CN110484471A - The method that one plant height produces the acidproof bacterial strain of bacteria cellulose and its produces bacteria cellulose - Google Patents
The method that one plant height produces the acidproof bacterial strain of bacteria cellulose and its produces bacteria cellulose Download PDFInfo
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Abstract
The present invention relates to microorganisms technical fields, and in particular to the method that a plant height produces the acidproof bacterial strain of bacteria cellulose and its produces bacteria cellulose.The present invention provides coltfoal shape bacillus (Komagataeibacter sp.) ATC301 of one plant of production bacteria cellulose, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.17875.The bacterial strain is resistant to acid condition, it grows in acid condition and efficiently produces bacteria cellulose, the bacterial strain can pass through the efficient production bacteria cellulose of either statically or dynamically culture using fermentation raw material cheap and easy to get simultaneously, the production efficiency of bacteria cellulose can be effectively improved, the production cost for reducing bacteria cellulose is applicable to industrialize extensive manufacture bacteria cellulose.
Description
Technical field
The present invention relates to microorganisms technical fields, and in particular to one plant of coltfoal shape bacillus (Komagataeibacter sp.)
ATC301, and the method using bacterial strain production bacteria cellulose.
Background technique
Bacteria cellulose (Bacterial cellulose) is the cellulose generated using bacterial fermentation, is that glucose is logical
Cross high molecular polymer made of β -1,4 glucosides key connection.Compared with plant cellulose, bacteria cellulose has high chemistry pure
Degree, high-crystallinity, high elastic modulus, high-hydroscopicity and excellent bio-compatible and biodegradability.Therefore, bacterial fibers
Element is expected to be widely used for the fields such as food, medicine, cosmetics, papermaking, weaving and chemical industry.
Being currently known the bacteriums of multiple kinds being capable of fermented-producing bacteria cellulose, comprising: glucose acetobacter
(Gluconacetobacter), acetobacter (Acetobacter), agrobacterium (Agrobacterium), pseudomonad
(Pseudomonas), enterobacteria (Enterobacter) etc..Wherein, most study surely belongs to acetobacter xylinum
(Gluconacetobacter xylinus) (Chen Jing etc., preparation and the application study progress of bacteria cellulose, Cellulose Science
With technology, 201422 (2): 58-63).Developed in recent years based on the metabolic engineering of recombinant DNA technology with synthetic biology technology fast
Suddenly, also for the genetic modification of bacteria cellulose producing strains provide theoretical basis and tool (such as: Chinese patent
CN108060112A discloses the bacteria cellulose Producing Strain of one plant of genetic modification).But directly screened from nature high yield and
Be not easy microbiological contamination, be suitble to the bacteria cellulose of industrialized production production bacterium to can yet be regarded as a kind of effective strain obtaining means.
Although known various bacteria can generate bacteria cellulose, the industrialization of current bacteria cellulose, it is extensive,
Low cost production is still faced with lot of challenges, such as: nutritional requirement height, low output, easy microbiological contamination etc..Industrialization is large-scale thin
Fungin production putting forward higher requirements for production bacterial strain, has excellent bacteria cellulose production performance, Neng Gouli
With cheap raw material (such as glucose, corn pulp), and the production bacterial strain for being not easy microbiological contamination in process of production can effectively improve carefully
The production efficiency of fungin reduces the production cost of bacteria cellulose.Therefore, it needs to develop efficient bacteria cellulose production
Bacterial strain.
Summary of the invention
To solve the technical problems existing in the prior art, the purpose of the present invention is to provide a plant heights to produce bacteria cellulose
And acidproof coltfoal shape bacillus (Komagataeibacter sp.) ATC301 and utilization strain fermentation production bacteria cellulose
Method.
To achieve the above object, technical scheme is as follows:
The present invention provides coltfoal shape bacillus (Komagataeibacter sp.) ATC301 of one plant of production bacteria cellulose, the bacterium
Strain be preserved on May 31st, 2019 China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC,
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode 100101), classification naming
For coltfoal shape bacillus Komagataeibacter sp., deposit number is CGMCC No.17875.
