CN113337425B - Gordoniella for producing long-carbon-chain mycolic acid and application thereof - Google Patents
Gordoniella for producing long-carbon-chain mycolic acid and application thereof Download PDFInfo
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Abstract
The invention discloses Gordonia for producing long-carbon-chain mycolic acid and application thereof, and belongs to the technical field of desulfurization microorganisms. The Gordonia producing long carbon chain mycolic acid is Gordonia (Gordonia sp.) JT-2, and the preservation number is CCTCCNO: m2021493. The Gordonia sp.JT-2 has strong hydrophobicity, and when the actual oil product is desulfurized, the bacteria can secrete more mycolic acid, so that the bacteria have stronger hydrophobicity. Therefore, the characteristic of extremely high hydrophobicity of Gordonia sp.JT-2 in an oil-water multiphase system can be utilized, the toxic action of the hydroxylated desulfurization product on bacteria is reduced, the problem of low oil-water mass transfer efficiency of sulfur-containing organic matters is solved, and the desulfurization efficiency is improved.
Description
Technical Field
The invention belongs to the technical field of desulfurization microorganisms, and particularly relates to Gordonia for producing long-carbon-chain mycolic acid and application thereof.
Background
Petroleum is one of the main energy sources in the world at present, and occupies an indispensable position in national economic development. Various forms of sulfur-containing organic matter exist in petroleum, which not only reduce the quality of the oil, but also release large amounts of Sulfur Oxides (SO) during combustion x ) Thereby having a great influence on the environment. In order to control such adverse effects, environmental protection agencies in many countries have proposed increasingly strict legislative regulations on the total sulfur content of oils, and currently, the standards for sulfur content in oils are set to 10ppm (5 ppm for some regions) in recent proposals in our country and in countries such as the united states, european union, and japan, and it is expected that these standards will become more stringent in the future, so that oil desulfurization will be under increasing pressure.
Biological desulfurizationThe technology is a new technology for desulfurizing sulfur-containing organic matters in oil products by using desulfurization bacteria at normal temperature and normal pressure, has the advantages of low investment, cleanness, no pollution, mild reaction conditions, easiness in desulfurizing organic sulfur and the like, is one of feasible technologies for obtaining ultra-low sulfur clean oil products at present, develops an approach and a method for deep desulfurization of the oil products due to the appearance of the technology, and has great application potential. However, the efficiency and stability of current biological desulfurization techniques have not yet met the needs of the industry. Wherein, factors such as poison inhibition of desulfurization products, oil-water mass transfer efficiency of sulfur-containing organic matters, viability of desulfurization microorganisms, desulfurization activity, desulfurization life and the like are key factors for realizing industrialization of the biological desulfurization technology. The prior art shows that the desulfurization product 2-HBP has strong lethality to desulfurization bacteria, and the prior art reports that extra MgSO (MgSO) is added 4 While sulfur sources or glucose are used as carbon sources to improve the resistance of the cells to 2-HBP, relatively few studies have been made on desulfurization bacteria that are resistant to the deleterious effects of desulfurization products 2-HBP.
Disclosure of Invention
The invention aims to provide Gordonia for producing long-carbon-chain mycolic acid and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
the Gordoniae capable of producing the long-carbon chain mycolic acid is Gordoniae (Gordonia sp.) JT-2 with the preservation number of CCTCC NO: m2021493.
The Gordoniae is applied to preparation of a biological desulfurizer which can resist the toxic action of hydroxylated desulfurization products.
The biological desulfurizer which can resist the toxic action of the hydroxylated desulfurization product comprises the following components with the preservation number of CCTCC NO: m2021493 Gordoniella.
A biological desulfurization method with little influence of the toxicity of hydroxylated desulfurization products is characterized in that a biological desulfurization method with the preservation number of CCTCC NO: m2021493 Gordoniella strain.
In a specific embodiment of the invention, the bacterium concentration of the Gordonia bacterium liquid is more than or equal to 1.6 x 10 8 CFU/mL; the addition amount is 4 percent(v/v); the desulfurization system is an oil-water multiphase system, and preferably, the oil-water multiphase system has an oil-water ratio of 1:10 (v/v).
