JPS6225033B2 - - Google Patents
Info
- Publication number
- JPS6225033B2 JPS6225033B2 JP15320984A JP15320984A JPS6225033B2 JP S6225033 B2 JPS6225033 B2 JP S6225033B2 JP 15320984 A JP15320984 A JP 15320984A JP 15320984 A JP15320984 A JP 15320984A JP S6225033 B2 JPS6225033 B2 JP S6225033B2
- Authority
- JP
- Japan
- Prior art keywords
- medium
- culture
- brettanomyces
- strain
- polyhydric alcohols
- 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
Links
- 244000005700 microbiome Species 0.000 claims description 18
- 150000005846 sugar alcohols Polymers 0.000 claims description 16
- 241000100312 Brettanomyces sp. Species 0.000 claims description 10
- 239000002609 medium Substances 0.000 description 27
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 13
- 230000012010 growth Effects 0.000 description 12
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 12
- 229920001817 Agar Polymers 0.000 description 11
- HEBKCHPVOIAQTA-QWWZWVQMSA-N D-arabinitol Chemical compound OC[C@@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-QWWZWVQMSA-N 0.000 description 11
- 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 11
- 239000008272 agar Substances 0.000 description 11
- 239000008103 glucose Substances 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 9
- 230000015572 biosynthetic process Effects 0.000 description 9
- 235000000346 sugar Nutrition 0.000 description 7
- 241000722885 Brettanomyces Species 0.000 description 6
- 239000013078 crystal Substances 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 5
- 229940041514 candida albicans extract Drugs 0.000 description 5
- 229910052799 carbon Inorganic materials 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000012138 yeast extract Substances 0.000 description 5
- HEBKCHPVOIAQTA-NGQZWQHPSA-N D-Arabitol Natural products OC[C@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-NGQZWQHPSA-N 0.000 description 4
- 150000001720 carbohydrates Chemical class 0.000 description 4
- 235000014633 carbohydrates Nutrition 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 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 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 2
- 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 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 241000235648 Pichia Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 244000061456 Solanum tuberosum Species 0.000 description 2
- 235000002595 Solanum tuberosum Nutrition 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-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
- PYMYPHUHKUWMLA-LMVFSUKVSA-N aldehydo-D-ribose Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 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
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 230000034303 cell budding Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 235000013379 molasses Nutrition 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000002699 waste material Substances 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
- AUTALUGDOGWPQH-YMRLNLKISA-N (2r,3s,4s,5r)-2,3,4,5,6-pentahydroxyhexanal;(2r,3s,4r)-2,3,4,5-tetrahydroxypentanal Chemical compound OC[C@@H](O)[C@H](O)[C@@H](O)C=O.OC[C@@H](O)[C@H](O)[C@H](O)[C@@H](O)C=O AUTALUGDOGWPQH-YMRLNLKISA-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
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- 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 1
- QWIZNVHXZXRPDR-UHFFFAOYSA-N D-melezitose Natural products O1C(CO)C(O)C(O)C(O)C1OC1C(O)C(CO)OC1(CO)OC1OC(CO)C(O)C(O)C1O QWIZNVHXZXRPDR-UHFFFAOYSA-N 0.000 description 1
- LKDRXBCSQODPBY-IANNHFEVSA-N D-sorbose Chemical compound OCC1(O)OC[C@@H](O)[C@H](O)[C@H]1O LKDRXBCSQODPBY-IANNHFEVSA-N 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-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
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 229920001202 Inulin Polymers 0.000 description 1
- HEBKCHPVOIAQTA-IMJSIDKUSA-N L-arabinitol Chemical compound OC[C@H](O)C(O)[C@@H](O)CO HEBKCHPVOIAQTA-IMJSIDKUSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 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