The 16S rDNA gene order of coltfoal shape bacillus (Komagataeibacter sp.) ATC301 provided by the invention is such as
Shown in SEQ ID NO.1.
The present invention also provides the microbial inoculums comprising the coltfoal shape bacillus ATC301.
In the present invention, the microbial inoculum comprising coltfoal shape bacillus ATC301 can be liquid bacterial agent or solid fungicide.The packet
The microbial inoculum of the bacillus of shape containing coltfoal ATC301 can be prepared into using the auxiliary material that conventional technical means, addition microbial preparation field allow
It arrives.
The present invention proves that coltfoal shape bacillus ATC301 is resistant to acid condition, and in acid condition by experimental verification
Efficiently production bacteria cellulose.
Further, the present invention provides the coltfoal shape bacillus ATC301 or the microbial inoculum containing the coltfoal shape bacillus ATC301 exists
Application in bacteria cellulose production.
Microbial inoculum the present invention also provides the coltfoal shape bacillus ATC301 or containing the coltfoal shape bacillus ATC301 is in acid item
Application under part, in dynamic fermentation or static fermentation production bacteria cellulose.
Above-mentioned application is to obtain bacteria cellulose by cultivating the coltfoal shape bacillus ATC301.
In the present invention, the coltfoal shape bacillus ATC301 can using the cheap carbon sources such as glucose, sucrose, corn pulp, nitrogen source into
Row growth and production bacteria cellulose.
Above-mentioned culture coltfoal shape bacillus ATC301 can contain the natural of carbon source, nitrogen source and inorganic ions or conjunction using conventional
At culture medium.
In the present invention, the cultivation temperature of the coltfoal shape bacillus ATC301 is 28~32 DEG C, and pH is 4.0~7.0.Preferably 29
~31 DEG C, pH is 4.5~5.5.
The present invention experiments prove that, coltfoal shape bacillus ATC301 is resistant to acid condition and ferment production of the pH down to 4.0
Bacteria cellulose can be produced more efficiently in the case where pH is 5.0 acid condition in endophytic bacteria cellulose.
Microbial inoculum the present invention also provides the coltfoal shape bacillus ATC301 or containing the coltfoal shape bacillus ATC301 is thin in building
Fungin produces the application in bacterial strain.
Above-mentioned application is to carry out bacteria cellulose by breeding methods such as mutagenesis, genetic modifications using coltfoal shape bacillus ATC301
Produce the breeding of bacterium.
The present invention also provides a kind of production methods of bacteria cellulose, thin to be produced using the coltfoal shape bacillus ATC301
Fungin.
Specifically, using the culture medium comprising glucose and/or sucrose as fermentation raw material, coltfoal shape bacillus ATC301 is carried out
Dynamic fermentation or static fermentation obtain the bacteria cellulose.
Preferably, the production method of the bacteria cellulose includes the following steps:
(1) actication of culture: the coltfoal shape bacillus ATC301 is inoculated on activation plate or inclined-plane, is cultivated in 28~32 DEG C
2~4 days；
(2) seed culture: the coltfoal shape bacillus ATC301 activated in step (1) is inoculated in seed culture medium, in 28~
32 DEG C, oscillation or stir culture are mature to seed；
(3) fermented and cultured: the mature seed of the coltfoal shape bacillus ATC301 is inoculated in fermentation medium, in 28~32
DEG C, oscillation, stirring or stationary culture.
It is further preferred that the culture medium of the actication of culture includes following component: 5~10g/L in above-mentioned steps (1)
Peptone, 2~5g/L yeast powder, 2~5g/L glucose, 0.2~0.5g/L epsom salt.
It is further preferred that the culture medium of the seed culture includes following component: 10~30g/L in above-mentioned steps (2)
Sugar source, 5~15g/L corn pulp, 1~2g/L yeast powder, 0.1~0.5g/L epsom salt, 0.1~0.5g/L biphosphate
Potassium, pH 5.0~5.5；The sugar source is selected from one or both of glucose, sucrose.
