CN101486984B - Gluconobacter oxydans and method for preparing ketoxylose using the same - Google Patents
Gluconobacter oxydans and method for preparing ketoxylose using the same Download PDFInfo
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- CN101486984B CN101486984B CN2009100245794A CN200910024579A CN101486984B CN 101486984 B CN101486984 B CN 101486984B CN 2009100245794 A CN2009100245794 A CN 2009100245794A CN 200910024579 A CN200910024579 A CN 200910024579A CN 101486984 B CN101486984 B CN 101486984B
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Abstract
The invention discloses a Gluconobacter oxydans, which is classified and named as Gluconobacter oxydans NH-10, and preserved in China General Microbiological Culture Center of Microbial Culture Collection Management Committee with the preservation number of CGMCC No.2709. The invention further discloses a D-xylulose preparation method that uses the Gluconobacter oxydans. The strain obtained by screening can transform D-Arabitol so as to generate D-xylulose in high efficiency, the D-xylulose preparation method reaches the highest D-Arabitol transformation ratio of 99.5 percent (w/w) and the D-xylulose yield ratio of 95 percent, the D-xylulose concentration of a conversion fluid can reach 95g/L, and the conversion fluid hardly contains other ingredients.
Description
Technical field
The invention belongs to fermentation engineering and technical field of enzyme engineering, relate to a kind of microbiological oxidation gluconobacter sp (Gluconobacter oxydans) of the D-of producing pectinose alcoholdehydrogenase, and it is used to produce the method for D-xylulose.
Background technology
Xylulose (xylulose) is a kind of ketopentose.The L-xylulose is discharged from the urine of pentosuria.The xylulose of L type and D type is reduced into Xylitol earlier in vivo, and that continues reoxidizes and conversion mutually.D-xylulose and ATP reaction generate D-xylulose-5-phosphoric acid and enter phosphopentose pathway.Grape glucose is through 6-phosphoric acid grape saccharic acid → D-ribulose-5-phosphoric acid forms D-xylulose-5-phosphoric acid, is the important intermediate of the phosphopentose pathway of another glycolytic pathway in addition.In addition, the D-xylulose can be used as the material of producing Xylitol, and Xylitol is one of the important sweeting agent in fields such as foodstuffs industry.Xylitol is natural five-carbon sugar alcohol, and its sugariness is 1.05 times of sucrose, and heat and sucrose are suitable, and its metabolism does not need Regular Insulin, and alternative sucrose is as the sweeting agent of patients with diabetes mellitus.Xylitol is by fermentation using bacteria, is used in the chewing gum as sweeting agent, has the function of keeping oral cavity acid base equilibrium, preventing dental caries.Along with the increase of people to healthy attention degree, the demand of Xylitol will be more and more higher, and therefore the demand of D-xylulose also can increase.
At present suitability for industrialized production D-xylulose mainly depends on the isomerization of D-wood sugar, and this method is produced the D-xylulose and had cost height, defective that yield is low.Utilize xylose isomerase to make xylose isomerase production D-xylulose as the medium people of beam generation, under 50 ℃, the average conversion of D-xylulose is 21.6%.Because transformation efficiency is low, must xylulose be separated with wood sugar by ion exchange treatment, the rate of recovery of D-xylulose only 43.8%.Cause the production cost of D-xylulose to increase like this.
In order to address this problem, many people are devoted to develop a kind of several different methods that adopts other pentitol that is easy to get to produce the D-xylulose as raw material.Form as utilizing D-pectinose alcoholdehydrogenase EC. 1.1.1.14 (D-Arabitoldehydrogenase) catalysis D-arabitol to change, known have some bacterial classifications can produce this enzyme.Obtain the D-arabitol as report in " applied microbiology " 18 (1969) 1031-1035 by using Han Xunde Ba Lishi yeast ATCC20121 glucose fermentation, adopt the weak oxide acetobacter to be converted into the D-xylulose then; Carry out fermentative production D-arabitol as EP403392A, EP421882A and CN1284564 by adopting height to ooze yeast, adopt the bacterium of genus acetobacter, Gluconobacter or klebsiella that the D-arabitol is converted into the D-xylulose then.In addition, obtain D-xylulose by adopting gluconobacter sp, acetobacter or Fu Late Bordetella from the glucose fermentation as CN1263944 and CN1269421.
