CN113444673B

CN113444673B - Rhodococcus gaucher YKSW-6 and application thereof

Info

Publication number: CN113444673B
Application number: CN202110940444.3A
Authority: CN
Inventors: 张宗源; 张英涛; 岳丹丹; 周留柱; 郭文阳; 潘梦诗; 亓兰达; 王雪妍; 李冠杰; 杨书豪; 徐宏光; 王秋菊
Original assignee: Henan Academy Of Sciences Institute Of Biology LLC; Henan Academy of Sciences
Current assignee: Henan Academy Of Sciences Institute Of Biology LLC; Henan Academy of Sciences
Priority date: 2021-08-17
Filing date: 2021-08-17
Publication date: 2022-06-07
Anticipated expiration: 2041-08-17
Also published as: CN113444673A

Abstract

The invention relates to Rhodococcus gordonii YKSW-6 and application thereof, which can effectively solve the degradation problem of high-concentration skatole and adopts the technical scheme that the Rhodococcus gordonii YKSW-6 is classified and named as Rhodococcus gordonii (Rhodococcus gordoniae), is preserved in the China general microbiological culture Collection center, has the preservation number of CGMCC No.22972, has the preservation date of 2021 year, 7 months and 28 days, has the preservation address of No. 3 of No.1 Homeh West Luo No.1 of the sunny region of Beijing, and is a microbial research institute of Chinese academy of sciences, has high-efficiency and rapid degradation capability on high-concentration skatole, has the degradation rate of the skatole in compost of 93.6 percent, has obvious deodorization effect, is an innovation on skatole degrading microbes, and has good application prospect.

Description

Rhodococcus gaucher YKSW-6 and application thereof

Technical Field

The invention relates to the technical field of environmental protection, in particular to a strain of Rhodococcus gaucher YKSW-6 and application thereof.

Background

With the rapid development of large-scale and intensive livestock and poultry breeding, the problem of environmental pollution caused by a large amount of discharged excrement and stink gradually becomes the focus of attention of people on the basis of meeting the requirements of residents on meat, eggs and milk. The emission and discharge of odor substances along with the discharge of a large amount of excrement have adverse effects on livestock and poultry production, resident health, air quality and the like.

The odor substances discharged by livestock and poultry are complex in components and various in types, and can be divided into four main types according to chemical properties, namely volatile nitrogen-containing compounds, volatile sulfur-containing compounds, volatile fatty acids and nitrogen heterocyclic compounds. The indole nitrogen heterocyclic organic substance is the main component of the odor pollution in the livestock and poultry compost. Wherein 3-methylindole (3MI) is commonly called as skatole, is considered as one of three harmful odor substances discharged by livestock and poultry, has strong dung odor, and has the odor threshold value lower than 0.003mg/m³Can be still perceived by people and livestock and poultry under the condition of extremely low concentration. Excessive skatole in the environment can cause harm to human and animal health. Studies show that skatole has cytogenetic toxicity and can cause tumorigenesis, acute pulmonary edema and emphysema of ruminants and death of the ruminants in severe cases; it also can cause pathological changes of neuroendocrine function and immunologic function of human, and has respiratory tract cell teratogenesis and carcinogenesis risk in severe cases. Therefore, the method has important significance for solving the environmental pollution of the skatole.

