g / L L-cysteine ​​hydrochloride monohydrate. In addition, 1 mL of vitamin solution and 1 mL of trace element solution are added to the unit volume (L) of solution, and sodium chloride is added in different proportions to adjust the pH to 7.0.

[0060] Example 1 Adaptive evolution strategy to cultivate salt-tolerant methanogenic sludge

[0061] This embodiment adopts the adaptive evolution strategy of microbial flora to cultivate salt-tolerant methanogenic sludge. The adaptive evolution is carried out in a fully mixed anaerobic reactor, using a continuous flow fermentation mode, and the initial inoculum is anaerobic methanogenic activated sludge. The components of the reactor influent are shown in Table 1, with peptone as the only carbon and nitrogen source, the initial inoculum sludge concentration is 33.0g-TS / L, the stirring speed is 100rpm, the fermentation temperature is 36°C, and the pH is controlled at 7.0. The initial influent salinity is 1.5%, and the hydraulic retention time is 10 days.

[0062] Table 1 Influent components of adaptive evolution reactor

[0063]

[0064] (1) Changes in COD removal rate during adaptive evolution

[0065] In the process of salt stress adaptive evolution, COD removal rate was used to characterize the change of salt tolerance of anaerobic bacteria. On the basis of COD removal rate higher than 70% in steady state, the salinity of reactor influent was gradually increased, and salt tolerance adaptive evolution of anaerobic methanogenic sludge was carried out to obtain methanogenic sludge that could tolerate salinity higher than 3.0%.

[0066] like Figure 2 As shown, the inoculated anaerobic methanogenic sludge was able to adapt quickly under 1.5% (1-6d) and 2.0% (7-14d) salt stress, with the effluent COD ranging from 1280 to 1920 mg / L and the COD removal rate maintained above 80%.

[0067] When the influent salinity was increased to 2.6% (15-21d), the removal rate dropped to 64.0% and then quickly recovered to 77.6%.

[0068] When the influent salinity was further increased to 2.9% (22-51d), under this stress, COD removal first decreased to 58.1%, and then gradually recovered to 72.3%, but the tolerance response time was significantly prolonged.

[0069] When the salinity was further increased to 3.2% (52nd-61st day), the effluent COD concentration did not change significantly, and the COD removal rate remained stable between 74.4% and 84.0%.

[0070] This shows that salt-tolerant microorganisms have become dominant through salinity gradient domestication. However, when the salinity increased to 3.6% (62-116d), the COD removal rate gradually decreased to about 55.0% and was difficult to recover. Therefore, the "top-down" adaptive evolution strategy of anaerobic bacteria can initially construct methanogenic sludge that tolerates 3.2% salinity.

[0071] (2) Changes in protein degradation rate during adaptive evolution

[0072] The degradation rate of proteins during adaptive evolution is Figure 3 As shown, under 1.5% low salt stress, the average protein degradation rate can reach 93.2%.

[0073] With the increase of the influent salinity gradient and the gradual adaptation of the anaerobic bacteria to the salt stress, protein degradation first decreased and then gradually recovered. Finally, under 3.2% salt stress, the protein degradation rate recovered to about 90%.

[0074] However, when the inlet salinity was further increased to 3.6%, the effluent protein concentration increased significantly, and the protein degradation rate decreased to about 71.2%, which was difficult to recover.

[0075] This indicates that the protease secretion capacity of anaerobic bacteria is insufficient under this salt stress, and protein hydrolysis is inhibited. Therefore, in this example, salt-tolerant methanogenic sludge obtained under 3.2% salt stress was used as a source to screen highly salt-tolerant protease-producing bacteria.

[0076] Example 2 Directed screening of highly salt-tolerant protease producing bacteria

[0077] (1) Isolation and purification of protease-producing bacteria and initial screening of enzyme production capacity

[0078] Although the "top-down" adaptive evolution strategy can initially construct a methanogenic bacterial community that tolerates 3.2% salinity, under higher salt stress, insufficient protease secretion leads to inhibition of protein hydrolysis. Therefore, in order to increase the protease production performance under high salt stress, salt-tolerant enzyme-producing microorganisms were screened from salt-tolerant methanogenic sludge.