More than in the mentioned production bacterium of these methods the transformation efficiency of principal product be 20～90%, transformation efficiency is lower, transformation time is 17～96 hours, and mainly adopt traditional zymotechnique, the main intermittent type that adopts ferments, these methods D-xylulose transformation efficiency and throughput are all lower, do not satisfy industrial requirement.This mainly be because: on the one hand cell be in unbound state can not recycle; Make downstream process become very difficult on the other hand, be unfavorable for the product separation purifying, production cost increases greatly.
Summary of the invention
Technical problem to be solved by this invention provides a kind of gluconobacter oxydans (Gluconobacter oxydans) of high yield D-pectinose alcoholdehydrogenase.
Another object of the present invention provides the method for utilizing above-mentioned gluconobacter oxydans to prepare the D-xylulose.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
The microorganism strains gluconobacter oxydans of seed selection of inventor laboratory and preservation (Gluconobacter oxydans) NH-10, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) at present, depositary institution address: China. Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.The numbering of registering on the books is CGMCC No.2709, and preservation date is on October 14th, 2008.With this bacterium is the production bacterial strain of D-xylulose.
CGMCC No.2709 bacterial strain has following character:
(1) colonial morphology feature:
On nutrient agar medium, cultivate 24h for 30 ℃ and occur
(2) physiology and biochemical characteristic:
A. culture temperature: 25～35 ℃, optimum temperuture is 30 ℃;
B. in pH2～8 scopes growth;
C. pigment produces: produce yellow pigment;
D. anti-NaCl concentration: can grow in 5% concentration.
(3) nutritional character:
Do not need to add somatomedin in the substratum of gluconobacter oxydans NH-10.Organonitrogen or inorganic nitrogen can use as nitrogenous source.
(4) 16S rDNA sequential analysis
Record the most of sequence 1409bp of 16S rDNA, specifically shown in SEQ ID No.1.
Institute's order-checking row must be correlated with from the GenBank database kind to be compared, and makes up with 16S rDNA total order and classifies basic phylogenetic tree as.The result shows: it is all very high that CGMCC2709 bacterial strain and gluconobacter suboxydans belong to (Gluconobacter) homology, with the G.oxydans homology be 99.5%, with the G.cerinus homology be 97.1%.The phylogeny result of study that combining form, cytochemistry composition and 16S rDNA complete series are analyzed can be orientated the classification of bacterial strain CGMCC2709 as the gluconic acid oxidation and bacillus, is specially G.oxydans CGMCC2709.
The method that the present invention utilizes gluconobacter oxydans CGMCC No.2709 to produce D-pectinose alcoholdehydrogenase and then production D-xylulose is that this inoculation is carried out aerobic cultivation in the aseptic culture medium of carbonaceous sources, nitrogenous source, inorganic salt and water, centrifugal or ultrafiltration obtains to contain the cell of D-pectinose alcoholdehydrogenase, and then directly adopts free to contain the cell of D-pectinose alcoholdehydrogenase or generate the D-xylulose through the cell transformation D-of the immobilized D-of containing pectinose alcoholdehydrogenase arabitol.
Condition of enzyme production:
Gluconobacter oxydans CGMCC No.2709 slant activation after one day (the slant culture based component also contains 2% agar except that containing carbon source, nitrogenous source etc.), is inoculated in the aseptic culture medium of carbonaceous sources, nitrogenous source, D-arabitol, inorganic salt and water.Each components contents is weightmeasurement ratio in the substratum, and promptly the g/L substratum is as follows.Ratio 100～the 800g/L of carbon source consumption and substratum, the ratio 100～400g/L of nitrogenous source consumption and substratum, the ratio 50～300g/L of D-arabitol consumption and substratum, the ratio 0.1～10g/L of inorganic salt consumption and substratum, all the other are water.Carbon source in the substratum is one or more in glucose, sucrose, fructose, maltose and the starch hydrolyzate; Nitrogenous source is extractum carnis, peptone, yeast extract paste, corn steep liquor, soybean cake powder, cottonseed meal, urea, (NH
4)
2SO
4And NH
4Among the Cl one or more; Inorganic salt are one or more in sodium salt, phosphoric acid salt and the dihydrogen phosphate.The initial pH scope of substratum is 4.0～8.0,25～35 ℃ of leavening temperatures, shaking speed 100～250r/min, cultivate 24～60h, wet cell weight reaches 20g/L, and fermentation broth enzyme work reaches 2.5～5.0U/mL, centrifugal then or ultrafiltration obtains to contain the cell of D-pectinose alcoholdehydrogenase, centrifugal condition: under the room temperature condition, 7000～12000r/min, 10～30min.When adopting ultrafiltration process, used ultra-filtration membrane aperture 0.01～0.1 μ m, working pressure 0.1～0.6MPa, the ultra-filtration membrane molecular weight that dams is 10
5～10
6Dalton.
