Rhodococcus fanqing sheng and application of rhodococcus fanqing sheng in efficient degradation of nicotine

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to rhodococcus fanqing sheng and application of the rhodococcus fanqing sheng to efficient degradation of nicotine. Specifically, a bacterium with the nicotine stable degradation capacity is screened from the rhizosphere environment of tobacco soil, is identified as rhodococcus qingshengii, is named as Cas430, grows rapidly, has high adaptability to the environment, can grow by using nicotine as a unique carbon source and a unique nitrogen source, has efficient degradation capacity, and can be used for preparing the rhodococcus qingshengii strain with the nicotine stable degradation capacity, so that the rhodococcus qingshengii strain can be used for preparing the rhodococcus qingshengii strain with the nicotine stable degradation capacity, and the rhodococcus qingshengii strain with the nicotine stable degradation capacity can be used for preparing the rhodococcus qingshengii strain. After the sprayed tobacco leaves are cured, the nicotine content is reduced, so that the quality of tobacco products can be effectively improved, and the harm of smoking to human bodies can be remarkably reduced; meanwhile, the strain can also be used for degrading nicotine-containing wastes generated in the production of tobacco products or treating nicotine in tobacco planting fields and water sources, so that the environment is improved, and the strain has important economic, social and ecological benefits.

Description

Technical Field

[0001] The present invention belongs to the field of microbial technology, and specifically relates to a strain of Rhodococcus fanqingsheng and its application in efficiently degrading nicotine. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance understanding of the overall background of the invention and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to those skilled in the art.

[0003] Nicotine (C 10 H 14 Nicotine (N2) is a naturally occurring pyridine alkaloid found in tobacco plants. As the primary addictive ingredient in tobacco products, its harmful effects have garnered widespread attention. Nicotine not only directly harms human health through smoking but also enters the environment through tobacco processing waste, discarded cigarette butts, and soil residues in tobacco fields, creating persistent pollution. From a health perspective, nicotine activates acetylcholine receptors in the central nervous system, leading to increased dopamine release and the development of physiological and psychological dependence. Long-term nicotine consumption significantly increases the risk of cardiovascular disease, respiratory illness, and various cancers (such as lung and oral cancer). Furthermore, nicotine can affect fetal and infant development through the placental barrier and breast milk, causing neurobehavioral abnormalities and cognitive impairment. Furthermore, due to its chemical stability, nicotine can persist in soil and water for long periods of time. Nicotine in the environment can inhibit soil microbial activity, interfere with plant root development, and accumulate in aquatic organisms through the food chain, ultimately threatening biodiversity and ecological security.

[0004] In response to the problem of nicotine pollution, existing technologies are mainly divided into two categories: physical and chemical methods and biological methods. Among them, physical and chemical methods include high-temperature incineration and acid-base treatment, the use of activated carbon, zeolite or polymer materials to adsorb nicotine, the use of catalysts such as TiO2 and ZnO to decompose nicotine under ultraviolet light, and the use of strong oxidants such as ozone and persulfate to degrade nicotine. However, the above methods have problems such as high degradation cost, poor adaptability to complex matrices (such as soil), and easy secondary pollution. Biodegradation has become a research hotspot due to its advantages such as environmental friendliness and low cost. Currently reported nicotine-degrading bacteria include Pseudomonas and Alcaligenes. However, existing strains generally have low degradation efficiency, high substrate dependence, and limited application scenarios. Summary of the Invention

[0005] Based on the above-mentioned deficiencies in the prior art, the present invention provides a strain of Rhodococcus qingshengii and its use in the efficient degradation of nicotine. Specifically, the present invention screened a bacterium with the ability to stably degrade nicotine from the rhizosphere environment of tobacco soil. The bacterium was identified as Rhodococcus qingshengii and named Cas430 (Rhodococcus qingshengii). This bacterium grows rapidly and has strong adaptability to the environment. It can use nicotine as the sole carbon and nitrogen source for growth, and has efficient degradation capabilities and research value. Based on the above research results, the present invention was completed.

