CN101942493A - Production method of pullulan without pigment - Google Patents
Production method of pullulan without pigment Download PDFInfo
- Publication number
- CN101942493A CN101942493A CN2009101487663A CN200910148766A CN101942493A CN 101942493 A CN101942493 A CN 101942493A CN 2009101487663 A CN2009101487663 A CN 2009101487663A CN 200910148766 A CN200910148766 A CN 200910148766A CN 101942493 A CN101942493 A CN 101942493A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- seed
- air flow
- concentration
- described method
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a fermentation production method of pullulan without pigment. Aureobasidium pullulans, which are used as the starting strain, are induced by microwaves to obtain the mutant strain with high-yield pullulan and little pigment secretion. The fermentation production method of the pullulan comprises the following steps: (1) preparing seeds and fermentation medium; (2) inoculating the activated strain to the seed culture medium according to the inoculum size of 2-10%, and culturing at 28+/-1 DEG C for 24-36 hours, wherein the concentration of seeds is 0.8-2.5*108/mL; (3) inoculating the seed liquid to the fermentation medium according to the inoculum size of 2-10%, pressurizing to 0.01-0.05 Mpa in a tank at 28+/-1 DEG C, adjusting the stirring rate and ventilation amount at different fermentation stages and fermenting for 60-72 hours; and (4) after-treatment of fermentation liquid: using decoloration of the fermentation liquid, plate-and-frame filter, ceramic membrane ultrafiltration ultrafiltering for the sugar solution and the other methods to carrying out ultrafiltration and concentratuion, and performing spray drying to obtain the pullulan product.
Description
Technical field
The present invention relates to microbial fermentation and produce the method for pulullan.
Background technology
Pulullan (Pullulan) is by a kind of microbial polysaccharide soluble in water of Aureobasidium pullulans excretory, and nineteen thirty-nine R.Baner at first finds a kind of emplastic, i.e. pulullan in the fermented liquid of aureobasidium pullulans (Aureobasidium pullulans).Nineteen fifty-nine Bender etc. has at first characterized the pulullan of Aureobasidium pullulans fermentation generation and has been made up of neutral glucose, it is that glucose is by α-1, the 4-glycosidic link is combined into trisaccharide maltose, two ends are again with α-1, the 6-glycosidic link macromolecule polysaccharide that is formed by connecting repeatedly, be a kind of soluble in water, safety non-toxic evil, edible, low-calorie natural polysaccharide, because of its plasticity-, good film-forming property, book film gas barrier properties good, so commonly used its as protecting look, protecting fragrant, fresh-keeping, anti-oxidant etc. wrapping material.Pulullan can be used as coating or wrapping material widely; As the replacer of starch, it can be as making low in calories and protective foods; Simultaneously can be widely used in cosmetic industry; In pharmaceutical industries, it can be used as tamanori, thickening material and capsule capsule material; Also can be used as biological flocculant and be used for sewage disposal; In addition, pulullan also has more extensively application at aspects such as petrochemical complex, agricultural, papermaking.The production of pulullan and use 30 years of researches history have been arranged, Japanese Lin Yuan company carries out the pilot scale suitability for industrialized production in the seventies, monopolize the world market so far always.Domestic also have bigger progress through research in 20 years, but still there is more problem, wherein produce the key issue of the transformation efficiency problem of melanochrome and substrate sugar in the Aureobasidium pullulans fermenting process for the restriction suitability for industrialized production, and the time of the present fermentative production pulullan that ferments is generally about 120h, cause fermentation efficiency to descend, lot of domestic and international scientific research institution is difficult to solve above several problems simultaneously at present, therefore makes course of industrialization quite slow.
The present invention uses the microwave irradiation method to obtain the low pigment mutant strain of high yield, utilize this bacterial strain to produce pulullan, by fermentation time stirring velocity and air flow are regulated and control during the fermentation simultaneously, increase the output of pulullan, reduce the pigment secretion, shorten fermentation time greatly, improved the production efficiency of pulullan.
