CN100451097C - Aspergillus niger - Google Patents
Aspergillus niger Download PDFInfo
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- CN100451097C CN100451097C CNB2006100488881A CN200610048888A CN100451097C CN 100451097 C CN100451097 C CN 100451097C CN B2006100488881 A CNB2006100488881 A CN B2006100488881A CN 200610048888 A CN200610048888 A CN 200610048888A CN 100451097 C CN100451097 C CN 100451097C
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- aspergillus niger
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- dioscin
- dioscorea panthaica
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Abstract
The invention discloses a microbe strain named Aspergillus niger YM33182 with reserving number at 206113, which is characterized by the following: transmitting Yunnan Dioscorea japonica dioscin material; improving extracting content by 40.
Description
Technical field
The present invention relates to microbial technology field, particularly relate to a kind of microorganism strains that can transform effective constituent kind and content in the Dioscorea panthaica Prain et Burkill of Yunnan.
Background technology
Yunnan Dioscorea panthaica Prain et Burkill (Dioscorea panthaica Yunnanensis) is Dioscoreaceae, yam.Its effective constituent is the steroid saponin material, and chemical ingredients comprises Progenin (A), Dioscin, Gracillin and pesudoproto-dioscin etc., and dioscin (dioscin) is the main component of Yunnan Dioscorea panthaica Prain et Burkill Chinese medicinal materials.The traditional Chinese medical science is thought that the Dioscorea panthaica Prain et Burkill root stock has and is activated blood circulation and dispersed blood clots the effect of promoting the circulation of QI to relieve pain.Modern pharmacology research, show that its root stock chemical ingredients dioscin (dioscin) can dilating coronary blood vessel, improve myocardial ischemia, has treatment myocardial ischemia disease, and the effect of anti-platelet aggregation and reducing blood-fat, clinical medicine is used for the treatment of cardiovascular and cerebrovascular diseases such as hyperlipidemia, stenocardia.Simultaneously, the diosgenin that obtains from the Dioscorea panthaica Prain et Burkill root stock also is the important lead compound of synthetic hormone medicine.
Summary of the invention
The object of the present invention is to provide a kind of separation from Chinese Xishuangbanna Nature Reserve soil, can impel microorganism--the aspergillus niger of Yunnan Dioscorea panthaica Prain et Burkill effective constituent steroidal saponin one class substance classes and content generation important change.
Aspergillus niger called after aspergillus niger Aspergillus niger YM33182 of the present invention now is deposited in specified depositary institution of State Intellectual Property Office, and deposit number is CCTCC NO:M206113.
Aspergillus niger of the present invention is to separate to obtain from Chinese Yunnan Province Xishuangbanna Nature Reserve soil.Now be deposited in the preservation of depositary institution of Patent Office of State Intellectual Property Office, depositary institution's title: Chinese typical culture collection center, depositary institution address: China, Wuhan University, postcode: 430072.Preservation date is on October 31st, 2006, and deposit number is M 206113.
Through morphologic observation, cultural characteristic and molecular biology 18SrDNA gene sequencing, determine its taxonomy status aspergillus niger (Aspergillus niger V.Tiegh) among imperfect fungi (Deuteromycotina) Aspergillus (Aspergillus link) the aspergillus niger group (Aspergillus niger) due to.
Aspergillus niger YM33182 is on the PDA substratum, and colony growth is very fast, cultivates 3-4 days colony diameters for 28 ℃ and just can surpass 20mm, and the bacterium colony outer rim is neat, and the surface is black, and like particulate state, middle mycelia densification is projection a little, and it is yellow that the reverse side center is, and is light black all around.Its representative configuration feature is that conidiophore is formed by the podocyte growth, obviously distinguishes with typical strain of the same race.
With 18SrDNA gene order homology is the phylogenetic tree that fundamental construction forms the relevant fungi of 10 strains that comprises bacterial strain YM34161, the result finds out, aspergillus niger YM33182 base sequence similarity is 100%, with this genus bacterial classification Aspergillus niger D63697 indifference.Referring to Fig. 5.