D-pectinose alcoholdehydrogenase enzyme activity determination method:
Fermented liquid is centrifugal 10min collecting cell under 8000r/min, uses the potassium phosphate buffer of the 100mM of pH 7.0 to wash 3 each washings afterwards at 8000r/min, 4 ℃ of centrifugal 10min.
Ultrasonication: the thalline after will washing is suspended in the damping fluid, ultrasonication 10min, and working strength is 40%, and work 4min ice bath 6min at 4 ℃ of centrifugal 15min of following 10000r/min, removes cell debris subsequently, and the gained supernatant liquor is crude enzyme liquid.
Reaction system: 0.025M Tris-HCl (pH8.4), 0.25mM MgCl
2, 0.1mM D-arabitol, 0.25mM NAD.Measure the change of absorbancy at the 340nm place.
Enzyme is lived and is defined:
An enzyme unit definition alive is the used enzyme amount of oxidation 1 μ mol arabitol in a minute.It is oxidized that the absorbancy of 3 units is equivalent to 1 μ mol arabitol.
The enzymatic conversion method method:
Free is contained the cell that contains D-pectinose alcoholdehydrogenase after the cell of D-pectinose alcoholdehydrogenase or the immobilization be filled in the retort, add the D-arabitol of 30～100g/L, 25～35 ℃ of temperature of reaction, transformation time 9～16h; Perhaps immobilized cell is packed in the filling type column type reactor, column volume is not limit, this reactor can 0.5～5mL/min way flow adds or the circulation Continuous Flow adds the D-pectinose alcoholic solution of 30～100g/L, under 25～35 ℃ condition, carry out enzymatic conversion reaction, every approving and forwarding time 12～16h, by adjusting flow velocity, the transformation efficiency of control D-arabitol is between 90～100%, collect effluent liquid, condensing crystal obtains the D-xylulose.
The cell that contains D-pectinose alcoholdehydrogenase carries out immobilized method and is: preparation 1～3% (the i.e. sodium alginate of 10～30g/L) concentration, the cell of adding 10～25% (being the cells that add 10～25 grams in every 100mL sodium alginate soln), and add 5～10% diatomite (being the diatomite that adds 50～100 grams in every liter of sodium alginate soln), be 1～4% CaCl again with concentration
2(10～40g/L) is fixed-type with the embedded material sodium alginate, reacts as the enzyme source with this for solution.Except sodium alginate as the fixation support embedding, can also use carrier embedding cells such as carrageenin, chitin.
Beneficial effect of the present invention:
The present invention's screening obtains a kind of bacterial strain that can efficiently transform D-arabitol generation D-xylulose, adopt preparation method of the present invention, D-pectinose alcohol conversion reaches as high as 99.5% (w/w), the yield of D-xylulose can reach 95%, D-xylulose concentration can reach 100g/L in the conversion fluid, contains other composition in the conversion fluid hardly.
Reference background technical literature material is produced bacterium to different D-xyluloses and is analyzed, and the result is as shown in table 1:
The different D-xyluloses of table 1 are produced the comparison of bacterium
It is few to found that oxidation grape bacillus CGMCC No.2709 transforms the by product that is generated, and is more conducive to further industrialization serialization and produces.
Embodiment:
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand that the described concrete material proportion of embodiment, processing condition and result thereof only are used to illustrate the present invention, and should also can not limit the present invention described in detail in claims.
Embodiment 1:
Slant medium: glucose 30g/L, yeast extract paste 10g/L, extractum carnis 5g/L, agar 20g/L.
Shake-flask culture base: glucose 30g/L, yeast extract paste 25g/L, extractum carnis 5g/L, D-arabitol 10g/L, KH
2PO
45g/L.