Summary of the invention
Microwave irradiation operation steps: the preparation of a. bacteria suspension: the Aureobasidium pullulans that will cultivate 24h is diluted to 10 with stroke-physiological saline solution
6Individual/mL, b. draw the bacteria suspension that makes, inject the smooth plate in bottom, the suspension amount of each plate is 10mL, the accent microwave power is 700W, pulse-repetition is 2450MHz, by different treatment times (being generally less than 1min), spore suspension is carried out radiation treatment, from each plate, take out the bacteria suspension of 0.1mL then respectively, suitably dilute, obtain different dilution bacteria suspensions, c. draws the bacteria suspension 0.3mL after the above-mentioned dilution, coating isolation medium flat board, place 30 ℃ of thermostat containers to cultivate 3d then, live bacterial count calculates lethality rate.As the primary dcreening operation sign, the bacterium colony that picking ratio is big and color is white goes down to posterity on the inclined-plane 3 times, carries out shake flask fermentation then and sieves again with the ratio of colony diameter on the separating plate and colony colour.
Seed culture medium and seed liquor preparation: a. seed culture medium consists of: sucrose 5%; Yeast extract paste 0.2%; NaCl 0.1%; MgSO
47H
2O 0.02%; K
2HPO
43H
2O 0.63%; (NH4)
2SO
40.06%, initial pH 6.5.115 ℃ of sterilization 20min; B. seed liquor preparation: gained Aureobasidium pullulans mutant strain in the step (1) is inoculated in the 500mL triangular flask that 1/5 seed culture medium is housed, 28 ± 1 ℃ of culture temperature, shaking speed 250r/min cultivates 24～36h, seed concentration 0.8～2.5 * 10
8Individual/mL, as first order seed, first order seed is inserted seed culture medium by 2%～10%, be extended to suitable corresponding inoculum size step by step, as fermentation seed liquid.
Fermention medium preparation and fermentation control: the used substratum of fermentative production is: sucrose 10%; Yeast extract paste 0.2%; NaCl0.1%; MgSO
47H
2O 0.02%; K
2HPO
43H
2O 0.63%; (NH4)
2SO
40.06%, initial pH 6.5.By 2%～10% inoculum size seed liquor is inserted fermention medium, 28 ± 1 ℃ of culture temperature, tank pressure 0.01～0.05Mpa.24h before the fermentation, stirring velocity 150～240r/min, air flow 1～2L/min.Cell concentration reaches 1.5～2.5 * 10 after the 24h
8Individual/mL, transfer stirring velocity to 400～650r/min, air flow 3～5L/min.After the 48h, stirring velocity is adjusted to 200～300r/min, and air flow 5～8L/min is until fermentation ends.
Fermented liquid downstream processing technology: the decolouring of (1) fermented liquid: fermented liquid adds 0.1～0.8% gac, transfers pH to 3.0～4.0, is heated to 65～85 ℃, stirs 15～30min; Leave standstill 10～20min after stirring end; (2) filtration sterilization: use 150～500 purpose screen plates to filter, wherein the flocculating aids of Jia Ruing can be perlite, diatomite or activated clay etc.; (3) ultrafiltration and concentration: use the ultrafiltration apparatus of types such as tubular type, spiral wound, tubular fibre formula to carry out ultrafiltration and concentration, obtain concentrating the back liquid glucose; (4) spraying drying: use spray drying devices such as rotary, pressure type, air-flowing type to carry out spraying drying, obtain Powdered pulullan polysaccharide product to concentrating the back liquid glucose.
Specific implementation method
Embodiment one: 30L ferment tank and downstream processing
Seed culture medium: sucrose 5%; Yeast extract paste 0.2%; NaCl 0.1%; MgSO
47H
2O 0.02%; K
2HPO
43H
2O0.63%; (NH4)
2SO
40.06%, initial pH 6.5.115 ℃ of sterilization 20min.
The used substratum of fermentative production is: sucrose 10%; Yeast extract paste 0.2%; NaCl 0.1%; MgSO
47H
2O 0.02%; K
2HPO
43H
2O 0.63%; (NH4)
2SO
40.06%, initial pH 6.5.
Seed liquor is cultivated: middle gained Aureobasidium pullulans mutant strain is inoculated in the 500mL triangular flask that 1/5 seed culture medium is housed, and 28 ± 1 ℃ of culture temperature, shaking speed 250r/min cultivates 24h, seed concentration 2.0 * 10
8Individual/mL, as first order seed, first order seed is equipped with the bacterial classification access in the 500mL triangular flask of 1/5 seed culture medium by 2% inoculum size, 28 ± 1 ℃ of culture temperature, shaking speed 250r/min cultivates 24h as secondary seed solution.
