CN104232498B - 一种纤维化纤维微细菌菌株及其应用 - Google Patents
一种纤维化纤维微细菌菌株及其应用 Download PDFInfo
- Publication number
- CN104232498B CN104232498B CN201310245307.3A CN201310245307A CN104232498B CN 104232498 B CN104232498 B CN 104232498B CN 201310245307 A CN201310245307 A CN 201310245307A CN 104232498 B CN104232498 B CN 104232498B
- Authority
- CN
- China
- Prior art keywords
- bacterial strain
- application
- fine
- hydrolysis
- strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 241000894006 Bacteria Species 0.000 title claims abstract description 24
- 239000000835 fiber Substances 0.000 title claims abstract description 23
- 206010016654 Fibrosis Diseases 0.000 title claims abstract description 21
- 230000004761 fibrosis Effects 0.000 title claims abstract description 21
- 230000007062 hydrolysis Effects 0.000 claims abstract description 34
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 34
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 claims abstract description 29
- 108090000790 Enzymes Proteins 0.000 claims abstract description 28
- 102000004190 Enzymes Human genes 0.000 claims abstract description 28
- 230000001580 bacterial effect Effects 0.000 claims abstract description 24
- DKVBOUDTNWVDEP-NJCHZNEYSA-N teicoplanin aglycone Chemical compound N([C@H](C(N[C@@H](C1=CC(O)=CC(O)=C1C=1C(O)=CC=C2C=1)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)OC=1C=C3C=C(C=1O)OC1=CC=C(C=C1Cl)C[C@H](C(=O)N1)NC([C@H](N)C=4C=C(O5)C(O)=CC=4)=O)C(=O)[C@@H]2NC(=O)[C@@H]3NC(=O)[C@@H]1C1=CC5=CC(O)=C1 DKVBOUDTNWVDEP-NJCHZNEYSA-N 0.000 claims abstract description 18
- 229930182478 glucoside Natural products 0.000 claims abstract description 17
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims abstract description 14
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000000855 fermentation Methods 0.000 claims abstract description 6
- 230000004151 fermentation Effects 0.000 claims abstract description 6
- 241000186221 Cellulosimicrobium cellulans Species 0.000 claims abstract description 5
- 230000003301 hydrolyzing effect Effects 0.000 claims abstract description 5
- 238000004321 preservation Methods 0.000 claims abstract description 4
- -1 glycoside compounds Chemical class 0.000 claims description 29
- 229930182470 glycoside Natural products 0.000 claims description 17
- 239000000126 substance Substances 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 5
- 150000008131 glucosides Chemical class 0.000 claims description 3
- 235000009392 Vitis Nutrition 0.000 claims 1
- 241000219095 Vitis Species 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 abstract description 20
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 abstract description 6
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 abstract description 6
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 abstract description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 54
- 238000000034 method Methods 0.000 description 23
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Natural products CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 17
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 15
- 229930182490 saponin Natural products 0.000 description 15
- 229930182494 ginsenoside Natural products 0.000 description 14
- 239000007788 liquid Substances 0.000 description 14
- 229960001592 paclitaxel Drugs 0.000 description 14
- 229940089161 ginsenoside Drugs 0.000 description 13
- 150000007949 saponins Chemical class 0.000 description 13
- OVSQVDMCBVZWGM-LQSBFMDOSA-N 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-3-[(2r,3s,4r,5r,6s)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxychromen-4-one Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@@H]1OC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O OVSQVDMCBVZWGM-LQSBFMDOSA-N 0.000 description 12
- 244000068988 Glycine max Species 0.000 description 12
- 235000010469 Glycine max Nutrition 0.000 description 12
- 229940123237 Taxane Drugs 0.000 description 11
- 229960005322 streptomycin Drugs 0.000 description 11
