CN110438028A - A kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose - Google Patents
A kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose Download PDFInfo
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Abstract
The present invention discloses a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose.It is deposited in China typical culture collection center (abbreviation CCTCC), the deposit date is on November 29th, 2018, deposit number was CCTCC NO:M 2018841, and classification naming is Bacillus velezensis GX-1.Bacterial strain of the invention can generate cellulase, can efficient degradation cellulose, the biodegrade for cellulose.GX-1 bacterial strain institute's cellulase-producing has preferable thermal stability, is able to maintain preferable enzyme activity within the scope of 40-70 DEG C；Bacterial strain GX-1 has fast-growth, yield of enzyme height, advantage of less demanding to condition of culture and condition of enzyme production, and being easy to colonize in chitling road；It is of great significance to animal husbandry and feed fermentation industrial expansion.
Description
Technical field
The invention belongs to agriculture herding application fields, and in particular to a kind of people pig source Bei Laisi gemma bar of degraded cellulose
Bacterium (Bacillus velezensis) GX-1.
Background technique
Cellulose is renewable resource most abundant, most cheap on the earth.Although China, which is one, possesses abundant cellulose
The large agricultural country of resource, but cellulose resource is not utilized rationally, and great waste is caused.Cellulose is in aquaculture
In reasonable utilization, it will greatly reduce the use of regular power feed (such as corn) in daily ration.It can be to knot in nature
The cellulose of structure complexity carries out mainly microorganism, including fungi, bacterium, actinomyces etc. that effectively degradation utilizes.Fungi easily produces
Raw extracellular cellulase easily extracts separation, enzymatic activity height, good degrading effect, but most of fungies are higher to temperature requirement.Such as
Saroj is from isolating 15 plants of thermophilic fungals that can be grown at 50 DEG C in soil --- and aspergillus fumigatus JCM 10253 has higher
Cellulase and β-glucuroide and zytase activity；Although and actinomyces are widely distributed in nature, put
The breeding of line bacterium is slow compared with bacterium, and enzymatic productivity does not have fungi strong.Ren great Ming filters out one plant of dull gray streptomycete (unwrapping wire from peat
Bacterium), the produced enzyme of the bacterium is good to the adaptability of pH and temperature.Bacterial strain is grown best in the environment of pH=7.5, is weak base
Property bacterium, but the most suitable enzymatic reaction pH=4.5 of its produced cellulase is acidic cellulase, and bacterial strain wants growing environment
It asks and the characteristic of the product of own metabolism has specific characteristics.Equally, bacterium is widely present in the natural environment, cellulose-decomposing bacterium
Strain is high to the resistance of environment.They are usually cold-resistant, heat-resisting, also have certain tolerance to soda acid.Therefore, these bacterial strains can
It exists in adverse conditions, they would generally generate stable enzyme under the conditions of extreme pressure, be used for biotransformation.Example
Such as, 3 plants of cellulose-degrading bacterias that Pourramezan (2012) is separated to from termite gut, belong to acinetobacter, vacation
Zygosaccharomyces and staphylococcus, wherein bacillus generates enzyme activity highest, and institute's producing enzyme has preferable thermal stability and acid
Alkaline stability.From the cellulose decomposing bacteria filtered out in nature compared to fungi and actinomyces for, have growth fast, fine
The advantage that plain enzymatic activity is high, not high to condition of culture and condition of enzyme production requirement is tieed up, current application of cellulase, fermentation are being coped with
Development of feed industry etc. is of great significance.
Summary of the invention
Based on practical problem and demand in the above livestock-raising production process, the purpose of the present invention is to provide a kind of degradations
The people pig source Bei Laisi bacillus (Bacillus velezensis) GX-1 of cellulose, can produce high-content cellulase, energy
Enough efficient degradation celluloses, sodium carboxymethylcellulose etc. facilitates animal to the digestibility of cellulose, to improve animal to fibre
Tie up the utilization of plain resource.