30L ferment tank: the seed liquor access is contained in the 30L fermentor tank of 20L fermention medium 28 ± 1 ℃ of culture temperature, tank pressure 0.03Mpa by 2% inoculum size.24h before the fermentation, stirring velocity 180r/min, air flow 1L/min.Cell concentration reaches 1.8 * 10 after the 24h
8Individual/mL, transfer stirring velocity to 500r/min, air flow 4L/min.After the 48h, stirring velocity is adjusted to 230r/min, and air flow 6.5L/min continues fermentation 12h, fermentation ends, and sugared transformation efficiency is 68%.
Fermented liquid downstream processing: a. is if the fermented liquid color is more then carried out the fermented liquid decolouring: fermented liquid adds 0.2% gac, transfers pH to 3.5, is heated to 70 ℃, stirs 20 minutes; Leave standstill 20min after stirring end; B. filtration sterilization: use 200 purpose screen plates to filter, wherein add massfraction and be 3% perlite and carry out filtration sterilization as flocculating aids; C. ultrafiltration and concentration: use the ceramic membrane ultrafitration system to carry out ultrafiltration and concentration, obtain concentrating the back liquid glucose; D. spraying drying: centrifugal spray drying equipment carries out spraying drying to concentrating the back liquid glucose, obtains Powdered pulullan polysaccharide product.The pulullan polysaccharide yield is 89%.
Embodiment two: 1m
3Ferment tank and downstream processing
Seed culture medium: sucrose 5%; Yeast extract paste 0.2%; NaCl 0.1%; MgSO
47H
2O 0.02%; K
2HPO
43H
2O0.63%; (NH4)
2SO
40.06%, initial pH 6.5.115 ℃ of sterilization 20min.
The used substratum of fermentative production is: glucose 10%; Yeast extract paste 0.2%; NaCl 0.1%; MgSO
47H
2O 0.02%; K
2HPO
43H
2O 0.63%; (NH4)
2SO
40.06%, initial pH 6.5.
Seed liquor is cultivated: middle gained Aureobasidium pullulans mutant strain is inoculated in the 500mL triangular flask that 1/5 seed culture medium is housed, and 28 ± 1 ℃ of culture temperature, shaking speed 250r/min cultivates 24min, seed concentration 2.0 * 10
8Individual/mL, as first order seed, first order seed is equipped with the bacterial classification access in the 500mL triangular flask of 1/5 seed culture medium by 2% inoculum size, 28 ± 1 ℃ of culture temperature, shaking speed 250r/min cultivates 24h as secondary seed solution.By 2% inoculum size the secondary seed solution access is contained in the 30L fermentor tank of 20L seed culture medium, 28 ± 1 ℃ of culture temperature, tank pressure 0.03Mpa, stirring velocity 180r/min, air flow 1L/min cultivates 24h, as three grades of seeds.
1m
3Ferment tank: seed liquor is inserted the 1m that contains the 700L fermention medium by 2% inoculum size
3In the fermentor tank, 28 ± 1 ℃ of culture temperature, tank pressure 0.03Mpa.24h before the fermentation, stirring velocity 180r/min, air flow 1L/min.Cell concentration reaches 1.8 * 10 after the 24h
8Individual/mL, transfer stirring velocity to 450r/min, air flow 4L/min.After the 48h, stirring velocity is adjusted to 250r/min, and air flow 6.0L/min continues fermentation 12h, fermentation ends, and sugared transformation efficiency is 67%.
Fermented liquid downstream processing: a. is if the fermented liquid color is more then carried out the fermented liquid decolouring: fermented liquid adds 0.2% gac, transfers pH to 3.5, is heated to 70 ℃, stirs 20min; Left standstill 20 minutes after stirring end; B. filtration sterilization: use 200 purpose screen plates to filter, wherein add massfraction and be 3% perlite and carry out filtration sterilization as flocculating aids; C. ultrafiltration and concentration: use the ceramic membrane ultrafitration system to carry out ultrafiltration and concentration, obtain concentrating the back liquid glucose; D. spraying drying: centrifugal spray drying equipment carries out spraying drying to concentrating the back liquid glucose, obtains Powdered pulullan polysaccharide product.The pulullan polysaccharide yield is 88%.
