JPH01199583A - Separation and recovery of erythritol from erythritol-containing culture fluid - Google Patents
Separation and recovery of erythritol from erythritol-containing culture fluidInfo
- Publication number
- JPH01199583A JPH01199583A JP63021347A JP2134788A JPH01199583A JP H01199583 A JPH01199583 A JP H01199583A JP 63021347 A JP63021347 A JP 63021347A JP 2134788 A JP2134788 A JP 2134788A JP H01199583 A JPH01199583 A JP H01199583A
- Authority
- JP
- Japan
- Prior art keywords
- erythritol
- exchange resin
- fraction
- supernatant
- column
- 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.)
- Granted
Links
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 title claims abstract description 71
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 title claims abstract description 71
- 239000004386 Erythritol Substances 0.000 title claims abstract description 70
- 235000019414 erythritol Nutrition 0.000 title claims abstract description 70
- 229940009714 erythritol Drugs 0.000 title claims abstract description 70
- 238000000926 separation method Methods 0.000 title claims description 39
- 238000011084 recovery Methods 0.000 title description 4
- 239000012531 culture fluid Substances 0.000 title 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 47
- 239000006228 supernatant Substances 0.000 claims abstract description 36
- 239000013078 crystal Substances 0.000 claims abstract description 29
- 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 claims abstract description 27
- 235000011187 glycerol Nutrition 0.000 claims abstract description 24
- 238000002425 crystallisation Methods 0.000 claims abstract description 23
- 239000003729 cation exchange resin Substances 0.000 claims abstract description 22
- 230000008025 crystallization Effects 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 18
- 230000002378 acidificating effect Effects 0.000 claims abstract description 13
- 241000894006 Bacteria Species 0.000 claims abstract description 9
- 239000007788 liquid Substances 0.000 claims description 38
- 238000000034 method Methods 0.000 claims description 31
- 239000011347 resin Substances 0.000 claims description 10
- 229920005989 resin Polymers 0.000 claims description 10
- 229910052783 alkali metal Inorganic materials 0.000 claims description 6
- 150000001340 alkali metals Chemical class 0.000 claims description 6
- 230000001580 bacterial effect Effects 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 6
- 239000012736 aqueous medium Substances 0.000 claims description 4
- 239000003456 ion exchange resin Substances 0.000 abstract description 6
- 229920003303 ion-exchange polymer Polymers 0.000 abstract description 6
- 238000010828 elution Methods 0.000 abstract description 4
- 239000003513 alkali Substances 0.000 abstract 2
- 239000000126 substance Substances 0.000 description 32
- 239000000243 solution Substances 0.000 description 26
- 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 19
- 239000008103 glucose Substances 0.000 description 19
- 239000007795 chemical reaction product Substances 0.000 description 18
- 238000007086 side reaction Methods 0.000 description 17
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 14
- 150000003839 salts Chemical class 0.000 description 14
- 229920001542 oligosaccharide Polymers 0.000 description 11
- 150000002482 oligosaccharides Chemical class 0.000 description 11
- 239000002994 raw material Substances 0.000 description 11
- 150000004043 trisaccharides Chemical class 0.000 description 11
- 150000004676 glycans Chemical class 0.000 description 10
- 229920001282 polysaccharide Polymers 0.000 description 10
- 239000005017 polysaccharide Substances 0.000 description 10
- 239000003795 chemical substances by application Substances 0.000 description 9
- 239000012535 impurity Substances 0.000 description 9
- 239000006103 coloring component Substances 0.000 description 7
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 6
- 239000003957 anion exchange resin Substances 0.000 description 6
- 239000012141 concentrate Substances 0.000 description 6
- 238000004042 decolorization Methods 0.000 description 5
- 238000010612 desalination reaction Methods 0.000 description 5
- 150000002500 ions Chemical class 0.000 description 5
- MYRTYDVEIRVNKP-UHFFFAOYSA-N 1,2-Divinylbenzene Chemical compound C=CC1=CC=CC=C1C=C MYRTYDVEIRVNKP-UHFFFAOYSA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 229920002472 Starch Polymers 0.000 description 4
- 240000008042 Zea mays Species 0.000 description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 4
- 235000005822 corn Nutrition 0.000 description 4
- 239000012452 mother liquor Substances 0.000 description 4
- 239000008107 starch Substances 0.000 description 4
- 235000019698 starch Nutrition 0.000 description 4
- 238000011282 treatment Methods 0.000 description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- -1 CaC42 Chemical compound 0.000 description 2
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 235000009754 Vitis X bourquina Nutrition 0.000 description 2
- 235000012333 Vitis X labruscana Nutrition 0.000 description 2
- 240000006365 Vitis vinifera Species 0.000 description 2
- 235000014787 Vitis vinifera Nutrition 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 238000005273 aeration Methods 0.000 description 2
- 150000001735 carboxylic acids Chemical class 0.000 description 2
- 238000013375 chromatographic separation Methods 0.000 description 2
- 238000004040 coloring Methods 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 229910001415 sodium ion Inorganic materials 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 150000005846 sugar alcohols Chemical class 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- JZRWCGZRTZMZEH-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 241000223651 Aureobasidium Species 0.000 description 1
- UNXHWFMMPAWVPI-QWWZWVQMSA-N D-Threitol Natural products OC[C@@H](O)[C@H](O)CO UNXHWFMMPAWVPI-QWWZWVQMSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 241000908267 Moniliella Species 0.000 description 1
- JVWLUVNSQYXYBE-UHFFFAOYSA-N Ribitol Natural products OCC(C)C(O)C(O)CO JVWLUVNSQYXYBE-UHFFFAOYSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 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
- 229930006000 Sucrose Natural products 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 229910052925 anhydrite Inorganic materials 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
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 238000005341 cation exchange Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000005465 channeling Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000925 erythroid effect Effects 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 238000005755 formation reaction Methods 0.000 description 1
- 150000002303 glucose derivatives Chemical class 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
- 150000002314 glycerols Chemical class 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000003317 industrial substance Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 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
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 229920000172 poly(styrenesulfonic acid) Polymers 0.000 description 1
- 229940005642 polystyrene sulfonic acid Drugs 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 description 1
- 229910052939 potassium sulfate Inorganic materials 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- HEBKCHPVOIAQTA-ZXFHETKHSA-N ribitol Chemical compound OC[C@H](O)[C@H](O)[C@H](O)CO HEBKCHPVOIAQTA-ZXFHETKHSA-N 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 235000019157 thiamine Nutrition 0.000 description 1
- 239000011721 thiamine Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Abstract
Description
【発明の詳細な説明】
（、）発明の目的
（産業上の利用分野）
本発明はエリスリトール生産菌の培養液から、培地に添
加された原料物質及び添加物質にもとづく各釉不純物及
び副反応生成物等、たとえば各種の塩類、着色成分及び
各種の多糖類等を簡単に分離・除去して、高純度のエリ
スリトールを高い晶析収率で容易に分離・回収する方法
に関する。なお、本明細書に記載する「エリスリトール
」とは、正確には「メソ−エリスリトール」を意味する
ものであり、このエリスＩＪ ）−ルは甘味料、医薬品
や工業薬品等の中間体などとして有用なものである。Detailed Description of the Invention (,) Purpose of the Invention (Field of Industrial Application) The present invention aims to eliminate various glaze impurities and side reaction formations based on the raw materials and additives added to the culture medium from the culture solution of erythritol-producing bacteria. The present invention relates to a method for easily separating and removing substances such as various salts, coloring components, and various polysaccharides, and easily separating and recovering high-purity erythritol at a high crystallization yield. In addition, "erythritol" described in this specification precisely means "meso-erythritol", and this erythritol is useful as a sweetener, an intermediate for pharmaceuticals, industrial chemicals, etc. It is something.