The culture medium culturing feature (see figure 1) of bacterial strain YM33182:
1. wort is cultivated after 4 days for dull and stereotyped last 28 ℃, and colony diameter can reach 36-40mm, and colony diameter can reach 44-50mm after 7 days.The bacterium colony outer rim is neat, and it is velvet-like to be white in color, and middle mycelia is deep green, protruding slightly, and aerial hyphae is flourishing.
2. PDA cultivates after 4 days for dull and stereotyped last 28 ℃, and colony diameter can reach 18-22mm, and colony diameter can reach 33-38mm after 7 days.The bacterium colony outer rim is neat, and mycelia is black, finely powdered, and middle mycelia densification is projection a little, and it is yellow that the reverse side center is, and is light black all around.
3. on the czapek's solution 28 ℃ cultivate colony growthing slows, cultivate that colony diameter is 12-16mm after 4 days, colony diameter reaches 20-22mm after 7 days.Whole bacterium colony projection, the edge green, middle endophyte silk is white in color, and aerial hyphae is black, and reverse side has fold, is light yellow.
The morphological specificity of YM33182 bacterial strain (seeing Fig. 2,3,4):
1. asexual generation mycelia have every, colourless, conidiophore expands hyphal cell (podocyte) by the heavy wall of specialization and bears, the conidiophore outer wall is smooth, conidiophore is two-layer, the top capsule is spherical or inferior spherical.The conidium fringe and is born in the globe-roof capsule, is black, and the edge splits, and forms the right cylinder of radial arrangement.Conidium is concatenated, and is spherical or circular, smooth, dead color, the about 2.6-2.8 μ of conidium diameter m.
2. sexual generation is not found.
The molecular biological characteristics of YM33182 bacterial strain:
Adopt the molecular biology round pcr, the dna sequencing instrument is analyzed, and this bacterium 18SrDNA genome is made up of 1861 bases (bp).Simultaneously based on 18S rDNA sequence homology, the be correlated with phylogenetic tree (see figure 5) of fungi of 10 strains that comprise bacterial strain YM33182 of structure.
YM33182 bacterial strain strain fermentation cultural characteristic:
1. substratum: Yunnan Dioscorea panthaica Prain et Burkill 50 (%), bright bean dregs 50 (%), (w/w) meter by weight, PH nature.
2. culture temperature: 28 ℃ ± 5 ℃.
3. fermentation time: 12-15 days.
4. fermentation culture feature: cultivated the 1st day, odd white hypha point growth is arranged.Cultivated the 2nd day, visible white hypha, mycelia is short, the fermented product caking.Cultivated the 4th day, mycelia is canescence, and the fermented product surface covers with the canescence mycelia, but shows sparse slightly.Cultivated the 5th day, the fermented product surface is canescence.Cultivated the 6th day, the fermented product surface covers with the beige mycelia, the fermented product densification.Cultivated the 7th day, fermented product is tight, is covered with the beige mycelia.Cultivated the 8th day, the fermented product surface covers with the black mycelia, and about 4/5ths parts are tawny on the fermented product, and about 1/5th parts are grey black down.Cultivated the 9th day, fermented product is grey black, and the yellow-green colour spore sporadicly appears in the surface, the fermented product densification.Cultivated the 10th day, fermented product is grey black, the fermented product densification, and the surface covers with the black spore.Cultivated the 11st day, fermented product was with the 10th day.After cultivating the 12nd day, be covered with the black mycelia, a large amount of grey yellow black spores in surface, fermented product is tight.
Yunnan Dioscorea panthaica Prain et Burkill (Dioscorea panthaica Yunnanensis) is the dry root stock of Dioscoreaceae plant, belong to Chinese herbal medicine yellow Chinese yam (Dioscorea panthaica) class medicinal material, main component has Progenin (A), Dioscin, Gracillin and pesudoproto-dioscin one class dioscin medicinal ingredients, Progenin (A) content 0.1-0.3% wherein, Dioscin is 1.5%-2.6%, the about 0.8-1.0% of Gracillin, pesudoproto-dioscin0.2-0.5%, starchy material 40-60%, protein, acids composition 10-16% is arranged, and other are 8.0% years old.With the Yunnan Dioscorea panthaica Prain et Burkill is fermented substrate, by aspergillus niger YM33182 microbial solid fermentation mode, can change the kind of Dioscorea panthaica Prain et Burkill dioscin effective constituent, form new natural dioscin metamember, the content that transforms Dioscorea panthaica Prain et Burkill dioscin medicinal ingredients is changed.