The prior methods for removing the skatole by adopting certain physics (such as adsorption, masking, dilution, diffusion and the like) and chemistry (such as chemisorption, washing, oxidation and the like) have certain effects, but the methods have high cost and large energy consumption and are easy to form secondary pollution. The biological degradation and transformation by using microorganisms is a feasible and eco-friendly method for removing skatole. At present, the activity of skatole-producing bacteria is interfered by influencing the structure of microbial flora in intestinal tracts of livestock and poultry by adding a microecological preparation, so that the content of skatole in excrement of livestock and poultry is reduced, but the method still has some problems at present: firstly, the cost of the feed is increased due to excessively high addition of the micro-ecological feed additive, and the nutrient components of the feed can be absorbed and digested; secondly, the residual skatole in the livestock manure which is discharged into the environment still has great pollution to the environment, and the problem how to degrade the livestock manure compost and pollute the skatole in the environment still needs to be solved. Since the 90 s of the 20 th century, various strains have been found to be able to degrade skatole, such as: kohda et al isolate Clostridium malayi (Clostridium maleininatum) A-3 from pig manure and chicken manure compost, the concentration of the bacterial strain capable of degrading skatole under anaerobic condition is 100-300 mg.L^-1The highest degradation rate of skatole when cultured in PYG culture solution for 4 weeks is 32.18%. Pseudomonas putida LPC24(Pseudomonas putida LPC24) completely degraded skatole by 2mmol/L within 30d under itching-limiting conditions. Under aerobic conditions, pseudomonas aeruginosa Gs (pseudomonas aeruginosa Gs) completely degrades 2mmol/L of skatole within 24d, and the strains of lactobacillus, Rhodopseudomonas (Rhodopseudomonas) and cupriasis (Cupriavidus) have degradation effect only on low-concentration skatole. Burkholderia (Burkehoderia), Achromobacter (Achromobacter) and Rhodococcus (Rhodococcus) are bacteria for degrading skatole in activated sludge, wherein Rhodococcus strains DMU1 and DMU2 can completely degrade 50mg/L skatole in 24h, but whether the strains have the degradation effect on skatole with higher concentration is not reported. In general, the time for degrading the skatole by the bacterial strains is long, the degradation concentration is low, and the bacterial strains with high-efficiency and rapid degradation capability on the high-concentration skatole are still the primary tasks of treatment of the livestock and poultry manure and environmental management.

Disclosure of Invention

Aiming at the situation, in order to solve the defects of the prior art, the invention aims to provide a strain of Rhodococcus gordonii YKSW-6 and application thereof, which can effectively solve the degradation problem of high-concentration skatole.

The technical scheme for solving the problem is that the Rhodococcus gordonii YKSW-6 is classified and named Rhodococcus gordonii (Rhodococcus gordoniae) and is preserved in the China general microbiological culture Collection center of the Committee for culture Collection of microorganisms, the preservation number is CGMCC No.22972, the preservation date is 2021, 7 and 28 days, the preservation address is No. 3 of the institute of microbiology, China institute of academy of sciences, North Kogyo-Yang district, Chaceway 1, Beijing.

The application of the Rhodococcus gordonii YKSW-6 in preparing a microbial preparation for degrading high-concentration skatole is provided.

The preparation method of the Rhodococcus gordonii YKSW-6 microbial inoculum comprises the following steps:

1) preparing shake flask seeds: inoculating the YKSW-6 strain into a seed culture medium, and culturing at 30 ℃ and 180r/min to a logarithmic phase;

2) fermentation culture: inoculating the seed solution obtained in the step 1) into a large amount of culture fermentation culture solution according to the inoculation amount of 5-10%, wherein the culture temperature is 30-42 ℃, the rotation speed is 180-240r/min, the initial pH is 6-9, the culture time is 24h, and the microbial deodorant is obtained after the fermentation is finished.

The viable count of Rhodococcus gordoniae YKSW-6 in the microbial deodorant can reach 1.0 × 10⁹CFU/mL。

The formula of the large-scale fermentation culture solution is as follows: 1.4 percent of cane sugar, 1.4 percent of beef extract, 0.6 percent of yeast extract powder and 0.15 percent of ammonium sulfate.

The Rhodococcus gordonii YKSW-6 has high-efficiency and rapid degradation capability on high-concentration skatole, has a degradation rate of 93.6 percent on skatole in compost, has an obvious deodorization effect, is an innovation on skatole degrading microorganisms, and has a good application prospect.

Drawings

FIG. 1 is a colony morphology chart of the strain YKSW-6 of the invention.

FIG. 2 is the gel electrophoresis chart of the strain YKSW-616 SrDNAPCR of the present invention.

FIG. 3 is a phylogenetic dendrogram of the strain YKSW-6 of the present invention.

FIG. 4 is a graph showing the growth-skatole degradation profile of strain YKSW-6 of the present invention.

FIG. 5 is a graph showing the effect of different factors on the growth of the strain YKSW-6 and the degradation of skatole.

FIG. 6 is a diagram showing the growth of strain YKSW-6 and its removal rate at different skatole concentrations.

Detailed Description

The following detailed description of embodiments of the present invention is provided in connection with the accompanying drawings and examples.

Example 1

In the specific implementation of the invention, the preparation method of the Rhodococcus deodouris YKSW-6 microbial inoculum comprises the following steps:

2) fermentation culture: inoculating the seed solution obtained in the step 1) into a large amount of culture fermentation culture solution according to the inoculation amount of 5%, wherein the culture temperature is 30 ℃, the rotation speed is 180r/min, the initial pH value is 7.2, the culture time is 24h, and the microbial deodorant is obtained after the fermentation is finished.

