CN114350539B - Microbial strain for lignin degradation and application thereof - Google Patents
Microbial strain for lignin degradation and application thereof Download PDFInfo
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- CN114350539B CN114350539B CN202111443709.5A CN202111443709A CN114350539B CN 114350539 B CN114350539 B CN 114350539B CN 202111443709 A CN202111443709 A CN 202111443709A CN 114350539 B CN114350539 B CN 114350539B
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Abstract
The invention discloses a microbial strain for lignin degradation and application thereof, and belongs to the field of waste treatment by a microbiological method. The separated cucurbitacin monospore strain can adapt to strong acid and strong alkaline wastewater conditions generated in industrial production, can well grow and propagate, has a good lignin degradation effect, and can reduce the lignin content in liquid wastewater by 5-25%. Meanwhile, the method can remove lignin in the agricultural and forestry wastes, reduce the content of the lignin by 3-10%, has a very wide application prospect in treating refractory substances in the industrial and agricultural wastes, and is also beneficial to the protection and sustainable development of the ecological environment.
Description
Technical Field
The invention relates to a microbial strain for lignin degradation and application thereof, belonging to the field of waste treatment by a microbiological method.
Background
Lignin is a complex aromatic polymer, widely distributed in plants, and is the second largest biomass resource with the second reserves of cellulose in the plant kingdom. Lignin is one of the components of plant cell walls, has the functions of enhancing the mechanical strength of the cell walls and bonding fibers, is widely applied to various industrial productions, and according to statistics, more than 95 percent of lignin is applied to the paper industry. The waste water discharged by paper mill includes black liquor of paper making, middle section water and white water, the pollutant in the black liquor accounts for more than 90% of total pollutant discharge amount, and the components are complex, belonging to 221-002-35 wastes in International hazardous wastes book (2021 edition). The strongly alkaline papermaking black liquor contains a large amount of lignin, the degradation is difficult, common treatment methods are an inorganic acid precipitation method, a flocculation precipitation method, an alkali recovery method and the like, however, the processes all need to add extra chemical reagents in the running process, the treatment cost is increased, the recovery efficiency is not high, and secondary pollution is easily caused. In addition, with the development of society, the artificial board industry in China is developed vigorously and becomes one of the forestry strut industries. However, a large amount of wastewater with complex components is generated in the production process of the artificial board, the wastewater is acidic, contains a large amount of organic matters which are difficult to degrade, such as lignin, and the like, and is treated by a flocculation method and the like, so that the effect is poor. Therefore, the biological method for treating lignin in dangerous wastes such as black liquor in paper making is an environment-friendly technology. However, in the field of biological treatment of lignin-containing waste liquid, a method capable of effectively degrading lignin is still lacking.
Disclosure of Invention
In view of the existing problems, the invention screens a strain capable of effectively degrading microorganisms from soil in a bamboo leaf growing area, and the strain can adapt to a large range of pH and different treatment scenes.
The technical scheme of the invention is as follows:
the invention provides a strain of Sphingomonas citrullini (Sphingomonas melonis), which is preserved in China general microbiological culture Collection center (CGMCC) at 18 days 5 and 18 months 2021, wherein the preservation number is CGMCC No.22550, and the preservation address is No. 3 of the national institute of sciences, institute of microbiology, north Cheng West Lu No. 1 of the sunward area in Beijing.
The invention provides a product containing the sphingomonas cucurbitae of claim 1.
In one embodiment, the product comprises a sewage treatment agent, a microbial agent, or an enzyme preparation.
In one embodiment, the microbial agent comprises other strains capable of degrading lignin, including but not limited to serratia marcescens, salvia pacifica, helminthosporium griseum, pantoea agglomerans, white rot fungi, basidiomycetes, rabdosia baileyi, ceriporiopsis carbonaria, and/or acinetobacter calcoaceticus.
The invention provides a method for culturing the sphingomonas cucurbitacearum, which comprises the steps of inoculating the sphingomonas cucurbitacearum into a culture medium according to the inoculation amount of 0.1-1% of the volume ratio, culturing for 10-48 h in a constant temperature environment of 30-70 ℃ at 100-200 r/min, and obtaining bacterial liquid OD 600 Reaching 0.8 to 1.2.
The invention provides a method for degrading lignin, which is characterized in that the sphingomonas cucurbitae or the product containing the lignin is added into a system containing the lignin for reaction.