Oxidation grape bacillus CGMCC No.2709 30 ℃ of cultivation 24h on slant medium, connect this bacterium of ring then in the shake-flask culture base, cultivate 10h for 30 ℃, shake a bottle rotating speed 200r/min, content of thalli is 20g/L in the fermented liquid that obtains, produce enzyme and reach 2.5～5.0U/mL, transform with free cell, taking by weighing the wet cell of 1g after centrifugal adds in the D-ara solution of 10mL 30g/L (the cell addition is by the D-ara solution of every 100g cell transformation 1L 30g/L), under 30 ℃ of conditions, stir vibration (rotating speed 120r/min) and transform production D-xylulose in shaking bottle, reaction times 9h surveys substrate conversion efficiency and production concentration after the stopping of reaction.The D-ara transformation efficiency is 99%, and D-xylulose yield reaches 95%, and D-xylulose concentration is 28.5g/L.
Embodiment 2:
With 30g/L maltose is carbon source, the 20g/L yeast extract paste, the 10g/L soybean cake powder is a nitrogenous source, other composition of substratum and yeast culture condition are with embodiment 1, the sodium alginate colloidal solution of preparation 1.5%, after mixing with the synthetic bacteria suspension of physiological saline, add 8% diatomite adsorption after, splash into 2% CaCl with syringe
2Solidify in the solution, make the spherical immobilized cell that diameter is about 3～4mm, in 4 ℃ of refrigerators, leave standstill a few hours, supernatant liquor then inclines, through distilled water wash 2 times, 1g is obtained the 5g immobilized cell through the centrifugal wet cell with sodium alginate to embed, under 30 ℃ of conditions, in shaking bottle, transform the D-pectinose alcoholic solution (rotating speed 200r/min) of 10mL 50g/L with this, every approving and forwarding time 13h transforms 30 batches, and the average conversion that obtains D-ara is 99%, the average yield of D-xylulose is 90%, and D-xylulose concentration is 45g/L.
Embodiment 3:
With glucose 225g, yeast extract paste 112.5g, extractum carnis 22.5g, D-arabitol 45g, KH
2PO
422.5g, pH5.0, water is settled to 4.5L, is made into substratum, in the 7.5 liters of stirred fermentors of packing into, 121 ℃ of steam sterilizing 15min.Preparation seed culture medium: glucose 30g/L, yeast extract paste 10g/L, extractum carnis 5g/L.Gluconobacter oxydans CGMCCNo.2709 is connect one to be encircled in seed culture medium, cultivate 8h for 30 ℃ and get seed liquor, the inoculum size of seed liquor by fermentating liquid volume 3% inserted in the cooled fermention medium 30 ℃ of cultivations (ventilation 0.7vvm, mixing speed are 600r/min) 48h.Fermented liquid is carried out bacteriological filtration handle in ultra-fine filter, the ultra-filtration membrane molecular weight that dams is 10
6Da, working pressure are 0.2MPa, 50 ℃ of controlled temperature, face speed 4m/s is after the method immobilization of 100g thalline by embodiment 2 that obtains of will damming, in the retort of the 2L that packs into, the D-ara solution 1000mL that adds 100g/L, 30 ℃ of temperature of reaction, mixing speed 200r/min, ventilation 1vvm, reaction times 13h, the D-ara transformation efficiency is 99%, and D-xylulose yield reaches 92%, and D-xylulose concentration reaches 92g/L.
Embodiment 4:
With the 50g/L starch hydrolyzate is carbon source, and 20g/L urea, 10g/L yeast extract paste are nitrogenous source, and other composition of substratum and yeast culture condition take by weighing the 100g wet thallus with embodiment 3, add sterilized water 100mL, in 30 ℃ of water bath heat preservations; Take by weighing the 10g carrageenin, add sterilized water 200mL, fully soak heating for dissolving.Both are mixed rapidly, pave plate, place 30min, be cut into the particle of size about 2 * 2 * 2cm after solidifying, in 2mol/L KCl solution, soak 30min again, it is fully solidified in 4 ℃ of refrigerators.Leach particle and wash 2-3 time with physiological saline, 4 ℃ of preservations are standby.The gained thalline forms immobilized cell 300g, packs into
In the bubbling filling bed type column type reactor of 4 * 45cm, this reactor adds the D-ara solution of 100g/L with the flow velocity way flow of 1mL/min, and ventilation 1vvm carries out enzymatic conversion under 30 ℃, and 14h is one batch.This device is operation 30d continuously, transforms D-ara solution 50L, and the average conversion that obtains D-ara is 99%, and the average conversion of D-xylulose is 91%, and the concentration of D-xylulose is 91g/L.
