CN111471625A - Sphingomonas spinosa and application thereof in garbage press filtrate flocculation - Google Patents
Sphingomonas spinosa and application thereof in garbage press filtrate flocculation Download PDFInfo
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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Abstract
The invention belongs to the field of biological environmental protection, and particularly relates to sphingomonas spinosa and application thereof in garbage press filtrate flocculation. The specific technical scheme is as follows: the 16S rDNA of the sphingomonas spinosa is shown as SEQ ID No. 1. When sphingomonas spinosa is used as the flocculant, the sphingomonas spinosa seed solution can be directly inoculated into a substance to be flocculated, cultured for more than 24h, and centrifuged to remove flocs. The invention provides a new sphingomonas spinosa, which has excellent adaptability, metabolic capability and degradation capability, and can be used as a flocculating agent to efficiently flocculate high-COD and high-BOD waste liquid such as garbage press filtrate under the conditions of no coagulant aid and no nutrient supplement. The sphingomonas spinosa can be used as an inoculum for repeated use; after primary inoculation, the floc obtained after treatment can be directly used as a new inoculum for new treatment without supplementary inoculation, so that the waste is treated by the waste.
Description
Technical Field
The invention belongs to the field of biological environmental protection, and particularly relates to sphingomonas spinosa and application thereof in garbage press filtrate flocculation.
Background
The garbage press filtrate is high-concentration sewage generated by the extrusion of garbage in the mechanical compression process. The garbage filter press has the characteristics of high COD and BOD, large change of water quality and water quantity, high ammonia nitrogen content, imbalance of nutrient element proportion and the like, heavy metals with high content even appear in part of the garbage filter press, the treatment cost is high, the treatment is difficult, and the standard discharge is difficult to reach. With the implementation of environmental protection policies and the improvement of environmental protection awareness of people, the problem of how to realize the low-cost treatment of garbage filter press liquid to reach the discharge standard needs to be solved urgently. In the prior art, the garbage filter press is subjected to flocculation treatment to remove most of easily-settled solids and suspended matters and reduce COD (chemical oxygen demand), and then the subsequent treatment is performed to reduce the difficulty of comprehensive treatment and finally achieve the aim of standard discharge.
The flocculants that are widely used at present mainly include inorganic polymeric flocculants typified by aluminum systems, and organic polymeric flocculants typified by polyacrylamide. The inorganic flocculant has the advantages of economy and simple use method; but also has the defects of large dosage, low flocculation effect, strong corrosivity and the like. Compared with the traditional flocculant, the organic polymer flocculant can improve the treatment efficiency by times, and has the advantages of small dosage, low scum yield, strong flocculation capacity, easy separation of flocs and good oil and suspended matter removing effects. However, most organic polymeric flocculants are toxic or hydrolysis and degradation products thereof, and acrylamide monomers for synthesis are toxic and can intoxicate the central nerve of people, and residual monomers have 'triple-effect' (teratogenicity, carcinogenicity and mutagenicity), so that the application range of the organic polymeric flocculants is limited. Meanwhile, when the chemical flocculant is used, the separated floc is also a toxic and harmful substance, and needs to be continuously treated, so that the floc is difficult to directly utilize or discharge.
Thus, microbial flocculants gradually walk into the field of vision of people. The microbial flocculant can overcome the inherent defects of inorganic polymer and synthetic organic polymer flocculants, is safe to use, and has been increasingly paid more attention to the research on the microbial flocculant in recent years. However, the existing flocculation microbial strains are deficient in resources, and the flocculation effect is still to be improved. Meanwhile, the application of the microbial flocculant is also severely limited due to the defects of high COD, high BOD, imbalance proportion of nutrient elements and the like in the garbage press filtrate.
Sphingomonas is a novel microorganism which is gradually recognized in recent years, has high metabolic capacity and degradation capacity, and has great potential in application of garbage press filtrate. However, the present research on this type of microorganism is relatively rare and is still in the first stage. It has been shown that when such microorganisms are used for flocculation, chemical coagulant aids are required, otherwise the flocculation effect is poor. For example, the screening, identification and flocculation test of microbial flocculant-producing bacteria published in agricultural science of Hubei in 2011, the culture of sphingomonas paucimobilis is used as a flocculant, and no flocculation effect is achieved when calcium chloride is not added as a coagulant aid. However, the addition of chemical coagulant aids increases the flocculation cost and introduces a new pollution source.
