CN105779473A - Synthesis gene cluster of rhamnose isoflavone and applications thereof - Google Patents
Synthesis gene cluster of rhamnose isoflavone and applications thereof Download PDFInfo
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Abstract
The invention relates to a field of microorganism gene engineering, and specifically discloses a synthesis gene cluster of rhamnose isoflavone and applications thereof. The synthesis gene cluster comprises seven genes: AORI_0586 gene encoded glucose 1-phosphoric acid thymidine transferase; AORI_0991 gen encoded deoxythymidine diphosphate-glucose-4,6-dehydrogenase; AORI_3188 gene encoded deoxythymidine diphosphate-glucose-4,6-dehydrogenase; AORI_0570 gene encoded deoxythymidine diphosphate-4-dehydrorhamnose-3,5-epimerase; AORI_1501 gene encoded deoxythymidine diphosphate-4-dehydrorhamnose-3,5-epimerase; AORI_6774 gene encoded deoxythymidine diphosphate-4-dehydrorhamnose-3,5-epimerase; and AORI_0992 gene encoded thymidine diphosphate-4-dehydrorhamnose reductase. The provided gene cluster can be used to prepare rhamnose isoflavone compounds.
Description
Technical field
The present invention relates to microbiological genetic engineering field, be specifically related to synthetic gene bunch and the application thereof of rhamnose isoflavone.
Background technology
The general low molecule polyphenols being distributed in plant of flavone compound (fiavonoids) is many with free state or be combined into the form of glycosides with sugar and exist in plant.This compounds generally has antioxidation, antitumor, anticancer, antiinflammatory, analgesia, hepatoprotective, antibacterial, antiviral, prevents multiple biological activity and the pharmacological actions such as arteriosclerosis.
Rhamnose, 6-deoxy-L-mannose, it is widely present in the polysaccharide of plant, glucosides, plant gum and bacterial polysaccharides.2000, Marie-FranceGiraud and JamesHNaismith elaborates rhamnose approach (GiraudMF, NaismithJH.Therhamnosepathway [J] .Currentopinioninstructuralbiology, 2000,10 (6): 687-696).2006, YoonTM etc. separate and obtain a kind of rhamnose genistein glycosides, proof has antifungal activity (YoonTM, KimJW, KimJG, etal.TalosinsAandB:newisoflavonolglycosideswithpotentant ifungalactivityfromKitasatosporakifunensisMJM341 [J] .JournalofAntibiotics, 2006,59:633-639.).And in 1993, KoyotoshiNishiyama etc. synthesize the isoflavone aglycone that a series of glycosyl replaces, prove that beta galactosidase is had stronger competitive inhibition (KiyotoshiN by the sugar-substituted isoflavone aglycone of Fructus rhamni (Rhamnus davurica Pall.), SachikoE, IkukoD, etal.Synthesesisoflavonesandisoflavoneglycosides, andtheirinhibitoryactivityagainstbovineliver β-galactosidase [J] .BiosciBiotechBiochem, 1993,57 (1): 107-114.).2009, Liu imitate steady wait in the Orient amycolatosis fermentation liquid to separate obtain small component rhamnose soybean isoflavone, thus it is speculated that for some composition in fermentation medium be Amycolatopsis orientalis bioconversion (Liu Xiaowen, Li Qiushuan, Ruan Lingao, etc..The isolation identification [J] of Amycolatopsis orientalis fermentation liquid small component. China's antibiotic magazine, 2009,34 (11): 654-658)
Up to now, but without the report about rhamnose isoflavone synthetic gene bunch correlational study.
Summary of the invention
In order to overcome the deficiencies in the prior art, it is an object of the invention to provide the synthetic gene bunch of a kind of rhamnose isoflavone and application thereof.
The present invention adopts the following technical scheme that
First aspect present invention provides the synthetic gene bunch of a kind of rhamnose isoflavone, the synthetic gene bunch of described rhamnose isoflavone is from Amycolatopsisorientalis Amycolatopsis orientalis CGMCCNo.6023, described Amycolatopsis orientalis, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preserving number CGMCCNo.6023 on April 20th, 2012.