The hydroxylated desulfurization product is a hydroxylated desulfurization compound generated after sulfur-containing organic matters (such as benzothiophene, benzothiophene derivatives, dibenzothiophene derivatives and the like) are desulfurized through a specific carbon-sulfur bond breaking way; for example: 2-hydroxybiphenyl generated after DBT desulfurization; 3-methyl-2-hydroxybiphenyl (3-methyl-2-hydroxybiphenyl) or 3'-methyl-2-hydroxybiphenyl (3' -methyl-2-hydroxybiphenyl) generated after 4-MDBT desulfurization; 2-hydroxy-3 ', 5-dimethyl-biphenyl (3',5-dimethyl-2-hydroxybiphenyl) generated after 2,8-DMDBT desulfurization; 2- (1-methylvinyl) phenol formed after 3-MBT desulfurization.
The technical scheme of the invention has the advantages that:
the invention separates and obtains a desulfurization bacterium Gordonia sp.JT-2 which can secrete mycolic acid with four unsaturated double bonds and 58 total carbon atoms on the cell wall. The presence of cell wall mycolic acid makes the bacteria strongly hydrophobic and when desulfurizing the actual oil, the bacteria can secrete more mycolic acid, making it more hydrophobic. Therefore, the characteristic of extremely high hydrophobicity of Gordonia sp.JT-2 in an oil-water multiphase system can be utilized, the toxic action of the hydroxylated desulfurization product on bacteria is reduced, the problem of low oil-water mass transfer efficiency of sulfur-containing organic matters is solved, and the desulfurization efficiency is improved.
Drawings
FIG. 1 a phylogenetic tree of Gordonia sp.JT-2;
FIG. 2 scanning electron micrograph of Gordonia sp.JT-2 and spectrum of mycolic acid ESI-HRMS;
FIG. 3 the mycolic acid signal intensity secreted by Gordonia sp.JT-2 cultured under different conditions;
fig. 4 Gordonia sp.jt-2 hydrophobicity under different conditions;
FIG. 5 surface contact angles of Gordonia sp.JT-2 cultured with different desulfurization systems;
FIG. 6 effect of hydroxylated desulfurization product concentration on biological desulfurization efficiency and growth of Gordonia sp.JT-2 under different systems;
7 Gordonia sp.JT-2 on the GC-SCD spectrum and desulfurization effect of real diesel oil sulfur metabolism.
Detailed Description
Terms used in the present invention have generally meanings as commonly understood by one of ordinary skill in the art, unless otherwise specified.
The present invention will be described in further detail with reference to the following data in conjunction with specific examples. The following examples are intended to illustrate the invention and are not intended to limit the scope of the invention in any way.
Example 1 isolation, purification and characterization of the strains
Obtaining a sample: and (4) in 2016, collecting petroleum polluted soil of a victory oil field from eastern Shandong.