- MUBMVGCGOYJTSS-FMTOCKGGSA-N OC[C@@H](O)[C@H](O)[C@H](O)[C@@H](O)C=O.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(O)O[C@@H]1CO Chemical compound OC[C@@H](O)[C@H](O)[C@H](O)[C@@H](O)C=O.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(O)O[C@@H]1CO MUBMVGCGOYJTSS-FMTOCKGGSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- NGFMICBWJRZIBI-JZRPKSSGSA-N Salicin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O1)c1c(CO)cccc1 NGFMICBWJRZIBI-JZRPKSSGSA-N 0.000 description 1
- 229920002472 Starch Polymers 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
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000005377 adsorption chromatography Methods 0.000 description 1
- 230000009603 aerobic growth Effects 0.000 description 1
- PNNNRSAQSRJVSB-BXKVDMCESA-N aldehydo-L-rhamnose Chemical compound C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 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
- NGFMICBWJRZIBI-UHFFFAOYSA-N alpha-salicin Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 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 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 229940008396 carrot extract Drugs 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 235000013312 flour Nutrition 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
- 239000006481 glucose medium Substances 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 229940093915 gynecological organic acid Drugs 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 229940029339 inulin Drugs 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 229910052742 iron Inorganic materials 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
- 230000014759 maintenance of location Effects 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- QWIZNVHXZXRPDR-WSCXOGSTSA-N melezitose Chemical compound O([C@@]1(O[C@@H]([C@H]([C@@H]1O[C@@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O)CO)CO)[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O QWIZNVHXZXRPDR-WSCXOGSTSA-N 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- HOVAGTYPODGVJG-ZFYZTMLRSA-N methyl alpha-D-glucopyranoside Chemical compound CO[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HOVAGTYPODGVJG-ZFYZTMLRSA-N 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 229910017464 nitrogen compound Inorganic materials 0.000 description 1
- 150000002830 nitrogen compounds Chemical class 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- NGFMICBWJRZIBI-UJPOAAIJSA-N salicin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UJPOAAIJSA-N 0.000 description 1
- 229940120668 salicin Drugs 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000003760 tallow Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
【発明の詳細な説明】
本発明は、新規微生物に関する。更に詳細に
は、ブレタノミセス（Brettanomyces）属に属す
る新規微生物に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel microorganisms. More specifically, the present invention relates to a novel microorganism belonging to the genus Brettanomyces.
従来より、発酵による多価アルコール類、就中
アラビトールの製造方法に於て使用される微生物
としては、例えばキヤンジダ属、サツカロミセス
属、トルロプシス属、ピヒア属などに属するもの
が知られている。 BACKGROUND ART Conventionally, microorganisms belonging to the genus Candida, Satucharomyces, Torulopsis, Pichia, etc. have been known as microorganisms used in the method for producing polyhydric alcohols, especially arabitol, by fermentation.
一般に、これら各属に属するアラビトール生産
能を有する公知の微生物を使用した発酵による多
価アルコール類の製造方法に於ては、例えば特公
昭47−20394号、同54−3949号、特開昭54−
145284号公報などに記載されているように、培地
中の主炭素源としてグルコース、グリセロールな
どの糖質を用いた場合に、微生物の耐糖性の問題
から主炭素源としての糖質濃度（基質濃度）に限
界があるばかりでなく、微生物による多価アルコ
ール類への変換率（対糖収率）が十分ではなく、
未だ工業的に利用されるに至つていない。例え
ば、ピヒア属に属する微生物をグルコースを主炭
素源とする培地を用いて好気的に培養した場合、
培養液中に生成するアラビトールの対糖収率は最
高でも50％に過ぎない（特公昭54−3949号）。 In general, methods for producing polyhydric alcohols by fermentation using known microorganisms capable of producing arabitol belonging to each of these genera include, for example, Japanese Patent Publications Nos. 47-20394, 54-3949, and JP-A-54. −
As described in Publication No. 145284, etc., when carbohydrates such as glucose and glycerol are used as the main carbon source in the culture medium, the concentration of carbohydrates as the main carbon source (substrate concentration ) Not only is there a limit to the conversion rate of polyhydric alcohols by microorganisms (yield to sugar),
It has not yet been used industrially. For example, when microorganisms belonging to the genus Pichia are cultured aerobically using a medium containing glucose as the main carbon source,
The yield of arabitol produced in the culture solution relative to sugar is at most only 50% (Special Publication No. 3949/1983).