Technical characteristic of the invention is as follows: a kind of people pig source Bei Laisi bacillus (Bacillus of degraded cellulose
Velezensis) GX-1 is preserved in China typical culture collection center, address: Wuhan City, Hubei Province Wuchang District Bayi Road 299
Number Wuhan University in the school, postcode: 430072, the deposit date is on November 29th, 2018, deposit number are as follows: CCTCC NO:
M 2018841, classification naming are Bacillus velezensis GX-1.
The present invention also has following technical characteristic:
1, a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose as described above, biological property packet
Include: White waxy, bacterium colony are white, and edge is irregular, dry, there is gauffer.
2, a kind of fermentation culture method of the people pig source Bei Laisi bacillus GX-1 of degraded cellulose as described above,
It is as follows: to take OD600The inoculum concentration of=1.0 seed liquor 1-5% into fermentation medium, 31-41 DEG C of temperature, 180-240rpm,
Shaking table shaken cultivation under the conditions of pH=4.0-7.0.Wherein, the formula of the fermentation medium are as follows: yeast powder 5g/L, tryptose arteries and veins
10g/L, sodium chloride 10g/L, CMC-Na 10g/L, pH=6 ± 0.5.
3, a kind of fermentation culture method of the people pig source Bei Laisi bacillus GX-1 of degraded cellulose as described above, it is excellent
35 DEG C of temperature, pH=5.0 are selected, inoculum concentration 2%.
4, a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose as described above, institute's cellulase-producing pair
Cellulose degradation condition is 40-70 DEG C of temperature, pH=4.0-7.0, reaction time 5-40min.
5, a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose as described above, institute's cellulase-producing pair
55 DEG C, pH=4.5, reaction time 25min of cellulose degradation condition preferable temperature, enzyme activity 41.18U/mL.
6, a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose as described above is preparing fibrous feedstuff
In application.
Advantages of the present invention and the utility model has the advantages that bacterial strain GX-1 provided by the present invention generate cellulase ability it is strong, and
Generated cellulase is able to maintain very high activity, can realize that efficient degradation, condition of enzyme production are of less demanding to cellulose, again
It is easy to the intestinal colonisation in pig；The bacterial strain is to the utilization of fibrous feedstuff resource and feed fermentation industry and animal husbandry
Development is of great significance.GX-1 bacterial strain institute's cellulase-producing has preferable thermal stability, be able to maintain within the scope of 40-70 DEG C compared with
Good enzyme activity；Bacterial strain GX-1 has fast-growth, yield of enzyme height, of less demanding to condition of culture and condition of enzyme production, and is easy to
In the advantage that chitling road colonizes；It is of great significance to animal husbandry and feed fermentation industrial expansion.
