CN101314788A - Method for bacteria cellulose high yield bacterial strain cultivation sifting motion - Google Patents
Method for bacteria cellulose high yield bacterial strain cultivation sifting motion Download PDFInfo
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- CN101314788A CN101314788A CNA2008100538254A CN200810053825A CN101314788A CN 101314788 A CN101314788 A CN 101314788A CN A2008100538254 A CNA2008100538254 A CN A2008100538254A CN 200810053825 A CN200810053825 A CN 200810053825A CN 101314788 A CN101314788 A CN 101314788A
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Abstract
The invention relates to a method for culturing and screening a high-yielding strain of bacteria cellulose, which comprises the following steps: (1) inoculating wood vinegar bacillus into a seed culture medium; simultaneously, adding cellulolytic enzyme in the culture medium for filtration and sterilization, with the amount of the cellulolytic enzyme in the culture medium per milliliter being 0.5U-8U; culturing at the temperature of 27 to 30 DEG C for 14 to 20hours; and obtaining bacterial suspension with the thallus concentration of 10<6>-10<8>/ml; (2) performing the mutagenic treatment to the bacterial suspension obtained; (3) diluting the thallus concentration of the bacterial suspension of the wood vinegar bacillus to 10<3>-10<5>/ml; coating the bacterial suspension on an agar medium with NaBr and NaBrO3, with the weight ratio of NaBr and NaBrO in the culture medium being 5:1, so that the concentration of NaBr in the solid culture reaches 1mmol/L-5mmol/L; culturing at the temperature of 27 to 30 DEG C; and performing the cellulose yield determination of live colony selected from the NaBrand and NaBrO3agar medium. The high-yielding strain is obtained after being cultured and screened by adopting the method. The method plays an important role in the commercial application of bacteria cellulose.
Description
Technical field
The present invention relates to the microorganism breeding technique, particularly a kind of method of bacteria cellulose high yield bacterial strain cultivation sifting motion.
Background technology
Mierocrystalline cellulose is a natural polymers the abundantest on the earth, extensively exists in the plants such as trees, cotton, and the annual Mierocrystalline cellulose that is produced by plant reaches hundreds of millions tons.But synthetic cellulose is not the plant specific function, and some bacterium also produces born of the same parents' outer fiber element in the heterotrophism mode than plant more efficiently, and we call " bacteria cellulose " to the Mierocrystalline cellulose of this bacterial origin.The bacterium synthetic cellulose 1886 by the Brown reported first, being wood vinegar bacterium (Acetobacter xylinum) is leaving standstill one deck white fiber shape material that forms in media surface when cultivating.Afterwards many gram negative bacteriums.As also having found the generation of bacteria cellulose in edaphic bacillus, tumorigenesis purulence bacillus and gram positive bacterium such as the sarcina.Bacteria cellulose and natural cellulose structure are closely similar, all be D-glucose with the chain polymer that β-1,4 glycosidic link is formed by connecting, have (C
6H
10O
5)
nComposition.But by comparison, bacteria cellulose has more superior characteristic, and it is that form with pure cellulose exists, and the existence form of plant cellulose is and hemicellulose and xylogen etc. are formed three grades of three-dimensional arrangements.Bacteria cellulose also has high-crystallinity, high strength and good hydrophilicity energy simultaneously.
The bacteria cellulose application of having achieved success at present at food, medicine, chemical industry, papermaking, high-end audio equipment, filter membrane permeable membrane and aspect such as spinning.But since it that low yield caused was expensive, limited its widespread use.Therefore be necessary that the Mierocrystalline cellulose generation bacterium that filters out high yield is applied to industrial production, to improve output, reduces cost.Acetobacter xylinum (Acetobacter xylinum) is to produce the strongest bacterial strain of Mierocrystalline cellulose ability, is the aerobic gram negative bacterium of obligate, and it can the secreting bacteria Mierocrystalline cellulose under normal metabolism condition.Usually can be by the mode breeding high-yield Mierocrystalline cellulose bacterial strain of various physical mutagenesis or chemomorphosis, to improve cellulosic output.
Acetobacter xylinum can be wrapped up by the Mierocrystalline cellulose that self generates in producing cellulosic process, so the somatic cells suspension of preparation high density is a difficult problem.Acetobacter xylinum is utilizing carbon source (sucrose, glucose) metabolism to produce the cellulosic while, can become gluconic acid and ketone gluconic acid etc. to conversion of glucose, and fermented liquid pH value is reduced.When pH is reduced to a certain degree, will limit the growth of acetobacter xylinum self, cause the reduction of cellulose output.