It is further preferred that the culture medium of the fermented and cultured includes following component: 20~40g/L in above-mentioned steps (3)
Sugar source, 10~25g/L corn pulp, 0.5~1g/L epsom salt, 0.5~1g/L potassium dihydrogen phosphate, pH 5.0~5.5；It is described
Sugar source is selected from one or both of glucose, sucrose.
It is further preferred that the revolving speed of shaken cultivation is 100-200rpm in above-mentioned steps (2).The dress of seed culture medium
Liquid measure is 10~20%.The seed culture time is 2~4 days.
It is further preferred that the revolving speed of shaken cultivation is 100-200rpm in above-mentioned steps (3).The inoculum concentration of seed is 5
~10%.The liquid amount of fermentation medium is 10~20%.
The beneficial effects of the present invention are: the present invention obtains coltfoal shape bacillus ATC301 by separation screening, which can
It is resistant to acid condition, grown under acid condition of the pH down to 4~5 and efficiently produces bacteria cellulose, life can be significantly reduced
Microbiological contamination risk during production；The bacterial strain can utilize fermentation raw material (such as glucose, sucrose, corn pulp) cheap and easy to get simultaneously
It is (high-cost organic without adding citric acid, acetic acid, lactic acid etc. by either statically or dynamically cultivating efficiently production bacteria cellulose
Acid is used as fermentation raw material), conversion ratio with higher.Coltfoal shape bacillus ATC301 provided by the invention can effectively improve bacterium fibre
The production efficiency for tieing up element, reduces the production cost of bacteria cellulose, is applicable to industrialize extensive manufacture bacteria cellulose.
Detailed description of the invention
Fig. 1 is the Blast comparison result of the 16S rDNA sequence of ATC301 bacterial strain in the embodiment of the present invention 2.
Fig. 2 is the infrared absorption pattern of bacteria cellulose in the embodiment of the present invention 3.
Specific embodiment
The preferred embodiment of the present invention is described in detail below in conjunction with embodiment.It will be appreciated that following real
Providing merely to play the purpose of explanation for example is applied, is not used to limit the scope of the present invention.The skill of this field
Art personnel without departing from the spirit and purpose of the present invention, can carry out various modifications and replace to the present invention.
Experimental method used in following embodiments is conventional method unless otherwise specified.
It in following embodiments unless otherwise specified, is the operation of microorganism normal sterile.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
The screening of 1 bacteria cellulose producing strains of embodiment
The present invention is largely screened the acidproof bacterial strain ATC301 of isolated bacteria cellulose high yield from vinegar fermented grain, is screened every time
Specific method include the following steps:
(1) fresh vinegar fermented grain 2g is taken, 5mL sterile water is added, mixes well, vinegar fermented grain suspension is made；
(2) 1mL suspension is taken, is added and 49mL liquid screening medium (20g/L glucose, 10g/L ethyl alcohol, 20g/L is housed
Corn pulp, 1g/L epsom salt, pH 5.0) 250mL triangular flask in, 30 DEG C stationary culture 3 days, until culture solution surface shape
At one layer of semi-transparent film；
(3) rinsed with sterile water semi-transparent film twice, is transferred to 50mL sterile centrifugation tube, and 10mL sterile water is added, and nothing is added
Bacterium bead, Vortex 5min, smashes semi-transparent film；
(4) it takes 200 μ L suspensions to carry out 10 times of gradient dilutions, is diluted to 100,000 times, it is solid that each dilution takes 100 μ L to be coated on
(15g/L agar powder, pH 5.0 is added) in body screening and culturing medium plate in aforesaid liquid screening and culturing medium；
(5) be inverted plate in 30 DEG C stationary culture 3 days, until grow single colonie.Select the bacterium colony that there is transparent region on periphery
20 cultures 3 times of crossing repeatedly, obtain single colonie；
(6) it takes last time scribing line to form 16 pieces of uniform plates of colonial morphology, chooses single bacterium and fall within equipped with 10mL liquid sieve
In the 50mL triangular flask for selecting culture medium (formula is same as above), 30 DEG C stationary culture 3~4 days, until culture solution surface formed one and half
Hyaline membrane；
(7) the time required to different strains film forming and film thickness has notable difference, take 5 plants of film forming fast, at film thickness bacterial strain into
Row shake flask culture (medium component: 30g/L glucose, 25g/L corn pulp, 1g/L epsom salt, 1g/L biphosphate
Potassium, pH 5.0), 30 DEG C are cultivated 3-4 days, until generating a large amount of snowflake group/blocks in culture solution；
(8) according to weight in wet base (going the weight in wet base of supernatant after centrifugation) bacterium.