Detailed description of the invention
Fig. 1 GX-1 thalli morphology；
The phylogenetic tree of Fig. 2 bacterial strain GX-1；
Fig. 3 glucose standard curve；
Influence of Fig. 4 pH to bacterium GX-1 producing enzyme；
Influence of Fig. 5 temperature to bacterium GX-1 producing enzyme；
Influence of Fig. 6 inoculum concentration to bacterium GX-1 producing enzyme；
Influence of Fig. 7 temperature to cellulose enzyme activity；
Influence of Fig. 8 pH to cellulose enzyme activity；
Influence of Fig. 9 reaction time to cellulase activity；
Specific embodiment
Embodiment 1: bacterial strain screening and identification
People pig is one of local eight big excellent pig kinds in China, and with good meat quality, resistance to crude feed, farrowing is more, winter resistance is strong, disease-resistant
The advantages that strong, the resistance to crude feed of people pig, fit for depasturing raising, energy large amounts of food green and rough feeds, largely search for food weeds, edible wild herbs.There is research
Show: in the case of three kinds of Diets of crude fiber content 9%, 12% and 15%, people pig and Large White glutamic-pyruvic transaminase, paddy
Careless transaminase, lactic dehydrogenase, creatine kinase content have increase, but people's pig increase degree is smaller；In addition high crude fibre is fed
The histoorgans such as people's pig liver, heart, muscle institute is impacted in the case where content daily ration is much smaller than Large White.Therefore, we push away
Survey the microorganism for settling down in people's chitling road and capable of largely utilizing plant cellulose.The present embodiment bacterial strain via adult people's pig manure
Just screen isolated, detailed process is as follows:
It weighs people's swine excrement 1.0g to be transferred in the triangular flask for filling 100mL sterile water, vibrates water in 80 DEG C of electric heating constant temperatures
30min is vibrated in bath.The LB+1%CMC plate using sodium carboxymethylcellulose as sole carbon source is applied to according to spread plate
On, 37 DEG C of cultures are for 24 hours.Dye liquor is discarded after appropriate 0.1% Congo red solution dyeing 1h is added into culture dish, is selected from plate
Bacterium colony with transparent circle carries out primary dcreening operation, therefrom continues to select the significant bacterium colony of transparent circle, and scribing line isolates and purifies on LB plate
To single colonie.Obtained single colonie is measured into cellulase activity respectively and carries out secondary screening, selects that cellulose degradation effect is best, fiber
The plain highest Strain Designation of enzyme activity is GX-1.
The biological property for the bacterial strain GX-1 that the present embodiment is found includes: White waxy, and bacterium colony is white, and edge is not whole
Together, dry, there is gauffer (see Fig. 1).The ability for generating cellulase is strong, and generated cellulase is able to maintain very high work
Property, efficient degradation can be realized to cellulose.
The physiological and biochemical property of bacterial strain of the present invention is following (see the table below 1):
1 bacterial strain GX-1 physiological and biochemical property of table
Note: "+" represents the positive, and "-" represents feminine gender
Molecular Identification process and result are as follows:
The total DNA that bacterial strain of the present invention is extracted using the raw work extracting genome DNA reagent SK8255 in Shanghai, is tried by SK8131
Agent box carries out 16S rDNA PCR amplification, obtains 1492bp16S rDNA segment.
Shanghai Sangon Biological Engineering Technology And Service Co., Ltd is entrusted to complete 16S rDNA sequencing, sequencing primer are as follows: upstream
Primer: 5'-CAGAGTTTGATCCTGGCTAGGAGGTGATCCAGCCGCA-3'；Downstream primer: 5'-AGTTTGATCMTGGCT
CAGGGTTACCTTGTTACGACTT-3'.Obtain DNA sequencing as a result, and in ribosomes database http: //
It compares on rdp.cme.msu.edu/index.jsp, is compared with the most similar bacterial strain 16S rDNA sequence of homology,
The phylogenetic tree for drawing bacterial strain, compares with the bacterial strain degree of approximation and reaches 99% (see Fig. 2).
Through morphologic observation, biochemical reactions and 16S rDNA Molecular Identification, determine that this bacterial strain is Bei Laisi bacillus
One novel species of kind (Bacillus velezensis) finds that Bei Laisi bacillus has fiber in people's swine alimentary canal for the first time
Plain degradation capability is named as Bei Laisi bacillus (Bacillus velezensis) GX-1.