The Rhodococcus deordorii YKSW-6 is obtained by separating and screening the soil in the pig manure compost of a certain pig breeding plant in Henan province, has high-efficiency and quick degradation capability on high-concentration skatole, can effectively degrade the high-concentration skatole in the compost, and has the following related test data:

1 materials and methods

1.1 sources of strains

The strain is obtained by separating and screening pig manure compost soil of a certain pig breeding factory in Henan province.

1.2 culture Medium

Inorganic salt medium formulation (g/L) for strain screening and degradation experiments: (NH)₄SO₄ 2.0g，KH₂PO₄2.0g， Na₂HPO₄·12H₂O 3.38，FeCl₃0.00025, pH is natural. LB culture medium formula (g/L) used for strain slant, seed liquid and scanning electron microscope observation experiment comprises NaCl 10.0, peptone 10.0, yeast powder 5.0, pH 7.0 (solid culture medium supplemented with agar 15.0); phosphate Buffered Saline (PBS) formulation (g/L): na (Na)₂HPO₄·12H₂O 10.75，NaH₂PO₄·2H₂O 2.34，pH7.0。

1.3 Experimental methods

1.3.1 preparation of skatole mother liquor

0.5g of skatole is dissolved in 100mL of absolute ethyl alcohol to prepare 5g/L of skatole. Filtering with 0.2 μm organic filter membrane to remove bacteria, and storing in a refrigerator at 4 deg.C.

1.3.2 screening and isolation of skatole degrading bacteria

Weighing 0.1g of collected pig manure compost soil, domesticating the soil by using an inorganic salt culture medium containing 100mg/L skatole at 30 ℃ for 180r/min for 3d as a domestication period, and inoculating the soil into a fresh inorganic salt culture medium containing skatole according to the inoculation amount of 5 percent for continuous domestication for about 4-5 periods. Then repeatedly coating the domesticated sample on a separation plate containing skatole, finally separating to obtain 1 strain which can grow by taking skatole as a unique carbon source and is named as YKSW-6, and inoculating the strain in a slant culture medium for preservation at 4 ℃.

1.3.3 sequencing of the 16S rDNA Gene of Strain YKSW-6

The 16S rDNA PCR amplification primer of the YKSW-6 strain adopts a universal primer: the forward primer 7F is 5'-CAGAGTTTGATCCTGGCT-3', the reverse primer 1540R is 5'-AGGAGGTGATCCAGCCGCA-3', the amplified product is sent to Beijing Optimalaceae Biotechnology Limited for sequencing, and the sequencing result shows that the strain YKSW-6 is finally determined to be rhodococcus (Rhodococcus sp.), and the 16S rDNA sequence gene sequence of the strain is shown as SEQ ID No. 1.

1.3.4 determination of skatole Standard Curve

Weighing skatole standard sample 10mg, placing in a 10mL volumetric flask, and preparing into 1mg/mL standard stock solution with ethanol. Sequentially diluting to obtain 0, 1, 5, 10, 15, 20, 25 μ g/mL standard series solution, filtering with 0.2 μm organic filter membrane, measuring peak area by HPLC, and repeating the measurement three times per concentration. And establishing a linear regression equation according to the sample concentration and the obtained peak area.

Chromatographic conditions are as follows: the chromatographic column is a Hypersil BDSC18 column (250mm × 4.6mm, 5 μm); column temperature: room temperature; ultraviolet detection wavelength: 254 nm; mobile phase: methanol: the water content is 80: 20; flow rate: 1 mL/min; the amount of the sample was 10. mu.L.

1.3.5 determination of the growth degradation Curve of the Strain YKSW-6

The growth and degradation conditions of the strain YKSW-6 in an inorganic salt culture medium with 100mg/L skatole as a unique carbon source under the conditions of 1 percent, 5 percent and 10 percent of inoculation amount are respectively explored. The culture conditions are 180r/min, the pH is natural, the samples are taken every 2h at 30 ℃, the absorbance (OD600) is measured at 600nm by using an ultraviolet-visible spectrophotometer to represent the growth condition of the strains, and simultaneously, the samples are extracted and subjected to liquid phase analysis to measure the concentration of the skatole.

Extracting skatole: adding equal volume of methanol into the culture solution, centrifuging at 5000r/min for 10min, sucking 1mL solution, filtering with 0.22 μm organic filter membrane, storing at 4 deg.C, and testing. If the concentration of the extracting solution is too high, HPLC detection is carried out after dilution, and the detection conditions are the same as 1.3.4.