It finally screens to obtain the acidproof bacterial strain that a plant height produces bacteria cellulose, ATC301 is named as, after carrying out to the bacterial strain
Continuous strain idenfication and shake flask fermentation production bacteria cellulose experiment.
The 16S rDNA of 2 strains A TC301 of embodiment is identified
Using Standard PCR technology, with primer 2 7F (AGAGTTTGATCCTGGCTC AG) and 1492R
(TACGGCTACCTTGTTACGACTT) it is primer pair, using the bacterium solution of strains A TC301 as template, PCR amplification is carried out, through agar
Sugared detected through gel electrophoresis is shown, amplifies the DNA fragmentation of 1 treaty 1.4kb.PCR reaction system and condition are as shown in table 1:
The pcr amplification reaction system and condition of 1 16S rDNA of table
It using Ago-Gel QIAquick Gel Extraction Kit, is operated according to specification, the DNA fragmentation of above-mentioned 1.4kb is recycled, with laggard
Row sequencing, after removing both ends low quality sequence, the 16S rDNA sequence for obtaining strains A TC301 is (complete as shown in SEQ ID NO.1
Long 1345bp).
The sequence of the 16S rDNA as shown in SEQ ID NO.1 is subjected to BLAST on NCBI, as a result as shown in Figure 1,
The 16S rDNA sequence of ATC301 bacterial strain is identical with the 16S rDNA sequence of the Komagataeibacter multiple species belonged to.
In consideration of it, ATC301 Strain Designation is Komagataeibacter sp.ATC301.ATC301 bacterial strain was protected on May 31st, 2019
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, address: Chaoyang District, Beijing City north
The institute 3 of occasion West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101), classification naming is coltfoal shape bacillus
Komagataeibacter sp., deposit number are CGMCC No.17875.
The shake flask fermentation that 3 ATC301 bacterial strain of embodiment produces bacteria cellulose tests (1)
In the present embodiment, investigate Komagataeibacter sp.ATC301 bacterial strain in acid condition (pH 5.0),
By fermentation raw material (glucose, sucrose, corn pulp etc.) cheap and easy to get, fermenting and producing is thin in the way of dynamic cultivation in shaking flask
The performance of fungin, the specific method is as follows:
(1) fresh plate activates ATC301 bacterial strain: the ATC301 bacterial strain that glycerol saves is crossed to the plate of Fresh
On (10g/L peptone, 5g/L yeast powder, 5g/L glucose, 0.5g/L epsom salt, pH 7.0), plate is inverted in 30 DEG C
Stationary culture 2-3 days to bacterium colony is grown.
(2) preparation of shake-flask seed: dress is inoculated into from 2 ring bacterium of picking on the activation plate that step (1) obtains with oese
There are 20mL seed culture medium (10g/L glucose, 10g/L sucrose, 15g/L corn pulp, 2g/L yeast powder, seven water sulfuric acid of 0.5g/L
Magnesium, 0.5g/L potassium dihydrogen phosphate, pH 5.0) 100mL triangular flask in；30 DEG C, 150rpm, shaken cultivation 3 days, contain in culture solution
There are a large amount of snowflake group/blocks.