In conclusion, the method provides a new microorganism of sphingomonas capable of effectively treating the garbage press filtrate without adding coagulant aids, and has important scientific research value and practical significance.
Disclosure of Invention
The object of the present invention is to provide a novel microorganism of the genus Sphingomonas which can effectively treat a refuse press filtrate without adding a coagulant aid.
In order to achieve the purpose of the invention, the technical scheme adopted by the invention is as follows: the 16S rDNA of the sphingomonas spinosa is shown as SEQ ID No. 1.
Correspondingly, the sphingomonas spinosa is preserved in China general microbiological culture Collection center (CGMCC) at 27 months 3 in 2020 with the preservation number as follows: CGMCC No. 19515.
Correspondingly, the sphingomonas spinosa is applied to serving as a flocculant.
Preferably, no coagulant aid is added during flocculation.
Preferably, the flocculant is sphingomonas spinosa seed liquid, or supernatant or floc obtained by flocculating the sphingomonas spinosa seed liquid or a mixture of the supernatant and the floc.
Preferably, the method comprises the following steps:
(1) Culturing the sphingomonas spinosa until the concentration of viable bacteria is more than or equal to 10 6CFU/m L, obtaining sphingomonas spinosa seed liquid;
(2) Adjusting the pH value of a substance to be flocculated to 7-7.5, and centrifuging to remove large-particle suspended matters and easily-precipitated solids;
(3) Inoculating the sphingomonas spinosa seed solution into the substance to be flocculated treated in the step (2), culturing for more than 24h, and centrifuging to remove flocs.
preferably, in step (1), the culture medium for culturing the Sphingomonas spinosum comprises 10 g/L of glucose, 0.5 g/L of peptone, 0.5 g/L of yeast extract, 0.5 g/L of urea, (NH) 4)2SO40.5g/L，KH2PO45g/L，MgSO4·7H2O 0.2g/L，NaCl 0.1g/L。
Preferably, the supernatant obtained after the treatment of the step (3), or the flocs, or the mixture of the supernatant and the flocs is used as an inoculum to replace the sphingomonas spinosa seed liquid in the step (3) and is inoculated into a new substance to be flocculated for circulation treatment.
Preferably, in the step (3), the culture conditions are 30 ℃ and 260 r/min.
Preferably, the centrifugation rate of the step (2) and/or the step (3) is 6000 to 7000 rpm.
The invention has the following beneficial effects: the invention provides a new sphingomonas spinosa, which has excellent adaptability, metabolic capability and degradation capability, and can be used as a flocculating agent to efficiently flocculate high-COD and high-BOD liquid such as garbage press filtrate under the conditions of no coagulant aid and no nutrient supplement. The sphingomonas spinosa is used as a flocculating agent to treat the garbage press filtrate, and has excellent flocculation effect: the turbidity removal rate reaches 90.71%, the COD removal rate reaches 64.89%, the suspended matter removal rate reaches 93.04%, and the ammonia nitrogen removal rate reaches 26.80%.
The sphingomonas spinosa provided by the invention can be repeatedly used as an inoculum; after the first inoculation, the subsequent supplementary inoculation is not needed, and the floc obtained after the treatment can be directly used as a new inoculum for new treatment to treat wastes with processes of wastes against one another.
Drawings
FIG. 1 is a schematic flow diagram of a flocculation treatment of the present invention;
FIG. 2 is a schematic diagram of flocculation effect of strains 4-7;
FIG. 3 is a schematic diagram showing the effect of different ways of treating the waste press filtrate on flocculation effect;
FIG. 4 is a schematic diagram showing the effect of different centrifugation rates on flocculation effect;
FIG. 5 is a graph showing the effect of flocculation at a centrifugation rate of 7000 rpm;
FIG. 6 is a schematic diagram showing flocculation effect of different inocula under different inoculum size;
FIG. 7 is a schematic diagram showing the flocculation effect of different inocula and different culture times;
FIG. 8 is a graph showing the flocculation effect of different strains;
FIG. 9 is a graph showing the comparison of flocculation effects of different Sphingomonas spinosus;
FIG. 10 is a graph showing the flocculation effect of strains 4-7 compared with that of a chemical flocculant.
