CN106978367B

CN106978367B - Bacillus ureafaciens strain TB42 and application thereof in promoting compost maturity

Info

Publication number: CN106978367B
Application number: CN201710184428.XA
Authority: CN
Inventors: 李群良; 徐佳琦; 陆彦宇
Original assignee: Guangxi University
Current assignee: Guangxi University
Priority date: 2017-03-24
Filing date: 2017-03-24
Publication date: 2020-07-28
Anticipated expiration: 2037-03-24
Also published as: CN106978367A

Abstract

The invention belongs to the technical field of development of plant growth-promoting bacteria, and particularly relates to a bacillus urealyticum strain TB42 and application thereof in promotion of compost maturity. A Bacillus ureafaciens strain TB42 is classified and named as Bacillus ureafaciens TB42(Ureibacillus subswenensis), and is preserved in China general microbiological culture Collection center (CGMCC) at 2017, 01-05 and with the preservation number of CGMCC 13529. The ureaplasma urealyticum strain TB42 is obtained by screening and separating a compost high-temperature period sample for the first time, is high-temperature resistant, can increase and reproduce quickly, and can produce lignocellulose degrading enzyme; the microbial inoculum containing the ureaplasma urealyticum strain TB42 can increase the composting temperature, accelerate the degradation of organic matters and water-soluble organic matters (DOM), increase the content of Kjeldahl nitrogen and promote compost maturity; also has the function of generating biosurfactant, so that the strain can be used for preparing the biosurfactant.

Description

Bacillus ureafaciens strain TB42 and application thereof in promoting compost maturity

Technical Field

The invention belongs to the technical field of development of plant growth-promoting bacteria, and particularly relates to a bacillus urealyticum strain TB42 and application thereof in promotion of compost maturity.

Background

The fertilizer plays a great role in modern agricultural production. But with the increase of the using amount of the fertilizer, the utilization rate of the fertilizer is reduced year by year. Pesticide and chemical fertilizer residues release a large amount of harmful substances to the environment, pollute soil, water sources and food, pose great threat to human health and living environment, and increasingly strong call for protecting ecological environment and producing safe food is required. Therefore, it has been an urgent necessity to develop a new fertilizer source using an alternative chemical fertilizer to meet the need for developing green agriculture and green food. The microbial fertilizer can make ineffective nutrition in soil effective, prevent and control crop diseases, reduce the use of pesticides and chemical fertilizers, is a fundamental way for solving the pollution of soil, water sources and foods, and is generally considered to be an environment-friendly, economic and effective method for improving the crop yield.

The microbial fertilizer is a specific product containing a microbial living body, is applied to agricultural production and can obtain a specific fertilizer effect. Wherein living microorganisms in the preparation play a critical role. Currently, microbial fertilizer products are generally divided into two main categories: one is a microbial fertilizer in the narrow sense, which means that the supply of plant nutrient elements, including the total supply of plant nutrient elements in soil and production environment, is increased by the life activities of microorganisms, resulting in improvement of the plant nutrient status and further increase of yield. Representative of this class of microbial fertilizers are rhizobia fertilizers; the other is a broad-sense microbial fertilizer, which means that the supply of plant nutrient elements can be increased through the life activities of microorganisms in the fertilizer, plant growth hormone can be generated, the absorption and utilization of the nutrient elements by plants are promoted or the pathogenic effect of certain pathogenic microorganisms is antagonized, the introduction of plant diseases and insect pests is reduced, and the crop yield is increased. Compared with chemical fertilizers, the microbial fertilizer has the following advantages: the soil structure is not damaged; the ecological environment is protected, the environment is not polluted, and the fertilizer is non-toxic and harmless to people and livestock; the fertilizer efficiency is durable; the yield of the crops is improved, and the quality of the crops is improved; low cost, economy and effectiveness.