（従来の技術）
培養法によりエリスＩＪ ）−ルを製造するためのエリ
スリトール産生菌としては、たとえばオーレオパシディ
ウム属（特開昭６１−３１０９１号公報、主な副反応生
成物はグリセリン）、モニリエラ・トメントサ・パール
・ポリニス（特開昭６０−１１０２９５〜８号公報、主
な副反応生成物はグリセリン及び炭素数５の糖アルコー
ルとしてのりビトール）、キャンシダ・ゼイライデス（
ＡＴＣＣ１５５８５）、トルロプシス・ファマタ（ＡＴ
ＣＣ１５８６） 等（％開昭４９−１１８８８９号公報
）、キャンシダ・リポリティカ（米国特許第３，７５６
，９１７号明細書）、及びトルロプシス属、キャンシダ
属（特公昭４７−４１５４９号公報）等が知られている
。(Prior Art) Erythritol-producing bacteria for producing Erys IJ)-l by a culture method include, for example, Aureopacidium (Japanese Patent Application Laid-Open No. 61-31091, main side reaction product is glycerin); Moniliella tomentosa pearl polynis (Japanese Unexamined Patent Publication No. 60-110295-8, the main side reaction products are glycerin and Noribitol as a sugar alcohol with 5 carbon atoms), Cansida zeilides (
ATCC15585), Torulopsis famata (AT
CC1586) etc. (%Kokai No. 49-118889), Cancida lipolytica (U.S. Pat. No. 3,756)
, No. 917), and the genus Torulopsis and Cancida (Japanese Patent Publication No. 47-41549).
従来、これらのエリスリトール産生菌を水性培地で培養
して得られた培養液からエリスリトールを高純度で分離
・回収するには、その前処理として、通常、培養液を活
性炭で脱色処理し、次いでイオン交換樹脂で脱塩、脱色
処理してから濃縮し、冷却・晶析させていた。Conventionally, in order to separate and recover erythritol in high purity from the culture solution obtained by culturing these erythritol-producing bacteria in an aqueous medium, the culture solution is usually decolorized with activated carbon and then ionized. After desalting and decolorizing with an exchange resin, it was concentrated, cooled, and crystallized.
ところが、エリスリトール産生菌の培養工程では、通常
、その培地にＫＨ２ＰＯ４、ＭｇＳＯ４、ＣａＣ４２、
Ｋ２ＳＯ４、ＣａＳＯ４、Ｆ ｅ ＳＯ４、ＭｎＳＯ４
、Ｚ ｎ ＳＯ４、（ＮＨ４）２Ｉ（ＰＯ４等の無機塩
類が多量に添加されるし、また窒素源として（ＮＨ４）
２８０４、尿素、ＮＨ４ｃｔ１（ＮＨ４）２Ｎ０６等が
、さらに栄養源としてコーン・ステイープリカー、大豆
粉、各種アミノ酸、被プトン、チアミン、酵母エキス等
が多量に添加される。However, in the culture process of erythritol-producing bacteria, KH2PO4, MgSO4, CaC42,
K2SO4, CaSO4, FeSO4, MnSO4
, Z n SO4, (NH4)2I (PO4 and other inorganic salts are added in large quantities, and (NH4) is used as a nitrogen source.
2804, urea, NH4ct1(NH4)2N06, etc., and furthermore, as nutritional sources, corn staple liquor, soybean flour, various amino acids, acetate, thiamin, yeast extract, etc. are added in large quantities.
特に、コーン・ステープリカーを用いた場合には、培養
液の着色が著しい。そのために、上記の従来法は、培養
液からこれらの各種の添加物にもとづく不純物を完全に
脱色・脱塩するのに、多量の活性炭やイオン交換樹脂や
、さらＫはそれらの再生用の薬剤全必要とする欠点があ
った。In particular, when corn staple liquor is used, the culture solution is significantly colored. For this reason, the conventional method described above requires a large amount of activated carbon, ion exchange resin, and regenerating agents to completely decolorize and desalinate impurities caused by these various additives from the culture solution. There were all the necessary shortcomings.