Yunnan of the present invention its processing step of Dioscorea panthaica Prain et Burkill effective constituent method for transformation is: bacterial classification → first order seed cultivation → secondary seed cultivation → fermented substrate → fermentation culture → oven dry → Dioscorea panthaica Prain et Burkill tunning → extraction → Dioscorea panthaica Prain et Burkill transforms extract → separation → dioscin.In this step, fermented substrate is Chinese medicinal materials Dioscorea panthaica Prain et Burkill (Dioscorea panthaica).This technology is suitable for other filamentous fungus quasi-microorganism of imperfect fungi.
In the above-mentioned Dioscorea panthaica Prain et Burkill zymotechnique, the fermentation condition of numbering YM33182 bacterial classification is as follows:
(1) fermentation mode: solid fermentation.
(2) seed culture: first order seed is cultivated and is adopted potato glucose solid medium (solid PDA).Secondary seed is cultivated and is potato glucose liquid nutrient medium (liquid PDA).
Potato glucose substratum (PDA) preparation: get fresh potato 200 grams, peeling is cut into small pieces, and adds 1 liter in water and boils 30 minutes, filters and abandons potato, and filter adds water and complement to 1 liter night, adds glucose 20 grams, the PH nature.
Add agar 15 grams in the above-mentioned potato glucose substratum, be solid PDA substratum.
(3) fermentation culture: fermention medium is Semen Ginkgo extrac (25-30%), bright bean dregs (30-35%), wheat bran (30-35%), (w/w) meter by weight.The PH order is right.
(4) incubation time: strain activation and culture 72-96 hour, the static incubation time 72-96 of first order seed hour, secondary seed shaking table incubation time 96-120 hour, fermented incubation time 12-15 days.
(5) culture temperature: 28 ℃ ± 2 ℃ of first order seed culture temperature, 28 ℃ ± 2 ℃ of secondary seed shaking table culture temperature, fermentation culture temperature are 28 ℃ ± 5 ℃.
(6) oven dry: fermentation finishes, and tunning is put 60 ℃ of oven dry down, and is standby.
(7) extract: tunning 70-90% alcohol extraction through concentrating, obtains Dioscorea panthaica Prain et Burkill and transforms extract.
(8) separate: Dioscorea panthaica Prain et Burkill transforms extract and adopts the silica gel G column chromatography for separation, through gel (Sephadex) post layer purifying, obtains the dioscin compounds respectively again.
The present invention is by the microbial fermentation approach, with the Chinese medicinal materials Dioscorea panthaica Prain et Burkill is matrix, by microbial conversion, excavate new medicinal part, new effective constituent or new texture (lead compound), promote the pharmaceutical use of Dioscorea panthaica Prain et Burkill, in the hope of obtaining anti-human cardiovascular and cerebrovascular diseases new type natural medicinal ingredients.
After bacterial strain YM33182 fermentation transforms 12-15 days, compare with former Yunnan Dioscorea panthaica Prain et Burkill, Dioscorea panthaica Prain et Burkill product after it transforms by β-daucosterol (Daucosterol) (I), diosgenin-α-L rhamnosyl (1 → 4)-β-D glycoside (Prosapogenin B) (II), diosgenin [α-L rhamnosyl (1 → 2)]-α-L rhamnosyl (1 → 4)-β-D glycoside (Dioscin) (III), diosgenin [α-L rhamnosyl (1 → 2)]-β-D glucose (1 → 3)-β-D glycoside (Gracillin) (IV) waits 4 kinds of main body dioscin materials to form.Former Dioscorea panthaica Prain et Burkill Progenin (A) and the degraded of pesudoproto-dioscin dioscin composition disappear, simultaneously, transform to form β-daucosterol (daucosterol) (I) and (II) new natural product of diosgenin-α-L rhamnosyl (1 → 4)-β-D glycoside (Prosapogenin B), its structural formula is known.