Detailed Description
The invention provides Sphingomonas spinosa 4-7(Sphingomonas echinoides) which is preserved in the common microorganism center of China Committee for culture Collection of microorganisms (address: Beijing, Chaoyang district, North Chen Xilu No.1, institute of microbiology, China academy of sciences, postal code: 100101) 3.27.3.2020, and the preservation numbers are as follows: CGMCC No. 19515.
The invention also provides a method for using sphingomonas spinosa 4-7 to flocculate the garbage press filtrate. The flow of the method is shown in fig. 1, and specifically comprises the following steps:
(1) Preparation of inoculum: inoculating the sphingomonas spinosa 4-7 to a seed culture medium, and culturing at 30 ℃ and 160r/min for one day to obtain a seed solution which is an inoculum. The concentration of viable bacteria in the inoculum is more than or equal to 10 6CFU/mL。
(2) Pre-treating garbage press filtrate: and adjusting the pH value of the garbage filter pressing liquid to 7-7.5 by using sodium hydroxide. Followed by centrifugation to remove large particle suspensions and easily settling solids.
(3) Inoculating the inoculum in the garbage press filtrate treated in the step (2) with an inoculum size of 10% (v/v). Culturing at 30 deg.C and 160r/min for more than 24 hr to form flocs in the garbage press filtrate, and centrifuging to remove flocs.
(4) In the step (3), the pressure filtrate after 24 hours of culture or flocs obtained by centrifugation or a mixture of the pressure filtrate and the flocs can be used as a new inoculum and is inoculated into the garbage pressure filtrate pretreated in the step (2) again, and then the step (3) is repeated, so that the circulating treatment can be carried out.
The present invention will be further explained with reference to specific examples.
The method for calculating the flocculation rate of the culture medium is as follows:
1. the seed culture medium comprises 10 g/L of glucose, 0.5 g/L of peptone, 0.5 g/L of yeast extract, 0.5 g/L of urea, (NH) 4)2SO40.5g/L，KH2PO45g/L，MgSO4·7H20.2 g/L of O, 0.1 g/L of NaCl, the pH value of 7-7.5, and the mixture is sterilized at 115 ℃ for 20min, and if a solid culture medium is needed, 20 g/L of agar is added.
2. A method for flocculating kaolin and a method for calculating the flocculation rate are characterized in that a spectrophotometer method is adopted to measure the change of absorbance values before and after kaolin treatment to calculate the flocculation rate, specifically, 4 g/L of kaolin solution 40m L is taken, a flocculating agent is added, the mixture is stirred uniformly, the flocculation effect is observed after the mixture is kept stand for 10min, liquid 2cm below the liquid level is taken, the absorbance value of the liquid at 550nm is measured, and the flocculation rate is obtained according to the change of the absorbance value between the treatment of adding the flocculating agent and the blank.
3. the garbage press filtrate flocculation method and the calculation method of the flocculation rate are characterized in that a turbidity meter is adopted to measure the change of turbidity before and after biogas slurry treatment, specifically, liquid 2cm below the liquid surface is respectively taken before and after the addition of an inoculum, the change of turbidity before and after treatment is measured, and the flocculation rate is calculated.
The first embodiment is as follows: screening and identifying Sphingomonas spinulosa
1. taking a water sample 10m L from a secondary sedimentation tank of a water treatment plant into a 150m L triangular flask filled with 50m L sterile water, shaking uniformly, standing for 1min, taking 5m L into a 150m L triangular flask filled with 100m L seed culture medium, carrying out shake culture for 24h at the temperature of 30 ℃ and the pH of 7 under 160r/min to obtain fermentation liquor, and then dividing into 10 parts by adopting a dilution coating method -1、10-2、10-3、10-4、10-5、10-6six gradient dilution fermentation liquor, then coating on a flat plate, selecting different single bacterial colonies, separating the single bacterial colonies for more than three times by using a plate marking method until the bacterial colonies are consistent in morphology to obtain purified bacterial strains, inoculating 5m L the purified bacterial strains into a 150m L triangular flask filled with 100m L seed culture medium, carrying out shake culture for 72 hours at the temperature of 160r/min and the pH value of 7 and 30 ℃, taking samples every 24 hours, measuring the flocculation rate of kaolin serving as a treatment object, selecting the bacterial strains with good flocculation effect, and naming the bacterial strains as 4-7.