Plant growth-promoting rhizobacteria (PGPR) is a kind of microorganism capable of high-density colonizing in Plant rhizosphere, and has the functions of inhibiting Plant pathogenic bacteria and harmful microorganism in rhizosphere, promoting Plant growth and increasing crop yield. The research and application of PGPR has played a very important role as an important resource pool for biofertilizers and biopesticides. The research and development of the microbial fertilizer from the aspect of resource recycling are of more realistic significance to the comprehensive utilization of resources and the environmental protection.

Bacillus belongs to the family Bacillaceae, the genus Bacillus, gram-positive bacilli capable of forming endospore-spores, aerobic or facultative aerobic, with flagella. The bacillus is widely distributed in soil, air, water and animal intestinal tracts, has strong stability, and once spores are formed, the bacillus can resist various adverse conditions, such as dry heat (the spores still survive for 1 hour at 150 ℃), moist heat, ultraviolet rays, strong acid, strong base, organic solvent, extreme dryness, vacuum drying, oxidation of oxidant and the like. Under certain conditions, the spores can be stored for a long time, and the effective period of the probiotic product can be ensured. Bacillus is the most desirable microbial additive in all genera.

A large number of tests prove that the bacillus additive can obviously improve the flora composition in organisms and has the effect of promoting growth, and the bacillus has higher stability in the processing process of granules and powder and in an acid environment, can play a role in proliferation in the environment in intestinal tracts and accords with the conditions of microbial additives. The bacillus has high stability in the acidic environment of the intestinal tract, can secrete protease and amylase with stronger activity, and promotes the digestion of nutrient substances of the feed. Bacillus can reduce the number of Escherichia coli in feces and digestive tract.

Currently, the Bacillus strains mainly used include Bacillus subtilis, Bacillus licheniformis, Bacillus cereus, Bacillus toyoi and the like, and research on Bacillus urealyticum is rarely reported, for example, the invention patent of application No. 201210222021.9, namely Bacillus thermophilus, microbial inoculum and application thereof, discloses a Bacillus ureophilus JD-50 with the preservation number of CGMCC No.5818, and discloses application thereof in oil exploitation.

The information disclosed in this background section is only for enhancement of understanding of the general background of the invention and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.

Disclosure of Invention

The invention aims to provide a Bacillus urealyticum strain TB42 which can be applied to preparation of a surfactant and promotion of compost maturity.

The purpose of the invention is realized by the following technical scheme:

the invention discloses a Bacillus ureafaciens strain TB42 which is classified and named as Bacillus ureafaciens TB42(Ureibacillus suwonensis) and is preserved in China general microbiological culture Collection center (CGMCC) at 2017, 01-05, and the preservation number is CGMCC 13529.

The invention also provides application of the Bacillus ureafaciens strain TB42 in degradation of lignocellulose.

The invention also provides application of the Bacillus ureafaciens strain TB42 in preparing a surfactant.

The invention also provides application of the Bacillus ureafaciens strain TB42 in preparation of laccase, lignin peroxidase, manganese peroxidase, cellulase and hemicellulase.

The invention also provides a microbial inoculum, which is prepared by inoculating the seed solution of the ureabacterium strain TB42 disclosed by the claim 1 into L B culture solution after 1% (v/v) of sterilization of 1L, culturing for 24h in a 120rmp shaking table at 50 ℃, and collecting the culture solution.

The invention also provides application of the microbial inoculum in promoting compost maturity.

Compared with the prior art, the invention has the following beneficial technical effects:

1. the ureaplasma urealyticum strain TB42 provided by the invention is obtained by screening and separating a compost high-temperature period sample for the first time, is high-temperature resistant, grows and breeds quickly, and can produce lignocellulose degrading enzyme.

2. The microbial inoculum containing the Bacillus ureafaciens strain TB42 provided by the invention can increase the composting temperature, accelerate the degradation of organic matters and DOM, increase the content of Kjeldahl nitrogen and promote the compost to be thoroughly decomposed.