The Dioscorea panthaica Prain et Burkill converted product of bacterial strain YM33182, compare with former Dioscorea panthaica Prain et Burkill, converted product diosgenin content generation noticeable change, promptly Prosapogenin B content 10%-11%, Dioscin are that 10%-12%, Gracillin are that 9%-10%, Daucosterol have also reached 7%-8% in the converted product.Simultaneously, Yunnan Dioscorea panthaica Prain et Burkill (Dioscorea panthaica Yunnanensis) is behind numbering YM33182 fermentation of Aspergillus niger, and the converted product dioscin extracts total content and reached 40%, and is the highest by 42%.Compare with Dioscorea panthaica Prain et Burkill Chinese medicinal materials dioscin total content before the conversion 36%, transform back dioscin total content and improved 6 percentage points, can reach 8 percentage points.This shows that this microorganism is the crude drug Microbial resources that transform Yunnan Dioscorea panthaica Prain et Burkill Chinese medicinal materials dioscin class substance classes and content.
Description of drawings
Fig. 1 is the bacterium colony figure of aspergillus niger YM33182 of the present invention on malt extract medium.
Fig. 2 is aspergillus niger YM33182 bacterial strain microscope figure of the present invention (* 200).
Fig. 3 is aspergillus niger YM33182 conidium fringe Electronic Speculum figure of the present invention (* 1800).
Fig. 4 is aspergillus niger YM33182 conidium Electronic Speculum figure of the present invention (* 5000).
Fig. 5 is the phylogenetic tree of YM33182 aspergillus niger of the present invention (Aspergillus niger V.Tiegh) 18SrDNA sequence homology.
Fig. 6 is the HPLC analysis chart of dioscin kind before and after Dioscorea panthaica Prain et Burkill of the present invention transforms.
Embodiment
Embodiment:
The Dioscorea panthaica Prain et Burkill fermentation culture of YM33182 bacterial strain:
Aspergillus niger YM33182 bacterial strain is fermentation conversion Yunnan Dioscorea panthaica Prain et Burkill (Dioscorea panthaicaYunnanensis) Chinese medicinal materials, raising or the natural microbial medicine source that changes anti-human cardiovascular disease's medicinal ingredients.
Get YM33182 (aspergillus niger Aspergillus niger V.Tiegh) bacterial classification, under aseptic condition (sterilisable chamber or Bechtop), with a little bacterial classification of inoculating needle picking, transfer and (in 15 * 150mm), put interior 28 ℃ ± 1 ℃ activation culture of incubator 96 hours in sterilized solid PDA (wort, Cha Shi, Ma Dingshi etc.) substratum test tube.Take out 96 hours bacterial classification of activation, under aseptic condition, insert the good bacterial classification of activation in sterilized first order seed solid PDA substratum, cultivated 72-96 hour down, be the YM33182 first order seed at 28 ℃ ± 2 ℃ with the same manner.
Adopt 500 milliliters of glass triangle bottles, 100 milliliters of the liquid PDA substratum that prepare of packing into, 15 pounds of sterilizations 30 minutes are standby.Get YM33182 bacterial strain first order seed, under aseptic condition, be inoculated in 500 milliliters of glass triangle bottles that 100 milliliters of liquid PDA substratum are housed, the first order seed inoculum size is that every inclined-plane connects three triangular flasks, put 28 ℃ ± 2 ℃ shaking tables and cultivate after 72-96 hour, use as the fermentation secondary seed.
According to the fermentation scale, by fermentation material weight ratio, take by weighing Dioscorea panthaica Prain et Burkill dry powder (fineness 100-200 order) 50% and add 50% bright bean dregs, fully mix control fermentation material moisture 62-68% thoroughly.After Semen Ginkgo extrac fermentation material prepares, pack in 500 milliliters of Cans, plastics film, kraft paper seal, and put in the Autoclave 15 pounds of sterilizations after 60 minutes, takes out to be cooled to 30 ℃, access liquid secondary seed, it is 2-5% (v/w) that secondary seed connects consumption.Shake up, place 28 ℃ ± 5 ℃ under, static fermentation 12-15 days.During fermentation, the control proving room 70-80% relative humidity of not preserving moisture low was shaken bottle respectively once on the 2nd day, and noticed that inspection has or not the microbiological contamination phenomenon.