In one embodiment, the sphingomonas cucurbitae is cultured to OD when the lignin-containing system is in a liquid state 600 =1.0 + -0.2, or adjusting the product containing sphingomonas citrullinis to sphingomonas citrullinis OD 600 The bacterial liquid of which the volume is 10-70% of that of the bacterial liquid is added into a lignin-containing liquid system, and the reaction is carried out at the temperature of 30-50 ℃ and the rpm of 100-150.
Preferably, the obtained bacterial liquid is added into a liquid system containing lignin according to 50-70% or 30-50% of the volume of the system.
Preferably, the reaction is carried out at a temperature of from 30 ℃ to 40 ℃.
In one embodiment, the liquid lignin system comprises black liquor and artificial board production wastewater.
In one embodiment, the sphingomonas cucurbitacearum is cultured to OD when the lignin-containing system is in a solid state 600 =1.0 + -0.2, or adjusting the product containing sphingomonas citrullinis to sphingomonas citrullinis OD 600 A bacterial solution of =1.0 ± 0.2, which is added to a solid system containing lignin in an amount of 10 to 50mL/g, and reacted at 30 to 50 ℃ and 100 to 200rpm.
Preferably, the bacterial liquid is added to the solid system containing lignin in an amount of 30 to 50 mL/g.
Preferably, the reaction temperature is 30-40 ℃; the rotating speed is 150-200 rpm.
In one embodiment, the solid system containing lignin comprises bamboo leaves, bamboo branches, straw, rice hulls, wood or other lignin-containing substances.
In one embodiment, the reaction is carried out at a pH of 3 to 13 for 0 to 96 hours.
Preferably, the reaction is carried out at pH7 to 9 or pH 7.
The invention provides application of the citrulline sphingomonas or the product containing the citrulline sphingomonas in degrading lignin or lignin-containing substances.
In one embodiment, the lignin-containing material comprises black liquor, wood-based board production wastewater, bamboo leaves, bamboo branches, straw, rice hulls, and/or wood.
Has the beneficial effects that: the sphingomonas cucurbitacearum is separated from soil, can effectively degrade lignin and can normally grow in the pH range of 3-13, and therefore can adapt to treatment scenes of various pollutants. The lignin content in complex environments such as papermaking black liquor, artificial boards and the like can be reduced by 5-25%, and the lignin content in agriculture and forestry solid wastes such as bamboo leaves, bamboo branches and the like can be reduced by 3-10%.
Drawings
FIG. 1 is a colony morphology and a cell morphology of sphingomonas citrullina according to the present invention.
FIG. 2 is the rDNA electrophoretogram of the sphingomonas citrullinis strain of the present invention.
Biological material preservation
The screened Sphingomonas citrullini is classified and named Sphingomonas citrullini Sphingomonas melonis, is preserved in China general microbiological culture collection center (CGMCC) No.22550 in 2021, 5 months and 18 days, and is preserved in the national institute of microbiology No. 1, 3 of the rising area of Beijing, north Chen province, china academy of sciences.
Detailed Description
The present invention is described in detail in the following description in conjunction with the embodiments illustrated in the figures, wherein like numerals indicate like features throughout. While specific embodiments of the invention are shown in the drawings, it should be understood that the invention may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
It should be noted that certain terms are used throughout the description and following claims to refer to particular components. As one skilled in the art will appreciate, various names may be used to refer to a component. This specification and claims do not intend to distinguish between components that differ in name but not function. In the following description and in the claims, the terms "include" and "comprise" are used in an open-ended fashion, and thus should be interpreted to mean "include, but not limited to. The description which follows is a preferred embodiment of the invention, however, the description is given for the purpose of illustrating the general principles of the invention and not for the purpose of limiting the scope of the invention.
The enrichment culture refers to that spores germinate, grow and propagate a large amount of mycelia, thalli grow robustly to form 'seeds' with strong activity, and the concentration of the thalli in a culture solution reaches a higher level.
The soil used in the following examples was derived from a soil sample rich in dead branch and fallen leaf depth of 15cm in the bamboo forest soil of the Special bamboo breeding center in Changzhou city.