2. the method comprises the steps of extracting the whole genome of a pure strain by adopting a bacterial whole genome rapid extraction kit, carrying out PCR amplification by selecting a bacterial 16SrRNA universal primer, then carrying out sequencing analysis, comparing a sequencing result with B L AST in an NCBI database, and identifying that the strain 4-7 has the homology of 97.22% with Sphingomonas echinoides (Sphingomonas echinoides), and the 16S rRNA gene sequence of the strain is shown as SEQ ID No. 1.
Is preserved in China general microbiological culture Collection center (CGMCC) at 27 months 3 and 2020; address: west road No.1, north zhou, chaoyang district, beijing, institute of microbiology, china academy of sciences, zip code: 100101. the preservation number is: CGMCC No. 19515.
Example two: influence of different garbage filter pressing liquid treatment modes on flocculation effect
Taking garbage press filtrate of a certain Hangzhou garbage disposal plant. The waste press filtrate in the embodiment of the invention comes from this. Dividing the garbage press filtrate into 5 equal parts, wherein the treatment modes of the parts are as follows: the method comprises the steps of directly adding a flocculating agent without pretreatment, performing centrifugal treatment on the garbage press filtrate, then adding the flocculating agent, performing centrifugal treatment after adding the flocculating agent, performing centrifugal treatment on the garbage press filtrate, then adding the flocculating agent, and then performing centrifugal treatment. The centrifugation rate was 7000 r/min. The flocculating agents are all inocula containing strains 4-7, the inoculum size is 10% (v/v), and the reactions are carried out for 24h under the conditions of 30 ℃ and 160 r/min.
As shown in FIG. 3, the flocculation rate of the flocculant group directly added was 15.21%, and the flocculation rate of the flocculant group added after the centrifugation was 35.37%; the flocculation rate of the group which is subjected to centrifugal treatment after the flocculant is added is 34.19 percent; after centrifugation, a flocculant is added and then the flocculation rate of the centrifugation group is 75.20%. The results show that: the garbage press filtrate is firstly subjected to centrifugal treatment, then a flocculating agent is added, and the flocculating effect after the centrifugal treatment is optimal.
Example three: effect of different centrifugation rates on flocculation Effect
The garbage press filtrate is divided into 7 equal parts, and the treatment mode of each part is as follows: firstly, carrying out centrifugal treatment on the garbage press filtrate, then adding a flocculating agent, and then carrying out centrifugal treatment. The centrifugation rates of the respective portions were 1000rpm, 2000rpm, 3000rpm, 4000rpm, 5000rpm, 6000rpm, 7000rpm, respectively. The remaining processing conditions were the same as in the examples. As shown in FIG. 4, the flocculation rates of the respective groups were-21.39%, -9.85%, 18.52%, 33.33%, 56.08%, 73.88%, and 75.20%, respectively. The reason for the negative flocculation rate at low rotation speeds may be: flocculants aggregate the previous suspensions and small particles to form larger suspensions and large particles, but do not sink, thereby affecting the turbidity of the solution.
wherein, when the centrifugal speed is 7000rpm, after the treatment, the COD of the garbage press filtrate is reduced from 28200 mg/L to 9900 mg/L, the removal rate reaches 64.89%, the suspended matters are reduced from 2228 mg/L to 155 mg/L, the removal rate reaches 93.04%, the ammonia nitrogen is reduced from 248.17 mg/L to 181.65 mg/L, and the flocculation effect pair of the group is shown in figure 5.
Example four: comparison of Effect of Recycling inoculum
The inoculum is divided into 2 groups, one group is sphingomonas spinosa 4-7 seed liquid, and the other group is floc sediment obtained by centrifuging after treating garbage press filtrate by using the sphingomonas spinosa 4-7 seed liquid. Two different sets of inocula were each treated with the waste press filtrate as in example three, at a centrifuge rate of 7000 rpm. The inoculum size of the inoculum was 10%, 20%, 30%, 40%, 50% (v/v), flocculation rate after 24h of culture, respectively, and the results are shown in FIG. 6. The flocculation rates of the cultures were measured at 10% (v/v) for 1, 2, 3, 4 and 5 days (24 h each and one more time after 24 h), respectively, and the results are shown in FIG. 7.