3. The Bacillus ureafaciens strain TB42 also has the function of producing a biosurfactant, so that the strain can be used for preparing the biosurfactant.

Description of preservation information

The bacillus ureafaciens TB42(Ureibacillus suwonensis) has the preservation number of CGMCC 13529, the preservation date of 2017, 01 and 05 days, the preservation unit is the common microorganism center of China Committee for culture Collection of microorganisms, and the preservation address is No. 3 of the Beijing university Hokkaido No. 1 of North Cheng of the sunward area, and the institute of microorganisms of the China academy of sciences.

Drawings

FIG. 1 is a graph of temperature versus composting time. RT: room temperature, blank group (CK), Experimental group (T)

FIG. 2 is a graph of pH as a function of composting time. Blank group (CK), Experimental group (T)

FIG. 3 is a graph of moisture as a function of composting time. Blank group (CK), Experimental group (T)

Figure 4 is a plot of organic matter content as a function of composting time. Blank group (CK), Experimental group (T)

FIG. 5 is a plot of Kjeldahl nitrogen content as a function of composting time. Blank group (CK), Experimental group (T)

FIG. 6 is a graph showing the variation of ammonia nitrogen content with composting time. Blank group (CK), Experimental group (T)

FIG. 7 is a curve of nitrate nitrogen content as a function of composting time. Blank group (CK), Experimental group (T)

FIG. 8 is a graph of DOM content versus composting time. Blank group (CK), Experimental group (T)

FIG. 9 is a photograph of a colony of Ureibacillus subwonensis TB 42.

FIG. 10 is a representation of Ureibacillus subwonensis TB42 on blood agar plates.

FIG. 11 is a glucose standard curve.

FIG. 12 is a xylose standard curve.

FIG. 13 is a gel electrophoresis pattern of the selected strains;

description of the reference numerals:

M-2000bpMake；1-TB21；2-TB22；3-TB23；4-TB24；5-TB25；6-TB41；7-TB42；8-TB43；9-TB44；10-TB45；11-TB61；12-TB62；13-TB63；14-TB64；15-TB65；16-TA65；17-TB46。

Detailed Description

The present invention will now be described in further detail with reference to specific examples, which are intended to be illustrative, but not limiting, of the invention.

The materials and reagents used in the following examples are commercially available, unless otherwise specified. The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Example 1: screening of Bacillus ureafaciens TB42(Ureibacillus suwonensis)

1. Experimental materials and apparatus

1.1 Primary reagents

TABLE 1 Main reagents

Name of medicine	Molecular formula	Specification of
			ABTS	C₁₈H₂₄N₆O₆S₄	AR
Hydrogen peroxide	H₂O₂	AR
			Citric acid	C₆H₈O₇	AR
Citric acid sodium salt	Na₃C₆H₅O₇	AR
			Glacial acetic acid	CH₃COOH	AR
Anhydrous sodium acetate	CH₃COONa	AR
			Xylose	C₅H₁₀O₅	AR
Xylan	(C₅H₈O₄)_n	AR
			Tartaric acid sodium salt	C₄H₅Na₂O₆	AR
Tartaric acid	C₄H₆O₆	AR
			Veratryl alcohol	C₈H₁₂O₃	AR
Lactic acid	C₃H₆O₃	AR
			Sodium lactate	C₃H₅O₃Na	AR
Manganese sulfate	MnSO₄	AR
			Sodium hydroxide	NaOH	AR
Hydrochloric acid	HCl	AR
			DNS reagent

1.2 Main instruments and devices

TABLE 2 Main instruments and Equipment

1.3 Medium

The bacteria culture medium comprises 33g of nutrient agar and 1000m of deionized water L;

l B culture solution, 10g peptone, 5g yeast powder, 10g sodium chloride and 1000m L deionized water.

