Some content of this application is unavailable at the moment.
If this situation persist, please contact us atFeedback&Contact
1. (WO2017148163) METHOD FOR EXTRACTING 2',3'-CYCLIC NUCLEOSIDE MONOPHOSPHATES
Latest bibliographic data on file with the International Bureau    Submit observation

Pub. No.: WO/2017/148163 International Application No.: PCT/CN2016/103058
Publication Date: 08.09.2017 International Filing Date: 24.10.2016
IPC:
C12P 19/30 (2006.01) ,C12P 19/32 (2006.01) ,C07K 14/245 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19
Preparation of compounds containing saccharide radicals
26
Preparation of nitrogen-containing carbohydrates
28
N-glycosides
30
Nucleotides
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19
Preparation of compounds containing saccharide radicals
26
Preparation of nitrogen-containing carbohydrates
28
N-glycosides
30
Nucleotides
32
having a condensed ring system containing a six-membered ring having two nitrogen atoms in the same-ring, e.g. purine nucleotides, nicotineamide-adenine dinucleotide
C CHEMISTRY; METALLURGY
07
ORGANIC CHEMISTRY
K
PEPTIDES
14
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
195
from bacteria
24
from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
245
Escherichia (G)
Applicants:
山东大学 SHANDONG UNIVERSITY [CN/CN]; 中国山东省济南市 历城区山大南路27号 No.27 Shanda Nan Rd., Licheng District Jinan, Shandong 250199, CN
Inventors:
陈秀兰 CHEN, Xiulan; CN
张玉忠 ZHANG, Yuzhong; CN
刘昂 LIU, Ang; CN
于洋 YU, Yang; CN
解彬彬 XIE, Binbin; CN
秦启龙 QIN, Qilong; CN
宋晓妍 SONG, Xiaoyan; CN
石梅 SHI, Mei; CN
Agent:
济南金迪知识产权代理有限公司 JINAN JINDI INTELLECTUAL PROPERTY AGENCY CO., LTD.; 中国山东省济南市 历城区山大南路27号 No.27 Shanda Nan Rd., Licheng District Jinan, Shandong 250199, CN
Priority Data:
201610112458.529.02.2016CN
Title (EN) METHOD FOR EXTRACTING 2',3'-CYCLIC NUCLEOSIDE MONOPHOSPHATES
(FR) PROCÉDÉ D'EXTRACTION DE NUCLÉOSIDES MONOPHOSPHATES 2',3'-CYCLIQUES
(ZH) 一种提取2',3'-环形核苷单磷酸的方法
Abstract:
(EN) The present invention provides a method for extracting 2',3'-cyclic nucleoside monophosphates, the method comprising the following steps: (1) extracting a genomic DNA, and amplifying the extracted genomic DNA by PCR to obtain a gene, if3; (2) ligating the gene, if3, with an expression plasmid, and performing transformation and fermentation to obtain a recombinant bacterial fermentation broth; (3) isolating the bacteria from the recombinant bacterial fermentation broth, disrupting the isolated bacterial cells, performing nickel column affinity chromatography purification and allowing the resulting product to stand, then performing solid-liquid separation to harvest a supernatant; (4) using ultrafiltration to remove protein impurities from the supernatant, and performing high-performance liquid-phase separation using a C18 reversed-phase chromatography column to obtain a solution containing four 2',3'-cyclic nucleoside monophosphates.
(FR) La présente invention concerne un procédé d'extraction de nucléosides monophosphates 2',3'-cycliques, le procédé comprenant les étapes suivantes, consistant à : (1) extraire de l'ADN génomique et amplifier l'ADN génomique extrait par PCR pour obtenir un gène if3; (2) ligaturer le gène, if3, avec un plasmide d'expression et réaliser une transformation et une fermentation pour obtenir un bouillon de fermentation bactérien recombinant; (3) isoler les bactéries du bouillon de fermentation bactérien recombinant, désintégrer les cellules bactériennes isolées, réaliser une purification par chromatographie d'affinité sur colonne au nickel et laisser le produit résultant au repos, puis réaliser une séparation solide-liquide pour récolter une phase surnageante; (4) utiliser l'ultrafiltration pour enlever les impuretés protéiniques de la phase surnageante et réaliser une séparation haute performance en phase liquide à l'aide d'une colonne C18 de chromatographie en phase inverse pour obtenir une solution contenant quatre monophosphates nucléosides 2',3'-cycliques.
(ZH) 本发明提供了一种提取2',3'-环形核苷单磷酸的方法,包括如下步骤:(1)提取基因组DNA,PCR扩增,制得基因if3;(2)将基因if3与表达质粒连接,转化后,经发酵培养,制得重组菌发酵液;(3)分离重组菌发酵液中的菌体,破碎细胞,镍柱亲和层析提纯,静置,固液分离,取上清液;(4)将上清液经超滤去除蛋白杂质,经C18反向色谱柱进行高效液相分离,分别制得含四种2',3'-环形核苷单磷酸溶液。
Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: Chinese (ZH)
Filing Language: Chinese (ZH)