Fermentation finishes, and tunning is put 60 ℃ of oven dry down, obtains the Dioscorea panthaica Prain et Burkill tunning.Handle through chemical extraction, isolation technique again, can obtain transforming Dioscorea panthaica Prain et Burkill dioscin compound.
Aspergillus niger of the present invention (Aspergillus niger V.Tiegh) YM33182 18SrDNA genome is made up of 1861 bases (bp), and its series is as follows:
1 GCTATGCATC?AACGCGTTTG?GGAGCTCTCC?CATATGGTCG?ACCTGCAGGC?GGCCGCACTA
61 GTGATTGTAG?TCATATGCTT?GTCTCAAAGA?TTAAGCCATG?CATGTCTAAG?TATAAGCACT
121 TTATACTGTG?AAACTGCGAA?TGGCTCATTA?AATCAGTTAT?CGTTTATTTG?ATAGTACCTT
181 ACTACATGGA?TACCTGTGGT?AATTCTAGAG?CTAATACATG?CTGAAAACCT?CGACTTCGGA
241 AGGGGTGTAT?TTATTAGATA?AAAAACCAAT?GCCCTTCGGG?GCTCCTTGGT?GAATCATAAT
301 AACTTAACGA?ATCGCATGGC?CTTGCGCCGG?CGATGGTTCA?TTCAAATTTC?TGCCCTATCA
361 ACTTTCGATG?GTAGGATAGT?GGCCTACCAT?GGTGGCAACG?GGTAACGGGG?AATTAGGGTT
421 CGATTCCGGA?GAGGGAGCCT?GAGAAACGGC?TACCACATCC?AAGGAAGGCA?GCAGGCGCGC
481 AAATTACCCA?ATCCCGACAC?GGGGAGGTAG?TGACAATAAA?TACTGATACG?GGGCTCTTTT
541 GGGTCTCGTA?ATTGGAATGA?GTACAATCTA?AATCCCTTAA?CGAGGAACAA?TTGGAGGGCA
601 AGTCTGGTGC?CAGCAGCCGC?GGTAATTCCA?GCTCCAATAG?CGTATATTAA?AGTTGTTGCA
661 GTTAAAAAGC?TCGTAGTTGA?ACCTTGGGTC?TGGCTGGCCG?GTCCGCCTCA?CCGCGAGTAC
721 TGGTCCGGCT?GGACCTTTCC?TTCTGGGGAA?TCTCATGGCC?TTCACTGGCT?GTGGGGGGAA
781 CCAGGACTTT?TACTGTGAAA?AAATTAGAGT?GTTCAAAGCA?GGCCTTTGCT?CGAATACATT
841 AGCATGGAAT?AATAGAATAG?GACGTGCGGT?TCTATTTTGT?TGGTTTCTAG?GACCGCCGTA
901 ATGATTAATA?GGGATAGTCG?GGGGCGTCAG?TATTCAGCTG?TCAGAGGTGA?AATTCTTGGA
961 TTTGCTGAAG?ACTAACTACT?GCGAAAGCAT?TCGCCAAGGA?ATGTTTTCAT?TAATCAGGGA
1021 ACGAAAGTTA?GGGGATCGAA?GACGATCAGA?TACCGTCGTA?GTCTTAACCA?TAAACTATGC
1081 CGACTAGGGA?TCGGACGGTG?TTTCTATTAT?GACCCGTTCG?GCACCTTACG?AGAAATCAAA
1141 GTTTTTGGGT?TCTGGGGGGA?GTATGGTCGC?AAGGCTGAAA?CTTAAAGAAA?TTGACGGAAG
1201 GGCACCACCA?GGCGTGGAGC?CTGCGGCTTA?ATTTGACTCA?ACACGGGGAA?ACTCACCAGG
1261 TCCAGACAAA?ATAAGGATTG?ACAGATTGAG?AGCTCTTTCT?TGATCTTTTG?GATGGTGGTG
1321 CATGGCCGTT?CTTAGTTGGT?GGAGTGATTT?GTCTGCTTAA?TTGCGATAAC?GAACGAGACC
1381 TCGGCCCTTA?AATAGCCCGG?TCCGCATTTG?CGGGCCGCTG?GCTTCTTAGG?GGGACTATCG