かかる実情に鑑み、本発明者らは更に多価アル
コール変換率の高い実用性のある発酵法による多
価アルコール類の製造を目的として、種々の起源
による多価アルコール類の生産性について研究、
検索を行なつたところ、沖縄県の製糖工場の廃糖
密から分離されたブレタノミセス属に属する微生
物が、意外にも多価アルコール類、就中アラビト
ールを特異的に生成することを見出し、本発明に
到達したものである。 In view of these circumstances, the present inventors conducted research on the productivity of polyhydric alcohols from various sources, with the aim of producing polyhydric alcohols by a practical fermentation method with a high conversion rate of polyhydric alcohols.
As a result of a search, it was unexpectedly discovered that a microorganism belonging to the genus Brettanomyces, isolated from the waste molasses of a sugar factory in Okinawa Prefecture, specifically produces polyhydric alcohols, especially arabitol. has been reached.
本発明は上記のごとき新知見に基づいて完成さ
れたもので、新規微生物、多価アルコール類生産
能を有するブレタノミセスsp.SN−88菌株に関す
る。 The present invention was completed based on the above new findings, and relates to a new microorganism, the Brettanomyces sp. SN-88 strain, which has the ability to produce polyhydric alcohols.
尚、本発明に係わるブレタノミセス属に属する
微生物の多価アルコール類生産能については未だ
知られておらず、本発明に於て分離、確認された
ブレタノミセスsp.SN−88菌株が最初のものであ
る。 It should be noted that the ability of the microorganisms belonging to the genus Brettanomyces related to the present invention to produce polyhydric alcohols is not yet known, and the Brettanomyces sp. SN-88 strain isolated and confirmed in the present invention is the first. .
当該ブレタノミセスsp.SN−88菌株は、沖縄県
の製糖工場の廃糖密から分離された新規な微生物
であるが、このものは常法に従い純粋分離するこ
とが出来る。即ち、分離源から希釈法によりYM
培地を基準とした寒天培地（グルコース50％
（ｗ／ｗ）、酵母エキス0.3％、麦芽エキス0.3％、
ペプトン1.5％、寒天1.5％；PH5.5）を用いて30
℃、１〜２週間平板培養することにより分離可能
である。 The Brettanomyces sp. SN-88 strain is a new microorganism isolated from the waste molasses of a sugar factory in Okinawa Prefecture, and it can be purified in a conventional manner. That is, YM is extracted from the separated source by dilution method.
Agar medium based on culture medium (glucose 50%
(w/w), yeast extract 0.3%, malt extract 0.3%,
30 using peptone 1.5%, agar 1.5%; PH5.5)
It can be isolated by plating at 1 to 2 weeks at ℃.
本発明に係わる新規微生物ブレタノミセスsp.
SN−88菌株は、次のような菌学的性質を有す
る。 The novel microorganism Brettanomyces sp.
The SN-88 strain has the following mycological properties.
１ 培地上の生育状況
１ 顕微鏡的所見
栄養細胞の大きさ（＊１）３〜７×４〜９μ
栄養細胞の形状 （＊１）
楕円形又は尖頭楕円形
栄養細胞の増殖法 （＊１） 多極出芽
菌 糸 体 （＊２） 形成せず
（註）＊１ YM寒天培地に27℃、５日間培
養
＊２ ポテトグルコース寒天によるス
ライド培養
２ 寒天斜面（＊３）
生 育 良 好
光 沢 白色（光沢）
色 素 ２週間以上培養すると橙
黄色のコロニーとなる。1 Growth status on medium 1 Microscopic findings Size of vegetative cells (*1) 3-7 x 4-9μ Shape of vegetative cells (*1)
Oval or pointed oval vegetative cell growth method (*1) Multipolar budding mycelium (*2) Not formed (Note) *1 Cultured on YM agar medium at 27℃ for 5 days *2 Using potato glucose agar Slide culture 2 Agar slope (*3) Growth Good Gloss White (glossy) Pigment When cultured for more than 2 weeks, orange-yellow colonies will form.