Therefore, providing a kind of method of bacteria cellulose high yield bacterial strain cultivation sifting motion, effectively solve the problem that prior art exists, is one of this technical field scientific research personnel new problem of being badly in need of developing.
Summary of the invention
The objective of the invention is for overcoming the weak point of prior art, provide that a kind of implementation step is simple, the method for the bacteria cellulose high yield bacterial strain cultivation sifting motion of effect highly significant.
Implementation step of the present invention for achieving the above object is as follows:
(1) adds cellulase and prepare the acetobacter xylinum bacteria suspension
Acetobacter xylinum is inoculated in the seed culture medium, the amount of cellulase is cultivated down at 27-30 ℃ then at 0.5U-8U in the cellulase that adds simultaneously filtration sterilization in substratum, every milliliter of substratum, incubation time 14-20h, the cell concentration of the bacteria suspension of acquisition is 10
6-10
8Individual/ml;
(2) bacteria suspension that obtains is carried out mutagenic treatment
The bacteria suspension that makes is changed in the culture dish of the bacterium of going out, be placed on the magnetic stirring apparatus, carry out ultraviolet mutagenesis, mutation time is 1-5min;
(3) by NaBr and NaBrO
3Nutrient agar proton suicide method is screened
The cell concentration of the acetobacter xylinum bacteria suspension after the mutagenesis is diluted to 10
3-10
5Individual/ml, coat and contain NaBr and NaBrO
3Nutrient agar on, NaBr and NaBrO
3Join in the substratum with 5: 1 ratio of weight ratio, make the concentration of NaBr in solid medium reach 1mmol/L-5mmol/L; Cultivate down at 27-30 ℃, from NaBr and NaBrO
3The bacterium colony of picking survival carries out cellulosic determination of yield on the nutrient agar.
Add bromide and bromate in substratum, when bacterial strain utilized carbohydrate to produce acid, the pH value will descend, and substratum is acid to be increased.Under higher acidic conditions, bromide and bromate react generation bromine, i.e. BrO
3 -+ 2Br
-+ 2H
+→ Br
2+ BrO
2 -+ H
2O.Deleterious bromine can kill bacterial strain, proton suicide method that Here it is.And those are not killed and on the substratum survival under bacterial strain, be exactly gluconic acid defective bacterial strain.
The present invention adopts the mode that adds cellulase in substratum, finds that cellulase can not suppress the growth of cell, and can not produce cellulose membrane, so just can obtain the bacteria suspension of high density.Diluted or removed from nutrient solution when the concentration of cellulase, thalline still can synthesize bacteria cellulose.
The invention has the beneficial effects as follows: the invention provides a kind of method of bacteria cellulose high yield bacterial strain cultivation sifting motion, enzyme-added preparation acetobacter xylinum bacteria suspension, and the bacteria suspension of the high density that makes according to this method carried out follow-up mutagenesis or other processing.Simultaneously, the enzyme-added method for preparing the acetobacter xylinum bacteria suspension also can be used for the fermentor tank inoculation of large volume, has solved the difficult problem that acetobacter xylinum can't obtain a large amount of seeds.By adopting enzyme-added legal system to be equipped with the acetobacter xylinum bacteria suspension, bacteria suspension is carried out mutagenic treatment, and with the inoculation after the mutagenesis and NaBr and NaBrO
3The enterprising row filter of screening culture medium.The superior strain that screening obtains, cellulose output has improved more than 54%.The screening method that adopts proton of the present invention to commit suiside, can rapidly and efficiently screen high yield bacteria cellulose bacterial classification, obtain glucose defective type acetobacter xylinum, what make the medium pH value can not drop to acid range, help the raising of bacteria cellulose output, significant to the business-like successful Application of bacteria cellulose.