1441 GCTCAAGCCG?ATGGAAGTGC?GCGGCAATAA?CAGGTCTGTG?ATGCCCTTAG?ATGTTCTGGG
1501 CCGCACGCGC?GCTACACTGA?CAGGGCCAGC?GAGTACATCA?CCTTGGCCGA?GAGGTCTGGG
1561 TAATCTTGTT?AAACCCTGTC?GTGCTGGGGA?TAGAGCATTG?CAATTATTGC?TCTTCAACGA
1621 GGAATGCCTA?GTAGGCACGA?GTCATCAGCT?CGTGCCGATT?ACGTCCCTGC?CCTTTGTACA
1681 CACCGCCCGT?CGCTACTACC?GATTGAATGG?CTCGGTGAGG?CCTTCGGACT?GGCTCAGGAG
1741 GGTTGGCAAC?GACCCCCCAG?AGCCGGAAAG?TTGGTCAAAC?CCGGTCATTT?AGAGGAAGTA
1801 AAAGTCGTAA?CAAGGTTTCC?GTAGGTGAAC?CTGCGGAAAT?CCCGCGGCCA?TGGCGGCCGG
1861 AGC。
Claims (2)
1, a kind of aspergillus niger is characterized in that, the specific name of this aspergillus niger is aspergillus niger Aspergillusniger YM33182, now is deposited in specified depositary institution of State Intellectual Property Office, and deposit number is CCTCCNO:M 206113.
2, according to the described aspergillus niger of claim 1, it is characterized in that its 18SrDNA genome by 1861 based compositions, its sequence is as follows:
1 GCTATGCATC?AACGCGTTTG?GGAGCTCTCC?CATATGGTCG?ACCTGCAGGC?GGCCGCACTA
61 GTGATTGTAG?TCATATGCTT?GTCTCAAAGA?TTAAGCCATG?CATGTCTAAG?TATAAGCACT
121 TTATACTGTG?AAACTGCGAA?TGGCTCATTA?AATCAGTTAT?CGTTTATTTG?ATAGTACCTT
181 ACTACATGGA?TACCTGTGGT?AATTCTAGAG?CTAATACATG?CTGAAAACCT?CGACTTCGGA
241 AGGGGTGTAT?TTATTAGATA?AAAAACCAAT?GCCCTTCGGG?GCTCCTTGGT?GAATCATAAT
301 AACTTAACGA?ATCGCATGGC?CTTGCGCCGG?CGATGGTTCA?TTCAAATTTC?TGCCCTATCA
361 ACTTTCGATG?GTAGGATAGT?GGCCTACCAT?GGTGGCAACG?GGTAACGGGG?AATTAGGGTT
421 CGATTCCGGA?GAGGGAGCCT?GAGAAACGGC?TACCACATCC?AAGGAAGGCA?GCAGGCGCGC
481 AAATTACCCA?ATCCCGACAC?GGGGAGGTAG?TGACAATAAA?TACTGATACG?GGGCTCTTTT
541 GGGTCTCGTA?ATTGGAATGA?GTACAATCTA?AATCCCTTAA?CGAGGAACAA?TTGGAGGGCA
601 AGTCTGGTGC?CAGCAGCCGC?GGTAATTCCA?GCTCCAATAG?CGTATATTAA?AGTTGTTGCA
661 GTTAAAAAGC?TCGTAGTTGA?ACCTTGGGTC?TGGCTGGCCG?GTCCGCCTCA?CCGCGAGTAC
721 TGGTCCGGCT?GGACCTTTCC?TTCTGGGGAA?TCTCATGGCC?TTCACTGGCT?GTGGGGGGAA
781 CCAGGACTTT?TACTGTGAAA?AAATTAGAGT?GTTCAAAGCA?GGCCTTTGCT?CGAATACATT
841 AGCATGGAAT?AATAGAATAG?GACGTGCGGT?TCTATTTTGT?TGGTTTCTAG?GACCGCCGTA
901 ATGATTAATA?GGGATAGTCG?GGGGCGTCAG?TATTCAGCTG?TCAGAGGTGA?AATTCTTGGA
961 TTTGCTGAAG?ACTAACTACT?GCGAAAGCAT?TCGCCAAGGA?ATGTTTTCAT?TAATCAGGGA