- 229930012538 Paclitaxel Natural products 0.000 description 10
- 239000000047 product Substances 0.000 description 10
- OVSQVDMCBVZWGM-IDRAQACASA-N Hirsutrin Natural products O([C@H]1[C@H](O)[C@H](O)[C@H](O)[C@@H](CO)O1)C1=C(c2cc(O)c(O)cc2)Oc2c(c(O)cc(O)c2)C1=O OVSQVDMCBVZWGM-IDRAQACASA-N 0.000 description 9
- FVQOMEDMFUMIMO-UHFFFAOYSA-N Hyperosid Natural products OC1C(O)C(O)C(CO)OC1OC1C(=O)C2=C(O)C=C(O)C=C2OC1C1=CC=C(O)C(O)=C1 FVQOMEDMFUMIMO-UHFFFAOYSA-N 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 9
- GXMWXESSGGEWEM-UHFFFAOYSA-N isoquercitrin Natural products OCC(O)C1OC(OC2C(Oc3cc(O)cc(O)c3C2=O)c4ccc(O)c(O)c4)C(O)C1O GXMWXESSGGEWEM-UHFFFAOYSA-N 0.000 description 9
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 9
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 8
- 241000208340 Araliaceae Species 0.000 description 7
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 7
- 235000003140 Panax quinquefolius Nutrition 0.000 description 7
- 235000008434 ginseng Nutrition 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- PYXFVCFISTUSOO-UHFFFAOYSA-N betulafolienetriol Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC(C(C)(O)CCC=C(C)C)C4C(O)CC3C21C PYXFVCFISTUSOO-UHFFFAOYSA-N 0.000 description 6
- 125000003147 glycosyl group Chemical group 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 239000000376 reactant Substances 0.000 description 6
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 6
- TYLVGQKNNUHXIP-MHHARFCSSA-N 10-Deacetyltaxol A Natural products O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)C=4C=CC=CC=4)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 TYLVGQKNNUHXIP-MHHARFCSSA-N 0.000 description 5
- SMDOOINVMJSDPS-UHFFFAOYSA-N Astragaloside Natural products C1=C(O)C(OC)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)OC2C(C(OC3C(C(O)C(O)C(CO)O3)O)C(O)C(CO)O2)O)=C1 SMDOOINVMJSDPS-UHFFFAOYSA-N 0.000 description 5
- QMNWISYXSJWHRY-XWJCTJPOSA-N astragaloside Chemical compound O1[C@H](C(C)(O)C)CC[C@]1(C)[C@@H]1[C@@]2(C)CC[C@]34C[C@]4(CC[C@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)CO4)O)C4(C)C)C4[C@@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)CC3[C@]2(C)C[C@@H]1O QMNWISYXSJWHRY-XWJCTJPOSA-N 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000013067 intermediate product Substances 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- WENNXORDXYGDTP-UOUCMYEWSA-N cycloastragenol Chemical compound O1[C@H](C(C)(O)C)CC[C@]1(C)[C@@H]1[C@@]2(C)CC[C@]34C[C@]4(CC[C@H](O)C4(C)C)[C@H]4[C@@H](O)C[C@H]3[C@]2(C)C[C@@H]1O WENNXORDXYGDTP-UOUCMYEWSA-N 0.000 description 4
- WENNXORDXYGDTP-UHFFFAOYSA-N cyclosiversigenin Natural products O1C(C(C)(O)C)CCC1(C)C1C2(C)CCC34CC4(CCC(O)C4(C)C)C4C(O)CC3C2(C)CC1O WENNXORDXYGDTP-UHFFFAOYSA-N 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 125000000969 xylosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 description 4
- 229930182986 10-Deacetyltaxol Natural products 0.000 description 3
- ZUXNULGHCOXCFL-UHFFFAOYSA-N 2-(4-tert-butyl-2,6-dimethylphenyl)acetonitrile Chemical compound CC1=CC(C(C)(C)C)=CC(C)=C1CC#N ZUXNULGHCOXCFL-UHFFFAOYSA-N 0.000 description 3
- UFNDONGOJKNAES-UHFFFAOYSA-N Ginsenoside Rb1 Natural products CC(=CCCC(C)(OC1OC(COC2OC(CO)C(O)C(O)C2O)C(O)C(O)C1O)C3CCC4(C)C3C(O)CC5C6(C)CCC(OC7OC(CO)C(O)C(O)C7OC8OC(CO)C(O)C(O)C8O)C(C)(C)C6CC(O)C45C)C UFNDONGOJKNAES-UHFFFAOYSA-N 0.000 description 3
- QMNWISYXSJWHRY-YLNUDOOFSA-N astragaloside IV Chemical compound O1[C@H](C(C)(O)C)CC[C@]1(C)[C@@H]1[C@@]2(C)CC[C@]34C[C@]4(CC[C@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)CO4)O)C4(C)C)[C@H]4[C@@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)C[C@H]3[C@]2(C)C[C@@H]1O QMNWISYXSJWHRY-YLNUDOOFSA-N 0.000 description 3
- QMNWISYXSJWHRY-BCBPIKMJSA-N astragaloside IV Natural products CC(C)(O)[C@@H]1CC[C@@](C)(O1)[C@H]2[C@@H](O)C[C@@]3(C)[C@@H]4C[C@H](O[C@@H]5O[C@H](CO)[C@H](O)[C@@H](O)[C@H]5O)[C@H]6C(C)(C)[C@H](CC[C@@]67C[C@@]47CC[C@]23C)O[C@@H]8OC[C@@H](O)[C@H](O)[C@H]8O QMNWISYXSJWHRY-BCBPIKMJSA-N 0.000 description 3
- 239000007795 chemical reaction product Substances 0.000 description 3