Embodiment 2: cellulase activity measurement
Method is as follows: crude enzyme liquid preparation: by the OD of activation culture bacterium solution600Being adjusted to 1.0 is the seed liquor tested；Then
Seed liquor OD=1.0 is inoculated in 50mL Medium of shaking flask fermentation with 1.0% inoculum concentration, is issued in 37 DEG C, 220rpm condition
Ferment culture for 24 hours, is centrifuged fermentation liquid 15min, supernatant is crude enzyme liquid under the conditions of 5000rpm, 4 DEG C；Glucose standard curve
Drafting: prepare 1mg/mL glucose mother liquid, take 0 respectively, 0.2,0.4,0.6,0.8,1.0,1.2,1.4ml is in clean ratio
In colour tube, is then mended with ultrapure water to 2mL less than 2mL, 2.5mL DNS solution is then added and shakes up, boiling water bath 5min, fast quickly cooling
But 10mL is settled to purified water afterwards to mix.100 μ L are taken to measure OD value under microplate reader 540nm wavelength, to measure at 540 nm
Absorbance OD value be ordinate, with glucose containing unit be (mg) be abscissa drafting standard curve (see Fig. 3).Measure fiber
Plain enzyme (CMCA) enzyme activity: being added the 1%CMC-Na phosphate buffer of 1mL crude enzyme liquid and 1.5mL in clean colorimetric cylinder,
30min is kept the temperature in 50 DEG C of water-baths；Then it is rapidly added 2.5mL DNS solution and shakes up, make reaction terminating, boiling water bath
5min；10mL is settled to purified water after rapid cooling to mix, and measures OD value in the case where wavelength is 540nm.In standard glucose song
Value on line according to absorbance finds corresponding glucose content.Enzyme amount needed for substrate generates 1 μ g glucose per hour is one
Enzyme activity unit (U).The present invention carries out cellulase activity measurement to the bacterial strain GX-1, finds the culture for 24 hours of bacterial strain GX-1
The CMC enzyme activity of liquid is 37.24U/mL.
Embodiment 3: influence of the different condition of culture to bacterial strain GX-1 growth and breeding
The present invention provides the fermentation culture method of the bacterial strain GX-1, specifically: lab shaker cultural method: take
OD600The inoculum concentration of=1.0 seed liquor 1% is into fermentation medium, in 31-41 DEG C, 180-240rpm, pH=4.0-7.0 item
Shaken cultivation under part.Wherein, the formula of the fermentation medium are as follows: yeast powder 5g/L, tryptose arteries and veins 10g/L, sodium chloride 10g/
L, CMC-Na 10g/L, pH=6 ± 0.5.
(1) different initial influences of the pH to bacterial strain GX-1: present embodiment Bei Laisi bacillus GX-1 is with initial ph value
The producing enzyme curve graph of variation is as shown in Figure 4.Bacterial strain enzyme activity is more stable when initial ph value is between 4-7, the enzyme activity when pH is 5.0
Reach peak, thus originate pH be 5.0 be Bei Laisi bacillus GX-1 producing enzyme most suitable initial ph value.
(2) influence of the different temperatures to bacterial strain GX-1: present embodiment Bei Laisi bacillus GX-1 is varied with temperature
Producing enzyme curve graph is as shown in Figure 5.When temperature is 35 DEG C, enzyme activity reaches maximum value, enzyme activity 22.03U, so temperature is 35 DEG C
When for Bei Laisi bacillus GX-1 producing enzyme optimum temperature.
(3) influence of the different vaccination amount to bacterial strain GX-1: present embodiment Bei Laisi bacillus GX-1 is with inoculation quantitative change
The producing enzyme curve graph of change is as shown in Figure 6.When inoculum concentration is 2%, enzyme activity reaches maximum value, enzyme activity 26.39U, so 2%
It is the most suitable inoculum concentration of Bei Laisi bacillus GX-1 producing enzyme when inoculum concentration.
(4) the temperature tolerance of bacterial strain GX-1: being taken at -80 DEG C and freeze after strain inoculated activates 8h in LB liquid medium,
It takes 1% switching in LB liquid medium, is respectively placed in 70 DEG C, 80 DEG C and stands overnight, cultivate afterwards for 24 hours, as the result is shown bacterial strain GX-1
It can be survived under high temperature with sleep mode.