Embodiment three: 10m
3Ferment tank and downstream processing
Seed culture medium: sucrose 5%; Yeast extract paste 0.2%; NaCl 0.1%; MgSO
47H
2O 0.02%; K
2HPO
43H
2O0.63%; (NH4)
2SO
40.06%, initial pH 6.5.115 ℃ of sterilization 20min.
The used substratum of fermentative production is: sucrose 10%; Yeast extract paste 0.2%; NaCl 0.1%; MgSO
47H
2O 0.02%; K
2HPO
43H
2O 0.63%; (NH4)
2SO
40.06%, initial pH 6.5.
Seed liquor is cultivated: middle gained Aureobasidium pullulans mutant strain is inoculated in the 500mL triangular flask that 1/5 seed culture medium is housed, and 28 ± 1 ℃ of culture temperature, shaking speed 250r/min cultivates 24h, seed concentration 2.0 * 10
8Individual/mL, as first order seed, first order seed is equipped with the bacterial classification access in the 8L fermentor tank of 5L seed culture medium by 2% inoculum size, 28 ± 1 ℃ of culture temperature, tank pressure 0.03Mpa, stirring velocity 180r/min, air flow 1L/min cultivates 24h as secondary seed solution.By 2% inoculum size the secondary seed solution access is contained in the 300L fermentor tank of 200L seed culture medium, 28 ± 1 ℃ of culture temperature, tank pressure 0.03Mpa, stirring velocity 180r/min, air flow 1L/min cultivates 24h, as three grades of seeds.
10m
3Ferment tank: seed liquor is inserted the 10m that contains the 7000L fermention medium by 2% inoculum size
3In the fermentor tank, 28 ± 1 ℃ of culture temperature, tank pressure 0.03Mpa.24h before the fermentation, stirring velocity 150r/min, air flow 1L/min.Cell concentration reaches 2.0 * 10 after the 24h
8Individual/mL, transfer stirring velocity to 380r/min, air flow 4L/min.After the 48h, stirring velocity is adjusted to 200r/min, and air flow 6.0L/min continues fermentation 12h, fermentation ends, and sugared transformation efficiency is 69%.
Fermented liquid downstream processing: a. is if the fermented liquid color is more then carried out the fermented liquid decolouring: fermented liquid adds 0.2% gac, transfers pH to 3.5, is heated to 70 ℃, stirs 20min; Leave standstill 20min after stirring end; B. filtration sterilization: use 200 purpose screen plates to filter, wherein add massfraction and be 3% perlite and carry out filtration sterilization as flocculating aids; C. ultrafiltration and concentration: use the ceramic membrane ultrafitration system to carry out ultrafiltration and concentration, obtain concentrating the back liquid glucose; D. spraying drying: centrifugal spray drying equipment carries out spraying drying to concentrating the back liquid glucose, obtains Powdered pulullan product.The pulullan yield is 90%.
Claims (8)
1. pulullan non-pigment production method is characterized in that: be made up of following steps:
(1) will activate the back bacterial classification by 2%～10% inoculum size and receive in the seed culture medium and cultivate, obtain seed liquid;
(2) by 2%～10% inoculum size seed liquor is received fermention medium, regulate stirring velocity and air flow in the different steps of fermentation, fermentation 60～72h obtains fermented liquid;
(3) fermented liquid obtains pulullan through downstream processing technology.
2. the described method of claim 1, wherein, bacterial classification is that Aureobasidium pullulans is through the microwave irradiation gained after the activation in the step (1).
3. the described method of claim 1, wherein, fermention medium consists of carbon source 10%, yeast extract paste 0.2%, NaCl 0.1%, MgSO in the step (2)
47H
2O 0.02%, K
2HPO
43H
2O 0.63%, (NH4)
2SO
40.06%, all the other are water, initial pH 6.5.
4. the described method of claim 1, wherein, the culture temperature of step (1) is 28 ± 1 ℃, incubation time is 24～36h, seed concentration 0.8～2.5 * 10
8Individual/mL; The culture temperature of step (2) is 28 ± 1 ℃, tank pressure 0.01～0.05Mpa.