Separating and purifying strains:
respectively taking 4-methyl dibenzothiophene (4-MDBT) and 3-methyl benzothiophene (3-MBT) as unique sulfur sources, and screening the strains desulfurized in a specific carbon-sulfur bond breaking mode from petroleum contaminated soil of the Shengli oil field. The method comprises the following specific steps:
adding a proper amount of soil sample into a test tube containing sterile deionized water, fully shaking, standing for 10min, transferring a proper amount of upper-layer liquid into 50mL of liquid selection culture medium (respectively taking 4-MDBT and 3-MBT as unique sulfur sources), and culturing for 4 days at the rotation speed of 160rpm at the temperature of 30 ℃;
secondly, transferring a proper amount of culture solution obtained in the first step to 50ml of liquid LB culture medium, and culturing for 1 day at the rotating speed of 160rpm and 30 ℃;
coating a proper amount of the culture solution obtained in the step II on a solid LB culture medium by a dilution coating method, and culturing for 3 days in a constant-temperature biochemical incubator at 30 ℃;
picking single colony from the solid LB culture medium in the step (c), inoculating the single colony into 50mL liquid LB culture medium, and culturing for 1 day at 30 ℃ and 160 rpm; then transferring a proper amount of culture solution to 50mL of liquid selection culture medium, and culturing for 4 days at the rotation speed of 160rpm at 30 ℃;
fifthly, detecting whether a compound without a substituent at a hydroxyl para position is generated in the culture solution through a Gibbs reaction, selecting a sample with positive Gibbs reaction, carrying out repeated streaking separation and purification on a single bacterium on a solid LB culture medium, and inoculating the single bacterium in a liquid LB culture medium for amplification culture;
sixthly, respectively inoculating the culture solution after the separation and the amplification culture into 50mL of inorganic salt culture medium containing different sulfur-containing organic matters, culturing for 4 days at the rotating speed of 160rpm at the temperature of 30 ℃, and inspecting the desulfurization effect and the substrate adaptability, wherein the culture solution detects whether a compound without a substituent at the phenolic hydroxyl group para position is generated in a sample through Gibbs reaction;
and seventhly, inoculating the strain with good desulfurization effect and strong substrate adaptability into a liquid LB culture medium, culturing for 1 day at the rotating speed of 160rpm at the temperature of 30 ℃, transferring 5mL of bacterial liquid into a centrifugal tube containing 5mL of glycerol (after high-temperature and high-pressure sterilization), fully oscillating by using a vortex instrument, preserving in a refrigerator at the temperature of minus 80 ℃, inoculating the strain into a solid LB inclined plane culture medium by an inclined plane low-temperature preservation method, preserving in a refrigerator at the temperature of 4 ℃ for a short time, and transferring once every month.
Finally, a high-efficiency Gordonia sp.JT-2 capable of desulfurizing a plurality of sulfur-containing organic matters by specifically breaking carbon-sulfur bonds is obtained.
The culture medium adopted in the process is as follows:
liquid LB medium: 5.0g of yeast powder, 10.0g of peptone, 10.0g of NaCl and 1000mL of deionized water, and adjusting the pH value to 7.0.
Solid LB medium: 20g/L agar powder is added into LB culture medium.
Liquid selection medium: 0.5mL of 4-methyl dibenzothiophene-ethanol solution and 3-MBT-ethanol solution are respectively added into every 50mL of sulfur-free culture medium, and the concentration of sulfur-containing organic matters in the culture medium is 0.54 mmol/L.
Sulfur-free medium (mineral salts medium): 5.0g of glucose, KH 2 PO 4 1.0g，NH 4 Cl 1.0g，K 2 HPO 4 8.0g，MgCl 2 ·6H 2 0.2g of O, 10mL of metal solution, 1mL of vitamin complex, 1000mL of deionized water and pH 7.5.
Metal solution: CaCl 2 2g，NaCl 1g，FeCl 2 ·4H 2 O 0.5g，ZnCl 2 0.5g，MnCl 2 ·4H 2 O 0.5g，Na 2 MoO 4 ·2H 2 O 0.1g，CuCl 2 0.05g，Na 2 WO 4 ·2H 2 0.05g of O, 120mmol of HCl and 1000mL of deionized water.
Vitamin complex: 400mg of calcium pantothenate, 200mg of inositol, 400mg of nicotinic acid, 6400 mg of vitamin B, 200mg of p-aminobenzoic acid, 120.5 mg of vitamin B and 1000mL of deionized water.
Wherein glucose, MgCl 2 ·6H 2 Filtering O and various sulfur-containing organic matter-ethanol solutions with microporous membrane (pore diameter of 0.22 μm) for sterilization (to prevent denaturation), adding into the solution, preparing the rest culture medium components into solution, and sterilizing at 121 deg.C under 0.12MPa for 30 min.
Identification of the strains:
physical and chemical properties:
the results of physiological and biochemical experiments in Gordonia sp.JT-2 are shown in Table 1, and the bacterium was preliminarily identified as a microorganism belonging to the genus Gordonia (Corynebacterium-Nocardiaceae) of the order Corynebacterium, according to Bergey's Manual of systematic bacteriology.