The results show that the effect of the floc precipitation treatment on the waste press filtrate obtained after the treatment is equivalent to that of the seed solution. The strain 4-7 provided by the invention is proved to have excellent adaptability and reproductive and metabolic capability, can circularly treat garbage filter press liquid for multiple times after the seed liquid is added for one time, saves the cost, effectively utilizes the flocculation precipitation obtained by treatment, and avoids secondary pollution.
Example five: effect display of different flocculating agents for treating garbage filter press liquid
1. Comparison of flocculation effects of strains of different species: the method of example three was used to treat the landfill leachate at a centrifuge rate of 7000rpm with the inoculum: flocculant AS28 (produced from Alcaligenes faecalis, CGMCC No.17863), flocculant AS30 (produced from Brevundimonas diminuta, CGMCC No.17864), Bacillus cereus flocculant AS33 (produced from Bacillus cereus, CGMCC No.17865), and strain 4-7 (produced from Sphingomonas spinulosa, CGMCC No.19515) provided by the invention. After 24 hours of culture, the flocculation rates were 55.27%, 46.21%, 49.62%, and 74.87%, respectively, as shown in FIG. 8. The results show that: under the condition of not adding additional nutrient components, most strains have poor growth performance in the garbage press filtrate, and cannot exert flocculation effect or have poor flocculation effect.
2. Comparison of flocculation effects of different sphingomonas spinosa: the method of example three was used to treat the landfill leachate at a centrifuge rate of 7000rpm with the inoculum: CGMCC1.16096 Sphingomonas spinosus (Sphingomonas echinoides), CGMCC1.12798 Sphingomonas spinosus (Sphingomonas echinoides), CGMCC1.10184 Sphingomonas spinosus (Sphingomonas echinoides), and the strain 4-7 provided by the invention (Sphingomonas spinosus, CGMCC No. 19515). As a result, as shown in FIG. 9, the flocculation rates were 49.32%, 46.78%, 21.51%, and 75.87, respectively. The results show that: only the specific sphingomonas spinosa provided by the invention can adapt to the severe environment of the garbage press filtrate and play an excellent flocculation role.
3. And the flocculation effect of the chemical flocculant is compared: the method of the third embodiment is adopted to treat the garbage press filtrate, the centrifugation speed is 7000rpm, and the flocculating agents are respectively: polyacrylamide (PAM), the bacterial strain 4-7 (Sphingomonas spinulosa, CGMCC No.19515) provided by the invention. As shown in FIG. 10, the flocculation rate of PAM was 56.78%, and the flocculation rate of strains 4 to 7 was 75.17%.
Sequence listing
<110> Zhongwei environmental protection science and technology (Hangzhou) Co Ltd
CHENGDU INSTITUTE OF BIOLOGY, CHINESE ACADEMY OF SCIENCES
<120> sphingomonas spinosa and application thereof in garbage press filtrate flocculation
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>1395
<212>DNA
<213> Sphingomonas spinosus (Sphingomonas echinoides)
<400>1
gaactggcgc tgcctaccat gcagtcgaac gaaggcttcg gccttagtgg cgcacgggtg 60
cgtaacgcgt gggaatctgc cctcaggttc ggaataacag cgagaaattg ctgctaatac 120
cggatgatat cgcgagatca aagatttatc gcctgaggat gagcccgcgt aggattagct 180
agttggtgtg gtaaaggcgc accaaggcga cgatccttag ctggtctgag aggatgatca 240
gccacactgg gactgagaca cggcccagac tcctacggga ggcagcagtg gggaatattg 300
gacaatgggc gaaagcctga tccagcaatg ccgcgtgagt gatgaaggcc ttagggttgt 360
aaagctcttt tacccgggat gataatgaca gtaccgggag aataagctcc ggctaactcc 420
gtgccagcag ccgcggtaat acggagggag ctagcgttat tcggaattac tgggcgtaag 480
cgcacgtagg cggctttgta agttagaagg tgaaagcctg gagctcaact ccagaatttg 540
cctttaagac tgcattcgct tgaatccagg agaggtgagt ggaattccga gtgkwrragg 600
tgaaattcgt agatattygg aagaacacca gtggcgaagg cggctcactg gactggtatt 660
gacgcttgag gtgcgaaars sktgggggag caaacaggat tagataccct ggtagtccac 720
gccgtaaacg atgataacta gctgtcgggg gctcttagag cttcggtggc gcagcttaac 780