2. Separation and screening of lignin-degrading bacteria

2.1 sampling

The screened samples are compost samples of 2 days, 4 days and 6 days respectively, the compost samples are sampled at multiple points (generally 10-20cm below the surface of the compost) at a higher temperature of the compost, and the samples are uniformly mixed and then put into a sample bag to be stored in a refrigerator at the temperature of-4 ℃.

2.2 screening and purification of thermophilic bacteria

Weighing 10g of sample, adding into a triangular flask containing 90m L sterilized NaCl solution (0.9% w/v), placing on a shaking table, oscillating for 30min to break loose zoogloea and disperse bacteria in the solution in a single cell state, adding sterilized NaCl solution into 7 test tubes (sterilized in the same way), adding 9m L into each test tube, adding vibrated bacteria solution 1m L into the test tubes, respectively, namely diluting to 10^-1、10^-2、10^-3、10^-4、10^-5、10^-6、10^-7Then 0.1m L of the bacterial liquid at each dilution is coated on a bacterial culture medium plate and cultured at a constant temperature of 50 ℃, each plate with dilution gradient is parallel to 5 plates, and the bacterial colony can be observed to fall at 10^-5The growth vigor is better under the dilution degree. Larger colonies were selected in five parallel plates for isolation and streaking for multiple purifications.

As shown in FIGS. 9-10, the colony morphology of Bacillus ureafaciens TB42 is irregular, the colony is large, the surface is smooth and wet, translucent and milky white, and the edges of the colony are irregular.

3. Preparation of crude enzyme solution

L B culture solution is prepared, sterilized, poured into a serum bottle (25m L) sterilized in the same way, the strain obtained by separation is inoculated into the serum bottle, the serum bottle is put at 50 ℃ and 120rmp shake culture is carried out to prepare seed solution L B culture solution, 100m L is poured into a conical bottle, sterilized, the 1m L seed solution is absorbed into the conical bottle, the conical bottle is also put at 50 ℃ and 120rmp shake culture is carried out to prepare bacterial solution.

Liquid enzyme production culture, centrifuging the bacterial solution at 5000r/mim for 5min, collecting supernatant as crude enzyme solution, and using heat inactivated enzyme solution (0.6 m L crude enzyme solution in centrifuge tube, boiling for 10min) as control.

4. Method for measuring enzyme activity

4.1 determination of laccase (L ac) Activity

1. Reagent

(1) Determined by the rate at which the enzyme oxidizes ABTS;

(2)0.1mo L/L citric acid-sodium citrate buffer, which is prepared by weighing 2.941g of sodium citrate and 2.1014g of citric acid respectively, adding deionized water 100m L, adding citric acid into sodium citrate, and adjusting the pH value to 5.0 by using a pH meter;

(3)0.5mL 0.5moL/L ABTS；

2. procedure for the preparation of the

(1) At 25 ℃, 2m L citric acid-sodium citrate buffer (0.1mmo L/L, pH 5), and 0.5m L ABTS were added to the tube, and the mixture was poured into a cuvette;

(2) then, 1m L enzyme solution was added to start the reaction, and the change in absorbance was measured at 420 nm.

3. Calculation method

The enzyme activity is defined as that ABTS which oxidizes 1 mu mo L per minute is an enzyme activity unit,

extinction coefficient 3.6 x 10⁴[(moL/L)^-1cm^-1]Enzyme activity unit U/L

Laccase (L ac) activity Δ OD 10⁶/ΔT*V_{Enzyme solution}*∈

＝(ΔD/ΔT)*10⁶/0.5*3.6*10⁴

In the formula: Δ OD-the value at which the difference between adjacent absorbance values measured is greatest; the time difference value corresponding to the delta T-delta OD value;

v-volume of enzyme solution.