Embodiment
Below in conjunction with embodiment, to details are as follows according to embodiment provided by the invention:
Embodiment 1
A kind of method of bacteria cellulose high yield bacterial strain cultivation sifting motion, implementation step is as follows:
(1) adds cellulase and prepare the acetobacter xylinum bacteria suspension
Getting initial bacterial strain acetobacter xylinum Z5 200 μ l is inoculated in the triangular flask that the 40ml seed culture medium is housed.The composition of seed culture medium is 70g glucose, 10g peptone, 2gMgSO
4, the 1g Trisodium Citrate, be dissolved in the 1L distilled water, pH 5.0.Add filtering cellulase solution then, the amount that makes cellulase in every milliliter of substratum is at 0.5U, 27 ℃ of static cultivation 14h, and the concentration of the bacteria suspension that makes is 2.2 * 10
6Individual/ml.
(2) bacteria suspension that obtains is carried out mutagenic treatment
The 15ml Z5 bacteria suspension that makes is changed in the diameter 9cm culture dish of the bacterium of going out, be placed on the magnetic stirring apparatus.Ultraviolet lamp 254nm, 20W place the place of media surface 20cm, open magnetic stirrer and make bacteria suspension keep the homogeneous state.Open ultraviolet lamp preheating 20min.Open the culture dish lid then and carry out mutagenesis, mutation time 1min.
(3) by NaBr and NaBrO
3Nutrient agar proton suicide method is screened
At first prepare NaBr and NaBrO
3Nutrient agar is about to contain NaBr 5mol/L and NaBrO
3The solution of 2mol/L carries out high-temperature sterilization respectively, and 121 ℃, 20min is then with NaBr and NaBrO
3Add solid medium with 5: 1 ratio of weight ratio: 50g glucose, 10g peptone, 2gMgSO
4, 1g Trisodium Citrate and 15g agar is dissolved in the 1L distilled water, pH 5.0, the concentration that makes NaBr in the solid medium is 1mmol/L.The mutagenic treatment sample that takes out 1ml carries out 1: 10 gradient dilution with physiological saline 0.85%NaCl.Cell concentration is diluted to 10
3-10
5Individual/ml, get bacterium liquid 100 μ l and be tiled in NaBr and NaBrO
3On the nutrient agar.27 ℃ of lucifuges are cultivated 8d, observe survival strains.
If it is strong that bacterial strain produces acid, just have deleterious bromine accumulation, so have only the gluconic acid deficient strain to survive.From NaBr and NaBrO
3Provoke residual viable bacteria on the agar enrichment medium and fall, change test tube over to and carry out primary dcreening operation, change triangular flask mensuration output then over to and carry out multiple sieve, the results are shown in Table 1.Each mutagenesis bacterial classification is compared acetobacter xylinum Z5, and output is improved largely.Wherein No. 30 bacterial strain output are the highest, called after acetobacter xylinum mutant strain Z530 (A.xylinum Z530), and the output of A.xylinum Z530 is 4.32g/L, and terminal point pH is 4.5, and output is compared original strain and has been improved 54%.
With the high yield acetobacter xylinum mutant strain Z530 continuous passage that obtains 5 times, measure this 5 cellulose outputs, the result proves that this bacterium produces the Mierocrystalline cellulose stable performance.
The superior strain that table 1 proton method for disinfection screening of the present invention obtains
| Bacterium numbering | Cellulose output (g/L) | Terminal point pH |
| 4 | 3.88 | 4.0 |
| 8 | 4.16 | 4.4 |
| 20 | 4.05 | 4.2 |
| 21 | 3.93 | 4.2 |
| 30 | 4.32 | 4.5 |
| Z5 (initial bacterial strain) | 2.80 | 2.8 |
Embodiment 2
A kind of method of bacteria cellulose high yield bacterial strain cultivation sifting motion, implementation step is as follows:
(1) adds cellulase and prepare the acetobacter xylinum bacteria suspension
Getting initial bacterial strain acetobacter xylinum Z5 200 μ l is inoculated in the triangular flask that the 40ml seed culture medium is housed.The composition of seed culture medium is 70g glucose, 10g peptone, 2gMgSO
4, the 1g Trisodium Citrate, be dissolved in the 1L distilled water, pH 5.0.Add filtering cellulase solution then, the amount that makes cellulase in every milliliter of substratum is at 8U, 30 ℃ of static cultivation 20h, and the concentration of the bacteria suspension that makes is 8.0 * 10
7Individual/ml.