捷た、その培養工程では、副反応生成物として、たとえ
ばグリセリン、リビトール、多糖類等が生成するばかり
でなく、原料として結晶しょ糖や結晶ぶどう糖の代りに
、でん粉の酵素糖化法等で得られた精製ぶどう糖（これ
はぶどう糖含有率が９３〜９７％、残りが三糖類、三糖
類、及びそれ以上のオリゴ糖である。）を用いた場合に
は、原料糠中に含まれている三糖類以上のオリゴ糖がそ
のまま又はその反応生成物の形で培養液中に不純物とし
て含まれてくる。そして、これらの各種の不純物は、培
養液の活性炭処理やイオン交換樹脂による脱色・脱塩処
理工程でも除去されないので、エリスＩＪ ）−ルの晶
析収率を高めるための培養液の濃縮時に、これら不純物
濃度も上昇して、晶析をさせる濃縮液が水飴状になシ、
エリスリトールの晶析速度を著しく低下させることにな
る。そのために、上記の従来法はやむをえず、低濃度の
濃縮にとどめて晶析全行なわせざるをえず、ひいてはエ
リスリトールの晶析収率を著しく低下させる欠点があっ
た。In the cultivation process, not only glycerin, ribitol, polysaccharides, etc. are produced as side reaction products, but also products obtained by enzymatic saccharification of starch, etc., are produced instead of crystalline sucrose or glucose as raw materials. When using refined glucose (which has a glucose content of 93 to 97%, with the remainder being trisaccharides, trisaccharides, and higher oligosaccharides), the amount of trisaccharides contained in the raw material bran or higher These oligosaccharides are contained as impurities in the culture solution, either as they are or in the form of their reaction products. These various impurities are not removed even when the culture solution is treated with activated carbon or decolorized and desalted using an ion exchange resin, so when concentrating the culture solution to increase the crystallization yield of Eris IJ) The concentration of these impurities also increases, and the concentrated liquid used for crystallization becomes starch syrup-like.
This will significantly reduce the crystallization rate of erythritol. For this reason, the conventional method described above has the drawback that the entire crystallization must be carried out only at a low concentration concentration, and as a result, the crystallization yield of erythritol is significantly reduced.
そして、本発明者等の研究によれば、エリスリトール培
養液中のエリスリトールの晶析に影響を与える不純物は
、主として下記の成分であることが判明した。According to the research conducted by the present inventors, it has been found that the impurities that affect the crystallization of erythritol in the erythritol culture solution are mainly the following components.
（ｉ） 副反応で生成するグリセリン（１１） グ
リセリン以外の副反応生成物＜ ｉｒ ） でん粉の
酵素糖化法等で得られた精製ぶどう糖を原料として用い
た場合には、原料ぶどう糠中に含まれる三糖類以上のオ
リゴ糖及びその反応生成物
（１ｖ） ぶどう糖を主な構成成分とするβ−１，４
結合をもつ多糖類
このうち、（１）、（１１）及び＜ １ｒ： ）の成分
が多く含まれる場合には、常法により活性炭とイオン交
換樹脂を用いて脱色・脱塩した液を濃縮して晶析を行な
わせると、充分な晶析収率に達しない段階で母液が急激
な粘度上昇を起して水飴状になシ、晶析速度を著しく低
下するとともに、析出した結晶と母液の分離をも困難に
する。(i) Glycerin produced by side reactions (11) Side reaction products other than glycerin <ir) When purified glucose obtained by enzymatic saccharification of starch is used as a raw material, it is contained in the raw material grape bran. Oligosaccharides of trisaccharide or higher and their reaction products (1v) β-1,4 whose main component is glucose
Among polysaccharides with bonds, if the components (1), (11) and < 1r: When crystallization is carried out, the viscosity of the mother liquor rapidly increases before a sufficient crystallization yield is reached, resulting in a starch syrup-like consistency, which significantly reduces the crystallization rate and causes the separation between the precipitated crystals and the mother liquor. It also makes separation difficult.
また、上記（１■）の多糖類は分子量が数千〜致方のも
ので、その生成量が３０〜５００ ｐｐｍ程度と低いが
、活性炭やイオン交換樹脂を用いる通常の脱色・脱塩処
理では除去されず、エリスリトール結晶の晶析工程で析
出し、生成エリスリトール結晶に混入してくる。そのた
めに、得られたエリスＩＪ ）−ル結晶は、これを水に
溶解させたときに白濁し、品質低下の原因となる。In addition, the polysaccharides mentioned in (1) above have molecular weights ranging from several thousand to six hundred thousand, and the amount produced is as low as 30 to 500 ppm, but normal decolorization and desalination treatments using activated carbon and ion exchange resins It is not removed, but precipitates during the crystallization process of erythritol crystals, and is mixed into the produced erythritol crystals. For this reason, the obtained Eris IJ)-le crystals become cloudy when dissolved in water, causing quality deterioration.
なお、晶析収率を向上させる手段とし、原料糖に、でん
粉の酵素糖化法で得られた三糖類以上のオリゴ糖を多く
含むぶどう糖を用いずに、結晶ぶどう糖や結晶しょ糖を
用いて上記（ｉｉｌ ）の原料に由来するオリゴ糖及び
その反応生成物量を少なくする方法を採用することが可
能であるが、しかしこの方法も、（１１）グリセリン以
外の副反応生成物が依然として含まれてくるので、なお
充分に満足できる方法でなかった。In addition, as a means to improve the crystallization yield, the above ( It is possible to adopt a method of reducing the amount of oligosaccharides and their reaction products derived from the raw materials of (11) (iii), but this method still contains side reaction products other than (11) glycerin. However, this method was not completely satisfactory.
（発明が解決しようとする問題点）
本発明は、エリスリトール含有培養液からエリスリトー
ルを分離・回収する方法の改良法、詳しくはエリスリト
ール含有培養液より菌体を分離して得られた上澄液から
、それに含まれる各種の塩類、着色成分及び各種のオリ
ゴ糖及び多糖類等を同時に効率よく分離して除き、高純
度のエリスリトール結晶金高い晶析収率で分離・回収す
る方法を提供しようとするものである。(Problems to be Solved by the Invention) The present invention provides an improved method for separating and recovering erythritol from an erythritol-containing culture solution, specifically, from a supernatant obtained by separating bacterial cells from an erythritol-containing culture solution. , various salts, coloring components, and various oligosaccharides and polysaccharides contained therein are simultaneously and efficiently separated and removed, thereby providing a method for separating and recovering high-purity erythritol crystalline gold with a high crystallization yield. It is something.
