US3674642A - Novel microbial acidic heteropolysaccharide and production thereof - Google Patents
Novel microbial acidic heteropolysaccharide and production thereof Download PDFInfo
- Publication number
- US3674642A US3674642A US38780A US3674642DA US3674642A US 3674642 A US3674642 A US 3674642A US 38780 A US38780 A US 38780A US 3674642D A US3674642D A US 3674642DA US 3674642 A US3674642 A US 3674642A
- Authority
- US
- United States
- Prior art keywords
- heteropolysaccharide
- acidic
- acidic heteropolysaccharide
- acid
- glucose
- 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 - Lifetime
Links
- 230000002378 acidificating effect Effects 0.000 title abstract description 46
- 238000004519 manufacturing process Methods 0.000 title abstract description 10
- 230000000813 microbial effect Effects 0.000 title abstract description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 abstract description 27
- 239000001963 growth medium Substances 0.000 abstract description 16
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 abstract description 14
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 abstract description 13
- 244000005700 microbiome Species 0.000 abstract description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 12
- 229910052799 carbon Inorganic materials 0.000 abstract description 12
- 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 abstract description 11
- 239000008103 glucose Substances 0.000 abstract description 11
- 241000158504 Rhodococcus hoagii Species 0.000 abstract description 10
- 229930195733 hydrocarbon Natural products 0.000 abstract description 10
- 150000002430 hydrocarbons Chemical class 0.000 abstract description 10
- 150000001720 carbohydrates Chemical group 0.000 abstract description 9
- 239000004215 Carbon black (E152) Substances 0.000 abstract description 7
- 239000004310 lactic acid Substances 0.000 abstract description 7
- 235000014655 lactic acid Nutrition 0.000 abstract description 7
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 6
- 239000002253 acid Substances 0.000 abstract description 5
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 abstract description 5
- 235000015097 nutrients Nutrition 0.000 abstract description 4
- GZCGUPFRVQAUEE-KVTDHHQDSA-N aldehydo-D-mannose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O GZCGUPFRVQAUEE-KVTDHHQDSA-N 0.000 abstract 1
- 238000000034 method Methods 0.000 description 16
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 15
- 239000000203 mixture Substances 0.000 description 12
- 239000002609 medium Substances 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 238000011534 incubation Methods 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 6
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 6
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 6
- 239000002244 precipitate Substances 0.000 description 6
- 239000001888 Peptone Substances 0.000 description 5
- 108010080698 Peptones Proteins 0.000 description 5
- 239000000853 adhesive Substances 0.000 description 5
- 230000001070 adhesive effect Effects 0.000 description 5
- 239000007864 aqueous solution Substances 0.000 description 5
- 235000019319 peptone Nutrition 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 125000004432 carbon atom Chemical group C* 0.000 description 4
- 239000012141 concentrate Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 150000002823 nitrates Chemical class 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000001384 succinic acid Substances 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- 244000215068 Acacia senegal Species 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 3
- 229920000084 Gum arabic Polymers 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000000205 acacia gum Substances 0.000 description 3
- 235000010489 acacia gum Nutrition 0.000 description 3
- 239000012736 aqueous medium Substances 0.000 description 3
- 229910000019 calcium carbonate Inorganic materials 0.000 description 3
- 235000010216 calcium carbonate Nutrition 0.000 description 3
- 235000014633 carbohydrates Nutrition 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 235000011187 glycerol Nutrition 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- OFOBLEOULBTSOW-UHFFFAOYSA-N Malonic acid Chemical compound OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 238000005273 aeration Methods 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 239000003729 cation exchange resin Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 2
- DCAYPVUWAIABOU-UHFFFAOYSA-N hexadecane Chemical compound CCCCCCCCCCCCCCCC DCAYPVUWAIABOU-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 125000000311 mannosyl group Chemical group C1([C@@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 2
- LQERIDTXQFOHKA-UHFFFAOYSA-N nonadecane Chemical compound CCCCCCCCCCCCCCCCCCC LQERIDTXQFOHKA-UHFFFAOYSA-N 0.000 description 2
- RZJRJXONCZWCBN-UHFFFAOYSA-N octadecane Chemical compound CCCCCCCCCCCCCCCCCC RZJRJXONCZWCBN-UHFFFAOYSA-N 0.000 description 2
- -1 ranose Chemical compound 0.000 description 2
- 238000005185 salting out Methods 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- BGHCVCJVXZWKCC-UHFFFAOYSA-N tetradecane Chemical compound CCCCCCCCCCCCCC BGHCVCJVXZWKCC-UHFFFAOYSA-N 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- VOZKAJLKRJDJLL-UHFFFAOYSA-N 2,4-diaminotoluene Chemical compound CC1=CC=C(N)C=C1N VOZKAJLKRJDJLL-UHFFFAOYSA-N 0.000 description 1
- VBUYCZFBVCCYFD-JJYYJPOSSA-N 2-dehydro-D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C(=O)C(O)=O VBUYCZFBVCCYFD-JJYYJPOSSA-N 0.000 description 1
- NSFXVRRBGNORBD-UHFFFAOYSA-N 2h-benzo[f]benzotriazole Chemical compound C1=C2C=CC=CC2=CC2=NNN=C21 NSFXVRRBGNORBD-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 241000186216 Corynebacterium Species 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- YXHKONLOYHBTNS-UHFFFAOYSA-N Diazomethane Chemical compound C=[N+]=[N-] YXHKONLOYHBTNS-UHFFFAOYSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 1
- 229920001202 Inulin Polymers 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229910017621 MgSO4-7H2O Inorganic materials 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 description 1
- 229910001854 alkali hydroxide Inorganic materials 0.000 description 1