The lignin enrichment medium used in the following examples had the composition of 0.7g/L ammonium sulfate, 0.6g/L potassium nitrate, 0.4g/L potassium dihydrogen phosphate, 0.1g/L dipotassium hydrogen phosphate, 1g/L dealkalized lignin; to 1000mL of distilled water were added 0.7g of ammonium sulfate, 0.6g of potassium nitrate, 0.4g of potassium dihydrogenphosphate, 0.1g of dipotassium hydrogenphosphate, 1g of dealkalized lignin, sterilized at 121 ℃ for 20min, and then 2mL of filter sterilized vitamin and 1mL of trace element were added.
The lignin screening identification medium used in the following examples was Marine agar 2216 medium (mannitol salt agar), brand: BD Difco.
The components of the potato glucose culture medium used in the following examples are 200g/L potato, 20g glucose, 1.5g/L magnesium sulfate, 3g/L potassium dihydrogen phosphate, and 0.05g/L vitamin B1; specifically, 200g of potato, 20g of glucose, 1.5g of magnesium sulfate, 3g of potassium dihydrogen phosphate and 0.05g of vitamin B1 are added into 1000mL of distilled water and sterilized at 121 ℃ for 20min.
The activation of sphingomonas citrullinii described in the following examples refers to that sphingomonas citrullinii single colonies are cultured in a potato culture medium to enable the growth of the strain to the logarithmic growth phase, so that the strain has good growth performance and proliferation activity. The specific method comprises the steps of selecting the single colony inoculation amount of the sphingomonas citrullini, inoculating the single colony inoculation amount to 100mL of potato glucose culture medium, and activating at 30 ℃ and 150 r/min; or inoculating the bacterial liquid of sphingomonas cucurbitacearum with the inoculum size of 2 percent by volume into a potato glucose culture medium at the temperature of 30 ℃ and the speed of 150r/min for activation.
And (3) determination of lignin: the Klason method is used for determining the content of lignin, and the principle is that 72% sulfuric acid is used for removing cellulose, and insoluble lignin can be obtained through the steps of filtering, washing, drying, weighing, calculating and the like.
The lignin degradation rate was calculated as described in the following examples:
lignin degradation rate = lignin content after reaction/lignin content in the original material.
The black liquor used in the following examples is from a paper industry Co., ltd, and the black liquor mainly contains 20 to 30% (mass volume fraction) of lignin and has a pH of 12 to 13.
The artificial board used in the following examples is from Jiangsu Runji renewable resources technology limited, and the artificial board mainly contains 0.5-1 g/cm 3 The pH value of the lignin is 5-6.
The lignin content in the waste bamboo leaves used in the following examples was 25 to 50% (mass fraction).
Examples
The materials used in the tests and the test methods are generally and/or specifically described, and the reagents or instruments used are not indicated by manufacturers, but are all conventional reagent products commercially available.
Example 1: screening and identification of strains
1. Screening of strains
Weighing 4g of soil sample, placing the soil sample into a triangular flask containing 100mL of enrichment medium, placing the soil sample into glass beads, fully scattering and uniformly mixing, and culturing for 3d in a constant-temperature shaking incubator at 37 ℃ and 150r/min.
Diluting the cultured bacterial liquid with sterile water in gradient with dilution degree of 10 -5 ，10 -6 ，10 -7 100 mu L of the bacterial liquid is coated on a screening and identifying culture medium, each dilution is made into 3 parallels, and the obtained product is placed in a constant temperature incubator at 37 ℃ in an inverted mode for culture for 3d until obvious bacterial colonies grow out. And (3) selecting single colonies with obviously different colony characteristics, streaking the single colonies on a screening culture medium, placing the single colonies in a constant-temperature incubator at 37 ℃ for continuous culture, and repeating the step until pure bacteria are obtained.
2. Identification of strains
The separated bacterial strain of the invention has small bacterial colony on a lignin screening and identifying culture medium, is milk white, has smooth surface and regular edge, and the morphological characteristics of the bacterial strain are observed by using an optical microscope, and the cells are in short rod shape (as shown in figure 1).
Identifying the genus of the strain by 16s RNA from Biotechnology engineering (Shanghai) GmbH, extracting genomic DNA with SK9255 kit, performing PCR amplification with bacterial universal primer, and performing electrophoresis detection on the amplified band, wherein the electrophoresis chart is shown in FIG. 2.