(2) bacteria suspension that obtains is carried out mutagenic treatment
The 15ml Z5 bacteria suspension that makes is changed in the diameter 9cm culture dish of the bacterium of going out, be placed on the magnetic stirring apparatus.Ultraviolet lamp 254nm, 20W place the place of media surface 20cm, open magnetic stirrer and make bacteria suspension keep the homogeneous state.Open ultraviolet lamp preheating 20min.Open the culture dish lid then and carry out mutagenesis, mutation time 5min.
(3) by NaBr and NaBrO
3Nutrient agar screens
At first prepare NaBr and NaBrO
3Nutrient agar is about to contain NaBr 5mol/L and NaBrO
3The solution of 2mol/L carries out 121 ℃ of high-temperature sterilizations respectively, and 20min is then with NaBr and NaBrO
3Add solid medium with 5: 1 ratio of weight ratio: 50g glucose, 10g peptone, 2gMgSO
4, 1g Trisodium Citrate and 15g agar is dissolved in the 1L distilled water, pH 5.0, the concentration that makes NaBr in the solid medium is 5mmol/L.The mutagenic treatment sample that takes out 1ml carries out 1: 10 gradient dilution with physiological saline 0.85%NaCl.Cell concentration is diluted to 10
3-10
5Individual/ml, get bacterium liquid 100 μ l and be tiled in NaBr and NaBrO
3On the nutrient agar.30 ℃ of lucifuges are cultivated 8d, observe survival strains.
If it is strong that bacterial strain produces acid, just have deleterious bromine accumulation, so have only the gluconic acid deficient strain to survive.From NaBr and NaBrO
3Provoking residual viable bacteria on the agar enrichment medium falls, change test tube over to and carry out primary dcreening operation, change triangular flask mensuration output then over to and carry out multiple sieve, it is the highest that the result screens No. 72 bacterial strain output, called after acetobacter xylinum mutant strain Z572 (A.xylinum Z572), the output of A.xylinum Z572 is 4.36g/L, and terminal point pH4.2 compares original strain output and improved 56%.
With the high yield acetobacter xylinum mutant strain Z572 continuous passage that obtains 5 times, measure this 5 cellulose outputs, the result proves that this bacterium produces the Mierocrystalline cellulose stable performance.
Embodiment 3
A kind of method of bacteria cellulose high yield bacterial strain cultivation sifting motion, implementation step is as follows:
(1) adds cellulase and prepare the acetobacter xylinum bacteria suspension
Get initial bacterial strain acetobacter xylinum Z5 200 μ l and be inoculated in the test tube that the 7.5ml seed culture medium is housed, the composition of seed culture medium is 70g glucose, 10g peptone, 2gMgSO
4, the 1g Trisodium Citrate, be dissolved in the 1L distilled water, pH 5.0.Getting the 3ml seed liquor then is inoculated in the culture dish that contains 70ml mutagenesis liquid nutrient medium.The composition of mutagenesis liquid nutrient medium is 50g glucose, 10g peptone, 2gMgSO
4, the 1g Trisodium Citrate is dissolved in the 1L distilled water, pH 5.0.The culture dish diameter is 12cm, cultivates 9d for 30 ℃.Then the cellulose membrane that is shaped being moved into one under aseptic condition went out in the culture dish of bacterium, the culture dish diameter is 12cm, subsequently this culture dish is placed on the shaking table, 200 commentaries on classics/min, add the cellulase 1.5ml of 105U/ml, carry out enzymolysis, 30 ℃ of hydrolysis temperatures, enzymolysis time 12h discharges acetobacter xylinum from the fibre network that is degraded.Bacterium liquid in the culture dish behind the enzymolysis is sucked in the centrifuge tube of the bacterium of going out, in culture dish, add 15ml mutagenesis liquid nutrient medium at twice, thalline is fully washed.The centrifugal then supernatant of abandoning is suspended in thalline in the 15ml nutrient solution.Twice, 10000 rev/min of centrifuge washing, 20min, 4 ℃, with enzyme liquid flush away.At last thalline is suspended in the 15ml nutrient solution again, the concentration of the bacteria suspension that makes is 5.0 * 10
8Individual/ml.
(2) bacteria suspension that obtains is carried out mutagenic treatment
The 15ml bacteria suspension that makes is changed in the culture dish of the bacterium of going out, and culture dish diameter 9cm is placed on the magnetic stirring apparatus.Ultraviolet lamp 254nm, 20W places the place of media surface 20cm, opens magnetic stirrer and makes bacteria suspension keep the homogeneous state.Open ultraviolet lamp preheating 20min.Open the culture dish lid then and carry out mutagenesis, mutation time 3min.