- 150000008044 alkali metal hydroxides Chemical class 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- 230000001174 ascending effect Effects 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 150000004649 carbonic acid derivatives Chemical class 0.000 description 1
- 229940023913 cation exchange resins Drugs 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- HPXRVTGHNJAIIH-UHFFFAOYSA-N cyclohexanol Chemical compound OC1CCCCC1 HPXRVTGHNJAIIH-UHFFFAOYSA-N 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 125000004177 diethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000002036 drum drying Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000000174 gluconic acid Substances 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 229940029339 inulin Drugs 0.000 description 1
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 229940038384 octadecane Drugs 0.000 description 1
- 238000004816 paper chromatography Methods 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 102220305863 rs1015663503 Human genes 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000011343 solid material Substances 0.000 description 1
- 230000028070 sporulation Effects 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 229940032147 starch Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 150000003467 sulfuric acid derivatives Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Images
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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09J—ADHESIVES; NON-MECHANICAL ASPECTS OF ADHESIVE PROCESSES IN GENERAL; ADHESIVE PROCESSES NOT PROVIDED FOR ELSEWHERE; USE OF MATERIALS AS ADHESIVES
- C09J105/00—Adhesives based on polysaccharides or on their derivatives, not provided for in groups C09J101/00 or C09J103/00
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/8215—Microorganisms
- Y10S435/822—Microorganisms using bacteria or actinomycetales
- Y10S435/843—Corynebacterium
Definitions
- the present invention relates to a novel microbial acidic heteropolysaccharide and a, method for the production of the same.
- This invention is based on the following iindings:
- the so-accumulated acidic heteropolysaccharide can be recovered in a desired purity from the culture broth, utilizing its physicochemical properties;
- the acidic heteropolysaccharide shows a strong vis-cosity and is excellent as an adhesive, for instance.
- the principal object of the present invention is to provide novel acidic heteropolysaccharide which may -be used, e.g. as an excellent adhesive.
- Another object of the present invention is to provide an industrially feasible method for producing said acidic heteropolysaccharide.
- the saccharide units of the acidic heteropolysaccharide consist substantially of glucose and mannose, and the acid units thereof consist substantially of succinic acid, lactic acid and acetic acid.
- the acidic heteropolysaccharide shows a high viscosity.
- the viscosity of its 0.5% aqueous solution as measured employing Ubbelohdes viscometer is higher than about 5,000 centipoises.
- the intrinsic viscosity of the acidic heteropolysaccharide falls within the range of about 25 to about 28.
- the acidic heteropolysaccharide shows significant infrared absorption bands in KBr disc method at the wave length of 12.8, 11.6, 6.23 and 5.75 microns.
- the Weight ratio of the saccharide units in terms of their free form; the total of succinic acid and lactic acid in terms of their free forms; and acetic acid calculated as OHgCOO-group; is about 70-85; 8-12; 8-15.
- the saccharid'e units contain about 50% to about 75% of glucose 3,674,642 Patented July 4, 1972 ice (l) Morphological. characteristics (l) Vegetative cells:
- Shape Cocci to elongated rods. Depending upon type of medium and duration of incubation, rnost of the cells occur in irregular chains or branching, occasionally single or pair.
- Motility Nonmotile.
- this microorganism accumulates the above-mentioned acidic heteropolysaccharide to give a culture broth of a high viscosity when incubated in a culture medium containing a hydrocarbon as the main carbon source. Therefore, this microorganism has been recognized as a variety of Corynebacterium equi, and named Corynebacterium equi var. muclagnosus.
- Corynebacterium equi var. muclaginosus is incubated in a culture medium containing hydrocarbon, particularly normal parafiins, as the main carbon source.
- hydrocarbons are normal parafiins of carbon atoms of 13 to 19 and their typical examples include tetradecane, hexadecane, octadecane, nonadecane, and n-paraffin mixture which boils in the range of about 230 to about 330 C. and the like.
- a culture medium containing about 2 to about 15 percent by volume of normal paraflins having carbon atoms of 13 to 19.
- a suspending agent e.g. a surfactant
- hydrocarbons as carbon sources, but, if desired, any other conventional carbon sources such as carbohydrates (eg. glucose, mannose), acetic acid, glycerol, succinic acid and the like may be employed.
- the culture medium should contain nitrogen sources(s) together with the hydrocarbon as nutrients.
- the nitrogen sources may be any of conventional ones such as peptone, soybean powder, corn steep liquor, meat extract, yeast extract, ammonium salts, nitrates, urea and the like.
- the medium inorganic salts which are commonly used in the incubation of microorganisms, such as the phosphate of potassium or sodium, as well as the sulfates, hydrochloride and/or carbonates of metals, e.g. magnesium, iron, manganese, calcium, zinc, cobalt, etc.
- metals e.g. magnesium, iron, manganese, calcium, zinc, cobalt, etc.
- the culture medium may be used in any form, but a liquid medium is generally advantageous.
- a liquid medium any of shaking cultural method, stirring cultural method or the like may be adopted, though the so-called submerged aerobic cultural method is the most desirable for the industrial process.
- Incubation conditions such as the pH of the medium and the incubation temperature should be controlled so as to produce the objective acidic heteropolysaccharide in the maximum amount.
- the pH of the culture medium and the incubation temperature are respectively adjusted to pH about 5 to about ⁇ 8, especially to pH about 6-7, and about 25 to about 40 C.