- PFKIBRPYVNVMRU-UHFFFAOYSA-N cyclosieversioside F Natural products CC(C)(O)C1COC(C)(C1)C2C(O)CC3(C)C4CC(OC5OC(CO)C(O)C(O)C5O)C6C(C)(C)C(CCC67CC47CCC23C)OC8OCC(O)C(O)C8O PFKIBRPYVNVMRU-UHFFFAOYSA-N 0.000 description 3
- 230000002708 enhancing effect Effects 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- GZYPWOGIYAIIPV-JBDTYSNRSA-N ginsenoside Rb1 Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)O[C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(CC[C@H]4C(C)(C)[C@@H](O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)CC[C@]4(C)[C@H]3C[C@H]2O)C)(C)CC1)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GZYPWOGIYAIIPV-JBDTYSNRSA-N 0.000 description 3
- TXEWRVNOAJOINC-UHFFFAOYSA-N ginsenoside Rb2 Natural products CC(=CCCC(OC1OC(COC2OCC(O)C(O)C2O)C(O)C(O)C1O)C3CCC4(C)C3C(O)CC5C6(C)CCC(OC7OC(CO)C(O)C(O)C7OC8OC(CO)C(O)C(O)C8O)C(C)(C)C6CCC45C)C TXEWRVNOAJOINC-UHFFFAOYSA-N 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- UGJAEDFOKNAMQD-DVQDXYAYSA-N (-)-Falcarinol Natural products CCCCCCC\C=C\CC#CC#C[C@@H](O)C=C UGJAEDFOKNAMQD-DVQDXYAYSA-N 0.000 description 2
- PYXFVCFISTUSOO-HKUCOEKDSA-N (20S)-protopanaxadiol Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@H]([C@@](C)(O)CCC=C(C)C)[C@H]4[C@H](O)C[C@@H]3[C@]21C PYXFVCFISTUSOO-HKUCOEKDSA-N 0.000 description 2
- UGJAEDFOKNAMQD-MQNTZWLQSA-N (3S,9Z)-1,9-Heptadecadiene-4,6-diyn-3-ol Chemical compound CCCCCCC\C=C/CC#CC#C[C@@H](O)C=C UGJAEDFOKNAMQD-MQNTZWLQSA-N 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- DBXFAPJCZABTDR-KUEXGRMWSA-N Cephalomannine Natural products O=C(O[C@@H]1C(C)=C2[C@@H](OC(=O)C)C(=O)[C@]3(C)[C@@H](O)C[C@@H]4[C@](OC(=O)C)([C@H]3[C@H](OC(=O)c3ccccc3)[C@@](O)(C2(C)C)C1)CO4)[C@@H](O)[C@H](NC(=O)/C(=C\C)/C)c1ccccc1 DBXFAPJCZABTDR-KUEXGRMWSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- UGJAEDFOKNAMQD-UHFFFAOYSA-N Falcarinol Natural products CCCCCCCC=CCC#CC#CC(O)C=C UGJAEDFOKNAMQD-UHFFFAOYSA-N 0.000 description 2
- 230000001430 anti-depressive effect Effects 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 239000000935 antidepressant agent Substances 0.000 description 2
- 229940005513 antidepressants Drugs 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- OUZGCGZMZBOCBH-MNLIZOKASA-N chembl311365 Chemical compound O([C@@H]1[C@]2(O)C[C@@H](C(=C([C@@H](O)C(=O)[C@]3(C)[C@@H](O)C[C@H]4OC[C@]4([C@H]31)OC(C)=O)C2(C)C)C)OC(=O)[C@H](O)[C@@H](NC(=O)CCCCC)C=1C=CC=CC=1)C(=O)C1=CC=CC=C1 OUZGCGZMZBOCBH-MNLIZOKASA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 206010015037 epilepsy Diseases 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- PWAOOJDMFUQOKB-WCZZMFLVSA-N ginsenoside Re Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](O[C@@H]2[C@H]3C(C)(C)[C@@H](O)CC[C@]3(C)[C@@H]3[C@@]([C@@]4(CC[C@@H]([C@H]4[C@H](O)C3)[C@](C)(CCC=C(C)C)O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C)(C)C2)O[C@H](CO)[C@@H](O)[C@@H]1O PWAOOJDMFUQOKB-WCZZMFLVSA-N 0.000 description 2
- AOGZLQUEBLOQCI-UHFFFAOYSA-N ginsenoside-Re Natural products CC1OC(OCC2OC(OC3CC4(C)C(CC(O)C5C(CCC45C)C(C)(CCC=C(C)C)OC6OC(CO)C(O)C(O)C6O)C7(C)CCC(O)C(C)(C)C37)C(O)C(O)C2O)C(O)C(O)C1O AOGZLQUEBLOQCI-UHFFFAOYSA-N 0.000 description 2
- 150000002338 glycosides Chemical class 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- KKAHGGJBKUXDNQ-KRWDZBQOSA-N panaxynol Natural products CCCCCCCC=CC=CCC#C[C@@H](O)C=C KKAHGGJBKUXDNQ-KRWDZBQOSA-N 0.000 description 2
- SWQINCWATANGKN-UHFFFAOYSA-N protopanaxadiol Natural products CC(CCC=C(C)C)C1CCC2(C)C1C(O)CC1C3(C)CCC(O)C(C)(C)C3CCC21C SWQINCWATANGKN-UHFFFAOYSA-N 0.000 description 2
- SHCBCKBYTHZQGZ-DLHMIPLTSA-N protopanaxatriol Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2[C@@H](O)C[C@@]3(C)[C@]4(C)CC[C@H]([C@](C)(O)CCC=C(C)C)[C@H]4[C@H](O)C[C@@H]3[C@]21C SHCBCKBYTHZQGZ-DLHMIPLTSA-N 0.000 description 2
- BBEUDPAEKGPXDG-UHFFFAOYSA-N protopanaxatriol Natural products CC(CCC=C(C)C)C1CCC2(C)C1C(O)CC3C4(C)CCC(O)C(C)(C)C4C(O)CC23C BBEUDPAEKGPXDG-UHFFFAOYSA-N 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- XUWSHXDEJOOIND-YYDKPPGPSA-N (1s,4as,5r,7s,7ar)-7-methyl-1-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-1,5,6,7a-tetrahydrocyclopenta[c]pyran-4a,5,7-triol Chemical compound O([C@@H]1OC=C[C@@]2(O)[C@H](O)C[C@@]([C@@H]12)(O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O XUWSHXDEJOOIND-YYDKPPGPSA-N 0.000 description 1