[0006] In order to achieve the above technical objectives, the present invention relates to the following technical solutions:

[0007] The first aspect of the present invention provides a strain of Rhodococcus qingshengii Cas430, which was deposited on December 5, 2024 at the General Microbiology Center of the China Culture Collection Administration (address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China), and its biological deposit number is CGMCC No. 32934.

[0008] The second aspect of the present invention provides a fermentation production method of Rhodococcus Cas430, which comprises: inoculating the Rhodococcus Cas430 into a fermentation medium for fermentation culture.

[0009] In the present invention, the fermentation production method is not particularly limited, and any conventional bacterial fermentation culture method can be used for culture.

[0010] The fermentation medium can be a common bacterial culture medium. For example, in one embodiment of the present invention, NB liquid culture medium is used.

[0011] The third aspect of the present invention provides a microbial agent, which contains at least the Fanqingsheng Rhodococcus Cas430.

[0012] In the present invention, the microbial agent may further include excipients acceptable to the agent.

[0013] In another embodiment of the present invention, the excipient is selected from one or more of a dispersant, a wetting agent, a disintegrant, a binder, a defoaming agent, an antifreeze agent, a thickener, a filler, and a solvent. The present invention has no particular limitation on the source of the excipients acceptable to the microbial agent, and commercially available products can generally be used.

[0014] The dispersant is an anionic dispersant and / or a nonionic dispersant, which can be selected from one or more of sodium lignin sulfonate, sodium naphthalene sulfonate formaldehyde condensate, sodium methylene bisnaphthalene sulfonate, formaldehyde condensate sulfate, polycarboxylate, alkylphenol polyoxyethylene phosphate and fatty acid polyoxyethylene ester.

[0015] The wetting agent can be selected from one or more of sodium lauryl sulfate, sodium dodecylbenzene sulfonate, saponin powder, soapberry powder, tea kudzu powder and lakai powder BX.

[0016] The disintegrant may be selected from one or more of bentonite, ammonium sulfate, aluminum chloride, urea, magnesium chloride and glucose.

[0017] The binder may be selected from one or more of starch, diatomaceous earth, cyclodextrin, rosin, carboxymethyl cellulose, carboxyethyl cellulose and carboxymethyl cellulose salt.

[0018] The defoaming agent can be selected from one or more of C8-C20 fatty alcohol compounds, C10-C20 saturated fatty acid compounds, epoxidized soybean oil, ethanol, silicone compounds and organic silicone oil.

[0019] The antifreeze agent may be selected from one or more of sorbitol, ethylene glycol, polyethylene glycol, propylene glycol, glycerol, urea and sodium chloride.

[0020] The thickener may be selected from one or more of gelatin, xanthan gum, polyethylene glycol and polyvinyl alcohol.

[0021] The filler may be selected from one or more of light calcium carbonate, diatomaceous earth, bentonite, attapulgite and white carbon black.

[0022] The solvent may be selected from water (preferably deionized water) or methyl oleate.

[0023] A fourth aspect of the present invention provides the use of the above-mentioned Rhodococcus Cas430 and / or the above-mentioned microbial agent in nicotine degradation.

[0024] The fifth aspect of the present invention provides the use of the above-mentioned Rhodococcus Cas430 and / or the above-mentioned microbial agent in degrading nicotine in tobacco products, tobacco industrial waste, tobacco plants, soil in tobacco-growing areas and / or water sources in tobacco-growing areas.

[0025] In the present invention, the tobacco products include but are not limited to tobacco leaves, redried tobacco leaves, tobacco flakes, shredded tobacco, cigarettes and cigars; the tobacco industry waste includes but is not limited to tobacco stems and tobacco dust.

[0026] In the present invention, the shredded tobacco refers to the shredded, powdered or granular products made from tobacco leaves, redried tobacco leaves or tobacco flakes as raw materials.