4.2 measurement of Lignin peroxidase (L ip) Activity

1. Reagent

(1)0.24mo L/L sodium tartrate buffer solution, which is prepared by respectively weighing 3.60216g of tartaric acid and 5.52192g of sodium tartrate, adding deionized water 100m L, adding tartaric acid into the sodium tartrate, and adjusting the pH to 3.0;

(2)0.1m L24 mmo L/L veratryl alcohol;

(3)0.05m L6.0.6.0 mm o L/L H₂O₂；

2. Procedure for the preparation of the

(1) At 37 ℃, the total reaction system was 3m L, 1.85m L0.24 mo L/L sodium tartrate buffer (pH 3.0), and 0.1m L24 mmo L/L veratryl alcohol and 1.0m L enzyme solution were added to the tube;

(2) preheating to 37 deg.C, pouring the solution into a cuvette, and adding 0.05m L6.0.0 mm H L/L₂O₂The reaction was started and the change in absorbance was measured at a wavelength of 310 nm.

3. Calculation method

The enzyme activity is defined as that veratryl alcohol which oxidizes 1 mu mo L per minute is an enzyme activity unit,

extinction coefficient 9.3 x 10³[(moL/L)^-1cm^-1]Enzyme activity unit U/L

Peroxidase (L ip) activity Δ OD 10⁶/ΔT*V_{Enzyme solution}*∈

＝(ΔOD/ΔT)*10⁶/1*9.3*10³

v-volume of enzyme solution.

4.3 determination of manganese peroxidase (Mnp) Activity

1. Reagent

(1)0.11mo L/L sodium lactate buffer, weighing 1.10098g lactic acid and 2.05516g sodium lactate, adding deionized water 100m L respectively, adding lactic acid into sodium lactate, and adjusting the pH to 4.5;

(2)0.025M l40 mmo L/L manganese sulfate;

(3)0.025m L1.6 mm o L/L H₂O₂；

2. Procedure for the preparation of the

(1) According to the enzyme in H₂O₂In the presence of Mn²⁺Oxidized Mn³⁺Is determined by the speed of the motor;

(2) the total reaction volume was 1m L, 0.85m L0.11.11 mo L/L of sodium lactate buffer (pH 4.5) was added to the tube, and 0.025m L was added,40mmoL/L MnSO₄And 1m L of enzyme solution;

(3) after preheating at 37 ℃ the solution was poured into a cuvette to which 0.025m L1.6.6 mm H L/L was added₂O₂The reaction was started and the change in absorbance was measured at 240 nm.

3. Calculation method

Enzyme activity is defined as the oxidation of 1. mu. mo L Mn per minute²⁺Is Mn³⁺Is an enzyme activity unit, and is characterized in that,

extinction coefficient 6.5 x 10³[(moL/L)^-1cm^-1]Enzyme activity unit U/L

Manganese peroxidase (Mnp) activity Δ OD 10⁶/ΔT*V_{Enzyme solution}*∈

＝(ΔOD/ΔT)*10⁶/1*6.5*10³

v-volume of enzyme solution.

4.4 determination of cellulase Activity

1. Reagent

(1)0.1mo L/L sodium citrate-sodium citrate buffer, which is prepared by weighing 2.941g of sodium citrate and 2.1014g of citric acid respectively, adding deionized water 100m L, adding citric acid into sodium citrate, and adjusting the pH value to 4.8 by using a pH meter;

(2)1.5mLDNS；

(3) glucose standard solution 1.0000g glucose is dissolved in 1000ml deionized water to prepare 1mg/m L glucose standard solution.

2. Standard glucose Curve preparation

Taking 1mg/m L standard glucose solution 0,0.2,0.4,0.6,0.8,1.0 and 1.2m L respectively in a test tube, adding deionized water to 2.0m L, adding 2.0m L DNS reagent, plugging, carrying out boiling water bath for 10min, cooling, fixing the volume to 15m L, measuring an OD value by using a spectrophotometer at the wavelength of 550nm, repeating the experiment for 3 times, taking a mean value to draw a graph, and taking a glucose standard curve as shown in figure 11.