- HDXIQHTUNGFJIC-UHFFFAOYSA-N (25R)-spirost-5-en-3beta-ol 3-O-<O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranoside> Natural products O1C2(OCC(C)CC2)C(C)C(C2(CCC3C4(C)CC5)C)C1CC2C3CC=C4CC5OC1OC(CO)C(O)C(O)C1OC1OC(C)C(O)C(O)C1O HDXIQHTUNGFJIC-UHFFFAOYSA-N 0.000 description 1
- NABVFHUVYXEKSQ-UHFFFAOYSA-N 6-tuliposide A Natural products OCCC(=C)C(=O)OCC1OC(O)C(O)C(O)C1O NABVFHUVYXEKSQ-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 201000002909 Aspergillosis Diseases 0.000 description 1
- 208000036641 Aspergillus infections Diseases 0.000 description 1
- 241000193752 Bacillus circulans Species 0.000 description 1
- 235000018185 Betula X alpestris Nutrition 0.000 description 1
- 235000018212 Betula X uliginosa Nutrition 0.000 description 1
- VNONINPVFQTJOC-RXEYMUOJSA-N Collettiside III Natural products O([C@@H]1[C@@H](O)[C@H](O[C@H]2[C@H](O)[C@H](O)[C@@H](O)[C@H](C)O2)[C@H](CO)O[C@@H]1O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]5[C@H](C)[C@@]6(O[C@H]5C4)OC[C@H](C)CC6)CC3)CC=2)CC1)[C@H]1[C@H](O)[C@H](O)[C@@H](O)[C@H](C)O1 VNONINPVFQTJOC-RXEYMUOJSA-N 0.000 description 1
- DORPKYRPJIIARM-UHFFFAOYSA-N Decaffeoylacteoside Natural products OC1C(O)C(O)C(C)OC1OC1C(O)C(OCCC=2C=C(O)C(O)=CC=2)OC(CO)C1O DORPKYRPJIIARM-UHFFFAOYSA-N 0.000 description 1
- 235000004360 Dioscorea zingiberensis Nutrition 0.000 description 1
- 241001678283 Dioscorea zingiberensis Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 208000035126 Facies Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- PUQSUZTXKPLAPR-UJPOAAIJSA-N Gastrodin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C(CO)C=C1 PUQSUZTXKPLAPR-UJPOAAIJSA-N 0.000 description 1
- 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 description 1
- 108010056771 Glucosidases Proteins 0.000 description 1
- 102000004366 Glucosidases Human genes 0.000 description 1
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 1
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 1
- KVRQGMOSZKPBNS-FMHLWDFHSA-N Harpagoside Chemical compound O([C@@H]1OC=C[C@@]2(O)[C@H](O)C[C@]([C@@H]12)(C)OC(=O)\C=C\C=1C=CC=CC=1)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O KVRQGMOSZKPBNS-FMHLWDFHSA-N 0.000 description 1
- KVRQGMOSZKPBNS-BYYMOQGZSA-N Harpagoside Natural products C[C@@]1(C[C@@H](O)[C@@]2(O)C=CO[C@@H](O[C@@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@H]3O)[C@H]12)OC(=O)C=Cc4ccccc4 KVRQGMOSZKPBNS-BYYMOQGZSA-N 0.000 description 1
- 241000191936 Micrococcus sp. Species 0.000 description 1
- 241000588628 Moraxella sp. Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241001597008 Nomeidae Species 0.000 description 1
- 241000178960 Paenibacillus macerans Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241001149649 Taxus wallichiana var. chinensis Species 0.000 description 1
- DWCSNWXARWMZTG-UHFFFAOYSA-N Trigonegenin A Natural products CC1C(C2(CCC3C4(C)CCC(O)C=C4CCC3C2C2)C)C2OC11CCC(C)CO1 DWCSNWXARWMZTG-UHFFFAOYSA-N 0.000 description 1
- SQRUWMQAWMLKPR-DZEUPHNYSA-N Tuliposide A Chemical compound OCCC(=C)C(=O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O SQRUWMQAWMLKPR-DZEUPHNYSA-N 0.000 description 1
- DORPKYRPJIIARM-GYAWPQPFSA-N Verbasoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](O)[C@H](OCCC=2C=C(O)C(O)=CC=2)O[C@H](CO)[C@H]1O DORPKYRPJIIARM-GYAWPQPFSA-N 0.000 description 1
- 241000588902 Zymomonas mobilis Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 1
- 238000005852 acetolysis reaction Methods 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 239000003005 anticarcinogenic agent Substances 0.000 description 1
- 229940034982 antineoplastic agent Drugs 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 102000006995 beta-Glucosidase Human genes 0.000 description 1
