CN103013863A - Sphingomonas and application thereof in preparation of rhamsan - Google Patents
Sphingomonas and application thereof in preparation of rhamsan Download PDFInfo
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- CN103013863A CN103013863A CN2012105044613A CN201210504461A CN103013863A CN 103013863 A CN103013863 A CN 103013863A CN 2012105044613 A CN2012105044613 A CN 2012105044613A CN 201210504461 A CN201210504461 A CN 201210504461A CN 103013863 A CN103013863 A CN 103013863A
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- sphingomonas
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- carbohydrate gum
- sphingol single
- carbon source
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- 238000002360 preparation method Methods 0.000 title claims description 4
- 241000736131 Sphingomonas Species 0.000 title abstract 5
- 239000001963 growth medium Substances 0.000 claims abstract description 21
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 15
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 15
- 238000004321 preservation Methods 0.000 claims abstract description 4
- 150000001720 carbohydrates Chemical class 0.000 claims description 35
- 235000014633 carbohydrates Nutrition 0.000 claims description 35
- 238000000855 fermentation Methods 0.000 claims description 29
- 230000004151 fermentation Effects 0.000 claims description 29
- 239000002609 medium Substances 0.000 claims description 25
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 21
- 241001135759 Sphingomonas sp. Species 0.000 claims description 20
- 239000007788 liquid Substances 0.000 claims description 19
- 239000008103 glucose Substances 0.000 claims description 18
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 11
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 9
- -1 extractum carnis Substances 0.000 claims description 8
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 7
- 230000001580 bacterial effect Effects 0.000 claims description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- 229930006000 Sucrose Natural products 0.000 claims description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 6
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 6
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- 239000005720 sucrose Substances 0.000 claims description 6
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- 235000010469 Glycine max Nutrition 0.000 claims description 5
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- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 4
- 235000005822 corn Nutrition 0.000 claims description 4
- 235000012343 cottonseed oil Nutrition 0.000 claims description 4
- 235000012054 meals Nutrition 0.000 claims description 4
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 3
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims description 3
- 229930091371 Fructose Natural products 0.000 claims description 3
- 239000005715 Fructose Substances 0.000 claims description 3
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 3
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- SRBFZHDQGSBBOR-LECHCGJUSA-N alpha-D-xylose Chemical compound O[C@@H]1CO[C@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-LECHCGJUSA-N 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims description 3
- 239000004202 carbamide Substances 0.000 claims description 3
- 235000015165 citric acid Nutrition 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-M dihydrogenphosphate Chemical compound OP(O)([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-M 0.000 claims description 3
- 235000011187 glycerol Nutrition 0.000 claims description 3
- 239000001257 hydrogen Substances 0.000 claims description 3
- 229910052739 hydrogen Inorganic materials 0.000 claims description 3
- 239000004310 lactic acid Substances 0.000 claims description 3
- 235000014655 lactic acid Nutrition 0.000 claims description 3
- 239000008101 lactose Substances 0.000 claims description 3
- 235000013379 molasses Nutrition 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 3
- 150000003016 phosphoric acids Chemical class 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- 229960003487 xylose Drugs 0.000 claims description 3
- 238000011534 incubation Methods 0.000 claims description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 8
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 4
- 235000013305 food Nutrition 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 150000003839 salts Chemical class 0.000 abstract description 2
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 abstract 2
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 abstract 2
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 abstract 2
- 238000005553 drilling Methods 0.000 abstract 1
- 238000009629 microbiological culture Methods 0.000 abstract 1
- 239000003208 petroleum Substances 0.000 abstract 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 24
- 238000011218 seed culture Methods 0.000 description 18
- 238000000034 method Methods 0.000 description 8
- 230000001954 sterilising effect Effects 0.000 description 8
- 238000004659 sterilization and disinfection Methods 0.000 description 8
- 238000012546 transfer Methods 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000006052 feed supplement Substances 0.000 description 2
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000010899 nucleation Methods 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000001502 supplementing effect Effects 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 1
- 102000013275 Somatomedins Human genes 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- AEMOLEFTQBMNLQ-WAXACMCWSA-N alpha-D-glucuronic acid Chemical compound O[C@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-WAXACMCWSA-N 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000004568 cement Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 150000002475 indoles Chemical class 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000012882 sequential analysis Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M sodium chloride Inorganic materials [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses Sphingomonas, which is classified and named as Sphingomonas (Sphingomonas sp) PG-8, and is preserved in China general microbiological culture collection center, the number of a registration book is CGMCC NO.6833, and the preservation date is 11/15/2012. The invention also discloses application of sphingomonas in preparing Rhamsan Gum. The strain is cultured in a culture medium containing a carbon source, a nitrogen source and inorganic salts, and 5-50 g/L of rhamnose gum can be accumulated under optimized conditions. The strain can utilize various carbon sources and nitrogen sources, has extensive culture conditions, is convenient and simple to operate, has high rhamnose gum yield and low production cost, and can be widely applied to the fields of food addition, petroleum drilling and the like.