5. the described method of claim 1, wherein, stirring velocity and air flow in the step (2) are: 24h before the fermentation, stirring velocity 150～240r/min, air flow 1～2L/min; Cell concentration reaches 1.5～2.5 * 10 after the 24h
8Individual/mL, transfer stirring velocity to 400～650r/min, air flow 3～5L/min; After the 48h, stirring velocity is adjusted to 200～300r/min, and air flow 5～8L/min is until fermentation ends.
6. the described method of claim 1, wherein, step (3) middle and lower reaches treatment process comprises fermented liquid decolouring, filtration sterilization, ultrafiltration and concentration and spraying drying.
7. the described method of claim 6, wherein, the fermented liquid decoloring method is: fermented liquid adds 0.1～0.8% gac, transfers pH to 3.0～4.0, is heated to 65～85 ℃, stirs 15～30min; Leave standstill 10～20min after stirring end; The method of filtration sterilization is: use 150～500 purpose screen plates to filter, wherein add flocculating aids perlite, diatomite and/or activated clay 0～an amount of; The method of ultrafiltration and concentration is: use the ultrafiltration apparatus of types such as tubular type, spiral wound, tubular fibre formula to carry out ultrafiltration and concentration; Spray-dired method is: use spray drying devices such as rotary, pressure type or air-flowing type to carry out spraying drying.
8. the described method of claim 2, wherein, the method for microwave irradiation is:
(1) preparation of bacteria suspension: the Aureobasidium pullulans that will cultivate 24h is diluted to 10 with stroke-physiological saline solution
6Individual/mL;
(2) draw the bacteria suspension that makes, inject the smooth plate in bottom, the suspension amount of each plate is 10mL, the accent microwave power is 700W, pulse-repetition is 2450MHz, by the different treatment time (being generally less than 1min) spore suspension is carried out radiation treatment, takes out the bacteria suspension of 0.1mL then respectively from each plate, suitably dilute, obtain different dilution bacteria suspensions;
(3) the bacteria suspension 0.3mL after the above-mentioned dilution of absorption, coating isolation medium flat board, place 30 ℃ of thermostat containers to cultivate 3d then, with the ratio of colony diameter on the separating plate and colony colour as the primary dcreening operation sign, the bacterium colony that picking ratio is big and color is white goes down to posterity on the inclined-plane 3 times, carries out shake flask fermentation then and sieves again.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101487663A CN101942493A (en) | 2009-07-03 | 2009-07-03 | Production method of pullulan without pigment |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101487663A CN101942493A (en) | 2009-07-03 | 2009-07-03 | Production method of pullulan without pigment |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101942493A true CN101942493A (en) | 2011-01-12 |
Family
ID=43434698
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009101487663A Pending CN101942493A (en) | 2009-07-03 | 2009-07-03 | Production method of pullulan without pigment |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101942493A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102994395A (en) * | 2012-10-15 | 2013-03-27 | 苏州大学 | Aureobasidium pullulans and application thereof |
| CN103172757A (en) * | 2012-12-31 | 2013-06-26 | 天津北洋百川生物技术有限公司 | Extracting process of pulullan polysaccharide |
| CN103740785A (en) * | 2013-12-28 | 2014-04-23 | 天津北洋百川生物技术有限公司 | Method for producing pullulan through high-density fermentation |
| CN104448019A (en) * | 2014-12-05 | 2015-03-25 | 通辽梅花生物科技有限公司 | High-purity Pulullan extraction process |
| CN104479038A (en) * | 2014-12-05 | 2015-04-01 | 通辽梅花生物科技有限公司 | Pulullan purification technique |
| US10568839B2 (en) | 2011-01-11 | 2020-02-25 | Capsugel Belgium Nv | Hard capsules |
| US11319566B2 (en) | 2017-04-14 | 2022-05-03 | Capsugel Belgium Nv | Process for making pullulan |
| US11576870B2 (en) | 2017-04-14 | 2023-02-14 | Capsugel Belgium Nv | Pullulan capsules |