(3) shake flask fermentation: the shake-flask seed of step (2) is inoculated into ferment equipped with 100mL with 10% inoculation volume and is trained
Support base (10g/L glucose, 20g/L sucrose, 25g/L corn pulp, 1g/L epsom salt, 1g/L potassium dihydrogen phosphate pH 5.0)
In the fermentation shake flask of 500mL；30 DEG C, 150rpm, shaken cultivation 3 days.Contain a large amount of snowflake group/blocks in culture solution, as
Bacteria cellulose.
Bacteria cellulose is further identified using infrared absorption, the specific method is as follows:
Snowflake group/the block for taking 5g weight in wet base uses 0.1M HCl after cooling with 80 DEG C of heat treatment 2h of 0.1M NaOH solution
It neutralizes, is then rinsed well with tap water, 80 DEG C are dried overnight, and obtain the film of white translucent.
Appropriate white translucent film obtained above is taken to measure its infrared absorption, obtained infrared absorption pattern such as Fig. 2 institute
Show, with identical (Neera et al., the Appl Biochem of bacteria cellulose infrared absorption pattern reported in the literature
Biotechnol,2015,176(4):1162-73)。
The culture of shake flask fermentation is crossed into 300 mesh stainless steel cloths, trapped substance is dry to constant weight at 80 DEG C, measure bacterium
The yield of cellulose, the results show that strains A TC301 72 hours bacteria cellulose outputs of shake flask fermentation are 2.4g/L, bacterium is fine
Dimension element is 8% relative to the conversion ratio of sugar.
The shake flask fermentation that 4 ATC301 bacterial strain of embodiment produces bacteria cellulose tests (2)
In the present embodiment, investigate Komagataeibacter sp.ATC301 bacterial strain in acid condition (pH 5.0),
By fermentation raw material (glucose, sucrose, corn pulp etc.) cheap and easy to get, fermenting and producing is thin in the way of dynamic cultivation in shaking flask
The difference of the performance of fungin, specific method and embodiment 1 is only that the formula of seed culture medium in step (2) is as follows:
20g/L glucose, 15g/L corn pulp, 2g/L yeast powder, 0.5g/L epsom salt, 0.5g/L potassium dihydrogen phosphate, pH 5.0；
In step (3), the formula of fermentation medium is as follows: 30g/L glucose, 25g/L corn pulp, 1g/L epsom salt, 1g/L phosphorus
Acid dihydride potassium, pH 5.0.
The culture of shake flask fermentation is crossed into 300 mesh stainless steel cloths, by trapped substance after 80 DEG C of drying to constant weights, measurement is thin
The yield of fungin, the results show that strains A TC301 72 hours bacteria cellulose outputs of shake flask fermentation are 1.8g/L, bacterium
Cellulose is 6% relative to the conversion ratio of sugar.
The shake flask fermentation that 5 ATC301 bacterial strain of embodiment produces bacteria cellulose tests (3)
In the present embodiment, investigate Komagataeibacter sp.ATC301 bacterial strain in acid condition (pH 5.0),
By fermentation raw material (glucose, sucrose, corn pulp etc.) cheap and easy to get, fermenting and producing is thin in the way of dynamic cultivation in shaking flask
The difference of the performance of fungin, specific method and embodiment 1 is only that the formula of seed culture medium in step (2) is as follows:
20g/L sucrose, 15g/L corn pulp, 2g/L yeast powder, 0.5g/L epsom salt, 0.5g/L potassium dihydrogen phosphate, pH 5.0；Step
Suddenly in (3), the formula of fermentation medium is as follows: 30g/L sucrose, 25g/L corn pulp, 1g/L epsom salt, 1g/L di(2-ethylhexyl)phosphate
Hydrogen potassium, pH 5.0.
The culture of shake flask fermentation is crossed into 300 mesh stainless steel cloths, trapped substance is dry to constant weight measurement bacterium at 80 DEG C
The yield of cellulose, the results show that strains A TC301 72 hours bacteria cellulose outputs of shake flask fermentation are 2.5g/L, bacterium is fine
Dimension element is 8.3% relative to the conversion ratio of sugar.