TABLE 1 Gordonia sp.JT-2 Biochemical and physiological Experimental results
Molecular biological identification
Amplifying a 16S rRNA coding gene sequence of high-efficiency Gordonia sp.JT-2, and obtaining a result shown in SEQ ID NO: 1, and the following components:
DNA sequences were subjected to identity search using BLAST program "blastn" of the National Center for Biotechnology Information (NCBI) and the results were sorted in ascending order of E value (E value) as shown in Table 2. BLAST analysis of the sequence revealed that the 16S rRNA-encoding gene of JT-2 has 99% Identity (Identity) with the 16S rRNA-encoding gene of multiple strains of Gordonia (Gordonia) bacteria, and therefore preliminary identification of JT-2 as Gordonia (Gordonia sp.).
Table 216S rRNA coding gene sequence alignment analysis result
Phylogenetic analysis:
to determine the phylogenetic position of Gordonia sp.jt-2, a phylogenetic tree was constructed using the neighbor joining method based on the 16S rRNA encoding gene sequence of Gordonia sp.jt-2, the NCBI database BLAST results, as shown in fig. 1. Phylogenetic analysis showed that Gordonia sp.jt-2 clustered with other Gordonia (Gordonia) bacteria into a large branch. Combining the results of phylogenetic analysis and physiological and biochemical experiments of Gordonia sp.JT-2, the strain is finally determined to be in Gordonia and named Gordonia sp.JT-2.
Sending the Gordonia sp.JT-2 to the China center for preservation, wherein the preservation name is Gordonia sp.JT-2, the preservation date is 2021, 4 and 30 months, and the preservation number is CCTCC NO: m2021493, wherein the preservation address is China Center for Type Culture Collection (CCTCC), Wuhan university, China.
Example 2 hydrophobicity and Mycobacteria determination of Gordonia sp
Gordonia sp.
Observing the surface topography of the bacteria by a biological scanning electron microscope, as shown in fig. 2, discovering that the Gordonia sp.jt-2 thallus cell wall surface is uneven and is wrapped by a layer of waxy substance, extracting the waxy substance on the cell wall surface, and analyzing by ESI-high resolution mass spectrometry (ESI-HRMS) (fig. 2), wherein 851.57680 m/z is mycolic acid (mycolic acid) with four unsaturated double bonds and a total carbon number of 58. Mycolic acid is fatty acid containing alpha-alkyl chain-beta-hydroxyl, bacteria are mutually adhered to form aggregates under the wrapping of the mycolic acid, the aggregation form is favorable for resisting the toxic action of an oil phase organic solvent in an oil-water two-phase reaction system, meanwhile, the existence of the mycolic acid and the carbon chain length of the mycolic acid are related to the surface hydrophobicity of the bacteria, the bacteria containing the mycolic acid are considered to be more hydrophobic than the bacteria lacking the mycolic acid, and the contact angle of the surface of the bacteria is increased (namely the surface hydrophobicity of the bacteria is stronger) along with the increase of the carbon chain length of the mycolic acid. The first discovered bacterium Rhodococcus erythropolis IGTS8 with the function of desulfurizing by a specific carbon-sulfur bond breaking way contains mycolic acid with the total carbon number of 34-50, and compared with the bacterium Gordonia sp.JT-2 with the function of longer carbon chain and stronger hydrophobicity.
Hydrophobicity assay for Gordonia sp.
Gordonia sp.jt-2 was cultured in the following media, respectively:
(i) DBT-aqueous phase: an inorganic salt culture medium with DBT as the only sulfur source (the DBT content in the culture medium is 0.543 mmol/L);
(ii) LB culture medium;
(iii) DBT-mock oil: a medium containing inorganic salts (DBT content in the medium is 0.543mmol/L) with a simulated oil (DBT content is 173.76ppm) prepared by dissolving DBT in n-tetradecane as the only sulfur source, wherein the simulated oil/water is 1/10 (v/v);
(iv) diesel oil: inorganic salt medium with pure diesel oil (with a sulphur content of 167.7ppm) from ritujin petrochemical plant, Shandong as the only sulphur source, wherein the oil/water ratio is 1/10 (v/v).