1.3.6 Effect of different environmental factors on the degradation of skatole by the strain YKSW-6

1) Exploring the influence of different environmental factors on the bacterial strain degradation of skatole, wherein the basic conditions of the inorganic salt culture medium are skatole concentration of 100mg/L, temperature of 30 ℃, shaking table rotating speed of 180r/min, natural pH, inoculum size of 5%, and sampling at regular time to determine the growth condition (OD) of the bacterial strain₆₀₀) And the concentration of the residual skatole in the culture medium, and taking the skatole removal rate as an investigation target.

Skatole removal (%) as ═ skatole content in uninoculated medium-skatole content in inoculated medium)/skatole content in uninoculated medium × 100%

2) The influence of temperature, pH and rotating speed on the degradation of skatole by the bacterial strain is respectively explored, and the influence comprises different temperatures: 20. 25, 30, 37 and 42 ℃; different pH values: 4. 5, 6, 8, 9 and natural pH7.2; different rotating speeds: 0. 60, 120, 180 and 240r/min, and the sampling time is 24 h.

3) The influence of NaCl content on the growth of the strain and the degradation of skatole is researched, wherein the NaCl content is respectively 0%, 3%, 6%, 9%, 12% and 15%, and the sampling time is 24 h.

4) The influence of skatole with different concentrations on the growth of the bacterial strain and the degradation of skatole is researched, wherein the skatole content is 50, 100, 150, 200 and 300mg/L, and the sampling time is 24 h.

1.3.7 YKSW-6 microbial inoculum preparation and application thereof in pig manure composting

The microbial inoculum was prepared according to the invention example 1:

1) and (3) shaking the bottle to prepare seeds: the YKSW-6 strain stored on the slant was inoculated into a seed medium and cultured at 30 ℃ at 180r/min to logarithmic phase (about 20 hours).

2) Fermentation culture: inoculating the seed solution obtained in the step 1) into a large-scale culture fermentation culture solution according to the inoculation amount (volume ratio) of 5%, wherein the culture temperature is 30 ℃, the rotation speed is 180r/min, the initial pH value is 7.2, and the culture time is 24 h. And (3) obtaining the microbial deodorizing microbial inoculum after fermentation is completed, wherein the formula of a large amount of fermentation culture solution is as follows: 1.4 percent of cane sugar, 1.4 percent of beef extract, 0.6 percent of yeast extract powder and 0.15 percent of ammonium sulfate.

1.3.8 application of YKSW-6 microbial inoculum in pig manure composting:

fresh pig manure was used for the test. 500g of pig manure is taken in a 2L beaker, and the fermentation broth: sterile water 1: 10, and uniformly spraying the diluted solution into the pig manure, wherein the spraying amount is 10% of the mass of the pig manure. Each treatment was done in 3 replicates with pig manure compost as control. The compost odor grade is determined by adopting an odor functional detection method (in the experiment, the odor is sequentially divided into 6 grades, namely 1 grade and no odor, 2 grade and no odor can be sensed, 3 grade and no odor can be sensed, 4 grade and no odor can be sensed, 5 grade and strong pungent odor can be sensed, 6 grade and no intolerable strong odor can be sensed, and the degradation rate of the skatole is determined by adopting an HPLC method.

2 results and analysis

2.1 isolation, screening and identification of skatole degrading bacteria

After repeated plate coating, 1 strain which can grow by taking skatole as a unique carbon source is obtained by separation and named as YKSW-6. The colony morphology of the strain YKSW-6 on the LB solid medium is shown in figure 1(a), the colony is smooth and convex, the color is orange, and the edge is radial.

The 16S rDNA gene of the strain is subjected to PCR amplification, and the result of agarose gel electrophoresis shows that the length of the amplified product is about 1500 bp (figure 2). Through the construction of a phylogenetic tree of strains, as shown in FIG. 3, it was preliminarily identified that the YKSW-6 strain belongs to Rhodococcus gordonii (Rhodococcus gordoniae.), and the sequence similarity between the amplified 16S rDNA and Rhodococcus gordoniae reached 99.9%, and the sequence homology was 99%, so that the strain was preliminarily determined to be Rhodococcus gordonae.