[0027] A sixth aspect of the present invention provides a method for degrading nicotine, comprising the steps of treating tobacco products, tobacco plants, soil in tobacco-growing areas, and / or water sources in tobacco-growing areas using the above-mentioned Rhodococcus Cas430 and / or the above-mentioned microbial agent.

[0028] The seventh aspect of the present invention provides the use of the above-mentioned Rhodococcus Cas430, microbial agent and / or method in the preparation of cigarettes and / or tobacco oils.

[0029] In the present invention, the smoke oil can be the smoke oil of electronic cigarettes and / or other non-combustion cigarette products.

[0030] Beneficial technical effects of one or more of the above technical solutions:

[0031] The above technical solution reports for the first time a strain of Fan Qingsheng Rhodococcus that efficiently degrades nicotine. The strain has the ability to grow with nicotine as the sole carbon and nitrogen source. After culturing on BMM culture medium containing 1g / L nicotine for 24 hours, the nicotine content can be reduced by more than 80%. Nicotine can be completely degraded after 42 hours. The nicotine degradation efficiency is high. In the presence of other carbon and nitrogen sources, nicotine is preferentially used as a metabolite. It has a strong nicotine metabolism ability. After spraying the strain on tobacco plants for 12 hours, the nicotine content decreases by about 20.34%. The nicotine content of the sprayed tobacco leaves is reduced after curing, thereby effectively improving the quality of tobacco products and significantly reducing the harm of smoking to the human body. At the same time, the strain can also be used to degrade nicotine-containing waste generated in the production of tobacco products, realizing the comprehensive utilization of waste, or used to treat nicotine in tobacco planting areas and water sources, thereby improving the environment. It has important economic, social and ecological benefits and therefore has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The accompanying drawings, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0033] Figure 1 This is the colony morphology of the strain Cas430 (Rhodococcus qingshengii) in the examples of the present invention.

[0034] Figure 2 The Neighbour-Joining method was used to construct a phylogenetic tree of Cas430 (Rhodococcus qingshengii) and other standard species of Rhodococcus in the examples of the present invention.

[0035] Figure 3This is the nicotine degradation curve of the strain Cas430 (Rhodococcus qingshengii) in the embodiment of the present invention after being cultured in BMM medium containing 1 g / L nicotine. The peak area is the nicotine absorption peak area detected by HPLC (black is 18 h, blue is 24 h, green is 42 h, and cyan is 48 h).

[0036] Figure 4 The growth rate of the strain Cas430 (Rhodococcus qingshengii) in the embodiment of the present invention after being cultured in BMM medium containing 1, 2, 3, 4, and 5 g / L nicotine at 16 h, 20 h, 24 h, 30 h, 40 h, and 48 h.

[0037] Figure 5 The peak areas of the liquid phase detection at 16 h, 20 h, 24 h, 30 h, 40 h and 48 h after the strain Cas430 (Rhodococcus qingshengii) in the embodiment of the present invention was cultured in BMM medium containing 1, 2, 3, 4, and 5 g / L nicotine. DETAILED DESCRIPTION

[0038] It should be noted that the following detailed descriptions are exemplary and intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0039] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments of the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, they indicate the presence of features, steps, operations, devices, components and / or combinations thereof.

[0040] The present invention is further explained by the following examples, but is not intended to limit the present invention. It should be understood that these examples are only intended to illustrate the present invention and are not intended to limit the scope of the present invention.

[0041] Example

[0042] 1 Materials and Methods

[0043] 1.1 Materials

[0044] Experimental samples: soil samples from the root surface of tobacco in Hubei.

[0045] Required culture media: NA medium, NB liquid medium, BMM medium (KH2PO4, 3.6 g; (NH4)2SO4, 2 g; L-glutamic acid, 2.9 g; glucose, 0.18 g; FeSO4-7H2O, 0.5 mg (pH = 6.5)), and LB medium.