（ｂ）発明の構成
（問題点を解決するための手段）
本発明者等は、前記の問題点を解決するために、さらに
研究を重ねた結果、アルカリ金属型若しくはアンモニア
型の強酸性カチオン交換樹脂を分離剤とするクロマトグ
ラフ分離法によりその目的を達成することができたもの
である。(b) Structure of the Invention (Means for Solving the Problems) In order to solve the above-mentioned problems, the present inventors have conducted further research and found that alkali metal type or ammonia type strongly acidic cation exchange This objective was achieved using a chromatographic separation method using a resin as a separation agent.
すなわち、本発明のエリスリトール含有培養液からエリ
スリトールを分離・回収する方法は、エリスリトール生
産菌を水性培地中で好気的条件下で培養して得られた培
養液から菌体を除去した上澄液を、アルカリ金属庫型若
しくはアンモニウム型の強酸性カチオン交換樹脂を充填
した分離塔に通し、次いで水で溶離流出させ、その流出
液からエリスＩＪ ）−ルを主成分として含有する画分
を分取し、該画分よりエリスリトールを回収すること全
特徴とする方法である。That is, the method of separating and recovering erythritol from an erythritol-containing culture solution of the present invention involves culturing erythritol-producing bacteria in an aqueous medium under aerobic conditions, and then removing bacterial cells from the resulting culture solution. is passed through a separation column filled with an alkali metal storage type or ammonium type strongly acidic cation exchange resin, and then eluted with water, and a fraction containing Eris IJ) as a main component is separated from the effluent. This method is characterized in that erythritol is recovered from the fraction.
本発明の分離・回収方法は、エリスＩＪ ）−ル生産菌
を水性培地中で好気的条件下で培養し得られた培養液か
ら菌体を除去した上澄液に対して、アルカリ金属型若し
くはアンモニア型の強酸性カチオン交換樹脂を分離剤と
するクロマトグラフ分離法を適用する方法であるが、そ
の上澄液は、予め硬度成分を除去し、かつ所定の濃度ま
で濃縮しておくのが望ましい。The separation/recovery method of the present invention involves culturing Erys IJ)-le-producing bacteria in an aqueous medium under aerobic conditions, removing the bacterial cells from the resulting culture solution, and using the supernatant to remove alkali metal Alternatively, a chromatographic separation method using an ammonia-type strongly acidic cation exchange resin as a separating agent is applied, but it is recommended that the supernatant liquid be removed in advance to remove hard components and concentrated to a predetermined concentration. desirable.
その上澄液の硬度成分の除去法（軟化処理法）としては
、通常、スルホン酸型の強酸性カチオン交換樹脂のＮａ
型のものを用い、このカチオン交換樹脂を充填した塔に
上澄液を通して上澄液中のＣａイオンやＭｇイオンヲＮ
ａイオンと交換させて除き、Ｃａ型及び／又はＭｇ型に
変ったカチオン交換樹脂１Ｎａｃｔ水溶液でＮａ型に再
生させて繰返し使用する方法と、カルボン酸型の弱酸性
カチオン交換樹脂のＮａ型のものを用い、このカチオン
交換樹脂を充填した塔に上澄液を通して上澄液中の硬度
成分を同様にイオン交換させて除き、Ｃａ及び／又はＭ
ｇ型に変った同樹脂をＨＣｌ又はＨ２ＳＯ４等の強酸で
再生してＨ型にしたのち、ＮａＯＨ水溶液を流してＮａ
型に戻してから繰返し再使用する方法とがある。この二
つの方法のうち、上記上澄液中の全塩濃度が、通常、２
０〜３００ ｍｅｑ／を程度と高いので、このような全
塩濃度の高い上澄液から硬度成分を完全に除くには、カ
ルボン酸型弱酸性カチオン交換樹脂のＮａ型のものを用
いる方法の方がより好ましい。As a method for removing the hard components of the supernatant liquid (softening treatment method), Na
The supernatant liquid is passed through a column filled with this cation exchange resin to remove Ca ions and Mg ions in the supernatant liquid.
A method in which the cation exchange resin is removed by exchanging it with a ions and changed to the Ca type and/or Mg type, and then regenerated into the Na type with a Nact aqueous solution and used repeatedly, and the Na type of the weakly acidic cation exchange resin in the carboxylic acid type. The hardness components in the supernatant are removed by ion exchange by passing the supernatant through a column filled with this cation exchange resin, and Ca and/or M
The same resin that has changed to G-type is regenerated with HCl or a strong acid such as H2SO4 to make H-type, and then a NaOH aqueous solution is poured to
There is a method of returning it to the mold and then reusing it repeatedly. Of these two methods, the total salt concentration in the supernatant is usually 2.
Since the hardness component is as high as 0 to 300 meq/, in order to completely remove the hardness component from the supernatant liquid with such a high total salt concentration, it is better to use a Na type of carboxylic acid type weakly acidic cation exchange resin. is more preferable.
次に本発明の方法における分離剤のアルカリ金属型若し
くはアンモニウム型の強酸性カチオン交換樹脂としては
、ジビニルベンゼン架橋ポＩＪＩ−レンスルホン酸型の
強酸性カチオン交換樹脂（架橋度４〜１０チのもの）の
アルカリ金属型若しくはＮＨ４型のものが好ましく、特
にそのＮａ型のものが最も好ましい。Next, as the alkali metal type or ammonium type strongly acidic cation exchange resin used as the separating agent in the method of the present invention, a divinylbenzene crosslinked polyJI-lenesulfonic acid type strongly acidic cation exchange resin (with a crosslinking degree of 4 to 10 ) are preferred, and the Na type is most preferred.