Embodiment 4: the research to the enzymatic property of bacterial strain GX-1 institute cellulase-producing
(1) Bei Laisi bacillus GX-1 cellulase-producing heat stability test: the change curve of reaction temperature such as Fig. 7 institute
Show.It is 55 DEG C that CMCase, which reacts most suitable temperature,.When temperature rises to 70 DEG C, enzyme activity size is still optimum temperature enzyme activity
80% or more, illustrate that GX-1 institute's cellulase-producing has good thermal stability.
(2) Bei Laisi bacillus GX-1 cellulase-producing pH tolerance detects: present embodiment Bei Laisi bacillus
The change curve of GX-1 enzyme reaction pH is as shown in Figure 8.It is 4.5 that CMCase, which reacts most suitable pH,.
(3) Bei Laisi bacillus GX-1 cellulase-producing detects the effective time of fibrin reaction: Bei Laisi gemma
The change curve of bacillus GX-1 enzyme reaction time is as shown in Figure 9.CMCase reaction enzyme activity highest when carrying out 25min, hereafter,
With the extension of reaction time, enzyme activity gradually decreases for enzyme and substrate.
The preparation of 5: Bei Laisi bacillus GX-1 microbial inoculum of embodiment
The present embodiment freezes the preparation that bacterial strain carries out microbial inoculum, concrete operation method using Bei Laisi bacillus GX-1 are as follows:
Bacterial strain GX-1 is accessed into reaction system (formula: yeast powder 5g/L, tryptose arteries and veins 10g/L, sodium chloride 10g/L, CMC-Na 10g/
L, pH=6 ± 0.5) in carry out activation culture 8h, after activation culture object transferred with 2% inoculative proportion (built into reaction system
View system: 50ml) in, condition of culture are as follows: 35 DEG C of temperature, pH=5 can obtain GX-1 microbial inoculum, to described after shaking flask culture for 24 hours
GX-1 microbial inoculum carries out cellulase activity measurement, and enzyme activity is 41.18U/mL (microbial inoculum short term stored condition are as follows: 4 DEG C, it is proposed that storage
Time is 48h；Long-term storage condition are as follows: -40 DEG C).
Embodiment 6: bacterial strain GX-1 fermentation seed liquid is gavaged on the multifarious influence of SD Bacteria from Gl Tract of Rats in short term
The present invention is designed using bacterial strain GX-1 and using single-factor randomized block experiment, chooses that 3 week old, weight be close, SPF
Grade healthy SD female rats 24, is randomly divided into 2 groups, every group of 6 repetitions, 2 mouse of each repetition.It is pre- to raise 3 days, control group (A
Group) physiological saline 0.2ml/d is gavaged, test group (B group) gavages bacterium solution 0.2ml/d；Daily stomach-filling 1 time, continuous gavage 7d.From
It by searching for food, in test the 21st day, puts to death A group and B group mouse and takes caecum chyme, freeze for DNA extraction, build library and be sequenced, at data
Reason, OTU division, diversity analysis；
The Bei Laisi bacillus GX-1 influence horizontal to SD rat cecal microorganism door is gavaged in short term is shown in Table 2.As a result table
Bright: gavaging Bei Laisi bacillus GX-1 significantly reduces (P to the ratio of SD rat cecal microorganism actinomyces door and Proteobacteria
<0.05)。
Table 2 gavages the Bei Laisi bacillus GX-1 influence horizontal to SD rat cecal microorganism door in short term
Note: with data line shoulder mark, different letters indicate significant difference (P < 0.05), and no letter indicates the not significant (P of difference
>0.05)。
Embodiment 7: influence of the bacterial strain GX-1 fermentation seed liquid to SD intestine in rats morphological development is gavaged for a long time
The present embodiment is designed using bacterial strain GX-1 and using single-factor randomized block experiment, choose 3 week old, weight it is close,
Healthy SD rat 24, it is randomly divided into 2 groups, every group of 6 repetitions, 2 mouse of each repetition.Control group gavages physiological saline 0.2mL/
D, test group gavages bacterium solution 0.2mL/d, and (bacterial concentration is 1 × 108CFU/mL)；Daily stomach-filling 1 time, continuous gavage 20d.Freely
Feeding tests the 21st day whole rats and puts to death sampling.Abdominal cavity is quickly opened, jejunum, ileum are collected.Clip 2cm intestinal segment is in 10%
It is fixed for enteron aisle HE dyeing measurement Intestinal Morphology in formalin.Fixed intestinal segment in 10% formalin is taken out, through washing
It washs, is transparent, waxdip, paraffin embedding, after the processing such as repair block, slice, dewaxing, carrying out hematoxylin eosin stain, dehydration and mounting.In
It is observed under optical microscopy, chooses the typical visual field, i.e. complete 5 of fluff morphology in every jejunal tissue slice, carry out
It takes pictures.Jejunum villi height and Crypt depth are measured using 6.0 image analysis software of Image-Pro Plus, according to measurement
Height of naps and Crypt depth calculate its ratio.Height of naps is in villus top and both ends crypts villus joint line
The distance of point, Crypt depth is the midpoint of both ends crypts villus joint line at a distance from mucous membrane base.