3. Procedure for the preparation of the

(1) Adding Whatman NO1 quantitative test paper of 1cm × 2cm into the test tube, and adding 1.0cm sodium citrate-sodium citrate buffer (0.1mo L/L, pH 4.8) and 0.5m L enzyme solution;

(2) preserving heat at 50 ℃ for 1h, taking out, adding 1.5m L DNS to terminate the reaction, carrying out water bath for 5min, cooling with running water, and metering the volume to 25m L;

(3) the absorbance was measured at 540 nm.

4. Calculation method

Cellulase activity ═ y 1000ug/V_{Enzyme solution}T, where y is derived from the standard 0.6916x

In the formula: x-measured absorbance values; t-time of water bath; v_{Enzyme solution}Volume of enzyme solution.

4.5 hemicellulase Activity assay

1. Reagent

(1)1.8m L1% xylan solution prepared into 1% solution with xylan in acetic acid buffer solution with pH 4.8;

(2)1.8m L acetic acid buffer solution, weighing 0.82g sodium acetate, weighing 0.6m L acetic acid, respectively adding deionized water 100m L, adding acetic acid into the sodium acetate, and adjusting the pH value to 4.8;

(3)2mL DNS；

(4) 1.0000g of xylose standard solution, namely dissolving xylose in 1000m of L deionized water to prepare 1mg/m of L xylose standard solution.

2. Production of xylose curve

Taking standard xylose liquid of 1mg/ml, 0,0.2,0.4,0.6,0.8,1.0 and 1.2m L respectively, adding deionized water to 2.0m L, then adding 2.0m L DNS reagent, plugging and boiling water bath for 10min, cooling, fixing the volume to 15m L, measuring OD value with a spectrophotometer at the wavelength of 550nm, repeating the test for 3 times, taking the mean value to draw a graph, and taking a xylose standard curve as shown in figure 12.

3. Procedure for the preparation of the

(1) Adding enzyme solution 0.2m L into a 15m L graduated test tube, and absorbing xylan solution 1.8m L1% respectively;

(2) adding 0.2m L enzyme solution into 1.8m L acetic acid buffer solution without adding xylan solution, and shaking up;

(3) taking out after 60min of 50 ℃ water bath, absorbing 2m L DNS reagent, shaking up, plugging, immediately boiling water bath for 10min, cooling, adding water to constant volume of 15m L, and shaking up and down gently;

(4) the absorbance was measured at a wavelength of 550nm while adjusting the point to zero with a blank.

4. Calculation method

Calculating according to the formula: hemicellulase activity ═ y 1000ug/V_{Enzyme solution}(ii) T, wherein y is 1.1504x +0.008 from standard; in the formula: x-measured absorbance values; t-time of water bath; v_{Enzyme solution}Volume of enzyme solution.

5. Results

TABLE 3 determination of the enzyme-producing Activity of the Ureibacillus Suwonensis TB42 Strain

As shown in Table 3, the activity of manganese peroxidase produced by Bacillus licheniformis TB42 was highest at 207U/L, followed by cellulase at 78U/L, and the activities of laccase, lignin peroxidase and hemicellulase were 7U/L, 0U/L and 16U/L, respectively.

Example 2: identification of Bacillus licheniformis TB42(Ureibacillus Suwonensis)

2.1 Main Instrument of experiment

TABLE 4 Instrument for experiments

2.2 Experimental procedures

2.2.1 extraction of DNA

DNA was extracted using the "Ezup column type bacterial genome DNA extraction kit" of the Producer. The specific operation steps are detailed in the specification.

2.2.2PCR amplification

PCR amplification was performed using 2 × Es Taq MasterMix, and the reaction system was as follows:

reaction procedure:

2.2.3 electrophoresis results of PCR products

Using 1% agarose gel electrophoresis, voltage 120V, electrophoresis for 30min, each hole loading 1u L, the electrophoretogram is shown in FIG. 13.