[0046] Required sample: Nicotine (purity: HPLC ≥ 95%)

[0047] 1.2 Methods

[0048] 1.2.1 Isolation and culture of strains

[0049] In August 2022, rhizosphere soil samples of tobacco plants infected with bacterial wilt in Hubei were collected. After shaking off the soil attached to the root surface, the root tissue sample was placed in sterilized deionized water, and the root surface soil was collected using ultrasonic vibration. The centrifuge tube was placed in water and ultrasonicated for 20 seconds each time, for a total of two ultrasonications. The fine roots in the centrifuge tube were removed and discarded with sterile tweezers in the clean bench, 1 mL of soil suspension was added to 9 mL of sterilized deionized water, and shaken to 10 -1 times the bacterial solution, and prepare 10 -2 , 10 -3 , 10 -4 , 10 -5 and 10 -6 times of bacterial solution. Take the concentration of 10 -3 , 10 -4 , 10 -5 and 10 -6 Prepare 0.1 mL of each dilution, replicate three times for each dilution factor, and spread onto NA medium using a disposable spreader. Incubate at 28°C for 48 hours. Single colonies are selected based on their growth characteristics, including color, size, degree of prominence, transparency, hardness, and regularity of edges. These colonies are then streaked three times onto NA medium and purified three times to obtain a pure culture. The strain with the best growth was selected for identification and further study of its nicotine degradation activity.

[0050] 1.2.2 Identification of strain types

[0051] ① Streak the strain onto fresh NA medium and incubate at 28°C for 24 hours. Use a 100 μL pipette tip to pick up the purified bacterial lawn and place it in a 2 mL centrifuge tube. Add 100 μL of sterile water and boil in a water bath for 10 minutes. After cooling, centrifuge at 12,000 rpm for 10 minutes. The supernatant is the DNA of the strain.

[0052] ② PCR amplification was performed using genomic DNA as a template using the universal bacterial primers 27F (5′-AGAGTTTGATCATGGCTCAG-3′) and 1492F (5′-AAGGAGGTGATCCAACCGCA-3′). The PCR reaction system (50 μL) consisted of 2 μL DNA template, 25 μL 2× Taq Plus Master Mix II (Dye Plus), 1.5 μL each of 27F and 1492R (10 μmol / L), and ddH2O to make up to 50 μL. PCR reaction conditions were: 95°C for 3 min, followed by 30 cycles of 95°C for 30 s, 58°C for 30 s, and 72°C for 1.5 min, followed by 72°C for 5 min. The PCR amplification products were detected by 1% agarose gel electrophoresis and sent to NIO for sequencing. The spliced results after sequencing were compared with BLAST similarity sequences on NCBI. Multiple sequence homology analysis was performed using MEGA software, and a phylogenetic tree was constructed. The taxonomic status of the strain was finally determined based on the similarity and the position of the phylogenetic tree.

[0053] 1.2.3 Culture preservation

[0054] The purified strain was streaked on a slant, and the single colony grown after the streak was picked up with an inoculating loop and transferred to NB liquid medium. The bacteria were shaken at 28°C and 180 rpm until the exponential growth phase. The bacteria were stored in 50% glycerol aqueous solution, divided into 2 mL cryovials, and frozen in a -80°C refrigerator for use. They were then sent to the China General Microbiological Culture Collection Center (CGMCC) with the deposit number CGMCC No. 32934.

[0055] 1.2.4 Establishment of Nicotine Liquid Phase Standard Curve

[0056] Nicotine was prepared into seven concentration gradient standard solutions of 2 g / L, 0.5 g / L, 0.25 g / L, 0.125 g / L, 0.0625 g / L and 0.03125 g / L, and a nicotine concentration standard curve was established using high performance liquid chromatography.

[0057] The chromatographic column was Agilent TC-C18; the mobile phase was 40% acetonitrile; the column temperature was 30°C; the flow rate was 0.6 mL / min; the injection volume was 20 μL; and the detection wavelength was 259 nm.