- 108010047754 beta-Glucosidase Proteins 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 108010089934 carbohydrase Proteins 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 238000000658 coextraction Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000022811 deglycosylation Effects 0.000 description 1
- VNONINPVFQTJOC-ZGXDEBHDSA-N dioscin Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O[C@H]1[C@@H]([C@H](O)[C@@H](O)[C@H](C)O1)O)O[C@@H]1CC2=CC[C@H]3[C@@H]4C[C@H]5[C@@H]([C@]4(CC[C@@H]3[C@@]2(C)CC1)C)[C@@H]([C@]1(OC[C@H](C)CC1)O5)C)[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O VNONINPVFQTJOC-ZGXDEBHDSA-N 0.000 description 1
- CJNUQCDDINHHHD-APRUHSSNSA-N dioscin Natural products C[C@@H]1CC[C@@]2(OC1)O[C@H]3C[C@H]4[C@@H]5CC=C6C[C@H](CC[C@@H]6[C@H]5CC[C@]4(C)[C@H]3[C@@H]2C)O[C@@H]7O[C@H](CO)[C@@H](O[C@@H]8O[C@@H](C)[C@H](O)[C@@H](O)[C@H]8O)[C@H](O)[C@H]7O[C@@H]9O[C@@H](C)[C@H](O)[C@@H](O)[C@H]9O CJNUQCDDINHHHD-APRUHSSNSA-N 0.000 description 1
- WQLVFSAGQJTQCK-VKROHFNGSA-N diosgenin Chemical compound O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)CC[C@H](O)CC4=CC[C@H]3[C@@H]2C1)C)[C@@H]1C)[C@]11CC[C@@H](C)CO1 WQLVFSAGQJTQCK-VKROHFNGSA-N 0.000 description 1
- WQLVFSAGQJTQCK-UHFFFAOYSA-N diosgenin Natural products CC1C(C2(CCC3C4(C)CCC(O)CC4=CCC3C2C2)C)C2OC11CCC(C)CO1 WQLVFSAGQJTQCK-UHFFFAOYSA-N 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- FVIZARNDLVOMSU-UHFFFAOYSA-N ginsenoside K Natural products C1CC(C2(CCC3C(C)(C)C(O)CCC3(C)C2CC2O)C)(C)C2C1C(C)(CCC=C(C)C)OC1OC(CO)C(O)C(O)C1O FVIZARNDLVOMSU-UHFFFAOYSA-N 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000005858 glycosidation reaction Methods 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- IFPWDIMCTSSWCJ-UHFFFAOYSA-N harpagide Natural products CC1(CC(O)C2(O)C=COCC12)OC3OC(CO)C(O)C(O)C3O IFPWDIMCTSSWCJ-UHFFFAOYSA-N 0.000 description 1
- 108010030923 hesperidinase Proteins 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 210000003716 mesoderm Anatomy 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- VELYAQRXBJLJAK-UHFFFAOYSA-N myoporoside Natural products C12C(C)(O)CC(O)C2C=COC1OC1OC(CO)C(O)C(O)C1O VELYAQRXBJLJAK-UHFFFAOYSA-N 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- VNONINPVFQTJOC-UHFFFAOYSA-N polyphyllin III Natural products O1C2(OCC(C)CC2)C(C)C(C2(CCC3C4(C)CC5)C)C1CC2C3CC=C4CC5OC(C(C1O)OC2C(C(O)C(O)C(C)O2)O)OC(CO)C1OC1OC(C)C(O)C(O)C1O VNONINPVFQTJOC-UHFFFAOYSA-N 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- SBVBJPHMDABKJV-PGCJWIIOSA-N secoisolariciresinol diglucoside Chemical compound C1=C(O)C(OC)=CC(C[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)[C@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)CC=2C=C(OC)C(O)=CC=2)=C1 SBVBJPHMDABKJV-PGCJWIIOSA-N 0.000 description 1
- SBVBJPHMDABKJV-UHFFFAOYSA-N secoisolariciresinol diglycoside Natural products C1=C(O)C(OC)=CC(CC(COC2C(C(O)C(O)C(CO)O2)O)C(COC2C(C(O)C(O)C(CO)O2)O)CC=2C=C(OC)C(O)=CC=2)=C1 SBVBJPHMDABKJV-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012807 shake-flask culturing Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 229960001866 silicon dioxide Drugs 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- SQRUWMQAWMLKPR-UHFFFAOYSA-N tuliposide-A Natural products OCCC(=C)C(=O)OC1OC(CO)C(O)C(O)C1O SQRUWMQAWMLKPR-UHFFFAOYSA-N 0.000 description 1
- 239000005418 vegetable material Substances 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- 229920001221 xylan Polymers 0.000 description 1
- 150000004823 xylans Chemical class 0.000 description 1
- 150000008216 xylosides Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P33/00—Preparation of steroids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
一种纤维化纤维微细菌菌株及其应用,该菌株为纤维化纤维微细菌(Cellulosimicrobium cellulans)菌株F16,保藏号为CCTCC M2013201,保藏日期为2013年5月;该菌株F16或其发酵产生的水解酶,将含木糖苷键和葡糖苷键的化合物水解,得到的产物是相应的苷元,也可以是在制备相应苷元的过程中所产生的中间体化合物。
Description
技术领域
本发明属于生物化工领域,具体涉及一种纤维化纤维微细菌菌株及其应用。
背景技术
许多天然具有活性化合物往往以糖苷的形式存在于自然界中,将其糖基去除后,便可释放出有活性的化合物。如抗癌药物紫杉醇,其主要存在于红豆杉植物的树皮中。而在红豆杉的枝叶中则大量存在着木糖苷化的紫杉醇前体或其相同母核的类似物(称之为紫杉烷类化合物),如7-木糖紫杉醇和7-木糖-10去乙酰紫杉醇(10DAXT),其在如中华、云南及南方红豆杉的枝叶中的累积量可高出紫杉醇10倍以上,将7-位木糖基水解后再经一步乙酰化即可转化为紫杉醇。