(3) by NaBr and NaBrO
3The agar enrichment medium screens
At first prepare NaBr and NaBrO
3The agar enrichment medium is about to contain NaBr 5mol/L and NaBrO
3The solution of 2mol/L carries out high-temperature sterilization 21min respectively, and 121 ℃, then with NaBr and NaBrO
3Add solid medium with 5: 1 ratios: 50g glucose, 10g peptone, 2gMgSO
4, 1g Trisodium Citrate and 15g agar is dissolved in the 1L distilled water, pH 5.0, the concentration that makes NaBr in the solid medium is 4mmol/L.The mutagenic treatment sample that takes out 1ml carries out 1: 10 gradient dilution with physiological saline 0.85%NaCl.Cell concentration is diluted to 10
3-10
5Individual/ml, get bacterium liquid 100 μ l and be tiled in NaBr and NaBrO
3On the agar enrichment medium.30 ℃ of lucifuges are cultivated 8d, observe survival strains.
(4) bacterial classification primary dcreening operation
If it is strong that bacterial strain produces acid, just have the accumulation of deleterious Br, so have only the gluconic acid deficient strain to survive.From NaBr and NaBrO
3Provoke residual viable bacteria on the agar enrichment medium and fall, these bacterium colonies are forwarded in the test tube that contains 7.5ml mutagenesis liquid nutrient medium.30 ℃ of following static cultivation 8d, Z5 compares with acetobacter xylinum, and Mierocrystalline cellulose synthetic thickness can pass through visual determination in the test tube, selects the big mutagenic strain of rate ratio acetobacter xylinum Z5 to carry out next step multiple sieve.
(5) bacterial classification sieves again
Mutagenic strain behind the primary dcreening operation and acetobacter xylinum Z5 change in the 100ml triangular flask that 40ml mutagenesis liquid nutrient medium is housed together, and inoculum size is cultivated 8d for 5%, 30 ℃, measures cellulosic output.It is the highest that the result screens No. 48 bacterial strain output, called after acetobacter xylinum mutant strain Z548 (A.xylinum Z548), and the output of A.xylinum Z548 is 4.36g/L, terminal point pH4.5 compares original strain output and has improved 57%.
(6) stability test
With the high yield acetobacter xylinum mutant strain Z548 continuous passage that obtains 5 times, measure this 5 cellulose outputs, the result proves that this bacterium produces the Mierocrystalline cellulose stable performance.
This method can be cultivated quickly and efficiently and be filtered out the bacteria cellulose high-yield strains.The acetobacter gluconic acid defective strain that utilizes the proton suicide method to filter out, later stage pH is not reduced to acid range in fermentation, effectively improves cellulosic output.
Above-mentioned with reference to the method detailed description of embodiment to bacteria cellulose high yield bacterial strain cultivation sifting motion; be illustrative rather than determinate; can list several embodiment according to institute's limited range; therefore in the variation and the modification that do not break away under the general plotting of the present invention, should belong within protection scope of the present invention.
Claims (1)
1, a kind of method of bacteria cellulose high yield bacterial strain cultivation sifting motion, implementation step is as follows:
(1) adds cellulase and prepare the acetobacter xylinum bacteria suspension
Acetobacter xylinum is inoculated in the seed culture medium, the amount of cellulase is cultivated down at 27-30 ℃ then at 0.5U-8U in the cellulase that adds simultaneously filtration sterilization in substratum, every milliliter of substratum, incubation time 14-20h, the cell concentration of the bacteria suspension of acquisition is 10
6-10
8Individual/ml;
(2) bacteria suspension that obtains is carried out mutagenic treatment
The bacteria suspension that makes is changed in the culture dish of the bacterium of going out, be placed on the magnetic stirring apparatus, carry out ultraviolet mutagenesis, mutation time is 1-5min;
(3) by NaBr and NaBrO
3Nutrient agar proton suicide method is screened
The cell concentration of the acetobacter xylinum bacteria suspension after the mutagenesis is diluted to 10
3-10
5Individual/ml, coat and contain NaBr and NaBrO
3Nutrient agar on, NaBr and NaBrO
3Join in the substratum with 5: 1 ratio of weight ratio, make the concentration of NaBr in solid medium reach 1mmol/L-5mmol/L; Cultivate down at 27-30 ℃, from NaBr and NaBrO
3The bacterium colony of picking survival carries out cellulosic determination of yield on the nutrient agar.