- the pH of the medium changes to acidic region with the progress of incubation, the pH of the medium is practically maintained through the incubation period within said range with the previous incorporation of calcium carbonate into the medium or by adding an alkali hydroxide solution or aqueous ammonia to the medium.
- the period required for the maximum accumulation of the acidic heteropolysaccharide is changeable depending upon various factors, the substantial amount of the heteropolysaccharide accumulates in the culture broth usually between about 48 to about 120 hours from the start of the incubation.
- the viscous culture broth is diluted fivefold or more with water to lessen its viscosity and subjected to centrifugation or filtration by employing a filter press with a filter aid to remove the microorganism cells and other solid materials.
- a filter press with a filter aid to remove the microorganism cells and other solid materials.
- the amount of water as the diluent may be reduced and filtration facilitated.
- hydrophilic organic solvent such as acetone, methyl alcohol, ethyl alcohol up to a concentration of about 60 to about 80% by volume, or alternatively, a small amount of sodium chloride is added first and a hydrophilic organic solvent is subsequently added up to a concentration of about 50 to about 60% by volume, whereby the objective acidic heteropolysaccharide precipitates in the form of a white spongy sediment.
- a suitable organic solvent such as ammonium sulfate, sodium sulfate may also be employed for the separation of the acidic heteropolysaccharide from the above-mentioned concentrate.
- the broth filtrate after removal of the microorganism cells may be concentrated to a certain extent and subjected to spray drying or drum drying to give a crude preparation of the acidic heteropolysaccharide.
- the crude heteropolysaccharide may further be purified.
- the purification may be carried out by, for example, further subjecting the crude acidic heteropolysaccharide to the above-mentioned precipitation treatments with the hydrophilic organic solvent, and then to dialysis or to treatments with strongly acidic cation exchange resins, and finally to lyophilization.
- the acidic heteropolysaccharide of the present invention has many superior properties, particularly an increased viscosity, as compared with hitherto-known polysaccharides.
- the viscosity of its 0.5% aqueous solution as measured employing Ubbelohdes viscometer at 30 C. is higher than about 5,000 centipoises and occasionally reaches about 20,000 centipoises.
- the acidic heteropolysaccharide of the present invention may be employed, for example, as an adhesive. When employed as an adhesive, about 2 to about 5% aqueous solution is preferable.
- the acidic heteropolysaccharide is very excellent as an adhesive because it can exhibit a strong adhesion in a low concentration and it is not hygroscopic. For instance, the acidic heteropolysaccharide shows a strong adhesion in one-fourth concentration of gum arabic as shown in test described below.
- ml., 1., mcg., g. kg and C. mean milliliter(s), liter(s), microgram(s), gram(s), kilogram(s) and centigrade(s), respectively, and percent is weight per volume excepting the case Where the meaning is clearly otherwise from the context.
- the ATCC No. indicates the accession number of the microorganism deposited at the American Type Culture Collection in Rockville, Md., U.S.A.
- EXAMPLE l Corynebacterium equi var. mucilagnosus (ATCC No. 21521) is inoculated into 30 ml. of an aqueous medium comprising 1% of glucose, 0.5% of yeast extract, 0.5% of peptone, 0.5% of NaCl (pH 7.0) and is incubated under shaking at 28 C. for 24 hours.
- the resulting culture broth of 7.4 of relative viscosity is diluted tenfold with Water and the diluted broth is subjected to centrifugation at 10,000 r.p.m. for 2O minutes to remove the microorganism cells and the residual calcium carbonate.
- the supernatant is concentrated to onetwentieth of the original volume.
- To the concentrate is added acetone in 3 to 4 times the volume of the concentrate with stirring, whereby the heteropolysaccharide separates as occulent precipitates.
- the precipitates are recovered by filtration.
- the precipitates are dehydrated with 150 ml. of acetone and then dried to give white spongy crude heteropolysaccharide.
- the yield is 1.55 g. or 20.1% based on the weight of the n-parain mixture used.
- the crude heteropolysaccharide is dissolved in 1 litre of water and the solution is subjected to dialysis with deionized 'water at 20 C. for 48 hou-rs.
- Thus-treated solution is allowed to pass through a column (5 cm. x 50 cm.) packed with strongly acidic cation exchange resin (H- form, such as Amberlite IRO-120) and the eluent is lyophilized to give white spongy heteropolysaccharide.
- H- form such as Amberlite IRO-120
- Total weight of succinic acid and lactic acid is 8.99% relative to the Whole weight of the heteropolysaccharide and the weight ratio of lactic acid relative to succinic acid is about 0.18 in terms of their free form, when measured by methylatiug the diethyl ether-soluble fraction of the above-mentioned hydrolyzate with diazomethane and subjecting the methylated fraction to gas chromatography with N2 gas employing cyclohexanol as control.
- Acetic acid is '9.99% calculated as CHaiCOO-group relative to the whole Weight of the heteropolysaccharide, when measured by the method described in Methods in Carbohydrate Chemistry vol. V, pp. 187 and 188, published by Academic Press, New York, in 1965.
- Corynebacterium equi var. muclagz'nosus (ATCC 21521) is inoculated into 30 ml. of an aqueous medium comprising 1% of glucose, 0.5 of yeast extract, 0.5 of peptone and 0.5% of NaCl (pH 7.0) and is incubated under shaking at 28 C. for 24 hours.
- the resulting culture broth of a high viscosity is subjected to the same isolation procedures as described in Example l to give white spongy crude heteropolysaccharide.
- the yield is 1.72 g. or 22.2% based on the Weight of the n-parain mixture used.