gcattaagtt attccgcctg gggagtacgg cccgcaaggt taaaamyyma awkrrwwkws 840
rsgggssysy rcmasmrgyg gwgsawgyrk kkkrrttyra wkcramrssm asrmscwkam 900
cmkyryywgm swtkkmcrkr ysmkkkmsgr wkwcsgrwsw cykmyctyyk sssmytsgrr 960
sacwggwrcw scakgksykk crtsrsctcg tgtcgtgaga tgttgggtta agtcccgcaa 1020
cgagcgcaac cctcgtcctt agttgccatc atttagttgg gcactctaag gaaaccgccg 1080
gtgataagcc ggaggaaggt ggggatgacg tcaagtcctc atggccctta cgcgctgggc 1140
tacacacgtg ctacaatggc ggtgacagtg ggcagcaatc tcgcaagggt gagctaatct 1200
ccaaaagccg tctcagttcg gattgttctc tgcaactcga gagcatgaag gcggaatcgc 1260
tagtaatcgc ggatcagcat gccgcggtga atacgttccc aggccttgta cacaccgccc 1320
gtcacaccat gggagttgga ttcacccgaa ggtaagtgcg ctaaccgcaa ggaggcagct 1380
accacggtgt cagca 1395
Claims (10)
1. A sphingomonas spinosa strain is characterized in that: the 16S rDNA is shown in SEQ ID No. 1.
2. A sphingomonas spinosa strain is characterized in that: is preserved in China general microbiological culture Collection center on 27 th 3 th 2020, with the preservation number: CGMCC No. 19515.
3. Use of sphingomonas spinosa according to claim 1 or 2 as flocculant.
4. The use of sphingomonas spinosa as a flocculant according to claim 3, characterized in that: in the flocculation process, no coagulant aid is added.
5. The use of sphingomonas spinosa as a flocculant according to claim 3, characterized in that: the flocculant is sphingomonas spinosa seed liquid, or supernatant or floc obtained by flocculating the sphingomonas spinosa seed liquid or a mixture of the supernatant and the floc.
6. The use of sphingomonas spinosa as a flocculant according to claim 3, characterized in that: the method comprises the following steps:
(1) Culturing the sphingomonas spinosa until the concentration of viable bacteria is more than or equal to 10 6CFU/m L, obtaining sphingomonas spinosa seed liquid;
(2) Adjusting the pH value of a substance to be flocculated to 7-7.5, and centrifuging to remove large-particle suspended matters and easily-precipitated solids;
(3) Inoculating the sphingomonas spinosa seed solution into the substance to be flocculated treated in the step (2), culturing for more than 24h, and centrifuging to remove flocs.
7. The use of sphingomonas spinosa as a flocculant according to claim 3, characterized in that: in the step (1), the culture medium for culturing the sphingomonas spinosa comprises the following components 10 g/L glucose, 0.5 g/L peptone, 0.5 g/L yeast extract, 0.5 g/L urea, (NH) 4)2SO40.5g/L，KH2PO45g/L，MgSO4·7H2O 0.2g/L，NaCl0.1g/L。
8. The use of sphingomonas spinosa as a flocculant according to claim 6, characterized in that: and (4) taking the supernatant or the flocs obtained after the treatment in the step (3) or the mixture of the supernatant and the flocs as an inoculum to replace the sphingomonas spinosa seed solution in the step (3) and inoculating the inoculum into a new substance to be flocculated for circulation treatment.
9. The use of sphingomonas spinosa as a flocculant according to claim 6, characterized in that: when the culture in the step (3) is carried out, the culture condition is 30 ℃ and 260 r/min.
10. The use of sphingomonas spinosa as a flocculant according to claim 6, characterized in that: and (3) the centrifugation speed of the step (2) and/or the step (3) is 6000-7000 rpm.
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| CN112159775A (en) * | 2020-09-27 | 2021-01-01 | 浙江工业大学 | Sphingomonas zeae and application thereof in biological deodorization |
| CN113151050A (en) * | 2021-03-10 | 2021-07-23 | 南京工业大学 | Sphingomonas and application thereof |
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| CN112159775A (en) * | 2020-09-27 | 2021-01-01 | 浙江工业大学 | Sphingomonas zeae and application thereof in biological deodorization |
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| CN113151050A (en) * | 2021-03-10 | 2021-07-23 | 南京工业大学 | Sphingomonas and application thereof |
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