Preferably, the synthetic gene bunch of described rhamnose isoflavone comprises 7 genes:
(1) AORI_0586 gene, nucleotide sequence is such as shown in SEQIDNO.1, and length is 882 base pairs, and encoding amino acid sequence is glucose 1-phosphoric acid thymus pyrimidine transferring enzyme (293 aminoacid) shown in SEQIDNO.8 such as；
(2) AORI_0991 gene, nucleotide sequence is such as shown in SEQIDNO.2, and length is 984 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-glucose-4 shown in SEQIDNO.9 such as, 6-dehydrogenase (327 aminoacid)；
(3) AORI_3188 gene, nucleotide sequence is such as shown in SEQIDNO.3, and length is 960 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-glucose-4 shown in SEQIDNO.10 such as, 6-dehydrogenase (319 aminoacid)；
(4) AORI_0570 gene, nucleotide sequence is such as shown in SEQIDNO.4, and length is 549 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-4-dehydrogenation rhamnose-3 shown in SEQIDNO.11 such as, 5-epimerase (182 aminoacid)；
(5) AORI_1501 gene, nucleotide sequence is such as shown in SEQIDNO.5, and length is 618 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-4-dehydrogenation rhamnose-3 shown in SEQIDNO.12 such as, 5-epimerase (205 aminoacid)；
(6) AORI_6774 gene, nucleotide sequence is such as shown in SEQIDNO.6, and length is 606 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-4-dehydrogenation rhamnose-3 shown in SEQIDNO.13 such as, 5-epimerase (201 aminoacid)；
(7) AORI_0992 gene, nucleotide sequence is such as shown in SEQIDNO.7, and length is 891 base pairs, and encoding amino acid sequence is the thymidine diphosphate-4-dehydrogenation rhamnose reductase (296 aminoacid) shown in SEQIDNO.14 such as.
Second aspect present invention provides the albumen of the synthetic gene bunch coding of rhamnose isoflavone described in first aspect present invention.
Third aspect present invention discloses expression vector or the genetic engineering bacterium of the synthetic gene bunch containing rhamnose isoflavone of the present invention, belongs to protection scope of the present invention.
Fourth aspect present invention discloses the host cell of the synthetic gene bunch of the aforementioned rhamnose isoflavone being integrated with external source on foregoing expression vectors or chromosome.
Fifth aspect present invention discloses the construction method of the engineering bacteria of the synthetic gene bunch containing described rhamnose isoflavone, it is simply that the recombinant expression carrier of the synthetic gene bunch containing above-mentioned rhamnose isoflavone is imported host；Or directly the synthetic gene bunch of above-mentioned rhamnose isoflavone is inserted the chromosome of expressive host.
Preferably, described host is selected from any one in escherichia coli, Rhodococcus fascians, Nocard's bacillus, bacillus subtilis, lactic acid bar bacterium or yeast；Preferred host is escherichia coli or Rhodococcus fascians.
The expression vector of the above-mentioned synthetic gene bunch for inserting described rhamnose isoflavone refers to the plasmid vector expressed in described host, such as can at the pET of expression in escherichia coli, or in Rhodococcus fascians express shuttle vector Pnv18, or can Pichia sp. strain express pPIC9K etc..The recombinant expression carrier of the synthetic gene bunch containing above-mentioned rhamnose isoflavone can build by conventional method of the prior art.
The reconstitution cell of the synthetic gene bunch containing above-mentioned rhamnose isoflavone can build by conventional methods of homologous recombination of the prior art.
Sixth aspect present invention discloses a kind of Amycolatopsis orientalis, and containing the synthetic gene bunch of aforesaid rhamnose isoflavone, its deposit number is CGMCCNo.6023.
Seventh aspect present invention discloses a kind of method preparing rhamnose isoflavone and the like, including step: cultivate the host cell described in fourth aspect present invention or power Amycolatopsis orientalis described in sixth aspect present invention, thus expressing rhamnose isoflavone and the like, and from culture fluid, separate rhamnose isoflavone and the like.
Eighth aspect present invention discloses the encoding proteins described in the synthetic gene bunch of rhamnose isoflavone, second aspect present invention described in first aspect present invention, the expression vector of third aspect present invention is lived or the application in preparation rhamnose isoflavone and the like of Amycolatopsis orientalis described in sixth aspect present invention.
Ninth aspect present invention, it is provided that a kind of Amycolatopsis orientalis, in described Amycolatopsis orientalis, in the synthetic gene bunch of aforesaid rhamnose isoflavone, one or more genes are deactivated, thus not producing rhamnose isoflavone and the like.The non-deactivated bacterium of the same race of described gene can produce rhamnose isoflavone and the like.