- the crude heteropolysaccharide is further subjected to the same purification treatments as described in Example 1 to give purified heteropolysaccharide.
- the yield is 0.98 g. or 12.6% based on the weight of the n-parain mixture used.
- Colynebacterium equi var. mucilgnosus (ATCC 21521) is pre-cultured in a 50 litres tank containing 30 litres of an aqueous medium comprising 1% of glucose, 0.5% of yeast extract, 0.5% of peptone and 0.5% of NaCl (pH 7.0) for 24 hours under the conditions of 28 C., 100% aeration and agitation at 250 r.p.m.
- l0 litres of the resulting culture broth is inoculated in a 200 litres tank containing 100 litres of a culture medium of the composition listed in the following Table 3 and incubated for 96 hours under the conditions of 30 C., 50% aeration and agitation at 250 r.p.m., during which time the pH of the medium is maintained at about 6.5 with the addition of 30% sodium hydroxide.
- the n-parain mixture has the same composition as that of the n-paran mixture described in Table 1.
- the spongy precipitates recovered by filtration are dehydrated with 30 litres of acetone and dried to give white spongy crude heteropolysaccharide.
- the yield is 1.58 kg. or 29.4% based on the weight of the n-parafiin mixture used.
- Table 4 clearly demonstrates that the acidic heteropolysaccharide of the present invention exhibits a. strong adhesion in one-fourth concentration of gum arabic.
- a method for producing microbial acidic heteropolysaccharide of which the saccharide units consist substantially of glucose and mannose and the acid units consist substantially of succinic acid, lactic acid and acetic acid, which comprises incubating Carynebacterum equi var. mucz'laginosus ATCC No. 21521 in a culture medium containing hydrocarbon as the main carbon source together with digestible nitrogen source and other nutrients necessary for the growth of the microorganism until a substantial amount of the acidic hcteropolysaccharide is accumulated in the culture broth, and recovering the accumulated acidic heteropolysaccharide therefrom.
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
METHOD FOR PRODUCING MICROBIAL ACIDIC HETEROPOLYSACCHARIDE, OF WHICH THE SACCHARIDE UNITS CONSIST SUBSTANTIALLY OF GLUCOSE AND MANNOSE AND THE ACID UNITS CONSIST SUBSTANTIALY OF SUCCINIC ACID, LACTIC ACID AND ACETIC ACID, WHICH COMPRISES INCUBATING CORYNEBACTERIUM EQUI VAR. MUCILAGINOSUS IN A CULTURE MEDIUM CONTAINING HYDROCARBON AS THE MAIN CARBON SOURCE TOGETHER WITH DIGESTIBLE NITROGEN SOURCE AND OTHER NUTRIENTS NECESSARY FOR THE GROWTH OF THE MICROORGANISM UNTIL A SUBSTANTIAL AMOUNT OF THE ACIDIC HETEROPOLYSACCHARIDE IS ACCUMULATED IN THE CULTURE BROTH, AND RECOVERING THE ACCUMULATED ACIDIC HETEROPOLYSACCHARIDE THEREFROM.
Description
July 4, 1972 sABuRo YAMAToDANl ETAL 3,674,642
NOVEL MICROBIAL ACIDIC HETEROPOLYSACCHARIDE AND PRODUCTION THEREOF Filed May 19, 1970 (maoaad) aamvmwsNvai INVENTORS SABURO YAMATODANI TSUNEO KANAMURU ATTORNEYS United States Patent 3,674,642 NOVEL MICRBIAL ACIDIC HETEROPOLY- SACCHARIDE AND PRODUCTION THEREOF Saburo Yamatodani, Minoo, and Tsuneo Kanamaru,
Toyonaka, Iapan, assignors to Takeda Chemical Industries, Ltd., Osaka, Japan Filed May 19, 1970, Ser. No. 38,780 Claims priority, application Japan, May 19, 1969, 44/ 38,551 Int. Cl. C1211 13/04 U.S. Cl. 195-28 R 4 Claims ABSTRACT F THE DISCLOSURE Method for producing microbial acidic heteropolysaccharide, of which the saccharide units consist substantially of glucose and mannose and the acid units consist substantialy of succinic acid, lactic acid and acetic acid, which comprises incubating Cozynebacterium equi var. muclaginosus in a culture medium containing hydrocarbon as the main carbon source together with digestible nitrogen source and other nutrients necessary for the growth of the microorganism until a substantial amount of the acidic heteropolysaccharide is accumulated in the culture broth, and recovering the accumulated acidic heteropolysaccharide therefrom.
The present invention relates to a novel microbial acidic heteropolysaccharide and a, method for the production of the same.
This invention is based on the following iindings:
(l) Corynebacterium equi. var. mucilagnosus, a nefw Variety of Corynebacterium equi isolated from a sample of soil, accumulates a novel acidic heteropolysaccharide when the microorganism is incubated in a culture medium containing a hydrocarbon as the main carbon source;
(2) The so-accumulated acidic heteropolysaccharide can be recovered in a desired purity from the culture broth, utilizing its physicochemical properties; and
(3) The acidic heteropolysaccharide shows a strong vis-cosity and is excellent as an adhesive, for instance.
Thus, the principal object of the present invention is to provide novel acidic heteropolysaccharide which may -be used, e.g. as an excellent adhesive.
Another object of the present invention is to provide an industrially feasible method for producing said acidic heteropolysaccharide.