かかるアルカリ金属型若しくはアンモニウム型の強酸性
カチオン交換樹脂（以下、これを単に「分離剤」という
ことがちる。）全充填した分離塔に、上記の上澄液、好
ましくは硬度成分を除いた上澄液、より好ましくは硬度
成分を除き、かつ３０〜７０重量％の濃度（この濃度は
溶存物質濃度を意味する。）にまで濃縮した濃縮上澄液
を、分離剤体積に対して５〜２０容量係量供給し、次い
で水で溶離流出させ、その流出液からエリスリましくけ
６０〜８０℃に保持する。この場合の分離塔等の温度が
低くすぎると分離塔内で微生物が増殖し、通液圧力の損
失が増大するし、かつチャンネリングの原因となシ、分
離能力が低下する。The above-mentioned supernatant liquid, preferably after removing the hard components, is added to a separation column completely filled with the alkali metal type or ammonium type strongly acidic cation exchange resin (hereinafter simply referred to as "separation agent"). A clear liquid, more preferably a concentrated supernatant liquid after removing hardness components and concentrating to a concentration of 30 to 70% by weight (this concentration refers to the concentration of dissolved substances), is added at a concentration of 5 to 20% by weight based on the volume of the separating agent. The solution is fed at a volumetric rate, then eluted with water, and the effluent is kept in an erythroid tube at 60-80°C. In this case, if the temperature of the separation tower, etc. is too low, microorganisms will proliferate within the separation tower, increasing the loss of liquid passing pressure, causing channeling, and reducing the separation ability.
また、その温度が高すぎると培養液中に含まれる三糖類
以上のオリゴ糖の熱分解を起し、液の着色が著しくなる
。また、分離塔に供給する硬度成分を除いた濃縮液の濃
度が高すぎると、液粘度が上昇して分離性能が低下して
くるし、またその液濃度が低くすぎると、分離塔に供給
すべき液体積が増大し、それにともない溶離液としての
水の使用量が増大するので、分離塔に供給する上澄液濃
度は、上述のように溶存物質濃度として３０〜７０重量
％とするのが好ましい。さらに、分離塔に供給する濃縮
上澄液量が少ないと分離性能が向上するが、分離剤当り
の分離の生産性が低下してくるし、その濃縮上澄液量が
多すぎると分離性能が低下してくるので、分離塔へ供給
する１回当シの濃縮上澄液量は分離剤体積に対して、上
述のように５〜２０容量チとするのが望ましい。On the other hand, if the temperature is too high, oligosaccharides of trisaccharides or higher contained in the culture solution will thermally decompose, resulting in significant coloration of the solution. In addition, if the concentration of the concentrated liquid excluding hardness components supplied to the separation tower is too high, the liquid viscosity will increase and the separation performance will decrease. Since the liquid volume increases and the amount of water used as an eluent increases accordingly, the concentration of the supernatant liquid supplied to the separation column is preferably 30 to 70% by weight as the dissolved substance concentration as described above. . Furthermore, if the amount of concentrated supernatant liquid supplied to the separation column is small, separation performance will improve, but separation productivity per separation agent will decrease, and if the amount of concentrated supernatant liquid is too large, separation performance will decrease. Therefore, it is desirable that the amount of concentrated supernatant liquid supplied to the separation column at one time be 5 to 20 volumes per volume of separation agent, as described above.
なお、前記培地には、通常、前述のようににイオンや窒
素源としてのＮＨ４イオン等の多量の塩が加えられるた
め罠、これらのイオンは硬度成分の除去工程で一部がＮ
ａイオンに置き換えられるにしても、なおこれらイオン
の大部分は分離塔に流入してくることになる。そのため
に、分離塔に最初はＮａ型のカチオン交換樹脂を充填し
ておいても、分離操作を繰り返す間に、Ｎａ型樹脂の一
部かに型やＮＨ４型に変るが、分離性能上は格別の支障
を生じない。Note that, as mentioned above, a large amount of salt such as ions and NH4 ions as a nitrogen source is usually added to the medium, and some of these ions are converted to N during the hardness component removal process.
Even if they are replaced by a ions, most of these ions will still flow into the separation column. For this reason, even if the separation column is initially filled with Na-type cation exchange resin, during repeated separation operations, some of the Na-type resin will change to cation exchange resin or NH4 type, but the separation performance will be significantly lower. does not cause any hindrance.
このようにして、上澄液、好ましくは硬度成分を除き、
かつ所定の濃度に濃縮した上澄液を通し、次いでＭ離液
として水を通して、分離剤に吸着された溶存物質を逐次
に溶離流出させる。すると、まず塩類、着色成分及び分
子量の大きい多糖類が流出し、引続いて原料ぶどう糖中
に含まれていた三糖類以上のオリゴ糖及びグリセリン以
外の副反応生成物が流出し、さらに引続いてエリスリト
ール及びグリセリンが流出してくるから、その流出液を
エリスリトールの流出がはじまる時点の前後の二つの画
分に分けると、塩類、着色成分、副反応で生成する多糖
類、原料ぶどう糖中の三糖類以上のオリゴ糖及びその副
反応生成分等を含む前段の画分と、エリスリトールを主
成分としグリセリンを含む後段の画分とに、容易に分割
することができる。In this way, the supernatant, preferably the hardness component, is removed;
A supernatant liquid concentrated to a predetermined concentration is passed therethrough, and then water is passed therethrough as an M syneresis liquid, so that the dissolved substances adsorbed on the separation agent are successively eluted out. Then, salts, coloring components, and polysaccharides with large molecular weights flow out, followed by oligosaccharides containing trisaccharides or higher contained in the raw glucose and side reaction products other than glycerin, and then further. Erythritol and glycerin flow out, so if you divide the flow into two fractions, before and after the point at which erythritol begins to flow out, you will find salts, coloring components, polysaccharides produced by side reactions, and trisaccharides in the raw material glucose. It can be easily divided into a former fraction containing the above-mentioned oligosaccharides and their side reaction products, and a latter fraction containing erythritol as a main component and glycerin.