Description
Technical field
The invention belongs to the fermentation engineering field, relate to a kind of microorganism strains Sphingol single-cell (Sphingomonas sp.) and application thereof of high yield sandlwood carbohydrate gum.
Background technology
Sandlwood carbohydrate gum (Rhamsan Gum) is a kind of exocellular polysaccharide that is produced by Sphingol single-cell, is that gelling gum is saccharoidal a kind of.Gelling gum class polysaccharide has identical tetrose repeating unit backbone structure: D-Glucose, D-glucuronic acid, D-Glucose and L-rhamnosyl, just side chain or a bit trickle difference of other structure.The rhamnosyl plastic structure is shown in (1).The sandlwood carbohydrate gum is with its unique physics and rheological properties and safety in utilization, can be used as suspension agent, stablizer, thickening material, emulsifying agent, membrane-forming agent etc. and be widely used in food, oil, cement, coating industry, also be used for the industries such as printing ink, food, textile printing and dyeing, agricultural chemicals, makeup, medicine.
At present, the sandlwood carbohydrate gum is not yet by large-scale development, and only U.S. Kelco company has carried out among a small circle research to it, and it is blank that the research of China in this field still belongs to, and has no its industrialization and produce.
Summary of the invention
Technical problem to be solved by this invention provides the Sphingol single-cell that a strain can high yield sandlwood carbohydrate gum.
The technical problem that the present invention also will solve provides the application of above-mentioned Sphingol single-cell.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
Microorganism Sphingol single-cell (Sphingomonas sp.) PG-8 of contriver's seed selection in the soil sample that National Forest National parks, the Nanjing Laoshan is chosen, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) at present, preservation address: Datun Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, postcode: 100101.The numbering of registering on the books is CGMCCNO:6833, and preservation date is: on November 15th, 2012.With this bacterium as producing bacterial strain.
The CGMCCNO:6833 bacterial strain has following character:
1, colonial morphology feature:
Vegetative cell is the bacillus of 0.5 ~ 1.2 * 0.5 ~ 3 μ m sizes on the peptone nutrient agar, and 30 ° of C cultivated 2 ~ 3 days.30 ° of C cultivation 12h thalline can raised growth in above-mentioned substratum.It is faint yellow that bacterium colony is, circle, and smooth surface, sticking shape, intermediate projections is opaque.
2, physiology and biochemistry characteristic:
(1) culture temperature: 25 ~ 38 ° of C, optimum temperuture is 30-32 ° of C;
(2) growth in pH5.0 ~ 9.0 scopes;
(3) gramstaining: feminine gender;
(4) methyl red experiment: feminine gender;
(5) indoles production: feminine gender;
(6) H
2S produces: feminine gender;
(7) anti-NaCl concentration: can grow more than 7% concentration.
3,16S rDNA sequential analysis:
Record the most of sequence 1443bp of 16S rDNA, shown in SEQ ID No:1.The relevant kind of check order row from the GeneBank database compared, make up the phylogenetic tree that 16S rDNA total order is classified the basis as.The result shows: bacterial strain and Sphingol single-cell reach 99.9% homology.So what assert the present invention's use is Sphingol single-cell, is specially Sphingol single-cell (Sphingomonas sp.) PG-8.