The determination method of mycolic acid comprises the following steps: taking 80mL of cultured bacterial liquid, treating for 20min in an autoclave at 121 ℃, cooling, centrifuging for 30min at 20 ℃ under 4000rpm by a high-speed centrifuge, collecting thalli, then resuspending the thalli by using a prepared PBS phosphate buffer solution, and then repeating the centrifugation step again to collect the thalli. Collecting thallus, and using CHCl 2 Resuspending thallus in MeOH solution, ultrasonically dispersing thallus for 5min under an ultrasonic oscillator, then extracting for 1h under magnetic stirring at normal temperature, centrifuging for 20min at 4000rpm through a high-speed centrifuge, taking supernatant, repeating the steps on the lower-layer thallus, combining the two supernatants, putting the combined supernatants into a clean round-bottom flask, and evaporating to dryness through a rotary evaporator. For extracted mycolic acid sampleDissolving in anhydrous ether, ESI-high resolution mass spectrometry in negative ion mode, and analyzing the content of mycolic acid by the intensity of corresponding peak in mass spectrometry (m/z 851.57523).
The hydrophobicity measuring method comprises the following steps: gordonia sp.JT-2 cells cultured in four different media were collected and washed 3 times with sterile physiological saline; diluted with sterile saline to an OD600 of about 0.6 (D0); adding 1mL of n-dodecane into 4mL of the bacterial liquid, oscillating for 2min by using a vortex instrument to uniformly mix the mixture, and standing for 15 minutes at room temperature; the upper n-dodecane layer was completely absorbed by a gun, and then the OD600 value (D) of the lower aqueous phase was measured, and the proportion of the bacteria in the aqueous phase was calculated to find the hydrophobicity. The surface hydrophobicity is expressed by a hydrophobic distribution value, and the calculation formula of the hydrophobic distribution value H is shown in formula (1).
Wherein H is the hydrophobic distribution value (%), D 0 The initial OD600 value of the aqueous phase solution is shown, and D is the OD600 value of the aqueous phase solution after n-dodecane is added, shaken and kept stand.
Method for measuring cell surface contact angle: washing the separated thallus with sterile physiological saline for 3 times; the washed bacteria are intercepted on an acetate fiber filter membrane (the aperture is 0.22 mu m) in a suction filtration mode, the bacteria-bearing surface faces upwards and is placed on an agar plate (the agar plate: 20g of agar, 10 percent (v/v) of glycerol, 1000mL of distilled water and the pH value is adjusted to 7.0.), the filter membrane is placed on a superclean bench at room temperature overnight to enable the filter membrane to be wet uniformly, then the filter membrane is taken out and is placed on common filter paper for more than 12h, and after the filter membrane is completely dried, the bacteria surface contact angle is measured by a JY-82B type contact angle measuring instrument in a sitting drop method.
Contact angles between 16 ° and 40 ° are generally considered to be hydrophilic to moderately hydrophobic bacteria; the contact angle is between 55 degrees and 103 degrees, and the bacteria are hydrophobic; contact angles greater than 111 ° are extremely hydrophobic bacteria. The mycolic acid content, hydrophobicity experiment and surface contact angle measurement results are shown in FIGS. 3-5, respectively, and the mycolic acid peak intensity is 1.84X 10 in DBT water phase 7 Corresponding hydrophobic partition value of 61.9Percent, the contact angle of the surface of the thallus is 74.5 degrees; in LB medium, the peak intensity of mycolic acid was 2.45X 10 7 The corresponding hydrophobic distribution value is 78.2%, and the contact angle of the thallus surface is 83.1 degrees; mycolic acid peak intensity in DBT-mock oil is 5.27X 10 7 The corresponding hydrophobic distribution value is 100%, and the contact angle of the thallus surface is 115.0 degrees; in diesel fuel, the peak intensity of mycolic acid is 5.58X 10 7 The corresponding hydrophobicity distribution value is 100%, and the cell surface contact angle is 127.7 degrees Gordonia sp.JT-2, so that the cell surface shows extremely high hydrophobicity in an oil-water multiphase system.