[0058] 1.2.5 Effect of fermentation time on nicotine degradation efficiency of Cas430 (Rhodococcus qingshengii)

[0059] Cas430 (Rhodococcus qingshengii) was cultured in BMM medium containing 1 g / L nicotine. Samples were taken at 18 h, 24 h, 42 h, and 48 h, and the nicotine degradation rate was detected by liquid chromatography. 1 mL of fermentation broth was centrifuged at 15,000 rpm for 2 min, the bacteria were discarded, and the supernatant was taken. The organic phase was filtered through a 0.22 μm filter membrane and then placed in a liquid chromatography vial for HPLC detection.

[0060] The chromatographic column was Agilent TC-C18; the mobile phase was 40% acetonitrile; the column temperature was 30°C; the flow rate was 0.6 mL / min; the injection volume was 20 μL; and the detection wavelength was 259 nm.

[0061] 1.2.6 Effect of fermentation concentration on nicotine degradation efficiency of Cas430 (Rhodococcus qingshengii)

[0062] Strain Cas430 (Rhodococcus qingshengii) was inoculated at a 1% inoculum into nicotine liquid medium containing 1, 2, 3, 4, and 5 g / L of nicotine. Culture conditions were 28°C and 200 rpm. Cell growth was measured every 4 hours, and the degradation rate at each time point was determined at different nicotine concentrations.

[0063] 1.2.7 Degradation of nicotine on tobacco plants

[0064] 1) Preparation of bacterial solution

[0065] Cas430 was cultured in LB liquid medium overnight to an OD 600 =1.0, dilute the bacterial solution 10 times with deionized water for later use.

[0066] 2) Treatment methods

[0067] Select tobacco plants with consistent growth after topping, spray the whole plant twice, spray the control group with clean water, and treat 3 tobacco plants in each treatment. After spraying, take 4 leaves from each plant 12 hours later, put them in an envelope and dry them in a 72-degree oven, grind them into powder for later use.

[0068] 3) Detection method

[0069] 10 g of dried tobacco leaf samples were taken from each treatment, and their nicotine content was detected using a Fourier transform near-infrared spectrometer.

[0070] 2. Experimental Results

[0071] 2.1.1 Morphological characteristics of strains

[0072] Cas430 (Rhodococcus qingshengii) was inoculated on NA medium and cultured at 28℃ for 24h. The single colony of the strain was light yellow, slightly transparent, and had smooth edges ( Figure 1 ).

[0073] 2.1.2 Identification of strain species

[0074] By 16S rDNA sequencing, the nucleotide sequence shown in SEQ ID NO.1 was obtained. The obtained sequence was compared with the EZBioCloud database, and it was found that Cas430 (Rhodococcus qingshengii) had the highest similarity with Fan Qingsheng Rhodococcus (Rhodococcus qingshengii) JCM 15477, which was 99.78%. A phylogenetic tree was constructed by comparing Cas430 with the 16S rDNA sequences of other standard bacteria of the genus Rhodococcus ( Figure 2 ), found that Cas430 and Rhodococcus fanqingsheng were clustered together, and finally determined that Cas430 was Rhodococcus fanqingsheng.

[0075] 2.2 Effect of fermentation time on nicotine degradation efficiency of Cas430 (Rhodococcus qingshengii)

[0076] Cas430 (Rhodococcus qingshengii) was cultured in BMM medium containing 1g / L nicotine. Samples were taken at 18h, 24h, 42h and 48h, and the nicotine degradation rate was detected by liquid chromatography. The experimental results showed that the degradation rate of Cas430 was 52.49% at 18h; the degradation rate was 78.76% at 24h; at 42h, the liquid chromatography showed that nicotine was completely degraded ( Figure 3 ).