4, nutritional character:
Do not need to add somatomedin in the substratum of Sphingol single-cell, can utilize multiple compounds as carbon source, these materials both can use separately, also can the composite in the proper ratio carbon source of serving as.Organonitrogen or inorganic nitrogen can use as nitrogenous source.Each amounts of components is in the substratum: carbon source 10 ~ 100g/L, and nitrogenous source 1 ~ 40g/L, inorganic salt 0.01 ~ 50g/L, all the other are water.Carbon source generally adopt in glucose, sucrose, maltose, lactose, wood sugar, fructose, lactic acid, citric acid, glycerine, starch and the molasses any one or multiple; Nitrogenous source can adopt yeast powder, extractum carnis, peptone, yeast extract paste, corn steep liquor, soybean cake powder, cottonseed meal, urea, (NH
4)
2SO
4, NH
4Cl, (NH
4)
2HPO
4And NH
4NO
3In any one or multiple; Also comprise one or more inorganic salts commonly used such as sylvite, sodium salt, magnesium salts, vitriol, phosphoric acid salt, dihydrogen phosphate, phosphoric acid hydrogen disalt and hydrochloride in the substratum.
The application of above-mentioned Sphingol single-cell in preparation sandlwood carbohydrate gum.
Concrete method is: with bacterial strain CGMCC NO.6833 be inoculated in contain carbon source, nitrogenous source, inorganic salt and water without in the bacteria fermentation culture medium, under the condition of proper growth, carry out aerobic cultivation, the fermented liquid of generation can obtain the sandlwood carbohydrate gum through extracting.
Wherein, described fermention medium comprises following component: carbon source 10 ~ 100g/L, and nitrogenous source 1 ~ 40g/L, inorganic salt 0.01 ~ 50g/L, all the other are water, pH5.0 ~ 9.0.
Wherein, described carbon source is any one or a few the combination in glucose, sucrose, maltose, lactose, wood sugar, fructose, lactic acid, citric acid, glycerine, starch and the molasses; Nitrogenous source is yeast powder, extractum carnis, peptone, yeast extract paste, corn steep liquor, soybean cake powder, cottonseed meal, urea, (NH
4)
2SO
4, NH
4Cl, (NH
4)
2HPO
4And NH
4NO
3In any one or a few combination; Inorganic salt are one or more in vitriol, phosphoric acid salt, dihydrogen phosphate, phosphoric acid hydrogen disalt and the hydrochloride.
Wherein, the condition of described proper growth is: initial pH is 5.0 ~ 9.0, culture temperature is that 25 ~ 38 ° of C, incubation time are 24 ~ 72h.
Wherein, after the fermentation ends, the fermented liquid of removing thalline can extract the sandlwood carbohydrate gum.
Specifically:
The method for preparing the sandlwood carbohydrate gum provided by the invention, that bacterial strain CGMCC No.6833 is inoculated into the inclined-plane after 1 ~ 4 day (slant medium employing beef-protein medium), cultivate 24 ~ 72h at the substratum that contains carbon source, nitrogenous source and inorganic salt, can generate 5 ~ 50g/L sandlwood carbohydrate gum, usually fermentation condition is preferred: substratum is: glucose 10 ~ 100g/L, yeast extract paste 1 ~ 10g/L, (NH
4)
2SO
41 ~ 20g/L, K
2HPO
41 ~ 30g/L, MgSO
47H
2O0.2 ~ 3g/L.25 ~ 38 ° of C of leavening temperature, that better is 30 ~ 32 ° of C; The initial pH scope of substratum is 5.0 ~ 9.0, and better is 6.5 ~ 8.0; Aerobic cultivation.
Beneficial effect: the present invention has following advantage:
(1) the present invention screens a strain sandlwood carbohydrate gum and produces bacterium, and this bacterium can utilize several kinds of carbon source and nitrogen source fermentation to produce the sandlwood carbohydrate gum, and easy to operate simple, culture condition is very extensive.
(2) the present invention screens a strain sandlwood carbohydrate gum and produces bacterium, can obtain the sandlwood carbohydrate gum of higher concentration, and the highest extremely approximately 50g/L of sandlwood carbohydrate gum content is far above the approximately 15g/L[JerryA.Peik of present report, San Diego; SuzannaM.Steenbergen, Lakeside; Harold R.Hayden, Escondido, all of Calif.HETEROPOLYSACCHARIDE S-194, U.S. Patent Application No.: 313,440, open August 30 nineteen eighty-three].
(2) can use cellulose hydrolysis liquid glucose (place of glucose or sucrose), soybean cake powder (substituting yeast extract paste or peptone) as carbon source, the nitrogenous source of sandlwood carbohydrate gum fermention medium, the fermentation raw material cost to be declined to a great extent.
Embodiment
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand, the described content of embodiment only is used for explanation the present invention, and should also can not limit the present invention described in detail in claims.