The above results show that: the mycolic acid produced on the cell wall surface of Gordonia sp.JT-2 makes it more hydrophobic on the surface, and the bacterium is a highly hydrophobic bacterium. Hydrophobicity is related to the production of mycolic acid with 58 total carbon atoms on the cell wall surface of Gordonia sp. The surface hydrophobicity of Gordonia sp.jt-2 is positively correlated with the amount of mycolic acid produced. In the process of biological desulfurization of oil products, the mass transfer efficiency of hydrophobic sulfur-containing organic matters in an oil phase from oil to cells is one of the key factors influencing the biological desulfurization efficiency. For hydrophilic bacteria, the sulfur-containing organic mass transfer process comprises: (1) transfer of organic sulfur from the oil phase to the aqueous phase; (2) transfer of organic sulfur from the aqueous phase into the cells; among them, the process (1) is very slow due to the very low solubility of sulfur-containing organic compounds in water, and thus becomes a key factor limiting the biological desulfurization efficiency. However, for hydrophobic bacteria, due to the extremely high hydrophobicity of the surface, cells can adhere to an oil-water interface to form an oil-water-cell emulsion layer in the culture process, and the cells are in direct contact with oil. On the other hand, the hydroxylation desulfurization product with biotoxicity is easy to dissolve in oil, and after biological desulfurization is completed by bacteria, the hydroxylation desulfurization product with a complete carbon ring structure returns to the oil, so that the loss of the calorific value of the oil product is reduced, and the toxic effect on the bacteria is reduced. In the invention, the characteristic of extremely high hydrophobicity of Gordonia sp.JT-2 in an oil-water multiphase system is utilized, the problem of low mass transfer efficiency of oil-water of sulfur-containing organic matters can be solved, the toxic action of hydroxylated desulfurization products on bacteria is reduced, and the desulfurization efficiency is improved.
EXAMPLE 3 Effect of hydroxylated desulfurization products on Gordonia sp.JT-2 growth and desulfurization Effect in different systems
The hydroxylated desulfurization products can generate inhibition (toxic action) on the desulfurization activity and growth of the desulfurization bacteria in the biological desulfurization process. Biological desulfurization is respectively carried out in a water phase system and an oil-water multiphase system, and the influence of a hydroxylated desulfurization product on the biological desulfurization efficiency of Gordonia sp.JT-2 in different systems is researched.
An aqueous phase system: DBT is added into a sulfur-free culture medium to serve as a unique sulfur source (the concentration of DBT in the culture medium is 0.543mmol/L), and hydroxylated desulfurization products (2-hydroxybiphenyl) with different concentrations are respectively added to carry out biological desulfurization experiments;
oil-water multiphase system: DBT-simulated oil (DBT is dissolved in n-tetradecane, the sulfur content of the simulated oil is 173.76ppm) is added into a sulfur-free culture medium at an oil-water ratio of 1:10(v/v), the concentration of the DBT in the culture medium is 0.543mmol/L, and the DBT is used as a unique sulfur source and is added with hydroxylated desulfurization products with different concentrations respectively to carry out biological desulfurization experiments.
As shown in FIG. 6, in the oil-water multiphase system, due to the high surface hydrophobicity (hydrophobicity distribution value of 100%, surface contact angle of 115.0 ℃) of Gordonia sp.JT-2, the bacteria were suspended at the interface of the oil-water multiphase system, and the hydroxylated desulfurization product was dissolved in the upper oil phase, thereby reducing the adverse effect on the desulfurization bacteria, and it can be seen from FIG. 6 that Gordonia sp.JT-2 was greatly reduced by the inhibition (poisoning effect) of the hydroxylated desulfurization product in the oil-water multiphase system, and the influence on the concentration of the hydroxylated desulfurization product was very small regardless of the amount of bacterial growth or the desulfurization rate.