The acidic heteropolysaccharide of the present invention is characterized by the following properties:
The saccharide units of the acidic heteropolysaccharide consist substantially of glucose and mannose, and the acid units thereof consist substantially of succinic acid, lactic acid and acetic acid.
The specic rotation of the lacidic heteropolysaccharide is [a]D25=l-35i5 (c.\=0.5%, in water).
The acidic heteropolysaccharide shows a high viscosity. For instance, the viscosity of its 0.5% aqueous solution as measured employing Ubbelohdes viscometer is higher than about 5,000 centipoises. The intrinsic viscosity of the acidic heteropolysaccharide falls within the range of about 25 to about 28.
The acidic heteropolysaccharide shows significant infrared absorption bands in KBr disc method at the wave length of 12.8, 11.6, 6.23 and 5.75 microns.
The Weight ratio of the saccharide units in terms of their free form; the total of succinic acid and lactic acid in terms of their free forms; and acetic acid calculated as OHgCOO-group; is about 70-85; 8-12; 8-15. The saccharid'e units contain about 50% to about 75% of glucose 3,674,642 Patented July 4, 1972 ice (l) Morphological. characteristics (l) Vegetative cells:
Shape: Cocci to elongated rods. Depending upon type of medium and duration of incubation, rnost of the cells occur in irregular chains or branching, occasionally single or pair.
Motility: Nonmotile.
Size: 1.0 to 1.3 microns by 1.0 to 3.0 microns.
Sporulation: Nonsporulating.
(2) Grams stain: Positive and unchanged even after a long incubation period.
(Il) Cultural characteristics (l) Bouillon (24 to 48 hours at 28 C.): Light reddish brown pellicles are formed, with fair amounts of light reddish sediment. Medium growth.
(2) Bouillon slant (48 hours at 28 C.): Growth abundant filamentous and raised; glistening and smooth. Light reddish pink to light reddish brown. No diffusible pigment.
(3) Bouillon stab: Growth abundant on surface; papillose and uprising.
(4) Nutrient agar plate (48 hours at 28 C.): Entire circular (1.4 to 2.2 millimeters in diameter), convex to capitate. Smooth and glistening surface, reddish brown with a hint of light yellow to light reddish pink.
(5) Gelatin stab (30 days at 28 C.): Surface growth.
No liquefaction.
(III) Physiological characteristics (l) Temperature relations: Growth at 20 to 45 C.,
optimum about 37 C.
(2) pH relations: Growth at pH 5.0 to 9.0, optimum about 6.5 to 8.5
(3) O2 relations: Aerobic (4) Indol: Not produced (5) Nitrate reduction: lNitrites produced from nitrates (6) Hydrogen sulde: Produced (7) `Starch assimilation: Negative (8) Methyl red test: Negative (9) Voges-Proskauer reaction: Negative (10) Catalase: Produced (ll) Production of ammonia from peptone: Very slight (l2) Litmus milk: No change (13) Ammonium salts, nitrates and urea are utilized as nitrogen sources (14) Utilization of carbon sources:
Growth on glucose, succinic acid, glycerol, propionic acid, pyruvic acid, acetic acid, benzoic acid and normal paraiiins from 8 to 19 carbon atoms asl the sole carbon source. Gluconic acid, citric acid, 2-ketogluconic acid and malonic acid are not utilized as the sole carbon source. No acid and no gas are produced from glucose, fructose, galactose, mannose, Xylose, arabinose, sucrose, lactose, maltose, trehalose, ranose, mannitol, glycerol, dextrin, starch, inulin and glycogen.
Detailed comparison of the above-mentioned characteristics with the description in Bergeys Manual of Determinative Bacteriology, seventh edition reveals that this microorganism belongs to the genus Corynebacterium and that these characteristics are substantially the same as those of Corynebacterium equi. However, though the type culture of Corynebacterium equi (IFO No. 3730) gives good growth in a culture medium containing normal parafiins as the sole carbon source, the resultant culture broth is substantially not viscous, nor does it show signs of any substantial accumulation of mucillagenous material. In sharp contrast thereto, this microorganism accumulates the above-mentioned acidic heteropolysaccharide to give a culture broth of a high viscosity when incubated in a culture medium containing a hydrocarbon as the main carbon source. Therefore, this microorganism has been recognized as a variety of Corynebacterium equi, and named Corynebacterium equi var. muclagnosus.
According to the method of the present invention, Corynebacterium equi var. muclaginosus is incubated in a culture medium containing hydrocarbon, particularly normal parafiins, as the main carbon source. Especially advantageous hydrocarbons are normal parafiins of carbon atoms of 13 to 19 and their typical examples include tetradecane, hexadecane, octadecane, nonadecane, and n-paraffin mixture which boils in the range of about 230 to about 330 C. and the like.
-From the viewpoints of the growth of the microorganism and the yield of the objective acidic heteropolysaccharide, there may be advantageously employed a culture medium containing about 2 to about 15 percent by volume of normal paraflins having carbon atoms of 13 to 19.
As these hydrocarbons are scarcely soluble in water, the addition thereof to an aqueous culture medium is practically carried out under shaking to prepare a suspension. If desired, a suspending agent, e.g. a surfactant, may be employed.
It is sufficient for the present method to use these hydrocarbons as carbon sources, but, if desired, any other conventional carbon sources such as carbohydrates (eg. glucose, mannose), acetic acid, glycerol, succinic acid and the like may be employed.
The culture medium should contain nitrogen sources(s) together with the hydrocarbon as nutrients. The nitrogen sources may be any of conventional ones such as peptone, soybean powder, corn steep liquor, meat extract, yeast extract, ammonium salts, nitrates, urea and the like.