1021 ACGAAAGTTA?GGGGATCGAA?GACGATCAGA?TACCGTCGTA?GTCTTAACCA?TAAACTATGC
1081 CGACTAGGGA?TCGGACGGTG?TTTCTATTAT?GACCCGTTCG?GCACCTTACG?AGAAATCAAA
1141 GTTTTTGGGT?TCTGGGGGGA?GTATGGTCGC?AAGGCTGAAA?CTTAAAGAAA?TTGACGGAAG
1201 GGCACCACCA?GGCGTGGAGC?CTGCGGCTTA?ATTTGACTCA?ACACGGGGAA?ACTCACCAGG
1261 TCCAGACAAA?ATAAGGATTG?ACAGATTGAG?AGCTCTTTCT?TGATCTTTTG?GATGGTGGTG
1321 CATGGCCGTT?CTTAGTTGGT?GGAGTGATTT?GTCTGCTTAA?TTGCGATAAC?GAACGAGACC
1381 TCGGCCCTTA?AATAGCCCGG?TCCGCATTTG?CGGGCCGCTG?GCTTCTTAGG?GGGACTATCG
1441 GCTCAAGCCG?ATGGAAGTGC?GCGGCAATAA?CAGGTCTGTG?ATGCCCTTAG?ATGTTCTGGG
1501 CCGCACGCGC?GCTACACTGA?CAGGGCCAGC?GAGTACATCA?CCTTGGCCGA?GAGGTCTGGG
1561 TAATCTTGTT?AAACCCTGTC?GTGCTGGGGA?TAGAGCATTG?CAATTATTGC?TCTTCAACGA
1621 GGAATGCCTA?GTAGGCACGA?GTCATCAGCT?CGTGCCGATT?ACGTCCCTGC?CCTTTGTACA
1681 CACCGCCCGT?CGCTACTACC?GATTGAATGG?CTCGGTGAGG?CCTTCGGACT?GGCTCAGGAG
1741 GGTTGGCAAC?GACCCCCCAG?AGCCGGAAAG?TTGGTCAAAC?CCGGTCATTT?AGAGGAAGTA
1801 AAAGTCGTAA?CAAGGTTTCC?GTAGGTGAAC?CTGCGGAAAT?CCCGCGGCCA?TGGCGGCCGG
1861 AGC。
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| CN103173427A (en) * | 2012-07-12 | 2013-06-26 | 北京伟嘉人生物技术有限公司 | Method for producing beta-mannase by utilizing Aspergillus niger fermentation |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1397649A (en) * | 2002-08-19 | 2003-02-19 | 上海华冠生物芯片有限公司 | Biochip base on 16S rDNA gene for diagnosing frequently encountered pathogenic bacteria |
| CN1188425C (en) * | 2002-12-09 | 2005-02-09 | 吉林天药科技股份有限公司 | Novel process for preparing diosgenin |
| CN1228344C (en) * | 2002-10-21 | 2005-11-23 | 吉林天药科技股份有限公司 | Method for preparing yam saponin, medicinal preparation and new usage in medication |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1397649A (en) * | 2002-08-19 | 2003-02-19 | 上海华冠生物芯片有限公司 | Biochip base on 16S rDNA gene for diagnosing frequently encountered pathogenic bacteria |
| CN1228344C (en) * | 2002-10-21 | 2005-11-23 | 吉林天药科技股份有限公司 | Method for preparing yam saponin, medicinal preparation and new usage in medication |
| CN1188425C (en) * | 2002-12-09 | 2005-02-09 | 吉林天药科技股份有限公司 | Novel process for preparing diosgenin |
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