Embodiment 1:500mL shake flask fermentation Sphingomonas sp.PG-8 produces the sandlwood carbohydrate gum.
Seed culture medium: glucose 10g/L, extractum carnis 3g/L, peptone 5g/L, MgSO
47H
2O0.5g/L utilizes NaOH solution to transfer pH6.8.
Fermention medium: glucose 40g/L, yeast extract paste 5g/L, (NH
4)
2SO
45g/L, K
2HPO
43H
2O2g/L, MgSO
47H
2O0.5g/L utilizes NaOH solution to transfer pH6.8.
Sphingol single-cell Sphingomonas sp.PG-8 (CGMCC NO:6833) is inoculated in the seed culture medium, cultivate 15h under the shaking table condition of 30 ° of C, 200r/min, with seed liquor by 5%(v/v) grain weight be inoculated in the shaking flask that is preinstalled with the fermention medium after the 500mL sterilization and cultivate, culture condition: 30 ° of C, 200r/min, fermentation 72h, sandlwood carbohydrate gum content reaches 20.6g/L.
Embodiment 2:3L fermentor tank batch fermentation Sphingomonas sp.PG-8 produces the sandlwood carbohydrate gum.
Seed culture medium and fermention medium are with embodiment 1.
Sphingol single-cell Sphingomonas sp.PG-8 (CGMCC NO:6833) is inoculated in the seed culture medium, cultivate 15h under the shaking table condition of 30 ° of C, 200r/min, with seed liquor by 8%(v/v) grain weight be inoculated in the fermentor tank that is preinstalled with the fermention medium after the 2.1L sterilization and cultivate, culture condition: 30 ° of C, 400r/min, air flow 3.0L/min, open the pH automatic control device in the fermenting process, utilize 3mol/L HCl or 10% ammoniacal liquor (v/v) controlled fermentation liquid pH value between 6.5-7.5.Fermentation 48h, sandlwood carbohydrate gum content reaches 23.5g/L.
Embodiment 3:3L fermentor tank batch fermentation Sphingomonas sp.PG-8 produces the sandlwood carbohydrate gum.
Seed culture medium and fermention medium are with embodiment 1.
Sphingol single-cell Sphingomonas sp.PG-8 (CGMCC NO:6833) is inoculated in the seed culture medium, cultivate 15h under the shaking table condition of 32 ° of C, 200r/min, with seed liquor by 5%(v/v) grain weight be inoculated in the fermentor tank that is preinstalled with the fermention medium after the 2.1L sterilization and cultivate, culture condition: 30 ° of C, 600r/min, air flow 2.3L/min, open the pH automatic control device in the fermenting process, utilize 3mol/L HCl or 3mol/L NaOH solution controlled fermentation liquid pH value between 7.0-7.5.Fermentation 48h, sandlwood carbohydrate gum content reaches 28.9g/L.
Embodiment 4:3L fermentor tank gap fed-batch fermentation Sphingomonas sp.PG-8 produces the sandlwood carbohydrate gum.
Seed culture medium and fermention medium are with embodiment 1.Supplemented medium is glucose 500g/L, and the glucose total amount is 80g/L.
Sphingol single-cell Sphingomonas sp.PG-8 (CGMCC NO:6833) is inoculated in the seed culture medium, cultivate 16h under the shaking table condition of 30 ° of C, 200r/min, with seed liquor by 10%(v/v) grain weight be inoculated in the fermentor tank that is preinstalled with the fermention medium after the 2.1L sterilization and cultivate, culture condition: 30 ° of C, 400r/min, air flow 3.0L/min, open the pH automatic control device in the fermenting process, utilize 3mol/L HCl or 3mol/L NaOH controlled fermentation liquid pH value between 6.8-7.2.When residual total reducing sugar is at 10g/L in the fermented liquid, open feed supplementing device, (each 20g/L) is that the glucose of 40g/L fills in the fermented liquid with total amount at twice, fermentation 72h, sandlwood carbohydrate gum content reaches 38.9g/L.
Embodiment 5:100L fermentor tank constant current fed-batch fermentation Sphingomonas sp.PG-8 produces the sandlwood carbohydrate gum.
Seed culture medium and fermention medium are with embodiment 1.Supplemented medium is glucose 500g/L, and the glucose total amount is 80g/L.