Furthermore, there are added to the medium inorganic salts which are commonly used in the incubation of microorganisms, such as the phosphate of potassium or sodium, as well as the sulfates, hydrochloride and/or carbonates of metals, e.g. magnesium, iron, manganese, calcium, zinc, cobalt, etc. Aside from the foregoing, it is of course possible to add as required, such trace growth factors as vitamins and nucleic acid related compounds.
The culture medium may be used in any form, but a liquid medium is generally advantageous. When a liquid medium is employed, any of shaking cultural method, stirring cultural method or the like may be adopted, though the so-called submerged aerobic cultural method is the most desirable for the industrial process.
Incubation conditions such as the pH of the medium and the incubation temperature should be controlled so as to produce the objective acidic heteropolysaccharide in the maximum amount. Advantageously, the pH of the culture medium and the incubation temperature are respectively adjusted to pH about 5 to about `8, especially to pH about 6-7, and about 25 to about 40 C. As the pH of the medium changes to acidic region with the progress of incubation, the pH of the medium is practically maintained through the incubation period within said range with the previous incorporation of calcium carbonate into the medium or by adding an alkali hydroxide solution or aqueous ammonia to the medium.
Although the period required for the maximum accumulation of the acidic heteropolysaccharide is changeable depending upon various factors, the substantial amount of the heteropolysaccharide accumulates in the culture broth usually between about 48 to about 120 hours from the start of the incubation.
Generally-known means for recovering acidic heteropolysaccharides from the culture broth containing them can be applied to the recovery of the acidic heteropolysaccharide of the present invention from the culture broth. The acidic heteropolysaccharide can be absorbed on various adsorbents, or precipitated with some precipitants. Moreover, conventional means for recovery, such as salting out or dialysis or a combination thereof, may be employed for the purpose of recovery and purification.
Most practically, the viscous culture broth is diluted fivefold or more with water to lessen its viscosity and subjected to centrifugation or filtration by employing a filter press with a filter aid to remove the microorganism cells and other solid materials. In this step, by warming the culture broth at about 50 to about 95 C., the amount of water as the diluent may be reduced and filtration facilitated.
Thus-obtained clear solution is concentrated as much as possible. To the concentrate is added hydrophilic organic solvent such as acetone, methyl alcohol, ethyl alcohol up to a concentration of about 60 to about 80% by volume, or alternatively, a small amount of sodium chloride is added first and a hydrophilic organic solvent is subsequently added up to a concentration of about 50 to about 60% by volume, whereby the objective acidic heteropolysaccharide precipitates in the form of a white spongy sediment. Salting out with a suitable organic solvent such as ammonium sulfate, sodium sulfate may also be employed for the separation of the acidic heteropolysaccharide from the above-mentioned concentrate.
Alternatively, the broth filtrate after removal of the microorganism cells may be concentrated to a certain extent and subjected to spray drying or drum drying to give a crude preparation of the acidic heteropolysaccharide.
Though thus-obtained crude acidic heteropolysaccharide may be employed as it is, the crude heteropolysaccharide may further be purified. The purification may be carried out by, for example, further subjecting the crude acidic heteropolysaccharide to the above-mentioned precipitation treatments with the hydrophilic organic solvent, and then to dialysis or to treatments with strongly acidic cation exchange resins, and finally to lyophilization.
The acidic heteropolysaccharide of the present invention has many superior properties, particularly an increased viscosity, as compared with hitherto-known polysaccharides. For instance, the viscosity of its 0.5% aqueous solution as measured employing Ubbelohdes viscometer at 30 C. is higher than about 5,000 centipoises and occasionally reaches about 20,000 centipoises.
The acidic heteropolysaccharide of the present invention may be employed, for example, as an adhesive. When employed as an adhesive, about 2 to about 5% aqueous solution is preferable. The acidic heteropolysaccharide is very excellent as an adhesive because it can exhibit a strong adhesion in a low concentration and it is not hygroscopic. For instance, the acidic heteropolysaccharide shows a strong adhesion in one-fourth concentration of gum arabic as shown in test described below.
The following examples and test are merely for illustrative purposes and are not to be construed as the limitation of the present invention. Throughout the present specification, the abbreviations ml., 1., mcg., g.," kg and C. mean milliliter(s), liter(s), microgram(s), gram(s), kilogram(s) and centigrade(s), respectively, and percent is weight per volume excepting the case Where the meaning is clearly otherwise from the context. The ATCC No. indicates the accession number of the microorganism deposited at the American Type Culture Collection in Rockville, Md., U.S.A.
EXAMPLE l Corynebacterium equi var. mucilagnosus (ATCC No. 21521) is inoculated into 30 ml. of an aqueous medium comprising 1% of glucose, 0.5% of yeast extract, 0.5% of peptone, 0.5% of NaCl (pH 7.0) and is incubated under shaking at 28 C. for 24 hours.
l ml. of the resulting culture broth is inoculated into 100 ml. of the culture medium mentioned in the following Table 1, and is incubated under shaking at 28 C. for 96 hours.
The resulting culture broth of 7.4 of relative viscosity is diluted tenfold with Water and the diluted broth is subjected to centrifugation at 10,000 r.p.m. for 2O minutes to remove the microorganism cells and the residual calcium carbonate. The supernatant is concentrated to onetwentieth of the original volume. To the concentrate is added acetone in 3 to 4 times the volume of the concentrate with stirring, whereby the heteropolysaccharide separates as occulent precipitates. The precipitates are recovered by filtration. The precipitates are dehydrated with 150 ml. of acetone and then dried to give white spongy crude heteropolysaccharide. The yield is 1.55 g. or 20.1% based on the weight of the n-parain mixture used.