[0079] The separation and screening process includes: taking the salt-tolerant methanogenic sludge obtained under 3.2% salt stress as a sample, placing the anaerobic bacteria in an anaerobic tube and adding glass beads, oscillating and dispersing, and then gradient dilution with physiological saline. Then the diluted bacterial suspension is spread on a 4.0% salinity peptone solid culture medium plate and inverted in a 36°C anaerobic constant temperature incubator for 48 hours. The plate streak separation method is continued to purify the grown single colonies, and then the isolated and purified strains are stored on a 4.0% salinity solid slant culture medium.

[0080] A total of 16 strains (named A1-A16) were isolated and purified at 4.0% salinity. The isolated and purified strains were inoculated into 4.0% salinity transparent zone medium and 4.0% salinity gelatin liquefaction medium by three-point method and puncture inoculation method, respectively. The ratio of transparent zone diameter to colony diameter (>1.5) and gelatin liquefaction degree were used as evaluation criteria to screen out salt-tolerant strains with strong protease production ability. As shown in Table 2, strains with 1.5 < transparent zone diameter / colony diameter <2 accounted for 56.25%. Nine strains (A1, A2, A3, A4, A5, A7, A8, A10, A13) with ratio >1.5 and high gelatin liquefaction degree were selected for further analysis of their salt tolerance.

[0081] Table 2 Preliminary screening of enzyme production capacity of protease producing bacteria

[0082]

[0083] (2) Rescreening of salt tolerance of protease-producing bacteria

[0084] The protease-producing bacteria screened were subjected to salt tolerance rescreening. The specific process included: after the screened strains were cultured in LB liquid medium for 48 hours, the diluted bacterial suspensions were respectively applied to 8.0% salt tolerance rescreening solid medium, and then the protease-producing bacteria that could tolerate 8.0% salinity were screened (such as Figure 4 shown).

[0085] like Figure 5 As shown in the figure, after 48 h of batch fermentation in 8.0% salt-tolerant medium, the biomass of the fermentation broth of strain A1 was significantly higher than that of other strains, and the OD 600 Reached 0.95. Therefore, strain A1 has better salt tolerance.

[0086] (3) Identification of bacterial species

[0087] The 16S rDNA sequence of strain A1 obtained by screening using bacterial universal primers was amplified and sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing, and the 16S rDNA sequence was compared on GenBank. The results showed that the strain sequence length was 1437bp, and the similarity with the 16S rDNA sequence of Arthrobacter protophormiae (GenBank Number: GQ395612.1) was as high as 99.72%. Therefore, the highly salt-tolerant protease-producing strain was identified as Arthrobacter protophormiae, and was deposited in the China Center for Type Culture Collection on May 23, 2024, with a collection number of CCTCC NO: M 20241041, named Arthrobacter protophormiae A1, and its 16S rDNA sequence is shown in SEQ ID NO.01. Example 3 Salt tolerance of Arthrobacter protophormiae A1

[0088] (1) Effect of salinity on the growth of Arthrobacterium truncatum A1

[0089] The original glass fly Arthrobacter A1 was transferred to the basic medium for batch fermentation to investigate the effects of salt stress on its growth, protease secretion and protein degradation. The growth curves under different salinities (2.0%, 4.0%, 6.0%, 8.0%) are shown in Figure 2. Figure 6 As shown, it can grow under salinity conditions of 2.0%-8.0%. When the salinity reaches 4.0%, 6.0% and 8.0%, the strain has the highest biomass (OD 600 ) were 1.68, 1.32 and 0.99, respectively, compared with 2.0% salinity (maximum OD 600 1.76) were reduced by 4.5%, 25.0% and 43.7% respectively. Correspondingly, it can be concluded that at 4.0%, 6.0% and 8.0% high salinity, the maximum biomass retention rates reached 95.5%, 75.0% and 56.3% respectively. It can be seen that the original glass fly arthrobacter A1 has a high salt tolerance. Although high salinity has a certain inhibitory effect on its growth efficiency, it can still maintain more than half of the maximum biomass at a high salinity of 8.0%.