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| CNA2008100538254A CN101314788A (en) | 2008-07-11 | 2008-07-11 | Method for bacteria cellulose high yield bacterial strain cultivation sifting motion |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101967471A (en) * | 2009-11-10 | 2011-02-09 | 东华大学 | Method for preparing immobilized enzyme by taking bacterial cellulose bead/membrane as vector |
| CN102586135A (en) * | 2011-12-30 | 2012-07-18 | 南京理工大学 | Bacteria and breeding method and method for producing cellulase thereof |
| TWI408232B (en) * | 2010-05-24 | 2013-09-11 | Nympheas Internationalcorp | Bacterial cellulose film, and use thereof |
| CN103289977A (en) * | 2013-07-01 | 2013-09-11 | 上海尚志生物科技有限公司 | Preparation and compounding methods of low-temperature neutral cellulase |
| CN103690995A (en) * | 2013-12-10 | 2014-04-02 | 深圳先进技术研究院 | Bioabsorbable fiber, as well as preparation method and application thereof |
| CN105861622A (en) * | 2016-05-26 | 2016-08-17 | 天津科技大学 | Viable bacterium counting method for bacterial cellulose production strains |
| CN106497789A (en) * | 2016-10-21 | 2017-03-15 | 天津工业大学 | A kind of screening technique of the bacterium bacterial strain with bacteria cellulose high productive capacity |
| CN106609295A (en) * | 2015-10-21 | 2017-05-03 | 广东东阳光药业有限公司 | Online evaluation method of concentration of acetobacter xylinus capable of generating cellulose |
| CN112708617A (en) * | 2019-10-25 | 2021-04-27 | 华东师范大学 | Method for rapidly screening bacterial cellulose strains with high yield |
-
2008
- 2008-07-11 CN CNA2008100538254A patent/CN101314788A/en active Pending
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101967471A (en) * | 2009-11-10 | 2011-02-09 | 东华大学 | Method for preparing immobilized enzyme by taking bacterial cellulose bead/membrane as vector |
| CN101967471B (en) * | 2009-11-10 | 2012-06-13 | 东华大学 | Method for preparing immobilized enzyme by taking bacterial cellulose bead/membrane as vector |
| EP2390344B1 (en) * | 2010-05-24 | 2015-06-03 | Nympheas International Corporation | Bacterial cellulose film and preparation thereof |
| TWI408232B (en) * | 2010-05-24 | 2013-09-11 | Nympheas Internationalcorp | Bacterial cellulose film, and use thereof |
| CN102586135A (en) * | 2011-12-30 | 2012-07-18 | 南京理工大学 | Bacteria and breeding method and method for producing cellulase thereof |
| CN103289977A (en) * | 2013-07-01 | 2013-09-11 | 上海尚志生物科技有限公司 | Preparation and compounding methods of low-temperature neutral cellulase |
| CN103690995A (en) * | 2013-12-10 | 2014-04-02 | 深圳先进技术研究院 | Bioabsorbable fiber, as well as preparation method and application thereof |
| CN106609295A (en) * | 2015-10-21 | 2017-05-03 | 广东东阳光药业有限公司 | Online evaluation method of concentration of acetobacter xylinus capable of generating cellulose |
| CN106609295B (en) * | 2015-10-21 | 2019-10-18 | 广东东阳光药业有限公司 | A kind of on-line evaluation method producing cellulose wood acetobacter cell concentration |
| CN105861622A (en) * | 2016-05-26 | 2016-08-17 | 天津科技大学 | Viable bacterium counting method for bacterial cellulose production strains |
| CN105861622B (en) * | 2016-05-26 | 2019-07-09 | 天津科技大学 | A kind of viable count method of bacteria cellulose production bacterial strain |
| CN106497789A (en) * | 2016-10-21 | 2017-03-15 | 天津工业大学 | A kind of screening technique of the bacterium bacterial strain with bacteria cellulose high productive capacity |
| CN112708617A (en) * | 2019-10-25 | 2021-04-27 | 华东师范大学 | Method for rapidly screening bacterial cellulose strains with high yield |
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Application publication date: 20081203 |