Sphingol single-cell Sphingomonas sp.PG-8 (CGMCC NO:6833) is inoculated in the seed culture medium, cultivate 16h under the shaking table condition of 31 ° of C, 200r/min, with seed liquor by 10%(v/v) grain weight be inoculated in the 10L seeding tank that is preinstalled with the seed culture medium after the 6.3L sterilization, cultivate 8h under 31 ° of C, the 400r/min condition, its culture transferring to the 100L fermentor tank, is preinstalled with the fermention medium after 63L sterilizes in this fermentor tank.100L ferment tank condition is as follows: 31 ° of C, 300r/min, air flow 84L/min opens the pH automatic control device in the fermenting process, utilize 3mol/LHCl or 3mol/LNaOH controlled fermentation liquid pH value between 6.8-7.2, when glucose concn is low to moderate 5g/L in the fermented liquid, utilize the feed supplement pump that supplemented medium is pumped in the fermented liquid, keep in the fermentor tank between the sugared concentration 5-10g/L, fermentation 72h, the glucose total amount reaches 80g/L, and sandlwood carbohydrate gum content reaches 50.0g/L in the fermented liquid.
Embodiment 6:500mL shake flask fermentation Sphingomonas sp.PG-8 produces the sandlwood carbohydrate gum.
Seed culture medium is with embodiment 1.Fermention medium: glucose 50g/L, corn steep liquor 50g/L, NH
4Cl3g/L, K
2HPO
43H
2O4g/L, MgSO
47H
2O0.2g/L utilizes NaOH solution to transfer pH7.0.
Sphingol single-cell Sphingomonas sp.PG-8 (CGMCC NO:6833) is inoculated in the seed culture medium, cultivate 12h under the shaking table condition of 35 ° of C, 220r/min, with seed liquor by 8%(v/v) grain weight be inoculated in the shaking flask that is preinstalled with the fermention medium after the 500mL sterilization and cultivate, culture condition: 35 ° of C, 220r/min, fermentation 68h, sandlwood carbohydrate gum content reaches 29.7g/L.
Embodiment 7:3L fermentor tank batch fermentation Sphingomonas sp.PG-8 produces the sandlwood carbohydrate gum.
Seed culture medium is with embodiment 1.Fermention medium: sucrose 50g/L, cottonseed meal 25g/L, NH
4NO
38g/L, K
2HPO
43H
2O4g/L, MgSO
47H
2O5g/L utilizes NaOH solution to transfer pH8.5.
Sphingol single-cell Sphingomonas sp.PG-8 (CGMCC NO:6833) is inoculated in the seed culture medium, cultivates 12h under the shaking table condition of 38 ° of C, 220r/min.With seed liquor by 10%(v/v) grain weight be inoculated in the fermentor tank that is preinstalled with the fermention medium after the 2.1L sterilization and cultivate, culture condition: 38 ° of C, 800r/min, air flow 3.0L/min, open the pH automatic control device in the fermenting process, utilize 3mol/LHCl or 3mol/LNaOH controlled fermentation liquid pH value between 7.5-7.8.When residual total reducing sugar is at 10g/L in the fermented liquid, open feed supplementing device, (each 15g/L) is that the sucrose of 30g/L fills in the fermented liquid with total amount at twice, fermentation 72h, sandlwood carbohydrate gum content reaches 42.5g/L.
Embodiment 8:100L fermentor tank batch fermentation Sphingomonas sp.PG-8 produces the sandlwood carbohydrate gum.
Seed culture medium is with embodiment 1.Fermention medium: glucose 50g/L, soybean cake powder 20g/L, NH
4NO
38g/L, K
2HPO
43H
2O4g/L, MgSO
47H
2O5g/L utilizes NaOH solution to transfer pH8.5.
Sphingol single-cell Sphingomonas sp.PG-8 (CGMCC NO:6833) is inoculated in the seed culture medium, cultivate 10h under the shaking table condition of 34 ° of C, 220r/min, with seed liquor by 12%(v/v) grain weight be inoculated in the 10L seeding tank that is preinstalled with the seed culture medium after the 6.3L sterilization, cultivate 10h under 34 ° of C, the 220r/min condition, its culture transferring to the 100L fermentor tank, is preinstalled with the fermention medium after 63L sterilizes in this fermentor tank.100L ferment tank condition is as follows: 34 ° of C, 220r/min, air flow 84L/min opens the pH automatic control device in the fermenting process, utilize 3mol/LHCl or 3mol/L NaOH controlled fermentation liquid pH value between 7.5-7.8, when glucose concn is low to moderate 5g/L in the fermented liquid, utilize the feed supplement pump that supplemented medium is pumped in the fermented liquid, keep in the fermentor tank between the sugared concentration 5-10g/L, fermentation 72h, the glucose total amount reaches 80g/L, and sandlwood carbohydrate gum content reaches 48.5g/L in the fermented liquid.