かくして分割して得られたエリスリトールを主成分とす
る画分は、充分に着色成分及び塩類等が除かれているか
ら、そのまま濃縮して晶析を行なわせると、容易に高純
度のエリスリトールを高い晶析収率で析出させて回収す
ることができる。すなわち、上記のエリスリトールを主
成分とする画分は、通常、溶存物質濃度として３〜３０
重量％程度の低い濃度のものであシ、かつ副反応生成物
のグリセリンを含むものであるが、この画分は濃縮して
晶析を行なわせれば容易に高純度のエリスリ） −ル結
晶を析出させることができる。たとえば、同画分を溶存
物質濃度が３０〜８５重量％になるまで濃縮してから、
エリスＩＪ ）−ルの種晶を加えて徐冷しながら結晶を
析出させる冷却晶析法、或いは同画分を減圧下で３０〜
８０℃の範囲内の一定温度で濃縮させながら結晶を析出
させる方法等の方法を用いれば、容易に高純度のエリ、
％ ＩＪ ）−ル結晶を高い晶析収率で析出させること
ができる。Since the fraction containing erythritol as the main component obtained by dividing in this manner has sufficiently removed coloring components and salts, it is easy to concentrate and crystallize it as it is to obtain highly pure erythritol. It can be precipitated and recovered with a crystallization yield. That is, the fraction containing erythritol as the main component usually has a dissolved substance concentration of 3 to 30
Although it has a low concentration of about % by weight and contains glycerin as a side reaction product, this fraction can easily precipitate high-purity erythrile crystals by concentrating and crystallizing it. be able to. For example, after concentrating the same fraction until the dissolved substance concentration is 30 to 85% by weight,
A cooling crystallization method in which seed crystals of Ellis IJ) are added and crystals are precipitated while slowly cooling, or the same fraction is heated under reduced pressure for 30 to 30 minutes.
High purity Eri,
% IJ)-ol crystals can be precipitated with a high crystallization yield.
なお、特に高純度のエリスリトール結晶を収得したい場
合には、前記のエリスＩＪ ）−ルを主成分とする画分
を活性炭で脱色し、さらに常法にしたがってカチオン交
換樹脂塔、アニオン交換樹脂塔、及びカチオン交換樹脂
とアニオン交換樹脂との両樹脂の混床塔で処理して完全
に脱塩したのち、上記したと同様の方法で濃縮・晶析を
行なわせると、より高純度のエリスリトール結晶を析出
させることができる。In addition, when it is desired to obtain particularly high-purity erythritol crystals, the fraction containing the above-mentioned erythritol as a main component is decolorized with activated carbon, and then added to a cation exchange resin tower, an anion exchange resin tower, After complete desalination by treatment in a mixed bed tower of both cation exchange resin and anion exchange resin, higher purity erythritol crystals can be obtained by concentrating and crystallizing in the same manner as above. It can be precipitated.
上記したエリスリトール結晶を析出させた残シの母液中
には、副反応生成物のグリセリンが含まれているから、
このグリセリンは別途容易に回収することができる。The mother liquor of the residue from which the above-mentioned erythritol crystals were precipitated contains glycerin, which is a side reaction product.
This glycerin can be easily recovered separately.
このように、本発明の方法を用いれば、培地に添加した
各種塩類、原料ぶどう糠中に含まれる三糖類以上のオリ
ゴ糖、及び副反応等により生成したグリセリン以外の各
種副反応生成物等の上澄液中に含まれる不純物の大部分
は、着色成分をも含めで、溶離工程において分離塔から
初期に溶離流出する画分として、後期に溶離流出するエ
リスリトールを主成分とする画分とに容易に分離できる
ので、そのエリスリトールを主成分とする画分はそれら
の不純物の含有量の著しく少ない画分として得られる。As described above, by using the method of the present invention, various salts added to the culture medium, oligosaccharides containing trisaccharides or higher contained in the raw material grape bran, and various side reaction products other than glycerin generated by side reactions, etc. Most of the impurities contained in the supernatant liquid, including colored components, are separated into a fraction that elutes out of the separation tower in the early stage and a fraction mainly composed of erythritol that elutes and flows out in the later stage of the elution process. Since it can be easily separated, the fraction containing erythritol as a main component can be obtained as a fraction containing significantly less impurities.
したがって、この後期の画分からエリスリトールを晶析
させれば、母液としてパージされる糖類及び糖アルコー
ル量が著しく減少し、エリスリトールの晶析収率が高く
なるし、得られるエリスリトール結晶の純度も高い。Therefore, if erythritol is crystallized from this late fraction, the amount of sugars and sugar alcohols purged as a mother liquor will be significantly reduced, the crystallization yield of erythritol will be high, and the purity of the resulting erythritol crystals will be high.
上記説明においては、回分分離法について述べているが
、精製効果という点では一定量の濃縮上澄液を供給し、
次いで溶離水を供給することＫより塩類、多糖類及びグ
リセリン以外の副反応生成物
を含む画分とエリスリトール翫含む画分に分離すればよ
いわけであり、アルカリ金属型若しくはアンモニウム型
の強酸性カチオン交換樹脂を用いるかぎりにおいて、ど
のような分離操作法を用いても良い。即ち下記の実施例
１及び２に記載されている方法に限定されることなく、
たとえばコロニアルシュガー社出願の特開昭４５−２４
８０７号公報、参松工業株式会社出願の特開昭５３−１
４９８７０号公報、三菱化成工業株式会社出願の特開昭
５５−６１９０３号公報等に記載の回分分離法や米国Ｕ
。０゜Ｐ社出願の米国特許第２９８５５８９号明細１記
載の擬似移動床方式による連続分離法等のような分離操
作法を採用してもよい。In the above explanation, the batch separation method is described, but in terms of purification effect, a fixed amount of concentrated supernatant liquid is supplied,
Next, by supplying elution water, it is sufficient to separate the fraction into a fraction containing salts, polysaccharides, and side reaction products other than glycerin, and a fraction containing erythritol. Any separation method may be used as long as an exchange resin is used. That is, without being limited to the methods described in Examples 1 and 2 below,
For example, JP-A-45-24 filed by Colonial Sugar Co.