-
2009
- 2009-07-03 CN CN2009101487663A patent/CN101942493A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10568839B2 (en) | 2011-01-11 | 2020-02-25 | Capsugel Belgium Nv | Hard capsules |
| CN102994395A (en) * | 2012-10-15 | 2013-03-27 | 苏州大学 | Aureobasidium pullulans and application thereof |
| CN103172757A (en) * | 2012-12-31 | 2013-06-26 | 天津北洋百川生物技术有限公司 | Extracting process of pulullan polysaccharide |
| CN103172757B (en) * | 2012-12-31 | 2015-01-14 | 天津北洋百川生物技术有限公司 | Extracting process of pulullan polysaccharide |
| CN103740785A (en) * | 2013-12-28 | 2014-04-23 | 天津北洋百川生物技术有限公司 | Method for producing pullulan through high-density fermentation |
| CN104448019A (en) * | 2014-12-05 | 2015-03-25 | 通辽梅花生物科技有限公司 | High-purity Pulullan extraction process |
| CN104479038A (en) * | 2014-12-05 | 2015-04-01 | 通辽梅花生物科技有限公司 | Pulullan purification technique |
| CN104479038B (en) * | 2014-12-05 | 2016-09-28 | 通辽梅花生物科技有限公司 | Pulullan polysaccharide purifying technique |
| US11319566B2 (en) | 2017-04-14 | 2022-05-03 | Capsugel Belgium Nv | Process for making pullulan |
| US11576870B2 (en) | 2017-04-14 | 2023-02-14 | Capsugel Belgium Nv | Pullulan capsules |
| US11878079B2 (en) | 2017-04-14 | 2024-01-23 | Capsugel Belgium Nv | Pullulan capsules |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101942493A (en) | Production method of pullulan without pigment | |
| CN101215592B (en) | Fermentation method for producing pullulan polysaccharide | |
| CN100365128C (en) | Method for preparing bacteria cellulose | |
| CN101659924B (en) | Aspergillus niger strain and application thereof in preparing fructo-oligosaccharide by anaerobic fermentation | |
| CN100448979C (en) | Production method for xylose by enzyme process | |
| CN108441528B (en) | Culture medium for efficiently producing bacterial cellulose | |
| CA3121566C (en) | Aspergillus oryzae blcy-006 strain and application thereof in preparation of galactooligosaccharides | |
| CN101513161A (en) | Formula for culture medium of cordyceps militaris liquid strains and method for culturing same | |
| CN102154408B (en) | Sclerotium rolfssii scleroglucan online fermentation extraction method and system | |
| CN103805650A (en) | Fermentation production method for pigment-free pullulan | |
| CN101942492A (en) | Method for producing pullulan with different molecular weights | |
| CN104087531A (en) | Alcaligenes faecalis mutant strain and method for preparing curdlan by using alcaligenes faecalis mutant strain | |
| CN102296032B (en) | Transglucosidase, its preparation method and immobilization method | |
| CN107641634A (en) | A kind of production technology of Aureobasidium pullulans fermenting and producing pulullan polysaccharide | |
| CN102492752A (en) | Method for co-producing pullulan polysaccharide and melanin by Aureobasidium pullulan | |
| CN102559799A (en) | Preparation method for algae endophytic fungi exocellular polysaccharide | |
| CN101487033A (en) | Preparation of bacteria cellulose special-shaped product by microbial fermentation direct biosynthesis | |
| CN103045489A (en) | Aspergillus niger and method for catalytically producing fructo-oligosaccharide by virtue of whole-cells of aspergillus niger | |
| CN107801938A (en) | A kind of biometrics technology method of betel nut | |
| CN101012466A (en) | Method for preparing low-polyxylose by microwave treatment corn core enzymatical process | |
| CN105543312A (en) | Preparation method of microbial fermentation-based N-acetyle-D-glucosamine | |
| CN100529055C (en) | Alcaligenes and application thereof in preparation of welan gum | |
| CN103589758A (en) | Method for preparing gallic acid by using gallnut tannin as raw material and utilizing fermentation separation coupling technology | |
| CN105420311A (en) | Novel technology for producing extracellular polysaccharides and fructose by means of litchi juice or longan juice fermentation | |
| CN102851328A (en) | Method for preparing citric acid through fermenting corn sugar solution by immobilized Aspergillus niger |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20110112 |