[0090] (2) Effect of salinity on protease secretion by Arthrobacterium truncatum A1

[0091] Effects of salinity on the secretion of protease from Arthrobacterium glabrata A1 Figure 7As shown, the protease activity in the fermentation broth under different salt stresses reached the maximum value during the stable growth period of bacteria (the second day). At 2.0%, 4.0%, 6.0% and 8.0% salinity, the maximum enzyme activity was 55.2U / mL, 52.6U / mL, 35.9U / mL and 25.5U / mL, respectively. It can be concluded that when the salinity reached 4.0%, 6.0% and 8.0%, the enzyme activity remained at 95.3%, 65.0% and 46.2% compared with the salinity of 2.0%. This shows that the original glass fly arthrobacter A1 can still grow effectively and secrete proteases under high salt stress.

[0092] (3) Effect of salinity on protein degradation by Arthrobacterium truncatum A1

[0093] Effects of salinity on protein degradation by Arthrobacterium glabrata A1 Figure 8 As shown in the figure, at 2.0%, 4.0%, 6.0% and 8.0% salinity, after 3 days of anaerobic batch fermentation, the protein degradation rates reached 61.1%, 55.1%, 48.9% and 41.4%, respectively. This shows that Arthrobacter spp. A1 can effectively hydrolyze proteins under high salt stress.

[0094] Example 4 Construction and application effect of anaerobic degradation bacterial community of sodium heparin wastewater

[0095] (1) Construction of anaerobic bacterial community for heparin sodium wastewater degradation

[0096] The screened original glass fly Arthrobacter A1 was activated in LB culture medium and then compounded with salt-tolerant methanogenic sludge in a volume ratio of 1:10, and the salt stress acclimation pressure was further increased.

[0097] like Fig. 9 As shown in the figure, compared with salt-tolerant methanogenic sludge, the COD removal rate and protein degradation rate of the composite bacterial community at 3.6% salinity increased to 76.5% and 93.2%, respectively. When the salinity increased to 4.0% (36-45 days), the COD removal rate remained above 75%; and when the salinity was further increased to 4.6% (46-67 days), after a short period of adaptation, the COD removal rate of the bacterial community could still reach 72.6%. However, when the salinity increased to 5.2% (68-161 days), the COD removal rate dropped significantly to about 39.0%, and it was difficult to recover. During the entire salinity increase stage, the protein removal rate remained above 90%. In addition, the VFAs concentration increased significantly to above 4.0g / L under 5.2% salinity stress. It can be seen that under 5.2% salinity stress, protein hydrolysis was not affected, and the accumulation of VFAs was the main reason for the decrease in COD removal rate. By combining Arthrobacterium glabratum A1 and salt-tolerant methanogenic sludge, we successfully constructed an anaerobic degradation bacterial community that can tolerate a salinity of 4.6% and used it for anaerobic treatment of sodium heparin wastewater.

[0098] (2) Application effect of anaerobic degradation bacteria in sodium heparin wastewater

[0099] The anaerobic degrading bacteria of sodium heparin wastewater (tolerant to 4.6% salinity), salt-tolerant methanogenic sludge (tolerant to 3.2% salinity) and original methanogenic sludge were used to treat actual sodium heparin wastewater with a salinity of 4.3%, respectively.

[0100] like Fig.10 As shown in the figure, the COD removal rate, protein degradation rate and methane production of the original methanogenic activated sludge were only 28.7%, 32.8% and 67.3 mL / g-COD respectively; the COD removal rate, protein degradation rate and methane production of the salt-tolerant methanogenic sludge were 39.1%, 45.7% and 101.4 mL / g-COD respectively. In comparison, the COD removal rate, protein degradation rate and methane production of the anaerobic degradation bacteria of sodium heparin wastewater reached 76.3%, 92.5% and 201.6 mL / g-COD respectively. It can be seen that the method of the present invention can effectively enhance the anaerobic treatment effect of high-salt and high-protein wastewater such as sodium heparin wastewater.