Publication No. 807, JP-A-53-1 filed by Sanmatsu Kogyo Co., Ltd.
No. 49870, the batch separation method described in Japanese Patent Application Laid-Open No. 55-61903 filed by Mitsubishi Chemical Industries, Ltd., and the U.S.
. A separation operation method such as a continuous separation method using a simulated moving bed method described in US Pat. No. 2,985,589 filed by 0°P may be employed.
以下、省白 （実施例等） 以下に実施例及び比較例をあげてさらに詳述する。Below, Shoubai (Examples, etc.) Further details will be given below with reference to Examples and Comparative Examples.
実施例１
無水結晶ぶどう糖をぶどう糖として３００ ｇ／ｌ及び
酵母エススを６．７Ｅｌ／ノ含む培地に、オーレオバシ
ジウム５Ｎ−Ｇ４２株を加え、３０℃で７２時間振とう
培養して得た種培地１．５１を、無水結晶ぶどう糖を３
４０１ ／ ｌ ％及びコーン・ステイープ・リカーを
５５ Ｆｌ／ｌ、を含む培地（初期−４、２） ２５１
に加え、通気量２５１／朋、攪拌速度６００ｒｐｍ、温
度３５℃、圧力０．５ ｋｙ／ｍ” Ｇで９３時間培養
し、ぶどう糖が完全になくなった時点を確認して培養を
停止し、直ちに加熱殺菌したのう、遠心分離により菌体
を分離した。得られた上澄液はエリスリトールを１ ｓ
７ ｉ ／ Ａ 、グリセリンを２５ ｇ／ｌ含有し
ていた。Example 1 Seed medium obtained by adding Aureobasidium 5N-G42 strain to a medium containing 300 g/l of anhydrous crystalline glucose and 6.7 El/l of yeast S, and culturing with shaking at 30°C for 72 hours. 1.51, anhydrous crystalline glucose 3
401/l % and corn staple liquor at 55 Fl/l (Initial-4, 2) 251
In addition, the cells were cultured for 93 hours at an aeration rate of 251/m, a stirring speed of 600 rpm, a temperature of 35°C, and a pressure of 0.5 ky/m"G. When the glucose was completely exhausted, the culture was stopped and immediately heated. After sterilization, the bacterial cells were separated by centrifugation.The resulting supernatant was treated with erythritol for 1 s.
7 i/A and contained 25 g/l glycerin.
この上澄液（１９，９７りをカルボン酸型弱カチオン交
換樹脂（三菱化成工業株式会社商品名 ダイヤイオンＷ
Ｋ−２０）のＮａ型を充填した塔に通してＣａ及びＭｇ
等の硬度成分をＮａイオンと交換したのち、溶存物質濃
度が４５重量％になるまで濃縮した。This supernatant liquid (19,97 ri
Ca and Mg are passed through a column packed with Na type of K-20).
After exchanging hardness components such as Na ions with Na ions, it was concentrated until the concentration of dissolved substances became 45% by weight.
分離塔（直径１フインチ（呼び寸法）×高さ２００ ｃ
ｍ ）に、ジビニルベンゼン架橋ポリスチレンスルホン
酸のＮａ型樹脂（三菱化成工業株式会社商品名 ダイヤ
イオンＵＢＫ−５３０）を１．２４／充填し、７５℃に
保持しながら、上記の濃度４５重量％の濃縮液（温度７
５℃）０．１２４１を塔頂から供給速度０．４６１．／
ｈｒで供給し、次いで引続き同じ速度で塔頂から水を供
給した。その際の塔底流出液の分析結果は、第１図に示
すとおシであった。Separation tower (1 inch diameter (nominal dimensions) x height 200 cm
m) was filled with Na-type resin of divinylbenzene cross-linked polystyrene sulfonic acid (Mitsubishi Chemical Industries, Ltd. trade name: Diaion UBK-530) at 1.24/m), and while maintaining the temperature at 75°C, the above concentration of 45% by weight was added. Concentrate (temperature 7
5°C) 0.1241 from the top of the column at a rate of 0.461. /
hr followed by water from the top at the same rate. The analysis results of the bottom effluent at that time were as shown in FIG.
第１図のＢにおける不明物Ｉは、主として塩類、着色物
質及びグルコースを主たる構成成分とするβ−１，４結
合をもつ多糖類からなシ、同図面かられかるように、こ
れら不明物■は初期の流出画分中に含まれて流出してく
る。次いで流出してくる不明物■は、主としてグリセリ
ン以外の副反応生成物、三糖類以上のオリが糖及びその
反応生成物であるが、これらも比較的初期の流出液中に
含まれて流出してくる。なお、この実施例では原料ぶど
う糖として結晶ぶどう糖が用いられたので、不明物■に
は、原料ぶどう糖に由来する三糖類以上のオリゴ糖及び
その反応生成物が含まれていなく、その不明物■は、主
としてグリセリン以外の副反応生成物であった。そして
、後期の流出液としてエリスリトールとグリセリンが流
出してくる。Unknown substance I in B of Fig. 1 is mainly composed of polysaccharides with β-1,4 bonds whose main constituents are salts, coloring substances, and glucose. is contained in the initial effluent fraction and flows out. The unknown substances that flow out next are mainly side reaction products other than glycerin, trisaccharides and higher sugars, and their reaction products, but these are also contained in the effluent at a relatively early stage and flow out. It's coming. In addition, since crystalline glucose was used as the raw material glucose in this example, the unknown substance (2) does not include oligosaccharides higher than trisaccharides derived from the raw material glucose and their reaction products, and the unknown substance (2) is , which were mainly side reaction products other than glycerin. Erythritol and glycerin then flow out as effluent in the latter stage.
また、第１図のＡから明らかなように、着色成分は、そ
の大部分が前期の流出液中に含まれている。Moreover, as is clear from A in FIG. 1, most of the colored components are contained in the effluent from the first stage.