It gavages influence of the Bei Laisi bacillus GX-1 to SD jejunum in rats and ileum morphological development and is shown in Table 3.Full period gavages
Bei Laisi bacillus GX-1 significantly improves the height of naps (P < 0.05) of jejunum in rats, significantly reduce ileum Crypt depth (P <
0.05).Have the tendency that reduction (P > 0.05) to Jejunum Crypt depth, ileum height of naps has raised trend (P > 0.05).
Influence of the 3 Bei Laisi bacillus GX-1 of table to SD intestine in rats morphological development
Note: with data line shoulder mark, different letters indicate significant difference (P < 0.05), and no letter indicates the not significant (P of difference
>0.05)。
Embodiment 8: Bei Laisi bacillus M2 is gavaged in short term to 35 age in days SD rat bloods biochemistry and the haematogenic immunity factor
Influence
The present embodiment is designed using bacterial strain GX-1 using the random group experiment of single-factor, chooses that 3 week old, weight be close, SPF
Grade healthy SD female rats (tieing up tonneau China in Beijing) 24, are randomly divided into 2 groups, every group of 6 repetitions, 2 mouse of each repetition.It is right
Physiological saline 0.2ml/d is gavaged according to group (A group), test group (B group) gavages bacterium solution 0.2mL/d；Daily stomach-filling 1 time, continuous gavage
7d.It is freely eaten, pre-feeding period 3 days, the 11st day every cage is tested in the formal phase and randomly selects a rat execution, it is anticoagulant with heparin sodium
Pipe acquires Rat blood samples (use eyeball blood taking method), is centrifuged 10min in 3500r/min, separated plasma, total cholesterol, sweet
Oily three esters, low density lipoprotein cholesterol, high-density lipoprotein cholesterol use GPO-PAP enzyme process, and glucose assays use Portugal
Grape oxidase procedure, the measurement of the haematogenic immunity factor use enzyme linked immunosorbent assay (ELISA), measure IgA, IgG in blood plasma,
IgM.The concentration of TNF-α, IL-1 β, IL-6 and IL-8；Kit is purchased from Nanjing and builds up Bioengineering Research Institute, and operation is stringent
It is carried out according to kit specification.
Influence of the Bei Laisi bacillus M2 to 35 age in days SD rat bloods biochemistry and the haematogenic immunity factor is gavaged in short term such as
Shown in table 4,5: compared with the control group, total cholesterol in blood plasma, blood glucose, low-density can be reduced by gavaging Bei Laisi bacillus in short term
The concentration of lipoprotein cholesterol, but difference is not significant (P > 0.05)；The concentration of blood plasma middle-high density lipoprotein cholesterol is improved,
But otherness is not significant (P > 0.05)；Significantly reduce triglyceride concentration (P < 0.01).