（註）＊３ YM寒天培地
３ 液体培養（＊４）
表面生育 皮膜形成
濁 度 透 明
沈 査 大
（註）＊４ YM液体培地
２ 子のう胞子の形成
ポテトグルコース寒天培地 形成せず
コーンミル寒天培地 形成せず
YM寒天培地 形成せず
ニンジンエキス寒天培地 形成せず
V8寒天培地 形成せず
３ 生理的性質
酸素要求性 好気的
生育温度 20〜40℃
最適生育温度 36℃
生育PH 2.5〜10.5
最適生育PH 8.0〜10.0
KNO3資化性（＊５） 有り
（NH4）2SO4資化性（＊５） 有り
脂肪の分解（＊６） 無し
尿素の分解 無し
ゼラチンの液化 有り
スクロースの
生育可能最高濃度（＊７） 約50％
生育最適濃度（＊７） 約５％
塩化ナトリウムの
生育可能最高濃度（＊８） 3.0％
生育最適濃度（＊８） 0.5〜1.0％
カロチノイドの生成 無し
有機酸の生成 有り
デンプン様物質の生成 無し
ビタミンの要求性（＊５） 無し
（註）＊５ Wickerhamの合成培地を用い
たJ.Lodderらの方法により判
定
＊６ 牛脂を使用
＊７ 液体培地
＊８ 20％（ｗ／ｗ）グルコース培地
中での生育、
液 体
４ 糖の発酵性（＊５）
グルコース ＋
ラクトース −
ガラクトース −
メリビオース −
スクロール −
ラフイノール ＋
マルトース ＋
５ 糖の資化性（＊５）
グルコース ＋
Ｄ−キシロース −
ガラクトース ＋
エリスリトール −
Ｄ−アラビノース ＋
Ｌ−アラビノール ＋
Ｄ−ソルボース −
Ｌ−ソルボース ＋
Ｄ−リボース −
スクロース ＋
Ｌ−ラムノース −
マルトース ＋
エタノール −
セロビオース −
グリセロール ＋
トレハロース −
アドニトール −
ラクトース −
ズルシトール −
メリビオース ＋
Ｄ−マンニトール −
ラフイノース −
Ｄ−ソルビトール ＋
メレジトース ＋
α−メチル−Ｄ−グルコシド −
イヌリン ＋
サリシン −
イノシトール −
可溶性デンプン −
DL−乳酸 −
コハク酸 −
クエン酸 −
上記した菌学的性質からも明らかなごとく、本
菌株は栄養細胞が楕円形もしくは尖頭楕円形をし
ており、多極出芽により増殖、子のう胞子、菌
糸、偽菌糸は形成しないが酸の生成が見られる、
などの特徴を有していることからブレタノミセス
属に属するものと考えられ、更にTHE YEASTS
（J.Loddr et al；1970年版）、YEASTS（J.A.
Barnett et al；1983年版）及びＡ GUIDE TO
IDENTIFYING AND CLASSIFYING（J.A.