Embodiment 5:
After in embodiment 1, transforming the D-ara product D-xylulose stopping of reaction, in reaction system, add the 0.5mL dehydrated alcohol, under 30 ℃, leave standstill and cultivate 12h, survey substrate conversion efficiency and production concentration after the stopping of reaction, finally can obtain Xylitol 24.6g/L, Xylitol is 82% to the D-ara transformation efficiency.
Embodiment 6:
Transform the D-ara product D-xylulose stopping of reaction in embodiment 2 after, add 1g glucose in reaction system, leave standstill under 30 ℃ and cultivate 15h, finally can obtain Xylitol 44g/L, Xylitol is 81% to the D-ara transformation efficiency.
SEQUENCE?LISTING
＜110〉Nanjing University of Technology
＜120〉a kind of gluconobacter oxydans and utilize it to prepare the method for xylulose
<130>?njut090112
<160>?1
<170>?PatentIn?version?3.3
<210>?1
<211>?1409
<212>?DNA
<213>?Gluconobacter?oxydans
<400>?1
agcgaacgct?ggcggcatgc?ttaacacatg?caagtcgcac?gaaggtttcg?gccttagtgg 60
cggacgggtg?agtaacgcgt?agggatctat?ccacgggtgg?gggacaactt?cgggaaactg 120
gagctaatac?cgcatgatac?ctgagggtca?aaggcgcaag?tcgcctgtgg?aggaacctgc 180
gttcgattag?ctagttggtg?gggtaaaggc?ctaccaaggc?gatgatcgat?agctggtttg 240
agaggatgat?cagccacact?gggactgaga?cacggcccag?actcctacgg?gaggcagcag 300
tggggaatat?tggacaatgg?gcgaaagcct?gatccagcaa?tgccgcgtgt?gtgaagaagg 360
tcttcggatt?gtaaagcact?ttcgacgggg?acgatgatga?cggtacccgt?agaagaagcc 420
ccggctaact?tcgtgccagc?agccgcggta?atacgaaggg?ggctagcgtt?gctcggaatg 480
actgggcgta?aagggcgcgt?aggcggttgt?tacagtcaga?tgtgaaatcc?ccgggcttaa 540
cctgggaact?gcatttgata?cgtgacgact?agagttcgag?agagggttgt?ggaattccca 600
gtgtagaggt?gaaattcgta?gatattggga?agaacaccgg?tggcgaaggc?ggcaacctgg 660
ctcgatactg?acgctgaggc?gcgaaagcgt?ggggagcaaa?caggattaga?taccctggta 720
gtccacgctg?taaacgatgt?gtgctggatg?ttgggaaact?tagtttctca?gtgtcgaagc 780
taacgcgcta?agcacaccgc?ctggggagta?cggccgcaag?gttgaaactc?aaaggaattg 840
acgggggccc?gcacaagcgg?tggagcatgt?ggtttaattc?gaagcaacgc?gcagaacctt 900
accagggctt?gcatggggag?gaccggttca?gagatggacc?tttcttcgga?cctcccgcac 960
aggtgctgca?tggctgtcgt?cagctcgtgt?cgtgagatgt?tgggttaagt?cccgcaacga 1020
gcgcaaccct?tgtctttagt?tgccagcact?ttcaggtggg?cactctagag?agactgccgg 1080
tgacaagccg?gaggaaggtg?gggatgacgt?caagtcctca?tggcccttat?gtcctgggct 1140
acacacgtgc?tacaatggcg?gtgacagtgg?gaagctacat?ggtgacatgg?tgctgatctc 1200
taaaagccgt?ctcagttcgg?attgtactct?gcaactcgag?tacatgaagg?tggaatcgct 1260
agtaatcgcg?gatcagcatg?ccgcggtgaa?tacgttcccg?ggccttgtac?acaccgcccg 1320
tcacaccatg?ggagttggtt?cgaccttaag?ccggtgagcg?aaccgcaagg?acgcagccga 1380
ccacggacgg?gtcagcgact?ggggtgaag 1409
Claims (8)
1. gluconobacter oxydans, its classification called after gluconobacter oxydans (Gluconobacter oxydans) NH-10 has been preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, and its deposit number is: CGMCCNo.2709.
2. utilize the described gluconobacter oxydans of claim 1 to prepare the method for D-xylulose, it is characterized in that bacterial strain CGMCC No.2709 is inoculated in the aseptic culture medium of carbonaceous sources, nitrogenous source, D-arabitol, inorganic salt and water and cultivate, centrifugal or ultrafiltration obtains to contain the cell of D-pectinose alcoholdehydrogenase, and then adopts free or generate the D-xylulose through the immobilized cell transformation D-arabitol that contains D-pectinose alcoholdehydrogenase.