Claims (6)
1. its Classification And Nomenclature of Sphingol single-cell is Sphingol single-cell (Sphingomonas sp.) PG-8, be preserved in Chinese common micro-organisms culture presevation administrative center, the numbering of registering on the books is CGMCC NO.6833, and preservation date is on November 15th, 2012.
2. the application of Sphingol single-cell claimed in claim 1 in preparation sandlwood carbohydrate gum.
3. application according to claim 2, it is characterized in that, with bacterial strain CGMCC NO.6833 be inoculated in contain carbon source, nitrogenous source, inorganic salt and water without in the bacteria fermentation culture medium, under the condition of proper growth, carry out aerobic cultivation, the fermented liquid of generation can obtain the sandlwood carbohydrate gum through extracting.
4. application according to claim 3 is characterized in that, described fermention medium comprises following component: carbon source 10 ~ 100g/L, and nitrogenous source 1 ~ 40g/L, inorganic salt 0.01 ~ 50g/L, all the other are water, pH5.0 ~ 9.0.
5. application according to claim 4 is characterized in that, described carbon source is any one or a few the combination in glucose, sucrose, maltose, lactose, wood sugar, fructose, lactic acid, citric acid, glycerine, starch and the molasses; Nitrogenous source is yeast powder, extractum carnis, peptone, yeast extract paste, corn steep liquor, soybean cake powder, cottonseed meal, urea, (NH
4)
2SO
4, NH
4Cl, (NH
4)
2HPO
4And NH
4NO
3In any one or a few combination; Inorganic salt are one or more in vitriol, phosphoric acid salt, dihydrogen phosphate, phosphoric acid hydrogen disalt and the hydrochloride.
6. application according to claim 3 is characterized in that, the condition of described proper growth is: initial pH is 5.0 ~ 9.0, culture temperature is that 25 ~ 38 ° of C, incubation time are 24 ~ 72h.
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| CN105385174A (en) * | 2015-10-27 | 2016-03-09 | 武汉理工大学 | Asphalt thickener and asphalt cement |
| CN108456652A (en) * | 2018-03-14 | 2018-08-28 | 南京工业大学 | Sphingosine monad gene engineering bacterium and construction method and application thereof |
| CN109234180A (en) * | 2017-07-10 | 2019-01-18 | 伽蓝（集团）股份有限公司 | Sphingol single-cell, its extracellular products and its preparation method and application |
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|---|---|---|---|---|
| CN104651284A (en) * | 2015-03-12 | 2015-05-27 | 南开大学 | Sphingomonas sp. T-3 and method for producing biological polysaccharide and poly-beta-hydroxy-butanoic acid by co-fermentation |
| CN104651284B (en) * | 2015-03-12 | 2017-12-05 | 南开大学 | Sphingol single-cell T 3 and its method for common fermentation production biological polyoses and poly- β hydroxybutyric acids |
| CN105385174A (en) * | 2015-10-27 | 2016-03-09 | 武汉理工大学 | Asphalt thickener and asphalt cement |
| CN105385174B (en) * | 2015-10-27 | 2017-10-24 | 武汉理工大学 | A kind of pitch thickener and bituminous cements |
| CN109234180A (en) * | 2017-07-10 | 2019-01-18 | 伽蓝（集团）股份有限公司 | Sphingol single-cell, its extracellular products and its preparation method and application |
| CN109234180B (en) * | 2017-07-10 | 2022-10-14 | 伽蓝（集团）股份有限公司 | Sphingomonas, extracellular products thereof, and preparation method and application thereof |
| CN108456652A (en) * | 2018-03-14 | 2018-08-28 | 南京工业大学 | Sphingosine monad gene engineering bacterium and construction method and application thereof |
| CN108456652B (en) * | 2018-03-14 | 2021-02-05 | 南京工业大学 | Sphingosine monad gene engineering bacterium and construction method and application thereof |
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