2.2 growth and degradation characteristics of Strain YKSW-6

And (3) inspecting the growth and degradation characteristics of the strain YKSW-6 under different inoculation amounts. The results are shown in fig. 4, when the inoculum size is 5% or more, the skatole degradation rate of the strain is slow in the adaptation phase, and when the strain grows to the logarithmic phase, the skatole degradation rate is increased, which shows that skatole degradation and strain growth are positively correlated to a certain extent. With the increase of the inoculation amount, the growth lag phase of the strain is shortened, and the skatole degradation rate is increased. When the inoculation amount is 1%, the bacterial strain hardly grows for 30h, and the skatole removal rate is about 20%. When the inoculation amount is 5%, the lag phase is 8h, and the bacterial strain can completely reduce 100mg/L skatole in about 18 h; when the inoculation amount is 10 percent and the stagnation period is 6 hours, the bacterial strain can completely degrade 100mg/L skatole within 14 hours.

2.3 influence of different environmental factors on the growth of the strain YKSW-6 and the degradation of skatole

The effect of different factors on the growth of the strain YKSW-6 and the degradation of skatole is shown in FIG. 5.

2.3.1 temperature

As can be seen from fig. 5(a), the skatole removal rate reached 100% when the temperature exceeded 30 ℃ after inoculation for 24 hours. From these graphs, it is found that the strain YKSW-6 grows best at 37 ℃. When the temperature is 25 ℃, the removal rate of the skatole is about 70 percent, which shows that the bacterial strain has higher skatole degradation capability in the range of 25-42 ℃.

2.3.2 pH

The pH is important for the growth of microorganisms, and the change of the pH can change the supply state of nutrient substances, influence the charged property and stability of cell membranes of thalli, greatly influence the permeability of the cell membranes and intracellular enzymatic reaction and also influence the absorption capacity of substances. As can be seen from FIG. 5(b), the skatole removal rate reaches 100% when the initial pH is 6-9, and the growth condition of the strain is good; when the pH value is 4-5, the removal rate is less than 40%, and the growth of the strain is inhibited, so that the strain YKSW-6 keeps better activity and growth vigor in neutral and alkaline environments, and when the pH value is less than 6, the activity of the strain YKSW-6 in degrading skatole is obviously inhibited.

2.3.3 rotational speed

The rotation speed is related to dissolved oxygen, and as can be seen from fig. 5(c), skatole is completely removed when the rotation speed is 180-240 r/min. The rotating speed indirectly influences the growth and the biological activity of the strains by influencing the oxygen content, when the rotating speed is 0-180 r/min, the higher the dissolved oxygen is along with the increase of the rotating speed, the higher the skatole removal speed is, the better the strains grow, but when the rotating speed is increased to 240r/min, the growth of the strains is lower than 180 r/min.

2.3.4 salinity

As is clear from FIG. 5(d), when the NaCl concentration in the medium exceeded 3%, the skatole removal rate was less than 37%, and the growth was also inhibited, indicating that this strain was not resistant to NaCl.

2.3.5 skatole

The growth condition and the removal rate result of the YKSW-6 strain under different skatole concentrations are shown in figure 6, the growth condition of the YKSW-6 strain in an inorganic salt culture medium is different according to different skatole concentrations, when the skatole concentration is 50mg/L and 100mg/L, the strain grows better, the removal rate of skatole reaches 100%, and when the skatole concentration exceeds 150mg/L, the degradation rate is lower than 40%, and the growth is severely inhibited.

2.3.6 YKSW-6 fermentation liquid for degrading compost odor

The results show that: after 7 days of composting, the odor grade of a control group is grade 5, the odor grade of the compost with strong pungent odor treated by the bacterial liquid is reduced to grade 1, the odor release of the compost is obviously reduced, and the degradation rate of skatole in the compost can reach 93.6% by HPLC (high performance liquid chromatography).

The Rhodococcus deodara YKSW-6 has high-efficiency and rapid degradation capability on high-concentration skatole, the strain has high skatole degradation capability within the temperature range of 25-42 ℃, the skatole removal rate reaches 100% when the initial pH is 6-9, and the strain has good growth condition; when the rotating speed is 180-240r/min, the skatole is completely removed, when the concentration of the skatole is 50mg/L and 100mg/L, the bacterial strain grows well, meanwhile, the removal rate of the skatole reaches 100%, the degradation rate of the skatole in the compost can reach 93.6%, the method is an innovation on skatole degrading microorganisms, and has good economic and social benefits.