[0101] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. An anaerobic treatment method for heparin sodium wastewater, characterized in that: The method includes using highly salt-tolerant and protease-producing ArthrobacterprotophormiaeA1 to strengthen the anaerobic treatment of sodium heparin wastewater, wherein the ArthrobacterprotophormiaeA1 was deposited in the China Center for Type Culture Collection on May 23, 2024, with a deposit number of CCTCC NO: M20241041.

2. The method according to claim 1, characterized in that: The method specifically includes constructing a sodium heparin wastewater-degrading bacterial community for anaerobic treatment of sodium heparin wastewater, and performing anaerobic fermentation for 10-15 days at an inoculation amount of 5%-15%, a pH of 6.5-7.5, and 30-38° C., wherein the sodium heparin wastewater-degrading bacterial community includes salt-tolerant methanogenic sludge and Protohyalobacterium glabratum A1, and the salt-tolerant methanogenic sludge is methanogenic sludge with a salinity tolerance of 3.0%-3.6%.

3. The method according to claim 2, characterized in that: In the sodium heparin wastewater degrading bacterial community, the volume ratio of the salt-tolerant methanogenic sludge bacterial liquid to the activated liquid of the original glass fly Arthrobacter A1 is 5:1-15:1, preferably 10:

1.

4. The method according to claim 2, characterized in that: The salt-tolerant methanogenic sludge is methanogenic sludge with a salinity tolerance of 3.0%-3.2%; Preferably, the method for cultivating the salt-tolerant methanogenic sludge comprises using anaerobic methanogenic sludge as the initial inoculum and peptone as the sole carbon source and nitrogen source, and taming the salt-tolerant methanogenic sludge capable of tolerating a salinity of more than 3.0% by gradiently increasing the salt stress pressure.

5. The method according to claim 1, characterized in that: The original glass fly Arthrobacterium A1 produces protease under anaerobic conditions and has a salt tolerance of more than 4.0%. Preferably, the salt tolerance of the original Arthrobacterium vitae A1 is up to 8.0%. Preferably, the salinity tolerance of the original Arthrobacterium vitae A1 is 4.0%-8.0%.

6. The method according to claim 5, characterized in that: The enzymatic activity of the protease reaches 25U / mL or more; Preferably, when the salt tolerance of the original glass fly Arthrobacter A1 is 4.0%-6.0%, the enzyme activity of the protease reaches 35U / mL or more; Preferably, when the salt tolerance of the original glass fly Arthrobacter A1 is 6.0%-8.0%, the enzyme activity of the protease reaches more than 25U / mL.

7. The method according to claim 1, characterized in that: The salinity of the sodium heparin wastewater reaches above 4.0%.

8. The method according to claim 1, characterized in that: The protein degradation rate of the sodium heparin wastewater is above 70%, and the COD removal rate is above 50%; Preferably, the protein degradation rate of the sodium heparin wastewater is above 80%, and the COD removal rate is above 60%; Preferably, the protein degradation rate of the heparin sodium wastewater is above 90%, and the COD removal rate is above 70%; Preferably, the methane production reaches above 150 mL / g-COD.

9. A heparin sodium wastewater anaerobic degradation bacterial community, characterized in that: It includes salt-tolerant methanogenic activated sludge and original glass fly Arthrobacter A1. The salt-tolerant methanogenic sludge is methanogenic sludge with a salinity tolerance of 3.0%-3.6%. The original glass fly Arthrobacter A1 was preserved in the China Center for Type Culture Collection on May 23, 2024, and the preservation number is CCTCC NO: M20241041.

10. The anaerobic degrading bacterial community for sodium heparin wastewater according to claim 9, characterized in that: The volume ratio of the salt-tolerant methanogenic sludge bacteria liquid to the activated liquid of Arthrobacter glabratus A1 is 5:1-15:1, preferably 10:1.

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