[0077] 2.3 Effect of fermentation concentration on nicotine degradation efficiency of Cas430 (Rhodococcus qingshengii)

[0078] The strain Cas430 was inoculated at a 1% inoculum into nicotine liquid culture medium with nicotine contents of 1, 2, 3, 4, and 5 g / L, and the OD 600 The results showed that the strain Cas430 grew uninhibitedly in 1 and 2 g / L nicotine liquid culture media, but grew slower at 3, 4, and 5 g / L concentrations compared to 1 and 2. Liquid phase results showed that the strain Cas430 degraded nicotine rapidly in 1 and 2 g / L nicotine liquid culture media, with 1 g / L nicotine completely degraded within 48 hours, while the nicotine degradation rate was very slow at 3, 4, and 5 g / L concentrations.

[0079] 2.4 Degradation of nicotine on tobacco plants

[0080] The results of the treated tobacco leaf samples were analyzed using a Fourier transform near-infrared spectrometer. The results showed that the nicotine content of the tobacco plants decreased by 20.34% 12 hours after the tobacco plants were sprayed with a bacterial solution containing Cas430 and its fermentation products.

[0081] In summary, the present invention isolated a bacterial strain, named Cas430, from the root surface of diseased tobacco plants. 16S rRNA sequencing and phylogenetic tree construction confirmed that strain Cas430 is Rhodococcus qingshengii. Experimental results demonstrated that strain Cas430 can grow using nicotine as its sole carbon and nitrogen source. Cultivation for 30 hours in a carbon and nitrogen-depleted BMM medium containing 0.1 g / L nicotine reduced nicotine content by 95.16%. In the presence of other carbon and nitrogen sources, the bacteria will still preferentially use nicotine as a metabolite and has a strong nicotine metabolism ability. After culturing on BMM culture medium containing 1g / L nicotine for 24 hours, the nicotine content can be reduced by more than 80%. Nicotine can be completely degraded at 42 hours, and the nicotine degradation efficiency is high. The strain Cas430 was inoculated at a 1% inoculum size into nicotine liquid culture medium with nicotine contents of 1, 2, 3, 4, and 5g / L for culture. The results showed that the growth of strain Cas430 was not inhibited in 1 and 2g / L nicotine liquid culture medium, 1g / L nicotine was completely degraded within 48 hours, and the three concentrations of 3, 4, and 5g / L grew slowly compared to 1 and 2g / L nicotine.

[0082] The bacteria also has a significant ability to degrade nicotine on tobacco plants. Spraying tobacco plants with a bacterial solution containing Cas430 (Rhodococcus qingshengii) and its fermentation products for 12 hours reduced the nicotine content of the tobacco plants by about 20.34%.