The crude heteropolysaccharide is dissolved in 1 litre of water and the solution is subjected to dialysis with deionized 'water at 20 C. for 48 hou-rs. Thus-treated solution is allowed to pass through a column (5 cm. x 50 cm.) packed with strongly acidic cation exchange resin (H- form, such as Amberlite IRO-120) and the eluent is lyophilized to give white spongy heteropolysaccharide. The yield is 1.06 g. or 13.8% based on the weight of the nparaiin mixture used.
Thus-obtained preparation of the acidic heteropolysaccharide has the following properties:
(l) Total Weight of glucose units and mannose units is 81.09% by weight relative to the whole weight of the heteropolysaccharide and weight ratio of glucose units and mannose units is 3:2 in terms of their free form, when measured by hydrolyzing the heteropolysaccharide preparation with 2 N sulfuric acid at 55 C. for 5 hours and subjecting the hydrolyzate to paper chromatography (developer A: a mixture of n-butyl alcohol, pyridine and Water in 6:4:3, developer B: a mixture of n-butyl alcohol, acetic acid and water in 3:l:1, ascending method) and to gas chromatography Iwith N2 gas employing authentic D- glucose and D-mannose as control.
Total weight of succinic acid and lactic acid is 8.99% relative to the Whole weight of the heteropolysaccharide and the weight ratio of lactic acid relative to succinic acid is about 0.18 in terms of their free form, when measured by methylatiug the diethyl ether-soluble fraction of the above-mentioned hydrolyzate with diazomethane and subjecting the methylated fraction to gas chromatography with N2 gas employing cyclohexanol as control.
Acetic acid is '9.99% calculated as CHaiCOO-group relative to the whole Weight of the heteropolysaccharide, when measured by the method described in Methods in Carbohydrate Chemistry vol. V, pp. 187 and 188, published by Academic Press, New York, in 1965.
(2) Its specic rotation is [a]D25=-{35 (C=0.5%, in Water).
(3) The viscosity of its 0.5% aqueous solution as measured employing Ubbelohdes Viscometer is 17,500 centipoises.
(4) Its intrinsic viscosity is about 26.
(5) Its infrared absorption spectrum measured by KBr disc method is as shown in the figure of the accompanying drawing.
(6) It reacts with quaternary ammonium salts such as cetylpyridiniurn chloride and cetyltrimethylammonium bromide to make white complex according to the method described in the said Methods in Carbohydrate Chemistry vol. V, pp. 38 to 44, this clearly means that the heteropolysaccharide of the present invention is acidic one.
EXAMPLE 2 Corynebacterium equi var. muclagz'nosus (ATCC 21521) is inoculated into 30 ml. of an aqueous medium comprising 1% of glucose, 0.5 of yeast extract, 0.5 of peptone and 0.5% of NaCl (pH 7.0) and is incubated under shaking at 28 C. for 24 hours.
10 ml. of the resulting culture broth is inoculated into m1. of the culture medium mentioned in the followiny Table 2, and is incubated under shaking at 28 C. for
The resulting culture broth of a high viscosity is subjected to the same isolation procedures as described in Example l to give white spongy crude heteropolysaccharide. The yield is 1.72 g. or 22.2% based on the Weight of the n-parain mixture used.
The crude heteropolysaccharide is further subjected to the same purification treatments as described in Example 1 to give purified heteropolysaccharide. The yield is 0.98 g. or 12.6% based on the weight of the n-parain mixture used.
EXAMPLE 3 Colynebacterium equi var. mucilgnosus (ATCC 21521) is pre-cultured in a 50 litres tank containing 30 litres of an aqueous medium comprising 1% of glucose, 0.5% of yeast extract, 0.5% of peptone and 0.5% of NaCl (pH 7.0) for 24 hours under the conditions of 28 C., 100% aeration and agitation at 250 r.p.m.
l0 litres of the resulting culture broth is inoculated in a 200 litres tank containing 100 litres of a culture medium of the composition listed in the following Table 3 and incubated for 96 hours under the conditions of 30 C., 50% aeration and agitation at 250 r.p.m., during which time the pH of the medium is maintained at about 6.5 with the addition of 30% sodium hydroxide.
TABLE 3 Percent n-Parafn mixture3 (v./v.) 7 NH4NO3 0.2 K2HPO.,l 0.5 KH2PO4 0.2 MgSO4-7H2O 0.1 Peso.; MnSO4'7H2O 0.002 ZnClZ 0.001 COClZ 0.001 Corn steep liquor 0.03 CaCO3 0.3 pH 7.0.
3The n-parain mixture has the same composition as that of the n-paran mixture described in Table 1.
90 litres of the resulting culture broth of a high viscosity is diluted fourfold with water. The diluted broth is heated at 90 C. under stirring for 10 minutes and is quickly ltered through a lter press along with 3. kg. of a filter aid (Hyflo Super Cel produced by Johns-Manville Products Corp. U.S.A.), whereby a clear viscous fluid is obtained. The fluid is concentrated to about 70 litres and |1% by volume of the uid of sodium chloride is added thereto, followed by the addition of 2 to 3 times of acetone, whereby the heteropolysaccharide separates as white spongy precipitates. The spongy precipitates recovered by filtration are dehydrated with 30 litres of acetone and dried to give white spongy crude heteropolysaccharide. The yield is 1.58 kg. or 29.4% based on the weight of the n-parafiin mixture used.