Example 4 Gordonia sp.JT-2 desulfurization performance on real diesel
Real diesel oil with initial sulfur content of 167.7ppm is taken as a desulfurization object, and the desulfurization performance of Gordonia sp.JT-2 on the real diesel oil is researched. The specific operation is as follows: 50mL of mineral salts medium was added to a 150mL conical flask, and 5mL of real diesel was added as the sole sulfur source, as 4% (v/v)Inoculating Gordonia sp.JT-2 bacterial liquid (the bacterial concentration is 1.6 multiplied by 10) 8 CFU/mL), at 30 ℃, 160rpm in a constant temperature shaker for 5 days, and determining the diesel sulfur type and total sulfur content before and after desulfurization. The sulfur-containing organic species in diesel oil were analyzed by a gas chromatograph (GC-SCD, model 7890A-SCD) equipped with a sulfur chemiluminescence detector (SCD model: 355), and the results are shown in FIG. 7. In the biological desulfurization process, because of the high surface hydrophobicity of the desulfurization bacteria, the bacteria are adsorbed in an oil-water interface, and as analyzed above, the problem of low oil-water mass transfer efficiency of sulfur-containing organic matters can be solved by utilizing the characteristic of the extremely high hydrophobicity of Gordonia sp.JT-2 in an oil-water multiphase system. After 5 days of biological desulfurization, the strength of each main peak in the diesel oil is obviously reduced, which shows that Gordonia sp.JT-2 can effectively desulfurize the diesel oil, the residual sulfur content of the diesel oil after biological desulfurization is 19.7ppm, the desulfurization rate is 88.3 percent, and the benzothiophene derivative (C) with the substituent carbon chain length of 4 is used as the derivative 4 -BT), Dibenzothiophene (DBT), dibenzothiophene derivative (C) with substituent carbon chain length of 1 1 -DBT), dibenzothiophene derivative with substituent carbon chain length of 2 (C) 2 DBT) sulfur is completely removed, and the desulfurization rate of Gordonia sp.JT-2 on the sulfur-containing organic matters is 100 percent; the sulfur component remaining in the diesel after biological desulfurization is mainly a part of benzothiophene derivative and a part of C 3 -DBT. GC-SCD spectrograms before and after diesel oil desulfurization show that Gordonia sp.JT-2 has desulfurization capability on all sulfur-containing organic matters in diesel oil and has potential of being applied to desulfurization of actual diesel oil.
TABLE 3 comparison of desulfurization capacities of different desulfurization bacteria for diesel oil
The foregoing is directed to preferred embodiments of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.
Sequence listing
<110> Shandong university of science and technology
<120> Gordonia capable of producing long-carbon-chain mycolic acid and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1345
<212> DNA
<213> Gordonia sp
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gaccccaatc cgaactgaga ctggctttaa gggattcgct ccacctcacg gtatcgcagc 180
cctctgtacc agccattgta gcatgtgtga agccctggac ataaggggca tgatgacttg 240
acgtcatccc caccttcctc cgagttgacc ccggcagtct cctgcaagtc cccggcataa 300
cccgctggca atacaggaca agggttgcgc tcgttgcggg acttaaccca acatctcacg 360
acacgagctg acgacagcca tgcaccacct gtacaccaac cacaagggaa catgtatctc 420
tacatgcgtc tggtgtatgt caaacccagg taaggttctt cgcgttgcat cgaattaatc 480
cacatgctcc gccgcttgtg cgggcccccg tcaattcctt tgagttttag ccttgcggcc 540
gtactcccca ggcggggtac ttaatgcgtt agctacggca cggaactcgt gaaatgagcc 600
ccacacctag tacccaccgt ttacggcgtg gactaccagg gtatctaatc ctgttcgcta 660
cccacgcttt cgctcctcag cgtcagttac tacccagaga cccgccttcg ccaccggtgt 720
tcctcctgat atctgcgcat ttcaccgcta caccaggaat tccagtctcc cctgtagtac 780
tcaagtctgc ccgtatcgcc tgcacgccta caattgagtt gcagaatttc acagacgacg 840
cgacaaaccg cctacgagct ctttacgccc agtaattccg gacaacgctc gcaccctacg 900
tattaccgcg gctgctggca cgtagttggc cggtgcttct tctccaggta ccgtcacttg 960
cgcttcgtcc ctggtgaaag aggtttacaa cccgaaggcc gtcatccctc acgcggcgtc 1020
gctgcatcag gcttgcgccc attgtgcaat attccccact gctgcctccc gtaggagtct 1080
gggccgtgtc tcagtcccag tgtggccgat caccctctca ggtcggctac ccgtcgtcgc 1140
cttggtaggc cattacccca ccaacaagct gataggccgc gggcccatcc tgaaccgcaa 1200
aagctttcca ccccagagca tgcactccaa ggtcatatcc ggtattagac ccagtttccc 1260
aggcttatcc caaagttcag ggcagatcac ccacgtgtac tcacccgttc gccactcgag 1320
tacccagcaa gctggccttc cgtcg 1345
Claims (7)
1. The Gordoniae capable of producing the long-carbon-chain mycolic acid is Gordoniae (Gordonia sp.) JT-2 with the preservation number of CCTCC NO: m2021493.