Barnett et al；1979年版）に基づき本菌株の分
類学上の位置の詳細に検討したところ、本菌株と
一致する記載が見当たらず、またブレタノミセス
属に属する既知株とも同定すべき記載が見当たら
ないので新菌種であると判定し、これをブレタノ
ミセスsp.SN−88菌株と命名した。 (Note) *3 YM agar medium 3 Liquid culture (*4) Surface growth Film formation Turbidity Transparent Sediment Large (Note) *4 YM liquid medium 2 Ascospore formation Potato glucose agar medium No formation Corn mill agar medium No formation YM agar medium No formation Carrot extract agar medium No formation V 8 agar medium No formation 3 Physiological properties Oxygen requirement Aerobic Growth temperature 20-40℃ Optimum growth temperature 36℃ Growth PH 2.5-10.5 Optimal Growth PH 8.0-10.0 KNO 3 assimilation (*5) Yes (NH 4 ) 2 SO 4 assimilation (*5) Yes Fat decomposition (*6) No Urea decomposition No Gelatin liquefaction Yes Sucrose growth possible Maximum concentration (*7) Approx. 50% Optimum concentration for growth (*7) Approx. 5% Maximum concentration of sodium chloride for growth (*8) 3.0% Optimum concentration for growth (*8) 0.5-1.0% No carotenoid production Organic acids Formation Yes Formation of starch-like substances No Vitamin requirements (*5) None (Note) *5 Determined by J. Lodder et al.'s method using Wickerham's synthetic medium *6 Beef tallow used *7 Liquid medium *8 20% (w/w) Growth in glucose medium, liquid 4 Sugar fermentability (*5) Glucose + Lactose - Galactose - Melibiose - Scroll - Rafuinol + Maltose + 5 Sugar assimilation (*5) Glucose + D -xylose - galactose + erythritol - D-arabinose + L-arabinol + D-sorbose - L-sorbose + D-ribose - sucrose + L-rhamnose - maltose + ethanol - cellobiose - glycerol + trehalose - adonitol - lactose - dulcitol - melibiose + D-Mannitol - Raffinose - D-Sorbitol + Melezitose + α-Methyl-D-glucoside - Inulin + Salicin - Inositol - Soluble starch - DL-Lactic acid - Succinic acid - Citric acid - Also clear from the above mycological properties As shown, the vegetative cells of this strain are oval or pointed oval, and they multiply by multipolar budding, and although they do not form ascospores, hyphae, or pseudohyphae, acid production is observed.
It is thought that it belongs to the genus Brettanomyces because it has such characteristics as THE YEASTS.
(J.Loddr et al; 1970 edition), YEASTS (JA
Barnett et al; 1983 edition) and A GUIDE TO
IDENTIFYING AND CLASSIFYING (JA
When we examined the taxonomic position of this strain in detail based on Barnett et al. (1979 edition), we found no description that matched this strain, and no description that would identify it as a known strain belonging to the genus Brettanomyces. It was determined that it was a new bacterial species, and it was named Brettanomyces sp. SN-88 strain.
本菌株は工業技術院微生物工業技術研究所に
「微生物受託番号微工研菌寄第7595号」のもとに
微生物保管委託してある。 This strain has been entrusted to the Institute of Microbial Technology, Agency of Industrial Science and Technology for storage under the ``Microorganism Accession Number 7595''.
本発明の新規微生物、ブレタノミセスsp.SN−
88菌株の諸性質は常に一定したものではなく、自
然的又は人工的に変異しうるものであり、このよ
うな変異株であつても多価アルコール類の生産能
を有するブレタノミセス属に属する菌株であれば
全て本発明に包含される。 Novel microorganism of the present invention, Brettanomyces sp.SN−
The properties of 88 bacterial strains are not always constant and can vary naturally or artificially, and even such mutant strains are strains belonging to the genus Brettanomyces that have the ability to produce polyhydric alcohols. All of them are included in the present invention.
次に、本発明の新規微生物を用いて多価アルコ
ール類を製造する方法について述べる。 Next, a method for producing polyhydric alcohols using the novel microorganism of the present invention will be described.
本発明に於る培養方法は通常、液体培地を用い
て好気的条件下に撹拌培養により実施されること
が望ましい。 The culture method according to the present invention is preferably carried out by stirring culture under aerobic conditions using a liquid medium.