The above result shows that bacterial strain Komagataeibacter sp.ATC301 is resistant to acid condition (pH 5.0),
And in acid condition (pH 5.0), utilize fermentation raw material (glucose, sucrose, corn pulp etc.) cheap and easy to get, dynamic fermentation
High yield bacteria cellulose.
The shake flask fermentation that 6 ATC301 bacterial strain of embodiment produces bacteria cellulose tests (4)
In the present embodiment, investigate Komagataeibacter sp.ATC301 bacterial strain in acid condition (pH 5.0),
By fermentation raw material (glucose, sucrose, corn pulp etc.) cheap and easy to get, fermenting and producing is thin in the way of static culture in shaking flask
The performance of fungin, the specific method is as follows:
(1) fresh plate activates ATC301 bacterial strain: the ATC301 bacterial strain that glycerol saves is crossed to the plate of Fresh
On (10g/L peptone, 5g/L yeast powder, 5g/L glucose, 0.5g/L epsom salt, pH 7.0), plate is inverted in 30 DEG C
Stationary culture 2-3 days to bacterium colony is grown.
(2) preparation of shake-flask seed: dress is inoculated into from 2 ring bacterium of picking on the activation plate that step (1) obtains with oese
There are 20mL seed culture medium (10g/L glucose, 10g/L sucrose, 15g/L corn pulp, 2g/L yeast powder, seven water sulfuric acid of 0.5g/L
Magnesium, 0.5g/L potassium dihydrogen phosphate, pH 5.0) 100mL triangular flask in；30 DEG C, 150rpm, shaken cultivation 3 days, contain in culture solution
There are a large amount of snowflake group/blocks.
(3) shake flask fermentation: the shake-flask seed of step (2) is inoculated into ferment equipped with 100mL with 10% inoculation volume and is trained
It supports base (10g/L glucose, 20g/L sucrose, 25g/L corn pulp, 1g/L epsom salt, 1g/L potassium dihydrogen phosphate, pH 5.0)
500mL fermentation shake flask in；30 DEG C, stationary culture 3 days, the gel state film of one layer of 1cm thickness was contained on culture solution surface, as carefully
Fungin.
Bacteria cellulose is further identified using infrared absorption, the specific method is as follows:
The gel state film for taking 5g weight in wet base is neutralized after cooling with 0.1M HCl with 80 DEG C of heat treatment 2h of 0.1M NaOH solution,
Then it is rinsed well with tap water, 80 DEG C are dried overnight, and obtain the film of white translucent.
Appropriate white translucent film obtained above is taken to measure its infrared absorption, obtained infrared absorption pattern and document report
The bacteria cellulose infrared absorption pattern in road coincide (Neera et al., Appl Biochem Biotechnol, 2015,176
(4):1162-73)。
The culture of shake flask fermentation is crossed into 300 mesh stainless steel cloths, trapped substance is dry to constant weight measurement bacterium at 80 DEG C
The yield of cellulose, the results show that strains A TC301 72 hours bacteria cellulose outputs of shake flask fermentation are 1.2g/L, bacterium is fine
Dimension element is 4% relative to the conversion ratio of sugar.
The result shows that bacterial strain Komagataeibacter sp.ATC301 is resistant to acid condition (pH 5.0), and
(pH 5.0) under acid condition, utilize fermentation raw material (glucose, sucrose, corn pulp etc.) cheap and easy to get, static fermentation high yield
Bacteria cellulose.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, without departing from the technical principles of the invention, several improvements and modifications can also be made, these improvements and modifications
Also it should be regarded as protection scope of the present invention.
Sequence table
<110>Beijing Ai Fa Science and Technology Ltd.