另外一种典型的天然产物如来自名贵中草药人参的人参皂苷。人参皂苷是公认人参中的主要活性成分,其基本骨架主要是原人参二醇(PPD)和原人参三醇(PPT)。其中含量高的如Rb1、Rb2、Rb3、Rc、Rd、Re、Rg1等,都是基本骨架被高度“糖基化”了的化合物,其所含糖基从一个到五个不等,而具有很好抗肿瘤和抑菌活性的却是人参中一些含量很少的稀有人参皂苷如Rh1、Rh2、Rg3、Rg5、CK、PPT、PPD等,这些化合物往往含糖基较少,或直接是不带有任何糖基的苷元,是上述高含量人参皂苷去糖基化后的产物(J PharmPharmacol.1998Oct;50(10):1155-60.)。日本公开特许58-131999(JPO)58-131999公布了20(S)-原人参二醇、20(R)-原人参二醇、20(S)-原人参三醇、20(R)-原人参三醇都具有抑制肿瘤生长的活性,其中20(S)-原人参二醇(PPD)的活性最强。进一步的研究表明20(S)-PPD可以通过增强机体免疫和直接的细胞毒作用杀死肿瘤细胞,还可以抑制肿瘤间质血管生成,阻止肿瘤细胞的生长;在神经系统方面,20(S)-PPD可以抗癫痫、抗抑郁、增强学习能力等,是一个非常具有开发价值的化合物(Front Pharmacol.2012;3:25.ChineseMedicine2010,5:20)。
此外,还有很多天然化合物都存在类似情况,如黄芪甲苷,红链霉素糖苷,异槲皮苷,大豆皂苷,红景天苷,天麻苷,山慈菇苷A,薯蓣皂苷,亚麻木酚素,哈巴俄苷及哈巴苷等等。
由于人们往往对其苷元更感兴趣,但这类化合物一般骨架复杂,化学方法断裂糖苷键的选择性差,副产物多,环境污染等原因,制备相应苷元十分困难。相比之下生物转化方法由于具有高度的选择性和专一性,几乎没有副产物,反应条件温和,环境友好,成为制备此类化合物相应苷元的首选。目前已有很多关于酶法水解糖苷类化合物的报道,如韩国研究者公布了利用针尾曲霉水解红参中人参皂苷以提高其中人参皂苷苷元的含量(J.Korean Soc.Appl.Biol.Chem.53(5),553-558(2010)),Kohada报道以G-Rb1、Rb2、Rb3、Rc和Rd为原料,用醋酸水解脱去C-20位上的糖,再用粗橙皮苷酶等水解脱去3位上的糖,制得20(S)-PPD(JPO58-131999),中国专利CN03101549.2公开了利用乳酸菌或肠道细菌生物转化制备人参组合物的方法,中国专利CN102703329公开了一株能够高效转化黄姜中皂苷生产薯蓣皂苷元的菌株,Hanson RL(1997)利用细菌Moraxella sp.、Bacillus macerans、Bacillus circulans和Micrococcus sp.将C-7木糖紫杉烷转化为C-7羟基紫杉烷,中国专利CN102296053公开了一种来自真菌的β-木糖苷酶也具有水解C-7木糖紫杉烷的能力,并实现其在酵母中的胞内表达等,但往往存在使用菌体转化、产酶及转化效率低的情况,尤其对于一些水溶性差、空间位阻大的化合物,如10DAXT,普通糖苷酶往往难以水解。
发明内容
本发明的目的在于提供一种纤维化纤维微细菌菌株及其应用,该菌株可将含木糖苷键或葡糖苷键的化合物水解而获得相应的苷元。本发明使用一种纤维化纤维微细菌菌株或该菌株所产生的酶接触含有至少一种木糖苷键或葡糖苷键的黄芪皂苷类、人参皂苷类、紫杉烷木糖苷类、红链霉素糖苷、异槲皮苷、大豆皂苷类化合物,并水解相应的糖苷键。该方法可以制备至少一种上述糖苷化合物的苷元,也可以制备在转化上述苷元的过程中所产生的具有药理活性的中间产物。
本发明提供了一种纤维化纤维微细菌菌株,该菌株为纤维化纤维微细菌(Cellulosimicrobium cellulans)菌株F16,菌种保藏号为CCTCC M2013201,保藏日期为2013年5月14日,保藏单位为中国典型培养物保藏中心,保藏地址为中国武汉大学。
本发明还提供了所述纤维化纤维微细菌菌株的应用,该菌株应用于糖苷化合物苷元的制备。
本发明提供的所述纤维化纤维微细菌菌株的应用,该菌株或其发酵产生的水解酶,接触含糖苷键的糖苷化合物,将糖苷键水解,得到的产物是相应的苷元或中间体。
本发明提供的所述纤维化纤维微细菌菌株的应用,所述糖苷键为葡萄糖苷键和/或木糖苷键。
本发明提供的所述纤维化纤维微细菌菌株的应用,所述糖苷化合物结构如下:
本发明菌株制备的相应苷元结构是:
本发明提供的菌株的水解方法已考虑到具有上述分子结构的化合物的手性中心的所有立体构型,这些立体异构体或单独被水解,或与其它立体异构体混合在一起被水解。
本发明提供的菌株实用价值在于:环黄芪醇,原人参醇,紫杉烷类化合物,红链霉素,异槲皮素和大豆皂苷苷元,在药物制备领域有重要的应用价值。从植物中提取出的上述化合物通常是连接有大量糖基的糖苷化合物,且多为混合物,其中大部分糖苷化合物带有的是木糖基或葡糖基,而去糖基后的苷元才是最终想要的产品。一个或多个含有至少一种木糖苷键或葡糖苷键的黄芪甲苷、人参皂苷、紫杉烷木糖苷、红链霉素糖苷、异槲皮苷或大豆皂苷,在菌株F16或其发酵产生的水解酶的作用下被水解,得到相应苷元;所得化合物可以是具有药理活性的苷元,如环黄芪醇,原人参醇,紫杉烷类化合物,红链霉素,异槲皮素或大豆皂苷苷元,或其类似物,也可以是制备上述苷元的过程中所产生的未完全水解的中间体。
本发明提供的菌株使得被水解的黄芪甲苷、人参皂苷、紫杉烷木糖苷、红链霉素糖苷、异槲皮苷和大豆皂苷的立体构型优先保留于产品中,所得相应苷元或其水解中间产物的糖苷键的绝对立体构型与原糖苷化合物的糖苷键绝对立体构型相同。
本发明提供的菌株用于从带有木糖苷键或葡糖苷键的黄芪甲苷、人参皂苷、紫杉烷木糖苷、红链霉素糖苷、异槲皮苷或大豆皂苷制备相应的苷元或其水解中间产物时十分有效。用本发明提供的方法可以水解一种带有木糖苷键或葡糖苷键的上述糖苷化合物,也可以连续性地或同时水解不同上述糖苷化合物的混合物。由植物材料提取的人参皂苷混合物、紫杉烷木糖苷和大豆皂苷混合物用本发明提供的方法水解,效果很好。
具体实施方式
下面的实施例将对本发明予以进一步的说明,但并不因此而限制本发明。
起始材料:
本发明所采用的起始材料可以是任何带有木糖苷键或葡糖苷键的黄芪甲苷、人参皂苷、紫杉烷木糖苷、红链霉素糖苷、异槲皮苷或大豆皂苷类化合物。带有木糖苷键或葡糖苷键的上述糖苷化合物可以是自然形成的,也可以是化学半合成或全合成所得。
酶与微生物:
本发明水解方法中使用的酶或微生物可以是下面描述的任何能催化酶水解反应的酶或微生物。这些酶或微生物材料,不管其来源或纯度,都可在游离状态下使用,或者将其用物理吸附或诱捕方法固定在支持物上使用。生物学纯的纤维化纤维微细菌(Cellulosimicrobium cellulans)菌株F16是新发现的同时产β-木糖苷酶和β-葡萄糖苷酶的微生物。这种微生物的突变体,例如经化学的,物理的(如紫外辐射)或生物学方法(如分子生物学技术)改造以用于水解反应的突变菌株,也在本发明的考虑范围内。
纤维化纤维微细菌(Cellulosimicrobium cellulans)菌株F16分离自大连市甘井子区西山水库的土壤之中,其特征为:革兰氏阳性,可在好氧条件下生长,菌落呈圆形,直径0.9~2mm,黄白色突起,反光,边缘整齐。在显微镜下观察菌体为棒状,随着培养时间的延长逐渐转化成短杆,甚至球形。
本发明应用的酶是水解酶,尤其是葡萄糖苷酶和葡聚糖酶,以及木糖苷酶和木聚糖酶。本发明提供的菌株生产这些酶,可用提取和纯化的方法分离它们。本发明提供的微生物可以以完整湿细胞的形式,或者以冻干,喷雾干燥或加热干燥的形式,或者以经破碎抽提后的形式被应用于水解反应。
本发明提供的水解方法可以在微生物发酵后进行,也可以与发酵同时进行。
将菌株接种于含诱导剂的培养基中,好氧条件下发酵培养后,离心收集上清液,即为粗酶液。培养基pH在4-7之间,培养温度20-40℃之间。水解反应进行0.5-72小时,直至目的产品的产量达最大。水解反应时,pH保持在6-9,其中pH7-8效果最好。
产物的分离:
应用本发明提供的方法所产生的六类苷元或其未完全水解的中间产物,可被分离纯化,根据水解产物的特征,可用提取,蒸馏,结晶,柱层析或多种分离技术结合的方法进行纯化。
应用:
应用本发明提供的水解方法所获得的化合物如紫杉醇,是很好的抗癌药。7-木糖-10-去乙酰紫杉醇经本发明提供的方法水解后获得10-去乙酰紫杉醇,此化合物再经10位乙酰化,即可得到紫杉醇。
应用本发明提供的水解方法所获得的化合物如人参二醇,具有非常好的抗肿瘤活性,同时还可以抗癫痫、抗抑郁、增强学习能力等,是一个非常具有开发价值的化合物,近些年被各国研究者进行改造,希望能够将其开发为一种新的抗肿瘤药。
实施例1:粗酶液的制备
使用含1%淀粉,0.2%蛋白胨,0.2%酵母提取物,0.2%磷酸氢二钾的培养基(灭菌前pH7.0)作为种子培养基。
将上述种子培养基20ml分配到100ml锥形瓶中,在120度下灭菌15min,将纤维化纤维单胞菌F16菌株的琼脂斜面培养物接种到其中,并在30度下振荡培养2天作为种子液。
500ml Erlenmeyer培养瓶中放入100ml含1%桦木木聚糖,0.2%酵母浸膏,0.2%氯化铵,0.1%NaH2PO4和0.1%Na2HPO4的培养基,pH7,高压灭菌121度30min。将1ml种子液接种于此培养基,150rpm,30℃摇瓶培养5天。离心收集上清液,即为粗酶液。
实施例2:黄芪甲苷IV的水解粗酶液的制备
在0.9ml实施例1中的粗酶液中加入1ml50mM Tris-HCl缓冲液(pH7.5),再加入0.1ml黄芪甲苷IV的甲醇溶液(浓度为10mg/ml)。100rpm,35℃孵育8小时后,以环黄芪醇标准品作对照,TLC分析,黄芪甲苷IV几乎全部转化为环黄芪醇。