As shown in FIG. 13, the electrophoresis results of the PCR products showed that the 16S rDNA fragment of the bacterium was successfully amplified.

2.2.4 sequencing and alignment results

Sequencing was performed in both directions using primers 27F/1492R, and the sequence of Bacillus ureafaciens strain TB42 is shown in SEQ.1.

B L ASTN comparison was performed in NCBI gene bank to obtain the strain identification results, and the results are shown in Table 5.

TABLE 5 Bacillus ureafaciens strain TB42 identification data

As shown in Table 5, the strain TB42 showed 99% similarity to Ureibacillus subswensis, and therefore, the strain was named Bacillus ureafaciens TB42(Ureibacillus subswensis).

Example 3: application of microbial inoculum containing bacillus ureafaciens strain TB42 in promotion of compost maturity

3.1 preparation of the bacterial agent

Inoculating TB42 seed solution into L B culture solution sterilized by 1% (v/v) according to the inoculation amount of 1%, culturing for 24h in a 120rmp shaking table at 50 ℃, collecting the culture solution which is the microbial inoculum, and adding the microbial inoculum to obtain an experimental group (T), and adding L B culture solution sterilized by 1L to a blank group (CK).

3.2 compost test

The compost raw materials are cow dung and sugarcane leaves, and the mass ratio is 17:3, and the total weight is 20 kg. And (4) composting for 45d, sampling at 0 th, 5 th, 10 th, 16 th, 23 th, 30 th and 45 th d, and measuring parameters such as temperature, pH, water content, organic matters, Kjeldahl nitrogen, inorganic nitrogen, DOM content and the like. The microbial inoculum is inoculated twice in the whole composting process, wherein the inoculation time is 0d and 10d respectively. The stack was turned three times for 10d, 20d and 30d respectively.

3.3 results of the experiment

The Bacillus ureafaciens TB42 is high temperature resistant, can grow and propagate quickly, and can produce lignocellulose degrading enzyme.

Compared with Compost (CK) without added bacterial agent, compost (T) with added TB42 bacterial agent has the following advantages:

(1) from the temperature of the composting process, the temperature of the heap with the TB42 microbial inoculum is obviously improved compared with the temperature of the heap without the microbial inoculum. This is beneficial to killing pathogens in the compost mass and accelerating the high temperature degradation of the compost material, promoting the compost mass to become thoroughly decomposed (figure 1).

(2) As can be seen from FIG. 2 and FIG. 3, the pH of the compost to which the TB42 microbial inoculum is added is increased higher, and the water content is reduced faster, namely, the reaction of the compost to which the microbial inoculum is added is more violent, so that NH is caused₃The large amount of volatilization increases the pH value, generates more heat and takes away a large amount of moisture.

(3) From the viewpoint of the change of the content of the heap soluble organic matter (DOM), the DOM content without adding the bacterial agent is reduced from 69.6mg/g to 9.8mg/g by 85.9%, and the DOM content with adding the TB42 bacterial agent is reduced from 68.8mg/g to 6.7mg/g by 90.3% (fig. 8).

In conclusion, the TB42 microbial inoculum is added, so that the composting temperature can be increased, the water loss is accelerated, the organic matter degradation and DOM degradation are accelerated, and the compost maturity is promoted.

Example 4: surfactant production by Bacillus ureafaciens strain TB42

The surfactant production condition of the three strains is identified by a blood agar plate and a surface tension meter.

The blood agar plate is prepared by sterilizing nutrient agar (Beijing Luqiao), cooling to about 50 deg.C, adding sterilized defibered sheep blood 5-10m L per 100m L nutrient agar in sterile environment, shaking, pouring into plate, and making into slant.

And (3) measuring the surface tension of the fermentation liquor: the fermentation liquor surface tension is measured by adopting a ring method, and a full-automatic surface tension meter and an instrument model are adopted: BZY-1.

Experiments have shown that the Bacillus ureafaciens strain TB42 produces surfactants.