Preferably, described rhamnose isoflavone and the like specifically refers to: rhamnose fire-bote flavonoid glycoside, rhamnose daidzein glycosides or rhamnose Semen Glycines flavonoid glycoside.
The invention have the benefit that
(1) synthetic gene bunch of rhamnose isoflavone of the present invention, the nucleotide provided or partial nucleotide sequence, may utilize the method for polymerase chain reaction (PCR) or the DNA comprising sequence of the present invention carries out the method for Southern hybridization as probe and obtains the homologous genes of rhamnose isoflavone synthetic gene from other microorganisms；
(2) comprise the cloned DNA of gene cluster nucleotide sequence provided by the present invention or at least part of nucleotide sequence to may be used for eastwardly amycolatosis Amycolatopsisorientalis genomic library positioning more Library plasmid, these Library plasmid, including at least the partial nucleotide sequence in the present invention, also include the DNA that before in Amycolatopsisorientalis genome, adjacent domain is not cloned；
(3) clone gene of nucleotide sequence provided by the present invention and partial nucleotide sequence or DNA fragmentation can obtain precursor or the derivant of new rhamnose isoflavone by interrupting rhamnose isoflavone one or several step biosynthetic or other homologous geness of primer；
(4) clone gene of nucleotide sequence provided by the present invention or at least part of nucleotide sequence can express to obtain corresponding enzyme or other higher biological activity or yield by suitable expression system in foreign host；
(5) the nucleotide sequence provided by the present invention or at least partly gene of nucleotide sequence or gene cluster can build plasmid to obtain new bio route of synthesis by genetic recombination, it is also possible to by inserting, displacement, disappearance or inactivation and then acquisition new bio route of synthesis；
(6) nucleotide sequence provided by the present invention or at least part of nucleotide sequence can be modified or suddenly change.These approach include inserting, replacing or disappearance, polymerase chain reaction, mistake mediated polymerization polymerase chain reaction, mutation site-specific, not homotactic reconnect, the different piece of sequence or the homologous sequence with other sources are oriented evolution, or by ultraviolet or chemical reagent mutation etc.；
(7) nucleotide sequence provided by the present invention or partial nucleotide sequence can be used to regulate the yield of rhamnose isoflavone or derivatives thereof；
(8) nucleotide sequence provided by the present invention or at least partly nucleotide sequence gene or gene cluster can be expressed in heterologous host and by DNA chip technology they functions in host metabolism of understanding；
(9) nucleotide sequence provided by the present invention or multiple sequence can merge with carrier sequence and obtain recombination sequence and corresponding DNA molecular.
In a word, all gene informations that rhamnose isoflavone provided by the present invention synthesis is relevant contribute to illustrating and understand the biosynthetic molecule mechanism of rhamnose isoflavone, thus for providing fundamental basis and material further with genetic engineering means transformation.Gene provided by the present invention may also be used for finding and inventing may be used for medicine, industrial or agriculture compound or gene, albumen.
Bacterial strain preservation information of the present invention is as follows:
Strain name: Amycolatopsis orientalis Amycolatopsisorientalis；
Preserving number is: CGMCCNo.6023.
Preservation date: on April 20th, 2012；
Depositary institution's title: China Committee for Culture Collection of Microorganisms's common micro-organisms center；
Depositary institution is called for short: CGMCC；
Depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica.
Accompanying drawing explanation
Fig. 1 is the UPLC-MS ion flow graph of blank cultures and CGMCCNo.6023 fermentation liquid.
Fig. 2 is the mass spectrum of rhamnose genistein glycosides, rhamnose soybean isoflavone aglycones and rhamnose Semen Glycines flavonoid glycoside.
Fig. 3 is the biosynthetic schematic diagram of rhamnose isoflavone.
Detailed description of the invention
Before further describing the specific embodiment of the invention, it should be appreciated that protection scope of the present invention is not limited to following specific specific embodiments；It is also understood that the term used in the embodiment of the present invention is to describe specific specific embodiments, rather than in order to limit the scope of the invention.
When embodiment provides numerical range, it should be appreciated that unless the present invention is otherwise noted, between two end points and two end points of each numerical range, any one numerical value all can be selected for.Unless otherwise defined, the same meaning that all technology used in the present invention and scientific terminology and those skilled in the art of the present technique are generally understood that.Except the concrete grammar used in embodiment, equipment, material, record according to those skilled in the art's grasp to prior art and the present invention, it is also possible to use similar with the method described in the embodiment of the present invention, equipment, material or that be equal to any method of prior art, equipment and material to realize the present invention.