2. Use of Gordonia bacteria according to claim 1 for the preparation of a biological desulphurating agent which is resistant to the poisoning effect of hydroxylated desulphuration products.
3. The biological desulfurizer which can resist the toxic action of the hydroxylated desulfurization product is characterized by comprising the following components in percentage by weight: m2021493 Gordoniella.
4. A biological desulfurization method with little influence of the toxicity of hydroxylated desulfurization products is characterized in that a biological desulfurization method with the preservation number of CCTCC NO: m2021493 Gordoniella strain.
5. The hydroxylated derivative of claim 4The biological desulfurization method with small toxic and harmful effects of sulfur products is characterized in that the bacterial concentration of Gordoniae bacterial liquid is more than or equal to 1.6 multiplied by 10 8 CFU/mL; the amount added was 4% (v/v).
6. The biological desulfurization method with less toxic effect of hydroxylated desulfurization products according to claim 4, characterized in that the desulfurization system is an oil-water multiphase system.
7. The biological desulfurization method with less toxic effect of hydroxylated desulfurization products according to claim 6, characterized in that the oil-water multiphase system has an oil-water ratio of 1:10 (v/v).
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| CN202110599142.4A CN113337425B (en) | 2021-05-31 | 2021-05-31 | Gordoniella for producing long-carbon-chain mycolic acid and application thereof |
| ZA2021/10400A ZA202110400B (en) | 2021-05-31 | 2021-12-14 | Gordonia bacterium producing long-carbon-chain mycolic acid and application thereof |
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| CN202110599142.4A CN113337425B (en) | 2021-05-31 | 2021-05-31 | Gordoniella for producing long-carbon-chain mycolic acid and application thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1445361A (en) * | 2002-03-19 | 2003-10-01 | 中国科学院过程工程研究所 | Gordona nitida and application of removing sulfur element from sulfur compound |
| CN1699547A (en) * | 2005-05-24 | 2005-11-23 | 南开大学 | Gordona terrae C-6 and its desulfurization effect |
| CN104651269A (en) * | 2015-01-06 | 2015-05-27 | 江南大学 | Desulphurizing bacterium for efficiently degrading DBT and application of desulphurizing bacterium in desulfurization |
-
2021
- 2021-05-31 CN CN202110599142.4A patent/CN113337425B/en active Active
- 2021-12-14 ZA ZA2021/10400A patent/ZA202110400B/en unknown
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1445361A (en) * | 2002-03-19 | 2003-10-01 | 中国科学院过程工程研究所 | Gordona nitida and application of removing sulfur element from sulfur compound |
| CN1699547A (en) * | 2005-05-24 | 2005-11-23 | 南开大学 | Gordona terrae C-6 and its desulfurization effect |
| CN104651269A (en) * | 2015-01-06 | 2015-05-27 | 江南大学 | Desulphurizing bacterium for efficiently degrading DBT and application of desulphurizing bacterium in desulfurization |
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| CN113337425A (en) | 2021-09-03 |
| ZA202110400B (en) | 2022-02-23 |
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