[0083] Strain Cas430 16S rRNA gene sequence

[0084] TGCAGTCGAGCGGTAAGGCCTTTCGGGGTACACGAGCGGCGAACGGG

[0085] TGAGTAACACGTGGGTGATCTGCCCTGCACTTCGGGATAAGCCTGGGA

[0086] AACTGGGTCTAATACCGGATATGACCTCCTATCGCATGGTGGGTGGTGG

[0087] AAAGATTTATCGGTGCAGGATGGGCCCGCGGCCTATCAGCTTGTTGGTG

[0088] GGGTAATGGCCTACCAAGGCGACGACGGGTAGCCGACCTGAGAGGGT

[0089] GACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGG

[0090] CAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCGAC

[0091] GCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGCAGG

[0092] GACGAAGCGCAAGTGACGGTACCTGCAGAAGAAGCACCGGCTAACTA

[0093] CGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTTGTCCGGAAT

[0094] TACTGGGCGTAAAGAGTTCGTAGGCGGTTTGTCGCGTCGTTTGTGAAA

[0095] ACCAGCAGCTCAACTGCTGGCTTGCAGGCGATACGGGCAGACTTGAGT

[0096] ACTGCAGGGGAGACTGGAATTCCTGGTGTAGCGGTGAAATGCGCAGAT

[0097] ATCAGGAGGAACACCGGTGGCGAAGGCGGGTCTCTGGGCAGTAACTG

[0098] ACGCTGAGGAACGAAAGCGTGGGTAGCGAACAGGATTAGATACCCTG

[0099] GTAGTCCACGCCGTAAACGGTGGGCGCTAGGTGTGGGTTCCTTCCACG

[0100] GAATCCGTGCCGTAGCTAACGCATTAAGCGCCCCGCCTGGGGAGTACG

[0101] GCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCG

[0102] GCGGAGCATGTGGATTAATTCGATGCAACGCGAAGAACCTTACCTGGG

[0103] TTTGACATATACCGGAAAGCTGCAGAGATGTGGCCCCCCTTGTGGTCG

[0104] GTATACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGG

[0105] GTTAAGTCCCGCAACGAGCGCAACCCCTATCTTATGTTGCCAGCACGTT

[0106] ATGGTGGGGACTCGTAAGAGACTGCCGGGGTCAACTCGGAGGAAGGT

[0107] GGGGACGACGTCAAGTCATCATGCCCCTTATGTCCAGGGCTTCACACA

[0108] TGCTACAATGGCCAGTACAGAGGGCTGCGAGACCGTGAGGTGGAGCG

[0109] AATCCCTTAAAGCTGGTCTCAGTTCGGATCGGGGTCTGCAACTCGACC

[0110] CCGTGAAGTCGGAGTCGCTAGTAATCGCAGATCAGCAACGCTGCGGTG

[0111] AATACGTTCCCGGGCCTTGTACACACCGCCCGTCACGTCATGAAAGTC

[0112] GGTAACACCCGAAGCCGGTGGCTTAACCCCTTGTGGGAGGGAGCCGT

[0113] CGAA(SEQ ID NO.1)

[0114] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or make equivalent replacements for parts thereof. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention. Although the above describes the specific implementation methods of the present invention, it is not intended to limit the scope of protection of the present invention. Those skilled in the art should understand that, based on the technical solution of the present invention, various modifications or variations that can be made by those skilled in the art without creative work are still within the scope of protection of the present invention.

Claims

1. A strain of Rhodococcus qingshengii Cas430, which was deposited in the General Microbiology Center of China Culture Collection Administration on December 5, 2024, and its biological deposit number is CGMCC No. 32934.

2. The fermentation production method of Rhodococcus Cas430 according to claim 1, characterized in that The fermentation production method comprises: inoculating the Fanqingsheng Rhodococcus Cas430 into a fermentation medium for fermentation culture.

3. A microbial agent, characterized in that: It contains at least the Rhodococcus Cas430 described in claim 1.

4. The microbial agent according to claim 3, wherein The microbial agent also includes an adjuvant acceptable to the agent; further, the adjuvant is selected from one or more of a dispersant, a wetting agent, a disintegrant, a binder, a defoaming agent, an antifreeze agent, a thickener, a filler and a solvent.

5. Use of the Rhodococcus Cas430 of claim 1 and / or the microbial agent of any one of claims 3-4 in nicotine degradation.

6. Use of the Rhodococcus Cas430 described in claim 1 and / or the microbial agent described in any one of claims 3-4 in degrading nicotine in tobacco products, tobacco industry waste, tobacco plants, soil in tobacco-growing areas, and / or water sources in tobacco-growing areas.

7. The use according to claim 6, characterized in that The tobacco products include tobacco leaves, redried tobacco leaves, tobacco flakes, shredded tobacco, cigarettes and cigars; the tobacco industry waste includes tobacco stems and tobacco dust.

8. A method for degrading nicotine, characterized in that: The method comprises the steps of treating tobacco products, tobacco plants, soil in tobacco planting areas and / or water sources in tobacco planting areas using the Rhodococcus Cas430 described in claim 1 and / or the microbial agent described in any one of claims 3-4.

9. Use of the Rhodococcus Cas430 of claim 1 and / or the microbial agent of any one of claims 3-4 and / or the method of claim 8 in the preparation of cigarettes and / or tobacco oils.

10. The use according to claim 9, characterized in that The e-liquid is e-cigarette and / or other non-combustion cigarette products.

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