Unless otherwise indicated, the experimental technique that disclosed in this invention, detection method, preparation method all adopt the conventional molecular biology of the art, biochemistry, chromatin Structure and analysis, analytical chemistry, cell are cultivated, the routine techniques of recombinant DNA technology and association area.These technology existing improving in existing document illustrates, specifically can referring to the MOLECULARCLONING:ALABORATORYMANUAL such as Sambrook, Secondedition, ColdSpringHarborLaboratoryPress, 1989andThirdedition, 2001；Ausubel etc., CURRENTPROTOCOLSINMOLECULARBIOLOGY, john wiley & sons, NewYork, 1987andperiodicupdates；TheseriesMETHODSINENZYMOLOGY, AcademicPress, SanDiego；Wolffe, CHROMATINSTRUCTUREANDFUNCTION, Thirdedition, AcademicPress, SanDiego, 1998；METHODSINENZYMOLOGY, Vol.304, Chromatin (P.M.WassarmanandA.P.Wolffe, eds.), AcademicPress, SanDiego, 1999；And METHODSINMOLECULARBIOLOGY, Vol.119, ChromatinProtocols (P.B.Becker, ed.) HumanaPress, Totowa, 1999 etc..
The discovery of embodiment 1 rhamnose isoflavonoid
Bacterial strain: AmycolatopsisorientalisCGMCCNO.6023
Slant medium: Gause I culture medium.
Seed culture medium (g/L):
Fermentation medium (g/L):
Sweat and condition:
From fresh inclined-plane, shovel scrapes a certain amount of spore, accesses in seed culture medium, is placed on 220rpm rotary shaker shaken cultivation 44～48h, cultivation temperature 28 DEG C, relative humidity 40%～60%.
Aseptically, proceeding in fermentation shake flask by cultured seed liquor, inoculum concentration is 8%, is placed in shaken cultivation 120h on 220rpm rotary shaker, cultivation temperature 28 DEG C.
Fermentation liquid is added isopyknic methanol, soaks 1 hour, be centrifuged 20 minutes under 8000rpm, take supernatant.Carry out UPLC-MS analysis.UPLC-MS analysis condition is as follows:
UPLC-MS: chromatographic column: AcquityUPLCBEHC18column, 1.7 μm, 100mm × 2.1mm (Waters, Milford, USA), column temperature: 45 DEG C；
Mobile phase: A phase is 0.05% formic acid water, and B phase is 0.05% formic acid-acetonitrile；
Flow velocity: 0.4ml/min
Eluting flow process:
Fermentation liquid is found that m/z401.12 [M+H]+、417.12[M+H]+、431.13[M+H]+Compound, in conjunction with document, thus it is speculated that these three compound molecule formula respectively C21H20O8、C21H20O9、C22H22O9, for rhamnose isoflavone like substance, its aglycon respectively daidzein, genistein and daidzein.
The extraction of embodiment 2AmycolatopsisorientalisCGMCCNo.6023 STb gene
1) take fresh slant pore and be inoculated in equipped with in 20mlYMB fluid medium, 28 DEG C, 220rpm shaken cultivation 36hr.
2) it is centrifuged (3000g, 15min) and collects mycelium, wash once with sterilized water.
3) mycelium is resuspended in 3mlTE, shakes through whirlpool scattered.
4) adding 300ul lysozyme (10mg/ml) and 30ulRNase (10mg/ml), 37 DEG C of water-bath 90min, period constantly vibrates.
5) 210ul20%SDS and 30ul E.C. 3.4.21.64 (10mg/ml), 55 DEG C of water-bath 2hr are added.
6) 600ul5MNaCl and 480ul10%CTAB, 65 DEG C of water-bath 30min are added.
7), after being cooled to room temperature, equal-volume chloroform is added: phenol: isoamyl alcohol (25:24:1), mixing, centrifugal (8900g, 15min).
8) take supernatant, add equal-volume chloroform, mixing, centrifugal (8900g, 15min).
9) take upper strata phase, the adherent isopropanol being slowly added to 0.6 times of volume, reverse mixing, precipitate out to floccule.