当該液体培養の主炭素源としてはグルコース、
フルクトース、マンノースなどの糖質が使用され
るが、これら糖質の培地中に於る量的割合として
は、10〜40％（ｗ／ｗ）、特に好ましくは20〜30
％（ｗ／ｗ）の範囲で添加使用されることが好ま
しい。窒素源としては微生物が利用可能な窒素化
合物であればよく、例えば大豆粉、酵母エキス、
ペプトン、肉エキス、コーンスチープリカーなど
が使用される。また、培地に加える無機塩類とし
ては例えばリン酸、マグネシウム、カルシウム、
カリウム、鉄などの塩類が使用される。更に、必
要に応じて微生物の生育に必要な各種の有機物、
無機物などを培地に添加することができる。 The main carbon source of the liquid culture is glucose,
Carbohydrates such as fructose and mannose are used, and the quantitative proportion of these sugars in the medium is 10 to 40% (w/w), particularly preferably 20 to 30%.
% (w/w). The nitrogen source may be any nitrogen compound that can be used by microorganisms, such as soybean flour, yeast extract,
Peptone, meat extract, corn steep liquor, etc. are used. In addition, examples of inorganic salts added to the medium include phosphoric acid, magnesium, calcium,
Salts such as potassium and iron are used. Furthermore, various organic substances necessary for the growth of microorganisms are added as necessary.
Inorganics and the like can be added to the medium.
培養は、前記組成からなる液体培地に微生物菌
体を直接接種するか、又は、別に前培養によつて
得られる種培養液を接種して行なわれる。この種
培養液の調製は、例えば前記のごとき本培養と同
様な組成からなる液体培地に直接菌体を接種して
32〜40℃の温度で２〜５日間程度培養することに
より可能である。 Cultivation is carried out by directly inoculating microbial cells into a liquid medium having the above composition, or by separately inoculating a seed culture obtained by pre-culturing. This seed culture solution can be prepared, for example, by directly inoculating bacterial cells into a liquid medium with the same composition as the main culture described above.
This can be done by culturing at a temperature of 32 to 40°C for about 2 to 5 days.
培養温度は微生物が発育しうる範囲内で適宜設
定されるが、通常32〜40℃、好ましくは34〜37℃
である。 The culture temperature is appropriately set within the range where microorganisms can grow, but is usually 32 to 40°C, preferably 34 to 37°C.
It is.
また、培地のPHは通常6.0〜10.0、好ましく8.0
〜9.0の範囲で調節される。 In addition, the pH of the medium is usually 6.0 to 10.0, preferably 8.0.
It is adjusted in the range of ~9.0.
培養期間は使用する培地の種類及び主炭素源で
ある糖質の濃度により異なるが、通常14〜20日間
程度である。 The culture period varies depending on the type of medium used and the concentration of carbohydrates, which are the main carbon sources, but is usually about 14 to 20 days.
本発明に於る培養は、培地の栄養源が最大限に
利用され、かつ培養液中の多価アルコール類の生
成量が最高に達した時点で終了させることが望ま
しい。尚、培養液中の多価アルコール類の生成量
は、ガスクロマトグラフイー、高速液体クロマト
グラフイーなどの周知の方法を用いて速やかに測
定することが出来る。 It is desirable that the culture in the present invention be terminated when the nutrient source of the medium is fully utilized and the amount of polyhydric alcohol produced in the culture solution reaches its maximum. The amount of polyhydric alcohols produced in the culture solution can be quickly measured using a well-known method such as gas chromatography or high performance liquid chromatography.