实施例3:人参皂苷Rb1水解制备化合物CK
在0.9ml实施例1中的粗酶液中加入1ml50mM Tris-HCl缓冲液(pH7.5),再加入0.1ml人参皂苷Rb1的甲醇溶液(浓度为10mg/ml)。100rpm,35℃孵育4小时。加入2ml甲醇终止反应,15000转/分离心20分钟取上清液进行UPLC分析。反应液中人参皂苷Rb1几乎完全转化,产生化合物K(即CK,产率75%),未完全水解的中间产物以及少量终产物PPD。
UPLC方法:
色谱柱:Kromasil ODS(2.1×150mm,3μm)
流动相:乙腈:水(0→30min:20:80→95:5梯度洗脱)
流速:0.4ml/min
柱温:室温
检测波长:203nm
实施例4:人参皂苷Rb1水解制备原人参二醇(PPD)
在0.9ml实施例1中的粗酶液中加入1ml50mM Tris-HCl缓冲液(pH7.5),再加入0.1ml人参皂苷Rb1的甲醇溶液(浓度为10mg/ml)。100rpm,35℃孵育12小时。加入2ml甲醇终止反应,15000转/分离心20分钟取上清液进行UPLC分析,方法同实施例3。反应液中人参皂苷Rb1几乎完全转化为终产物原人参二醇(PPD),产率90%。
实施例5:人参皂苷Re水解制备原人参三醇(PPT)
在0.9ml实施例1中的粗酶液中加入1ml50mM Tris-HCl缓冲液(pH7.5),再加入0.1ml人参皂苷Re的甲醇溶液(浓度为10mg/ml)。100rpm,35℃孵育8小时。加入2ml甲醇终止反应,15000转/分离心20分钟取上清液进行UPLC分析,方法同实施例3。反应液中人参皂苷Re几乎完全转化为终产物原人参三醇(PPT),产率93%。
实施例6:7-木糖紫杉醇水解制备紫杉醇
在0.9ml实施例1中的粗酶液中加入1ml50mM Tris-HCl缓冲液(pH7.5),再加入0.1ml7-木糖-紫杉醇的甲醇溶液(浓度为5mg/ml)。100rpm,30℃孵育5小时。加入2ml甲醇终止反应,15000转/分离心20分钟取上清液进行HPLC分析。反应液中无7-木糖-紫杉醇残留,并产生0.41mg紫杉醇(产率94%)。
HPLC方法:
色谱柱:Kromasil ODS(4.6×200mm,5μm)
流动相:甲醇:水(60:40)
流速:1ml/min
柱温:室温
检测波长:227nm
实施例7:7-木糖-10-去乙酰紫杉醇水解制备10-去乙酰紫杉醇
将10ml7-木糖-10-去乙酰紫杉醇的甲醇溶液(浓度为5mg/ml)加入到90ml实施例1所述的粗酶液中,100rpm,30℃下反应20小时后,加入20ml乙酸乙酯萃取,共萃取3次,合并上层有机相,减压蒸干,获得固体物质102mg,上硅胶柱(20g)并用氯仿:甲醇(98:2)进行洗脱,得到39mg10-去乙酰紫杉醇(总收率91%)。
实施例8:7-木糖紫杉烷混合物的水解
将200ml含有7-木糖-10-去乙酰紫杉醇,7-木糖-10-去乙酰三尖杉宁碱,7-木糖-10-去乙酰紫杉醇C的混合物(从天然红豆杉提取物中分离,含量分别为65%,9.9%,3.3%)的甲醇溶液(5mg/ml)加入到2000ml如前所述的粗酶液中,100rpm,35℃下反应8小时。反应液用乙酸乙酯萃取三次,每次200ml,合并有机相蒸干溶解于100ml甲醇中,用实施例6中HPLC方法进行定量检测,发现其中无三种带木糖基的底物残留,得到的产物中10-去乙酰紫杉醇,10-去乙酰三尖杉宁碱,10-去乙酰紫杉醇C的含量分别504mg,75mg,28mg,回收率达到90%。
实施例9:红链霉素龙胆二糖苷的水解
在0.9ml实施例1中的粗酶液中加入1ml50mM Tris-HCl缓冲液(pH7.5),再加入0.1ml红链霉素龙胆二糖苷的甲醇溶液(浓度为12mg/ml)。100rpm,35℃孵育4.5小时。加入2ml甲醇终止反应,15000转/分离心20分钟取上清液进行UPLC分析。反应液中无红链霉素龙胆二糖苷残留,并产生0.52mg红链霉素(产率95%)。
UPLC方法:
色谱柱:Kromasil ODS(2.1×150mm,3μm)
流动相:乙腈:水(0→10min:20:80→95:5梯度洗脱)
流速:0.4ml/min
柱温:室温
检测波长:278nm
实施例10:异槲皮苷的水解
在0.9ml实施例1中的粗酶液中加入1ml50mM Tris-HCl缓冲液(pH7.5),再加入0.1ml异槲皮苷的甲醇溶液(浓度为7mg/ml)。100rpm,35℃孵育4小时后,以异槲皮素标准品作对照,TLC分析,异槲皮苷几乎全部转化为异槲皮素。
实施例11:大豆皂苷的水解
在0.9ml实施例1中的粗酶液中加入1ml50mM Tris-HCl缓冲液(pH7.5),再加入0.1ml大豆皂苷的甲醇溶液(浓度为5mg/ml)。100rpm,35℃孵育12小时后,以大豆皂苷苷元标准品作对照,TLC分析,大豆皂苷几乎全部转化为其苷元。
Claims (5)
1.一种纤维化纤维微细菌菌株,其特征在于:该菌株为纤维化纤维微细菌(Cellulosimicrobium cellulans)菌株F16,保藏号为CCTCC M2013201,保藏日期为2013年5月。
2.权利要求1所述纤维化纤维微细菌菌株的应用,其特征在于:该菌株应用于糖苷化合物苷元的制备。
3.按照权利要求2所述纤维化纤维微细菌菌株的应用,其特征在于:该菌株或其发酵产生的水解酶,接触含糖苷键的糖苷化合物,将糖苷键水解,得到的产物是相应的苷元或中间体。
4.按照权利要求3所述纤维化纤维微细菌菌株的应用,其特征在于:所述糖苷键为葡萄糖苷键和/或木糖苷键。
5.按照权利要求2或3所述纤维化纤维微细菌菌株的应用,其特征在于:所述糖苷化合物结构如下:
。
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310245307.3A CN104232498B (zh) | 2013-06-19 | 2013-06-19 | 一种纤维化纤维微细菌菌株及其应用 |
PCT/CN2013/001510 WO2014201596A1 (zh) | 2013-06-19 | 2013-12-06 | 一种纤维化纤维微细菌菌株及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310245307.3A CN104232498B (zh) | 2013-06-19 | 2013-06-19 | 一种纤维化纤维微细菌菌株及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN104232498A CN104232498A (zh) | 2014-12-24 |
CN104232498B true CN104232498B (zh) | 2016-08-10 |
Family
ID=52103772
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201310245307.3A Expired - Fee Related CN104232498B (zh) | 2013-06-19 | 2013-06-19 | 一种纤维化纤维微细菌菌株及其应用 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN104232498B (zh) |
WO (1) | WO2014201596A1 (zh) |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104342381A (zh) * | 2013-08-01 | 2015-02-11 | 东北师范大学 | 一种生物转化制备稀有人参皂苷Rg2的方法 |
CN106148471A (zh) * | 2015-04-08 | 2016-11-23 | 中国科学院大连化学物理研究所 | 一种纤维单胞菌科作为工业生物催化剂的应用 |
CN104830910B (zh) * | 2015-05-25 | 2017-11-07 | 江苏师范大学 | 利用纤维化纤维微细菌制备微生物絮凝剂的方法 |
CN106701856A (zh) * | 2015-07-28 | 2017-05-24 | 中国科学院大连化学物理研究所 | 一种酶法水解蒙花苷制备金合欢素的方法 |
CN105670962B (zh) * | 2016-01-22 | 2019-05-10 | 江南大学 | 一株芳香烃的高效降解菌及其应用 |
CN108504606A (zh) * | 2018-04-20 | 2018-09-07 | 大连理工大学 | 一种放线菌培养基及其应用 |
CN115247142B (zh) * | 2022-08-16 | 2023-05-12 | 安徽农业大学 | 一株纤维化纤维微细菌及其在秸秆田间堆肥中的应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005210977A (ja) * | 2004-01-30 | 2005-08-11 | Hitachi Kiden Kogyo Ltd | 細胞壁溶解酵素生産菌 |
CN101381708A (zh) * | 2007-09-04 | 2009-03-11 | 中国科学院大连化学物理研究所 | 纤维化纤维单胞菌、水解酶及其在紫杉烷转化方面的用途 |