(1) The primers used were:
7F：CAGAGTTTGATCCTGGCT，SEQ ID NO:1，
1540R：AGGAGGTGATCCAGCCGCA，SEQ ID NO:2。
(2) PCR amplification reaction System: 12.5 μ L of the mix (10 XPCR Buffer, dNTP, 62.5U Taq Plus DNA Polymerase, 50mM MgSO 4 ) 1. Mu.L of 10. Mu. Mol/L primer F, 1. Mu.L of μmol/L primer R, 1. Mu.L of Template (DNA), 9.5. Mu.L of ddH 2 O。
(3) And (3) PCR reaction conditions: respectively reacting at 95 ℃ for 5min,94 ℃ for 30s,57 ℃ for 30s,72 ℃ for 30s, circulating for 30 times, and reacting at 72 ℃ for 10min.
(4) Gel electrophoresis: electrophoresis was carried out on 1% agarose, 150V, 100mA, 20min.
The result is shown in FIG. 2, the size of the amplified band is 1403bp, and the amplified band is sent to a company for sequencing, the sequencing result is compared with that on NCBI, and after analysis, the bacterium is identified as Sphingomonas citrullinii (Sphingomonas melonis).
Example 2: optimization of culture conditions for sphingomonas citrullinis
(1) Effect of temperature on growth of sphingomonas cucurbitae
(1) Inoculating the screened sphingomonas citrullini with the inoculum size of 2 percent in volume ratio to 100mL of potato glucose culture medium, activating at 30 ℃ at 150r/min, inoculating the activated sphingomonas citrullini with the inoculum size of 0.2 percent in volume ratio to 100mL of fresh lignin-enriched culture medium, and enabling the OD of the activated sphingomonas citrullini in the culture medium 600 To 0.1;
(2) inoculating the bacterial liquid obtained in the step (1) into a fresh lignin enrichment medium in an inoculation amount of 0.2% by volume, and respectively inoculating the bacterial liquid into the lignin enrichment medium at different temperatures: culturing at 10 deg.C, 20 deg.C, 30 deg.C, 40 deg.C, 50 deg.C, 60 deg.C, 70 deg.C, 150r/min for 36 hr;
(3) determination of OD in media at different temperatures 600 。
As shown in Table 1, sphingomonas citrullini has a high OD at 30-50 ℃ 600 The optimum temperature is thus between 30 and 50 ℃.
TABLE 1 OD of sphingomonas citrullini at different temperatures 600
(2) Effect of rotational speed on growth of sphingomonas citrullinis
(1) Inoculating the screened sphingomonas citrullini with the inoculum size of 2 percent in volume ratio to 100mL of potato glucose culture medium, activating at the temperature of 30 ℃ at the speed of 150r/min, and inoculating the activated sphingomonas citrullini with the inoculum size of 0.2 percent in volume ratio to 100mL of fresh lignin enrichment culture mediumIn the medium, and making it OD in the medium 600 Up to 0.1;
(2) inoculating the bacterial liquid obtained in the step (1) into 100mL of fresh lignin-enriched culture medium in an inoculation amount of 0.2% by volume, and culturing at 0r/min, 50r/min, 100r/min, 150r/min, 200r/min and 30 ℃ for 36h respectively;
(3) determination of OD in media at different rotational speeds 600 。
As shown in Table 2, sphingomonas citrullini has a higher OD at a shaker rotation speed of 100r/min or more 600 Considering the cost factor, the optimum rotating speed is 150r/min.
TABLE 2 OD of sphingomonas cucurbitae at different rotation speeds 600
(3) Effect of pH on growth of sphingomonas citrullinis
(1) Inoculating the screened sphingomonas citrullini with the inoculum size of 2 percent in volume ratio to 100mL of potato glucose culture medium, activating at 30 ℃ at 150r/min, inoculating the activated sphingomonas citrullini with the inoculum size of 0.2 percent in volume ratio to 100mL of fresh lignin-enriched culture medium, and enabling the OD of the activated sphingomonas citrullini in the culture medium 600 Up to 0.1;
(2) preparing a fresh lignin enrichment medium, adjusting the pH value of the lignin enrichment medium to be 3, 5, 7, 9, 11 and 13 respectively, inoculating the bacterial liquid obtained in the step (1) into 100mL of the lignin enrichment medium with the adjusted pH value by an inoculum size of 0.2% of the volume ratio, and culturing at 30 ℃ and 150r/min for 36h;
(3) determination of OD in media at different pH 600 。
As shown in Table 3, OD was measured at pH7 to 9 for sphingomonas citrullini 600 The value is higher.