3. the method for utilizing gluconobacter oxydans to prepare the D-xylulose according to claim 2, it is characterized in that in the described substratum, each amounts of components is: the carbon source consumption is 100～800g/L, nitrogenous source consumption 100～400g/L, D-arabitol consumption 50～300g/L, the inorganic salt consumption is 0.1～10g/L, and all the other are water.
4. according to claim 2 or the 3 described methods of utilizing gluconobacter oxydans to prepare the D-xylulose, it is characterized in that in the substratum, described carbon source is one or more in glucose, sucrose, fructose, maltose and the starch hydrolyzate, and described nitrogenous source is extractum carnis, peptone, yeast extract paste, corn steep liquor, soybean cake powder, cottonseed meal, urea, (NH
4)
2SO
4And NH
4Among the Cl one or more, described inorganic salt are one or more in sodium salt, phosphoric acid salt and the dihydrogen phosphate.
5. the method for utilizing gluconobacter oxydans to prepare the D-xylulose according to claim 2, it is characterized in that bacterial strain CGMCC No.2709 is carried out culture condition is: the initial pH scope of substratum is 4.0～8.0,25～35 ℃ of leavening temperatures, shaking speed 100～250r/min cultivates 24～60h.
6. the method for utilizing gluconobacter oxydans to prepare the D-xylulose according to claim 2, it is characterized in that adopting the method for ultrafiltration to obtain to contain the cell of D-pectinose alcoholdehydrogenase, used ultra-filtration membrane aperture 0.01～0.1 μ m, working pressure is 0.1～0.6MPa, and the ultra-filtration membrane molecular weight that dams is 10
5～10
6Dalton.
7. the method for utilizing gluconobacter oxydans to prepare the D-xylulose according to claim 2, it is that the cell after free cell or the immobilization is filled in the retort that the cell transformation D-arabitol that it is characterized in that containing D-pectinose alcoholdehydrogenase is produced the D-xylulose, the D-arabitol that adds 30～100g/L carries out enzymatic conversion reaction, 25～35 ℃ of temperature of reaction, mixing speed 50～250r/min, transformation time 9～16h; Perhaps, immobilized cell is packed in the bubbling filling bed type column type reactor, and this reactor adds with the flow velocity way flow of 0.5～5mL/min or the circulation Continuous Flow adds the D-pectinose alcoholic solution of 30～100g/L, carries out enzymatic conversion reaction, 25～35 ℃ of temperature of reaction, every approving and forwarding time 12～16h.
8. according to claim 2 or the 7 described methods of utilizing gluconobacter oxydans to prepare the D-xylulose, it is characterized in that immobilized cell is to adopt alginate calcium or carrageenin as fixation support.
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| CN101880643B (en) * | 2010-06-03 | 2012-04-25 | 南京工业大学 | Gluconobater oxydans genetic engineering strain and construction method thereof |
| CN102250820B (en) * | 2010-06-03 | 2013-03-13 | 南京工业大学 | Genetic engineering bacteria of gluconobacter oxydans and construction methods thereof |
| CN101948878B (en) * | 2010-09-02 | 2012-04-25 | 南京工业大学 | Application of gluconobacter oxydans in preparing 1,3-dioxyacetone |
| CN102382868B (en) * | 2011-11-10 | 2013-09-11 | 中科医药行业生产力促进中心有限公司 | Method for producing dihydroxyacetone by using gluconobacter sp. |
| CN102978148B (en) * | 2012-12-18 | 2014-08-06 | 南京工业大学 | Gluconobacter oxydans engineering bacteria and preparation method and application thereof in preparing xylitol |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1247230A (en) * | 1998-07-30 | 2000-03-15 | 味之素株式会社 | Xylitol dehydrogenase of acetobacter and its gene |
| CN1269421A (en) * | 1999-01-20 | 2000-10-11 | 味之素株式会社 | Process for producing xylitol or D-xyluose |
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2009
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1247230A (en) * | 1998-07-30 | 2000-03-15 | 味之素株式会社 | Xylitol dehydrogenase of acetobacter and its gene |
| CN1269421A (en) * | 1999-01-20 | 2000-10-11 | 味之素株式会社 | Process for producing xylitol or D-xyluose |
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