CN101691554A (zh) * | 2009-06-30 | 2010-04-07 | 广西科学院 | 一种能产丙烯酸的纤维化纤维菌 |
CN102634470A (zh) * | 2012-04-10 | 2012-08-15 | 宋建民 | 一种纤维化纤维微细菌及渗透发酵生产海藻糖的方法 |
-
2013
- 2013-06-19 CN CN201310245307.3A patent/CN104232498B/zh not_active Expired - Fee Related
- 2013-12-06 WO PCT/CN2013/001510 patent/WO2014201596A1/zh active Application Filing
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005210977A (ja) * | 2004-01-30 | 2005-08-11 | Hitachi Kiden Kogyo Ltd | 細胞壁溶解酵素生産菌 |
CN101381708A (zh) * | 2007-09-04 | 2009-03-11 | 中国科学院大连化学物理研究所 | 纤维化纤维单胞菌、水解酶及其在紫杉烷转化方面的用途 |
CN101691554A (zh) * | 2009-06-30 | 2010-04-07 | 广西科学院 | 一种能产丙烯酸的纤维化纤维菌 |
CN102634470A (zh) * | 2012-04-10 | 2012-08-15 | 宋建民 | 一种纤维化纤维微细菌及渗透发酵生产海藻糖的方法 |
Non-Patent Citations (2)
Title |
---|
Cloning and Characterization of the Glycoside Hydrolases That Remove Xylosyl Groups from 7-β-xylosyl-10-deacetyltaxol and Its Analogues;Hai-Li Cheng 等;《Mol Cell Proteomics.》;20130831;第12卷(第8期);2236-2248 * |
一株纤维化纤维微细菌的生物学特性及其对几种苯环类化合物的利用研究;陈燕红 等;《微生物学报》;20080720;第35卷(第7期);1021-1027 * |
Also Published As
Publication number | Publication date |
---|---|
WO2014201596A1 (zh) | 2014-12-24 |
CN104232498A (zh) | 2014-12-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN104232498B (zh) | 一种纤维化纤维微细菌菌株及其应用 | |
JP4953547B2 (ja) | ジンセノサイド糖基を加水分解するジンセノサイドグリコシダーゼ及びその使用 | |
KR100329259B1 (ko) | 효소(酵素)로 인삼 사포닌 당기(糖基)를 변화시켜서 희소한 인삼사포닌을 제조하는 방법 | |
Cheng et al. | Conversion of major ginsenoside Rb 1 to ginsenoside F 2 by Caulobacter leidyia | |
Upadhyaya et al. | Enzymatic formation of compound-K from ginsenoside Rb1 by enzyme preparation from cultured mycelia of Armillaria mellea | |
CN105648021B (zh) | 人参稀有皂苷c-k、f1及四种异构体人参皂苷元的制备方法 | |
Kim et al. | Highly regioselective biotransformation of ginsenoside Rb2 into compound Y and compound K by β-glycosidase purified from Armillaria mellea mycelia | |
Quan et al. | Bioconversion of ginsenoside Rb1 into compound K by Leuconostoc citreum LH1 isolated from kimchi | |
Cheng et al. | Microbial Conversion of Ginsenoside $ Rb_1 $ to Minor Ginsenoside $ F_2 $ and Gypenoside XVII by Intrasporangium sp. GS603 Isolated from Soil | |
Jiang et al. | Biotransformation of ginsenoside Rb1 to ginsenoside CK by strain XD101: A safe bioconversion strategy | |
CN1982438A (zh) | 一株芽孢杆菌及用其制备单糖链人参皂苷及苷元的方法 | |
US10870857B2 (en) | Method of producing ginsenosides 20(S)-Rg3 and 20(S)-Rh2 using ginsenoside glycosidases | |
KR20020009756A (ko) | 효소적 방법에 의한 진세노사이드 컴파운드 케이의 제조방법 | |
Zhu et al. | Production of diosgenin from Dioscorea zingiberensis tubers through enzymatic saccharification and microbial transformation | |
Gao et al. | Efficient biotransformation for preparation of pharmaceutically active ginsenoside compound K by Penicillium oxalicum sp. 68 | |
Feng et al. | The microbiological transformation of steroidal saponins by Curvularia lunata | |
CN1793320A (zh) | 一株镰刀霉菌及其用于制备人参皂苷Rh2的方法 | |
An et al. | Gram-scale production of ginsenoside F1 using a recombinant bacterial β-glucosidase | |
KR20080028266A (ko) | 펙티네스 또는 비스코자임을 이용하여 인삼 사포닌으로부터장내 진세노사이드 대사물질인 화합물 케이, 화합물와이, 진세노사이드 에프 1 및 화합물 피지-2를 제조하는방법 | |
CN117089465B (zh) | 一种疣梗曲霉及应用 | |
KR20150055703A (ko) | 저분자 진세노사이드의 제조방법 | |
CN113897406A (zh) | 一种从红景天粉末中提取并纯化红景天苷的方法 | |
Quan et al. | Bioconversion of ginsenoside Rd into compound K by Lactobacillus pentosus DC101 isolated from Kimchi | |
CN102220274B (zh) | 巴氏微杆菌xj及应用该菌制备甜菊醇的方法 | |
KR101139508B1 (ko) | 생물전환된 화합물 k를 함유하는 인삼 발효물 및 이의 제조방법, 그리고 이를 이용한 기능성 인삼 막걸리 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20180301 Address after: 215600 A 207 room A building center of Zhangjiagang Free Trade Zone, Suzhou Free Trade Zone, Jiangsu Patentee after: ZHANGJIAGANG INDUSTRY TECHNOLOGY RESEARCH INSTITUTE CO.,LTD. DALIAN INSTITUTE OF CHEMICAL PHYSICS CHINESE ACADEMY OF SCIENCES Address before: 116023 Zhongshan Road, Liaoning, No. 457, Patentee before: Dalian Institute of Chemical Physics, Chinese Academy of Sciences |
|
CF01 | Termination of patent right due to non-payment of annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160810 |