TEST
0.5 ml. of the respective aqueous solutions of the acidic heteropolysaccharide obtained in Example 1 and gum arabic described in Japanese Pharmacopoeia VII in concentrations listed in Table 4 were daubed on each sheet of white paper (5 cm. x 7 cm.) and each sheet was pasted upon a fresh paper sheet of the same shape. After being kept at C. for 24 hours, it was tested whether or not thus-pasted paper sheets could be separated from each other. The results are summarized in Table 4.
Norm-Substantially no adhesion, or pasted paper sheets could bc separated from each other. -i- =Pasted paper sheets couldnot be separated from nach other. (N)=Samp1cs were unable to be obtalned due to very high viscosity.
Table 4 clearly demonstrates that the acidic heteropolysaccharide of the present invention exhibits a. strong adhesion in one-fourth concentration of gum arabic.
We claim:
1. A method for producing microbial acidic heteropolysaccharide, of which the saccharide units consist substantially of glucose and mannose and the acid units consist substantially of succinic acid, lactic acid and acetic acid, which comprises incubating Carynebacterum equi var. mucz'laginosus ATCC No. 21521 in a culture medium containing hydrocarbon as the main carbon source together with digestible nitrogen source and other nutrients necessary for the growth of the microorganism until a substantial amount of the acidic hcteropolysaccharide is accumulated in the culture broth, and recovering the accumulated acidic heteropolysaccharide therefrom.
2. A method according to claim 1, wherein the incubation is carried out at a temperature of about 25 to about 40 C. and at a pH of about 5 to about 8 under aerobic conditions.
3. A method according to claim 1, wherein the culture medium contains about 2 to about 15 percent by volume of normal paraflins.
4,. A method according to claim 3, wherein the normal parafiins have carbon atoms ranging from 13 to 19.
References Cited UNITED STATES PATENTS 3,406,114 10/1958 yGoren 195-31 P A. LOUIS MONACELL, Primary Examiner G. M. NATH, Assistant Examiner
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP3855169 | 1969-05-19 |
Publications (1)
Publication Number | Publication Date |
---|---|
US3674642A true US3674642A (en) | 1972-07-04 |
Family
ID=12528413
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US38780A Expired - Lifetime US3674642A (en) | 1969-05-19 | 1970-05-19 | Novel microbial acidic heteropolysaccharide and production thereof |
Country Status (5)
Country | Link |
---|---|
US (1) | US3674642A (en) |
CA (1) | CA922653A (en) |
DE (1) | DE2024065A1 (en) |
GB (1) | GB1314992A (en) |
NL (1) | NL7007241A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3856626A (en) * | 1972-06-09 | 1974-12-24 | Exxon Research Engineering Co | Fermentation process for the simultaneous production of protein and bio polymers |
-
1970
- 1970-05-16 DE DE19702024065 patent/DE2024065A1/en active Pending
- 1970-05-19 CA CA083006A patent/CA922653A/en not_active Expired
- 1970-05-19 NL NL7007241A patent/NL7007241A/xx unknown
- 1970-05-19 US US38780A patent/US3674642A/en not_active Expired - Lifetime
- 1970-05-19 GB GB2409570A patent/GB1314992A/en not_active Expired
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3856626A (en) * | 1972-06-09 | 1974-12-24 | Exxon Research Engineering Co | Fermentation process for the simultaneous production of protein and bio polymers |
Also Published As
Publication number | Publication date |
---|---|
NL7007241A (en) | 1970-11-23 |
CA922653A (en) | 1973-03-13 |
GB1314992A (en) | 1973-04-26 |
DE2024065A1 (en) | 1971-02-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US4575551A (en) | Acidic heteropolysaccharide AM-2 | |
US3933788A (en) | Polysaccharide and bacterial fermentation process for its preparation | |
US4186025A (en) | Aqueous polysaccharide composition | |
USRE30872E (en) | Process for producing 2-keto-L-gulonic acid | |
Takeda et al. | Production of 1-kestose by Scopulariopsis brevicaulis | |
US4514563A (en) | Highly viscous polysaccharides | |
De Vuyst et al. | Use of industrial medium components for xanthan production by Xanthomonas campestris NRRL-B-1459 | |
US3823070A (en) | Process for producing a straight chain dicarboxylic acid,an omega-hydroxy fatty acid,and an omega-1-keto fatty acid | |
EP0030393A1 (en) | Xanthanase enzyme and its production | |
US5071976A (en) | Novel heteropolysaccharide | |
US3674642A (en) | Novel microbial acidic heteropolysaccharide and production thereof | |
US4309505A (en) | Process for the production of fructose transferase enzyme | |
US3763008A (en) | Process for producing ribosides of heterocyclic organic bases by fermentation | |
US3658648A (en) | Method for the production of coenzyme q | |
US2978383A (en) | Method for the production of 1-glutamic acid | |
US3806421A (en) | Amylase inhibitor and method of producing the same | |
US3689359A (en) | Method for producing citric acid | |
US4391908A (en) | Method for producing citric acids | |
US3087863A (en) | Amino acid synthesis | |
US3642575A (en) | Process for producing sugars by fermentation | |
US4950604A (en) | Culture of a microorgansim of the genus klebsiella sp., having a high content of rhamnose | |
US3737383A (en) | Process for production of enzyme alkaline dextranase | |
CA1338471C (en) | Process for producing menaquinone-4 | |
US3751337A (en) | Process for producing cells of microorganisms | |
US3806414A (en) | Process for producing citric acid by fermentation |