Sequence listing

<110> institute of sciences of Henan province, Bioresearch institute of Henan province, Inc., department of sciences of Henan province

<120> Rhodococcus gaurea YKSW-6 and application thereof

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1345

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 1

gtggattagt ggcgaacggg tgagtaacac gtgggtgatc tgccctgcac tctgggataa 60

gcctgggaaa ctgggtctaa taccggatag gaccttcggc cgcatggctg ggggtggaaa 120

gtttttcggt gcaggatgag cccgcggcct atcagcttgt tggtggggta atggcctacc 180

aaggcgacga cgggtagccg gcctgagagg gcgaccggcc acactgggac tgagacacgg 240

cccagactcc tacgggaggc agcagtgggg aatattgcac aatgggcgca agcctgatgc 300

agcgacgccg cgtgagggat gacggccttc gggttgtaaa cctctttcac ccatgacgaa 360

gcgcaagtga cggtagtggg agaagaagca ccggccaact acgtgccagc agccgcggta 420

atacgtaggg tgcgagcgtt gtccggaatt actgggcgta aagagctcgt aggcgggttt 480

gtcgcgtccg tctgtgaaaa cccgcagctc aactgcgggc ttgcaggcga tacgggcaga 540

ctcgagtact gcaggggaga ctggaattcc tggtgtagcg gtgaaatgcg cagatatcag 600

gaggaacacc ggtggcgaag gcgggtctct gggcagtaac tgacgctgag gagcgaaagc 660

gtgggtagcg aacaggatta gataccctgg tagtccacgc cgtaaacggt gggcgctagg 720

tgtgggtttc cttccacggg atccgtgccg tagccaacgc attaagcgcc ccgcctgggg 780

agtacggccg caaggctaaa actcaaagga attgacgggg gcccgcacaa gcggcggagc 840

atgtggatta attcgatgca acgcgaagaa ccttacctgg gtttgacatg taccggacga 900

ctgcagagat gtggtttccc ttgtggccgg tagacaggtg gtgcatggct gtcgtcagct 960

cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca acccttgtcc tgtgttgcca 1020

gcacgtaatg gtggggactc gcaggagact gccggggtca actcggagga aggtggggac 1080

gacgtcaagt catcatgccc cttatgtcca gggcttcaca catgctacaa tggtcggtac 1140

agagggctgc gataccgtga ggtggagcga atcccttaaa gccggtctca gttcggatcg 1200

gggtctgcaa ctcgaccccg tgaagtcgga gtcgctagta atcgcagatc agcaacgctg 1260

cggtgaatac gttcccgggc cttgtacaca ccgcccgtca cgtcatgaaa gtcggtaaca 1320

cccgaagccg gtggcctaac ccctc 1345

Claims

1. The Rhodococcus gordonii YKSW-6 is characterized in that the Rhodococcus gordonii (Rhodococcus gordoniae) is classified and named, and is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.22972, and the 16S rDNA sequence gene sequence of the strain is shown as SEQ ID No. 1.

2. Use of the Rhodococcus gordonii YKSW-6 according to claim 1 for the preparation of a microbial preparation for degrading skatole at high concentration.

3. The use of Rhodococcus gordonii YKSW-6 in the preparation of a microbial preparation for degrading high-concentration skatole according to claim 2, wherein the preparation method of the microbial preparation for treating skatole comprises the following steps:

1) and (3) shaking the bottle to prepare seeds: inoculating the YKSW-6 strain into a seed culture medium, and culturing at 30 ℃ and 180r/min to a logarithmic phase;

2) fermentation culture: inoculating the seed solution obtained in the step 1) into a fermentation culture solution according to the inoculation amount of 5-10% of the volume ratio, wherein the culture temperature is 30-42 ℃, the rotation speed is 180-240r/min, the initial pH is 6-9, the culture time is 24h, and the microbial deodorant is obtained after the fermentation is completed.

4. The use of Rhodococcus gordonii YKSW-6 in the preparation of a microbial preparation for degrading high-concentration skatole according to claim 3, wherein the preparation method of the microbial preparation for treating skatole comprises the following steps:

2) fermentation culture: inoculating the seed solution obtained in the step 1) into a fermentation culture solution according to the inoculation amount of 5% of the volume ratio, wherein the culture temperature is 30 ℃, the rotation speed is 180r/min, the initial pH value is 7.2, the culture time is 24h, and the microbial deodorant is obtained after the fermentation is finished.

5. The use of the microorganism of claim 3 or 4, wherein the viable count of the microorganism is 1.0 x 10, and the microorganism is the microorganism of the genus Rhodococcus gordonii YKSW-6⁹CFU/mL。