TABLE 3 OD of sphingomonas citrullini at different pH 600
Example 3: degradation effect of bacterial strain on lignin in papermaking black liquor
Inoculating activated sphingomonas citrullinis to 100mL of fresh lignin-enriched culture medium with the inoculum size of 2% by volume, and culturing in a shaking table at 30 ℃ and 150r/min until OD is reached 600 Up to 1.0. Then inoculating the bacterial liquid into papermaking black liquor with the temperature of 30 ℃ (the pH value is 12-13), reacting for a period of time at the temperature of 30 ℃ and at the speed of 150r/min, collecting the reaction liquid, measuring the content of lignin in the reaction liquid, and calculating the lignin degradation rate, wherein the result is shown in table 4.
TABLE 4 Lignin degradation Rate in papermaking Black liquor
Example 4: degradation effect of bacterial strain on lignin in artificial board production wastewater
Inoculating activated sphingomonas citrullinis to 100mL of fresh lignin-enriched culture medium with the inoculum size of 2% by volume, and culturing in a shaking table at 30 ℃ and 150r/min until OD is reached 600 Up to 1.0. Then inoculating the bacterial liquid into artificial board production wastewater (pH is 5-6) at 30 ℃, reacting for a period of time at 30 ℃ and 150r/min, collecting the reaction liquid, measuring the content of lignin in the reaction liquid, and calculating the lignin degradation rate, wherein the results are shown in Table 5.
TABLE 5 degradation rate of lignin in artificial board production wastewater
Example 5: degradation effect of bacterial strain on lignin in agricultural and forestry waste
Inoculating the activated sphingomonas cucurbitacearum to 100mL of fresh lignin enrichment medium by the inoculum size of 2 percent of the volume ratio, and culturing the lignin enrichment medium in a shaker at 30 ℃ and 150r/min to OD 600 Up to 1.0. Then, the bacterial solution and 1g of waste bamboo leaves were mixed uniformly, reacted at 30 ℃ for a period of time at 150r/min, and the reaction solution was collected at 24h, 48h, and 72h of the reaction, and the content of lignin in the reaction solution was measured, and the lignin degradation rate was calculated, and the results are shown in table 6.
TABLE 6 Lignin degradation Rate in waste bamboo leaves
Example 6: application of sphingomonas cucurbitae in treating lignin-containing substances
Adding the sphingomonas cucurbitacearum into a system containing lignin, and reacting for 0-96 h at the pH of 3-13, the temperature of 30 ℃ and the speed of 150r/min to finish the degradation of the lignin.
Example 7: preparation of microbial agent containing sphingomonas cucurbitacearum
(1) Preparation of single bacterium agent
Inoculating 400 μ L of sphingomonas citrullinii liquid into 20mL of potato glucose culture medium, activating at 30 deg.C for 2-3 generations until sphingomonas citrullini reaches 10 8 When the viable count is more than cfu/mL, centrifuging for 10-20 min at 5000-10000 rpm, removing supernatant, sequentially adding buffer solution and freeze-drying protective agent in an aseptic environment until the cell concentration is not less than 10 6 And (5) performing vacuum freeze drying treatment when cfu/mL is reached to obtain the solid microbial inoculum.
(2) Preparation of compound microbial agent
Mixing the sphingomonas citrullini with other strains capable of degrading lignin and culturing to obtain a mixed bacterial liquid, or respectively culturing the strains and then mixing to obtain the mixed bacterial liquid, and freeze-drying the mixed bacterial liquid to obtain the microbial preparation.
The strain capable of degrading lignin comprises serratia marcescens, salvia pacifica, helminthosporium griseum, pantoea agglomerans, white rot fungi, wood rot basidiomycetes, white capsule rabdosia, ceriporiopsis carbowax, acinetobacter calcoaceticus and other strains capable of degrading lignin.
Example 8: preparation of products containing sphingomonas citrullinis
Adding the solid microbial inoculum obtained in the embodiment 6 into a product capable of degrading lignin to prepare a product containing the sphingomonas citrullini; the product comprises sewage treatment agent, enzyme preparation and other preparations capable of being mixed with microorganism.