かくして、培養液中に蓄積された多価アルコー
ル類は、引き続き常法に従つて培養液から分離さ
れる。即ち、かかる場合に当該分野において通常
使用されている周知の手段、例えばろ過、遠心分
離、イオン交換又は吸着クロマトグラフイー、溶
媒抽出、蒸留、結晶化などの操作が必要に応じて
適宜組み合わせて用いられる。一例を挙げれば、
培養液からろ過、遠心分離などによつて菌体を除
去し、次いでこの液を活性炭で処理して着色物質
などを除き、更にイオン変換樹脂により脱イオン
した後、液を濃縮乾固する。これに熱アルコール
などの有機溶媒を加えてアラビトール、グリセロ
ールなどの多価アルコール類を抽出し、抽出液を
そのままもしくは適度に濃縮して室温又は冷却下
に放置すると白色のアラビトール結晶が析出す
る。これを更にエタノールなどの有機溶媒を用い
て再結晶を行なうことにより、純粋なアラビトー
ルが白色結晶として得られる。 The polyhydric alcohols thus accumulated in the culture medium are then separated from the culture medium in a conventional manner. That is, in such cases, well-known means commonly used in the field, such as filtration, centrifugation, ion exchange or adsorption chromatography, solvent extraction, distillation, crystallization, etc., may be used in appropriate combinations as necessary. It will be done. For example,
The bacterial cells are removed from the culture solution by filtration, centrifugation, etc., then the solution is treated with activated carbon to remove colored substances, and further deionized with an ion conversion resin, and then the solution is concentrated to dryness. An organic solvent such as hot alcohol is added to this to extract polyhydric alcohols such as arabitol and glycerol, and when the extract is left as it is or after being appropriately concentrated and left at room temperature or under cooling, white arabitol crystals are precipitated. By further recrystallizing this using an organic solvent such as ethanol, pure arabitol is obtained as white crystals.
次に、実施例により本発明を説明する。 Next, the present invention will be explained by examples.
実施例 １
グルコース20％（ｗ／ｗ）、酵母エキス（Difco
製）0.5％を含む培地50mlを綿栓した500mlの三角
フラスコに入れ、120℃、15分間滅菌した。放冷
後、培地のPHを無菌的に9.0に調整した。このフ
ラスコにブレタノミセスsp.SN−88菌株「微工研
菌寄第7595号」を接種し、36℃，220rpmで14日
間振とう培養を行なつた。その結果、この培養液
中に6.2ｇのＤ−アラビトール及び微量のグリセ
ロールが蓄積した。Example 1 Glucose 20% (w/w), yeast extract (Difco
50 ml of a culture medium containing 0.5% (manufactured by Nippon Paper Industries, Ltd.) was placed in a 500 ml Erlenmeyer flask with a cotton stopper and sterilized at 120°C for 15 minutes. After cooling, the pH of the medium was aseptically adjusted to 9.0. This flask was inoculated with Brettanomyces sp. SN-88 strain "Feikoken Bacteria No. 7595", and cultured with shaking at 36° C. and 220 rpm for 14 days. As a result, 6.2 g of D-arabitol and a trace amount of glycerol were accumulated in this culture solution.
得られた培養液を遠心分離して菌体を除去し、
活性炭処理及びイオン交換樹脂（IRA−410：IR
−12OB＝２：１）を通して脱塩したのち、50℃
で減圧下濃縮乾固した。これに熱エタノールを加
えて抽出し、過度に濃縮した後５℃で保存した。
生成した結晶をエタノールより再結晶し、収量
4.4ｇの白色結晶を得た。 The obtained culture solution is centrifuged to remove bacterial cells,
Activated carbon treatment and ion exchange resin (IRA-410: IR
-12OB=2:1), then 50℃
The mixture was concentrated to dryness under reduced pressure. This was extracted with hot ethanol, concentrated excessively, and then stored at 5°C.
The generated crystals were recrystallized from ethanol, and the yield
4.4 g of white crystals were obtained.
この結晶の融点は103℃であつた。高速液体ク
ロマトグラフイー、TMS誘導体によるガスクロ
マトグラフイーの保持時間による同定及び旋光度
の測定結果からこの結晶はＤ−アラビトールと判
定された。 The melting point of this crystal was 103°C. This crystal was determined to be D-arabitol based on the retention time results of high performance liquid chromatography and gas chromatography using a TMS derivative, and the measurement results of optical rotation.