10) slowly stir with the Glass rod of sealing, DNA is wound on it, by 70% washing with alcohol twice, drying at room temperature, it is dissolved in 100ulTE buffer, 4 DEG C of preservations
Embodiment 3 high-flux sequence
1) by the DNA fragmentation of embodiment 2 gained, sequencing library is built；
2) the GSFLX sequenator of Roche 454 Corp. is used to carry out high-flux sequence: to obtain 561423 and read sequence (reads), after the assembled software carried with 454 Corp. splices, contig number is 56 (> 500bp), coverage is 25.6 times of full-length genome.
3) assembling is spliced further, complete filling-up hole work (gapclosing): common mistake designs 548 specific PCRs and 2 fosmid clone's shotgun order-checkings, and final assembly obtains the complete genomic sequence of Amycolatopsis orientalis CGMCCNo.6023.Wherein, every 100,000 base error rates, less than 0.5, exceed international requirement every 100,000 the base error rates level less than 1.What full-length genome assembly adopted is phred/phrap/consed packet.
Embodiment 4 genomeannotation
1) determination of open reading frame (ORF): first predict with Glimmer3.02, be then corrected with Genemark and Z-curve.
2) protein function annotation: to the CDSs predicted, carry out preliminary genetic annotation by BLASTP comparison KEGG and NR data base, all annotation results are each through artificial last manual correction；And ortholog (clustersoforthologousgroups, COGs) and the prediction of albumen conserved domain then obtained by the CDD data base of comparison NCBI, the confirmation parameter of COGs is identity >=30%, lengthdiversity≤20%.
Table 1, rhamnose isoflavone biological synthesis gene cluster bioinformatic analysis
The synthetic gene bunch functional verification of embodiment 5 rhamnose isoflavone
Amycolatosis CGMCCNo.6023 clones the synthetic gene bunch of described rhamnose isoflavone eastwardly, the recombinant vector pET28a plasmid of the synthetic gene bunch containing described rhamnose isoflavone is imported in host e. coli, then cultivating the escherichia coli containing target plasmid, whether checking gained engineering bacteria can express rhamnose isoflavonoid.
Engineering bacterium fermentation liquid is added isopyknic methanol, soaks 1 hour, be centrifuged 20 minutes under 8000rpm, take supernatant.Carry out UPLC-MS analysis.UPLC-MS analysis condition is as follows:
UPLC-MS: chromatographic column: AcquityUPLCBEHC18column, 1.7 μm, 100mm × 2.1mm (Waters, Milford, USA), column temperature: 45 DEG C；
Mobile phase: A phase is 0.05% formic acid water, and B phase is 0.05% formic acid-acetonitrile；
Flow velocity: 0.4ml/min
Eluting flow process:
Fermentation liquid is found that m/z401.12 [M+H]+、417.12[M+H]+、431.13[M+H]+Compound, in conjunction with document, thus it is speculated that these three compound molecule formula respectively C21H20O8、C21H20O9、C22H22O9, for rhamnose isoflavone like substance, its aglycon respectively daidzein, genistein and daidzein.Show that this engineering bacteria bacterium creates rhamnose isoflavonoid.
The above; it is only presently preferred embodiments of the present invention; not to any formal and substantial restriction of the present invention; should be understood that; for those skilled in the art; under the premise without departing from the inventive method, also can making some improvement and supplement, these improve and supplement and also should be regarded as protection scope of the present invention.All those skilled in the art, without departing from the spirit and scope of the present invention, the equivalent variations of a little change, modification and the differentiation made when available disclosed above technology contents, it is the Equivalent embodiments of the present invention；Meanwhile, all change of any equivalent variations, modification and differentiation above-described embodiment made according to the substantial technological of the present invention, all still fall within the scope of technical scheme.