Example 9: application of microbial agent in treatment of papermaking black liquor
Adding microbial agent containing sphingomonas cucurbitacearum or product containing sphingomonas cucurbitacearum into potato glucose culture medium for activation, inoculating into 100mL fresh lignin enrichment culture medium by inoculum size of 2% in volume ratio after activation, and culturing in a shaker at 30 ℃ and 150r/min to OD 600 To 1.0; inoculating the bacterial liquid into papermaking black liquor (pH is 12-13) at 30 ℃, and reacting for 0-96 h at 30 ℃ and 150r/min.
Example 10: application of microbial agent in treatment of artificial board production wastewater
Adding a microbial agent containing citrulline monad or a product containing citrulline monad into a potato glucose culture medium for activation, and inoculating the activated product into 100mL of fresh lignin enrichment culture medium by an inoculation amount of 2% by volume; inoculating the bacterial liquid into artificial board production wastewater (pH is 5-6) at 30 ℃, and reacting for 0-96 h at 30 ℃ and 150r/min.
Example 11: application of microbial agent in forest waste treatment
Adding a microbial agent containing citrulline monad or a product containing citrulline monad into a potato glucose culture medium for activation, and inoculating the activated product into 100mL of fresh lignin enrichment culture medium by an inoculation amount of 2% by volume; the bacterial liquid and 1g of waste bamboo leaves are mixed evenly and reacted for 0 to 96 hours at 30 ℃ and 150r/min.
Example 12: application of product containing sphingomonas citrullinis in treatment of lignin-containing substance
Adding the product containing sphingomonas citrullini into a system containing lignin, and reacting for 0-96 h at the pH of 3-13, the temperature of 30 ℃ and the speed of 150r/min.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
SEQUENCE LISTING
<110> Jiangsu institute of technology and industry
<120> microbial strain for lignin degradation and application thereof
<130> GBAA211589A
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 18
<212> DNA
<213> Artificial sequence
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cagagtttga tcctggct 18
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<212> DNA
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aggaggtgat ccagccgca 19
Claims (12)
1. Sphingomonas citrullina (Sphingomonas melonis) It has been preserved in China general microbiological culture Collection center (CGMCC) at 18.5.2021 with the preservation number of CGMCC No.22550.
2. A product comprising the sphingomonas cucurbitae of claim 1.
3. The product of claim 2, wherein the product comprises a sewage treatment agent, a microbial agent, or an enzyme preparation.
4. The product according to claim 3, wherein the microbial agent comprises other lignin-degrading strains including but not limited to Serratia marcescens, salvia pacifica, helminthosporus griseus, pantoea agglomerans, white rot fungi, basidiomycetes xylosus, irpex alba, ceriporiopsis carbonarius and/or Acinetobacter calcoaceticus.
5. A method for degrading lignin, characterized in that sphingomonas citrullini according to claim 1, or the product according to claim 2 or 3 is added to a system containing lignin and reacted.
6. The method according to claim 5, wherein the sphingomonas citrullini is cultured to OD when the lignin-containing system is in a liquid state 600 =1.0 + -0.2, or adjusting the product containing sphingomonas citrullinis to sphingomonas citrullinis OD 600 The bacterial liquid of which the volume is 10-70% of that of the bacterial liquid is added into a lignin-containing liquid system, and the reaction is carried out at the temperature of 30-50 ℃ and the rpm of 100-150.
7. The method according to claim 6, wherein the liquid lignin system comprises black liquor and artificial board production wastewater.
8. The method of claim 5, wherein the sphingomonas citrullinis cultured to OD when the lignin-containing system is in a solid state 600 ＝1.0±0.2，Or regulating product containing sphingomonas citrullinii to sphingomonas citrullinii OD 600 A bacterial solution of =1.0 ± 0.2, which is added to a solid system containing lignin in an amount of 10 to 50mL/g, and reacted at 30 to 50 ℃ and 100 to 150 rpm.
9. The method of claim 8, wherein the solid system containing lignin comprises bamboo leaves, bamboo branches, straw, rice hulls, wood or other lignin-containing substances.
10. The process according to any one of claims 5 to 9, wherein the reaction is carried out at a pH of 3 to 13 for 24 to 96 hours.
11. Use of a sphingomonas citrullina according to claim 1 or a product according to any one of claims 2 to 4 for degrading lignin or lignin-containing materials.
12. The use according to claim 11, wherein the lignin-containing substance comprises black liquor from paper making, waste water from the production of artificial boards, bamboo leaves, bamboo branches, straw, rice hulls and/or wood.
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