Table 4 gavages influence of the Bei Laisi bacillus M2 to 35 age in days SD rat blood biochemistry in short term
Note: with data line shoulder mark, different letters indicate significant difference (P < 0.05), and no letter indicates the not significant (P of difference
>0.05)。
Table 5 gavages influence of the Bei Laisi bacillus M2 to 35 age in days SD rat blood immune factors in short term
Note: with data line shoulder mark, different letters indicate significant difference (P < 0.05), and no letter indicates the not significant (P of difference
>0.05)。
Embodiment 9: full period gavages influence of the Bei Laisi bacillus M2 to SD rat diet nutriment apparent digestibility
Test receives excrement method using complete, starts to receive excrement in test 18d.The feeding of daily accurate recording each group test SD rat
It measures and accurately collects the excrement that every group of test SD rat drains daily, continuously collect 3d.The excrement collected daily is weighed, is added
Enter the hydrochloric acid solution that quality is excrement fresh weight 10% to mix well, is saved in -20 DEG C, after the test, same group will be picked up from
Excrement is sufficiently mixed uniformly, and every group is about chosen 100g or so fresh excrement sample, after baking 15min enzyme deactivation is living first in 120 DEG C of baking ovens,
65 DEG C of drying are down to, are weighed after sufficiently getting damp again for 24 hours, until constant weight, crushes, sample to be tested is made.Crude protein content uses kelvin
Nitriding measurement；Crude fat content is measured using ether extraction method；Energy is measured using 6300 type oxygen bomb calorimeter of Parr；Ca
Permanganimetric method and colorimetric method for determining is respectively adopted with P.
The results are shown in Table 6: compared with the control group, full period gavages Bei Laisi bacillus to crude protein, crude fat, thick fibre
The apparent digestibility of dimension is improved trend, but difference is not significant (P > 0.05)；The apparent digestibility of calcium has decreasing trend, but difference
Property not significant (P > 0.05)；Total energy, ash content and phosphorus apparent digestibility significantly improve (P < 0.05).
6 full period of table gavages influence of the Bei Laisi bacillus M2 to SD rat diet nutriment apparent digestibility
Note: with data line shoulder mark, different letters indicate significant difference (P < 0.05), and no letter indicates the not significant (P of difference
>0.05)。
Claims (7)
1. a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose, is deposited in China typical culture collection center,
Deposit number is CCTCC NO:M 2018841, and classification naming is Bacillus velezensis GX-1.
2. a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose according to claim 1, which is characterized in that
Its biological property includes: White waxy, and bacterium colony is white, and edge is irregular, dry, there is gauffer.
3. the fermented and cultured side of the people pig source Bei Laisi bacillus GX-1 of degraded cellulose according to claim 1 a kind of
Method, which is characterized in that method is as follows: OD is taken600The inoculum concentration of=1.0 seed liquor 1-5% is into fermentation medium, temperature 31-
41 DEG C, shaking table shaken cultivation under the conditions of 180-240rpm, pH=4.0-7.0.Wherein, the formula of the fermentation medium are as follows: ferment
Female powder 5g/L, tryptose arteries and veins 10g/L, sodium chloride 10g/L, CMC-Na 10g/L, pH=6 ± 0.5.
4. the fermented and cultured side of the people pig source Bei Laisi bacillus GX-1 of degraded cellulose according to claim 3 a kind of
Method, which is characterized in that 35 DEG C of temperature, pH=5.0, inoculum concentration 2%.
5. a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose according to claim 1, which is characterized in that
Its cellulase-producing is 40-70 DEG C of temperature, pH=4.0-7.0, reaction time 5-40min to cellulose degradation condition.
6. a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose according to claim 5, which is characterized in that
55 DEG C of temperature, pH=4.5, reaction time 25min.
7. a kind of people pig source Bei Laisi bacillus GX-1 of degraded cellulose according to claim 1 is in preparation fibroid
Application in feed.
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