Claims (10)
1. the synthetic gene bunch of a rhamnose isoflavone, it is characterised in that the synthetic gene bunch of described rhamnose isoflavone comprises 7 genes:
(1) AORI_0586 gene, nucleotide sequence is such as shown in SEQIDNO.1, and length is 882 base pairs, and encoding amino acid sequence is glucose 1-phosphoric acid thymus pyrimidine transferring enzyme (293 aminoacid) shown in SEQIDNO.8 such as；
(2) AORI_0991 gene, nucleotide sequence is such as shown in SEQIDNO.2, and length is 984 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-glucose-4 shown in SEQIDNO.9 such as, 6-dehydrogenase (327 aminoacid)；
(3) AORI_3188 gene, nucleotide sequence is such as shown in SEQIDNO.3, and length is 960 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-glucose-4 shown in SEQIDNO.10 such as, 6-dehydrogenase (319 aminoacid)；
(4) AORI_0570 gene, nucleotide sequence is such as shown in SEQIDNO.4, and length is 549 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-4-dehydrogenation rhamnose-3 shown in SEQIDNO.11 such as, 5-epimerase (182 aminoacid)；
(5) AORI_1501 gene, nucleotide sequence is such as shown in SEQIDNO.5, and length is 618 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-4-dehydrogenation rhamnose-3 shown in SEQIDNO.12 such as, 5-epimerase (205 aminoacid)；
(6) AORI_6774 gene, nucleotide sequence is such as shown in SEQIDNO.6, and length is 606 base pairs, and encoding amino acid sequence is deoxythymidine diphosphate-4-dehydrogenation rhamnose-3 shown in SEQIDNO.13 such as, 5-epimerase (201 aminoacid)；
(7) AORI_0992 gene, nucleotide sequence is such as shown in SEQIDNO.7, and length is 891 base pairs, and encoding amino acid sequence is the thymidine diphosphate-4-dehydrogenation rhamnose reductase (296 aminoacid) shown in SEQIDNO.14 such as.
2. the albumen that the synthetic gene bunch of rhamnose isoflavone encodes according to claim 1.
3. contain the expression vector of the synthetic gene bunch of rhamnose isoflavone described in claim 1.
4. contain expression vector described in claim 3 or chromosome is integrated with the host cell of synthetic gene bunch of the rhamnose isoflavone described in claim 1 of external source.
5. contain the construction method of the genetic engineering bacterium of the synthetic gene bunch of rhamnose isoflavone described in claim 1, it is characterised in that the recombinant expression carrier of the synthetic gene bunch containing the rhamnose isoflavone described in claim 1 is imported host.
6. method according to claim 5, it is characterised in that described host is selected from any one in escherichia coli, Rhodococcus fascians, Nocard's bacillus, bacillus subtilis, lactic acid bar bacterium or yeast.
7. an Amycolatopsis orientalis, it is characterised in that containing the synthetic gene bunch of the rhamnose isoflavone described in claim 1, its deposit number is CGMCCNo.6023.
8. the method preparing rhamnose isoflavonoid, it is characterized in that, including step: cultivate the host cell described in claim 4 or the Amycolatopsis orientalis described in claim 7, thus expressing rhamnose isoflavone and the like, and from culture fluid, separate rhamnose isoflavone and the like.
9. the synthetic gene bunch of rhamnose isoflavone according to claim 1, encoding proteins described in claim 2, the expression vector described in claim 3 or the application in preparation rhamnose isoflavone and the like of the Amycolatopsis orientalis described in claim 7.
10. an Amycolatopsis orientalis, it is characterised in that in described Amycolatopsis orientalis, in the synthetic gene bunch of rhamnose isoflavone described in claim 1, one or more genes are deactivated, thus not producing rhamnose isoflavone and the like.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1416470A (en) * | 2000-02-11 | 2003-05-07 | 默克专利有限公司 | method for producing monoglycosidated flavonoids |
| CN102549155A (en) * | 2009-08-07 | 2012-07-04 | 国立大学法人东京农工大学 | Novel glycosyltransferase, novel glycosyltransferase gene, and novel glycosyl donor compound |
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2014
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1416470A (en) * | 2000-02-11 | 2003-05-07 | 默克专利有限公司 | method for producing monoglycosidated flavonoids |
| CN102549155A (en) * | 2009-08-07 | 2012-07-04 | 国立大学法人东京农工大学 | Novel glycosyltransferase, novel glycosyltransferase gene, and novel glycosyl donor compound |
Non-Patent Citations (4)
| Title |
|---|
| KIYOTOSHI NISHIYAMA ET AL.: "Syntheses of isoflavones and isoflavone glycosides,and their inhibitory activity against bovine liver beta-galactosidase", 《BIOSCI.BIOTECH.BIOCHEM》 * |
| LI XU.ET AL.: "Complete genome sequence and comparative genomic analyses of the vancomycin-producing Amycolatopsis orientalis", 《BMC GENOMICS》 * |
| 刘效稳等: "东方拟无枝酸菌发酵液小组分的分离鉴定", 《中国抗生素杂志》 * |
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