The method that<120>one plant heights produce the acidproof bacterial strain of bacteria cellulose and its produce bacteria cellulose
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cgcctgtgga ggaacctgcg ttcgattagc tagttggtgg ggtaaaggcc taccaaggcg 180
atgatcgata gctggtctga gaggatgatc agccacactg ggactgagac acggcccaga 240
ctcctacggg aggcagcagt ggggaatatt ggacaatggg cgcaagcctg atccagcaat 300
gccgcgtgtg tgaagaaggt tttcggattg taaagcactt tcagcgggga cgatgatgac 360
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gagggttgtg gaattcccag tgtagaggtg aaattcgtag atattgggaa gaacaccggt 600
ggcgaaggcg gcaacctggc tcatgactga cgctgaggcg cgaaagcgtg gggagcaaac 660
aggattagat accctggtag tccacgctgt aaacgatgtg tgctggatgt tgggtgactt 720
tgtcattcag tgtcgtagtt aacgcgataa gcacaccgcc tggggagtac ggccgcaagg 780
ttgaaactca aaggaattga cgggggcccg cacaagcggt ggagcatgtg gtttaattcg 840
aagcaacgcg cagaacctta ccagggcttg acatgcggag gccgtgtcca gagatgggca 900
tttctcgcaa gagacctcca gcacaggtgc tgcatggctg tcgtcagctc gtgtcgtgag 960
atgttgggtt aagtcccgca acgagcgcaa ccctcgcctt tagttgccag cacgtctggg 1020
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ctcatggccc ttatgtcctg ggctacacac gtgctacaat ggcggtgaca gtgggaagcc 1140
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Claims (10)
1. coltfoal shape bacillus (Komagataeibacter sp.) ATC301 of one plant of production bacteria cellulose, which is characterized in that it is protected
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.17875.
2. including the microbial inoculum of coltfoal shape bacillus ATC301 described in claim 1.
3. application of the microbial inoculum described in coltfoal shape bacillus ATC301 or claim 2 described in claim 1 in bacteria cellulose production.
4. application according to claim 3, which is characterized in that the application is by cultivating the coltfoal shape bacillus ATC301
Obtain bacteria cellulose；The condition of the culture is 28~32 DEG C, pH 4.0~7.0.
5. application according to claim 4, which is characterized in that the condition of the culture is 29~31 DEG C, pH4.5~5.5.
6. microbial inoculum described in coltfoal shape bacillus ATC301 or claim 2 described in claim 1 produces bacterial strain in building bacteria cellulose
In application.
7. a kind of production method of bacteria cellulose, which is characterized in that it is to utilize coltfoal shape bacillus described in claim 1
ATC301 produces bacteria cellulose.
8. production method according to claim 7, which is characterized in that make comprising the culture medium of glucose and/or sucrose
For fermentation raw material, coltfoal shape bacillus ATC301 is subjected to dynamic fermentation or static fermentation, obtains the bacteria cellulose.
9. production method according to claim 7 or 8, which comprises the steps of:
(1) actication of culture: the coltfoal shape bacillus ATC301 is inoculated on activation plate or inclined-plane, cultivates 2~4 in 28~32 DEG C
It；
(2) seed culture: the coltfoal shape bacillus ATC301 activated in step (1) is inoculated in seed culture medium, in 28~32 DEG C,
Oscillation or stir culture are mature to seed；
(3) fermented and cultured: the mature seed of the coltfoal shape bacillus ATC301 is inoculated in fermentation medium, in 28~32 DEG C,
Oscillation, stirring or stationary culture.
10. production method according to claim 9, which is characterized in that the culture medium of the fermented and cultured includes such as the following group
Point: 20~40g/L sugar source, 10~25g/L corn pulp, 0.5~1g/L epsom salt, 0.5~1g/L potassium dihydrogen phosphate, pH
5.0~5.5；The sugar source is selected from one or both of glucose, sucrose；
And/or
The culture medium of the seed culture includes following component: 10~30g/L sugar source, 5~15g/L corn pulp, 1~2g/L yeast
Powder, 0.1~0.5g/L epsom salt, 0.1~0.5g/L potassium dihydrogen phosphate, pH 5.0~5.5；The sugar source is selected from grape
One or both of sugar, sucrose.
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| CN116925974A (en) * | 2023-08-18 | 2023-10-24 | 江苏省农业科学院 | Bacterial cellulose-producing strain and application thereof |
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