The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.

SEQUENCE LISTING

<110> Guangxi university

<120> Bacillus urealyticum strain TB42 and application thereof in promoting compost maturity

<130>ZYWS

<160>1

<170>PatentIn version 3.3

<210>1

<211>1458

<212>DNA

<213> Artificial sequence

<400>1

cgggggggga tgcctataca tgcaagtcga gcggaccaat tagaaagctt gctttttaat 60

tggttagcgg cggacgggtg agtaacacgt gggtaacctg ccctatagac cgggataact 120

cgcggaaacg cgtgctaata ccggataaca caccgaagcg catgcttcgg ggttgaaaga 180

tggttctgct atcactatag gatgggcccg cggcgcatta gctggttggt ggggtaacgg 240

cctaccaagg cgacgatgcg tagccgacct gagagggtga tcggccacac tgggactgag 300

acacggccca gactcctacg ggaggcagca gtagggaatc ttccacaatg ggcgaaagcc 360

tgatggagca acgccgcgtg agcgaagaag gtcttcggat cgtaaagctc tgttgtaagg 420

gaagaacaag cgcagcagtc actggctgcg ccctgacggt accttactag aaagccacgg 480

ctaactacgt gccagcagcc gcggtaatac gtaggtggca agcgttgtcc ggaattattg 540

ggcgtaaagc gcgcgcaggc ggtctcttaa gtctgatgtg aaagcccccg gctcaaccgg 600

ggagggtcat tggaaactgg gagacttgag tgcaggagag ggaagyggaa ttccatgtgt 660

agcggtgaaa tgcgtagaga tatggaggaa caccagtggc gaaggcggct tcctggcctg 720

taactgacgc tgaggcgcga aagcgtgggg agcaaacagg attagatacc ctggtagtcc 780

acgccgtaaa cgatgagtgc taggtgttag ggggtttccg ccccttagtg ctgcagctaa 840

cgcattaagc actccgcctg gggagtacgg tcgcaagact gaaactcaaa ggaattgacg 900

ggggcccgca caagcggtgg agcatgtggt ttaattcgaa gcaacgcgaa gaaccttacc 960

aggtcttgac atcccgctga ccgccatgga gacatggctt tcccttcggg gacagcggtg 1020

acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1080

gagcgcaacc cttgtcctta gttgccatca ttcagttggg cactctaagg agactgccgt 1140

acaaatacgg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acctgggcta 1200

cacacgtgct acaatgggtg gtacaaaggg cggcaaaccc gcgaggggga gcgaatccca 1260

aaaagccact ctcagttcgg attgcaggct gcaactcgcc tgcatgaagc cggaatcgct 1320

agtaatcgcg gatcagcatg ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg 1380

tcacaccacg agagtctgta acacccgaag tcggtgaggt aaccctccgg gagccagccg 1440

ccgaaaggtg acccgagt 1458

Claims

1. Bacillus ureafaciensUreibacillus suwonensis) The strain TB42 is preserved in the China general microbiological culture Collection center on the 05 th month 01 in 2017,the preservation number is CGMCC number 13529.

2. Use of the bacillus ureafaciens strain TB42 according to claim 1 for degrading lignocellulose.

3. Use of the bacillus ureafaciens strain TB42 according to claim 1 in the preparation of a surfactant.

4. Use of the bacillus ureafaciens strain TB42 according to claim 1 for the preparation of laccase, lignin peroxidase, manganese peroxidase, cellulase and hemicellulase.

5. A microbial inoculum is characterized in that the seed solution of the Bacillus ureafaciens strain TB42 of claim 1 is inoculated into L B culture solution after 1% (v/v) sterilization of 1L, cultured for 24h in a shaking table at 50 ℃ and 120rpm, and the culture solution is collected, thus obtaining the microbial inoculum.

6. Use of a microbial inoculum according to claim 5 in promoting compost maturity.