実施例 ２
グルコース20％（ｗ／ｗ）、酵母エキス0.5％、
CaCO30.05％からなる培地80mlを500mlの三角フ
ラスコに入れ滅菌した。放冷後、培地のPHを無菌
的に9.0に調整した。このフラスコにブレタノミ
セスsp.SN−88菌株「微工研菌寄第7595号」を接
種し、30℃、180rpmで14日間振とう培養を行な
つた。Example 2 Glucose 20% (w/w), yeast extract 0.5%,
80 ml of a medium consisting of 0.05% CaCO 3 was placed in a 500 ml Erlenmeyer flask and sterilized. After cooling, the pH of the medium was aseptically adjusted to 9.0. This flask was inoculated with Brettanomyces sp. SN-88 strain "Feikoken Bacteria No. 7595" and cultured with shaking at 30° C. and 180 rpm for 14 days.
その結果、この培養液中には8.2ｇのＤ−アラ
ビトール及び1.2ｇのグリセロールが蓄積した。 As a result, 8.2 g of D-arabitol and 1.2 g of glycerol were accumulated in this culture solution.
実施例 ３
グルコース20％（ｗ／ｗ）、酵母エキス0.5％、
KH2PO40.02％からなる培地50mlを500mlの三角フ
ラスコに入れ滅菌した。放冷後、培地のPHを9.0
に調整した。このフラスコにブレタノミセスsp.
SN−88菌株「微工研菌寄第7595号」を接種し、
30℃、220rpmで20日間振とう培養を行なつた。Example 3 Glucose 20% (w/w), yeast extract 0.5%,
50 ml of a medium consisting of 0.02% KH 2 PO 4 was placed in a 500 ml Erlenmeyer flask and sterilized. After cooling, adjust the pH of the medium to 9.0.
Adjusted to. In this flask, Brettanomyces sp.
Inoculated with SN-88 strain "Feikoken Bacteria No. 7595",
Shaking culture was performed at 30°C and 220 rpm for 20 days.
この結果、この培養液中に6.5ｇのＤ−アラビ
トール及び0.6ｇのグリセロールが蓄積した。 As a result, 6.5 g of D-arabitol and 0.6 g of glycerol were accumulated in this culture solution.
Claims (1)
ブレタノミセスsp.SN−88菌株（微工研菌寄第
7595号）。1. A new microorganism Brettanomyces sp.SN-88 strain that has the ability to produce polyhydric alcohols.
No. 7595).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP15320984A JPS6131080A (en) | 1984-07-25 | 1984-07-25 | Novel microorganism and preparation of polyhydric alcohol by fermentation using same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP15320984A JPS6131080A (en) | 1984-07-25 | 1984-07-25 | Novel microorganism and preparation of polyhydric alcohol by fermentation using same |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP24955986A Division JPS6296090A (en) | 1986-10-22 | 1986-10-22 | Production of polyhydric alcohol by fermentation using novel microorganism |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6131080A JPS6131080A (en) | 1986-02-13 |
| JPS6225033B2 true JPS6225033B2 (en) | 1987-06-01 |
Family
ID=15557428
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP15320984A Granted JPS6131080A (en) | 1984-07-25 | 1984-07-25 | Novel microorganism and preparation of polyhydric alcohol by fermentation using same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6131080A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4939091A (en) * | 1986-09-09 | 1990-07-03 | Director Of National Food Research Institute, Ministry Of Agriculture, Forestry And Fisheries | Novel auerobasidium sp. microorganisms, method for obtaining the same and method for preparing erythritol with the same |
-
1984
- 1984-07-25 JP JP15320984A patent/JPS6131080A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6131080A (en) | 1986-02-13 |
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