したがって、第１図から明らかなように、その流出液を
、流出液床容量０．５４を境にして前段と後段の二つの
画分に分けると、前段の画分には着色物質の大部分と不
明物Ｉ及び■の殆んど全部とが含まれ、後段の画分には
エリスリトールとグリセリンの殆んど全部が含まれてく
るから、この後段の画分をエリスリトールを主成分とす
る画分として収得すれば、これには着色成分や塩類や多
糖類が殆んど含まれていない。換言すれば、上澄液から
エリスリトール及びグリセリンを含み、不純物を殆んど
含まない画分を後段の画分として容易に分離できるので
ある。Therefore, as is clear from Fig. 1, when the effluent is divided into two fractions, the former and the latter, with the effluent bed volume of 0.54 as the border, most of the colored substances are contained in the former fraction. and almost all of Unknown Substances I and If obtained as a fraction, it contains almost no coloring components, salts, or polysaccharides. In other words, a fraction containing erythritol and glycerin and almost no impurities can be easily separated from the supernatant as a subsequent fraction.
前記の流出液床容量０．５４を境にして前段と後段の二
つの画分に分けた場合の各段の画分の液量は、前段が０
．６７１であシ、後段が０．４７１であった。When dividing into two fractions, the front stage and the rear stage, with the above-mentioned effluent bed volume of 0.54 as the boundary, the liquid volume of the fraction in each stage is 0.
．． The score was 671, and the second half was 0.471.
次いで、この操作を７０回繰シ返し、後段流出液として
合計３３ノを得る。その液組成はエリスリトール濃度１
０６１ｊ／ｌ、グリセリン濃度１４．２ ／１／ｌ、不
明物■濃度０−９８＆／Ｍであった。この液に粉末活性
炭６６、Ｐを加え３０分間攪拌後、活性炭を濾過し、Ｐ
液を常法に従ってＨ型強酸性カチオン交換樹脂（三菱化
成工業株式会社商品名ダイヤイオン５ＫＩＢ）を充填し
た塔、ＯＨ型の弱塩基性アニオン交換樹脂（三菱化成工
業株式会社商品名 ダイヤイオンＷＡ３０）を充填した
塔、及び前記のＨ型強酸性カチオン交換樹脂とＯＨ型強
塩基性アニオン交換樹脂（三菱化成工業株式会社商品名
ダイヤイオンＰＡ４０８ ）を充填した混床塔に順次
通して、さらに脱塩・脱色したのち、減圧下８０℃で溶
存物質濃度が５３重量％になるまで濃縮した。Next, this operation was repeated 70 times to obtain a total of 33 pieces of effluent as the latter stage effluent. The liquid composition is erythritol concentration 1
061j/l, glycerin concentration 14.2/1/l, unknown substance concentration 0-98&/M. Powdered activated carbon 66, P was added to this liquid and after stirring for 30 minutes, the activated carbon was filtered and P
The solution was transferred to a column filled with H-type strongly acidic cation exchange resin (Mitsubishi Chemical Industries, Ltd., trade name: Diaion 5KIB), and an OH-type weakly basic anion exchange resin (Mitsubishi Chemical Industries, Ltd., trade name: Diaion WA30). and a mixed-bed column filled with the above-mentioned H-type strongly acidic cation exchange resin and OH-type strongly basic anion exchange resin (trade name: Diaion PA408, manufactured by Mitsubishi Chemical Industries, Ltd.) for further desalination. - After decolorization, it was concentrated under reduced pressure at 80°C until the concentration of dissolved substances was 53% by weight.
次いで、この８０℃の濃縮液を２時間かけて１５℃まで
徐冷し、さらに１５℃に２時間保存して結晶を成長させ
た。生成結晶を炉別し、得られた湿潤結晶をその０．６
倍量の１５℃の水で洗浄し、純度１００％のエリスリト
ール結晶が１．６６ｋｙ得られた。この場合の前記の５
３重量％の濃縮液中のエリスリトールに対する晶析収率
は４７．５％であった。また得られた結晶を水に溶解し
、１０重量％の溶液をつくったが、にごシは認められな
かりだ。Next, this 80°C concentrated solution was slowly cooled to 15°C over 2 hours, and further stored at 15°C for 2 hours to grow crystals. The produced crystals are separated by furnace, and the obtained wet crystals are 0.6
By washing with twice the amount of water at 15°C, 1.66 ky of erythritol crystals with 100% purity were obtained. In this case, the above 5
The crystallization yield for erythritol in the 3% by weight concentrate was 47.5%. I also dissolved the obtained crystals in water to make a 10% by weight solution, but no cloudiness was observed.
実施例２
原料ぶどう糖として純度９６．８重量％（残９３．２重
量％が三糖類以上のオリゴ糖）の精製ぶどう糖を用い、
このぶどう糖を純ぶどう糖として４００９／１１コーン
・ステイープ・リカーを６７１７１１含み、−が４．２
の初期培地２５Ａ’に、実施例１におけると同一の種菌
培養液１．５１を加え、温度３５℃、通気量１３１／”
ｚ圧力０．５ ｋｇ７ｃｍ２、攪拌速度６００ ｒｐｍ
の条件で培養を行なったところ、９５時間の培養でぶど
う糖が全く々くなったので、直ちに培養を停止し、加熱
殺菌後、菌体を分離した。Example 2 Using purified glucose with a purity of 96.8% by weight (the remaining 93.2% by weight is oligosaccharides of trisaccharides or higher) as the raw material glucose,
This glucose is pure glucose and contains 671,711 4009/11 corn steep liquor, - is 4.2
To 25 A' of initial culture medium, 1.5 l of the same inoculum culture solution as in Example 1 was added, and the temperature was 35°C and the aeration rate was 131/''.
Z pressure 0.5 kg7cm2, stirring speed 600 rpm
When culturing was carried out under the following conditions, the amount of glucose decreased completely after 95 hours of culture, so the culture was immediately stopped, and after heat sterilization, the bacterial cells were isolated.
得られた上澄液（約１９．９７）の分析結果は、エリス
リトールが２１３１１７Ａ’、グリセリンが１７．５ｇ
／ノ、不明物Ｉが１８ ｇ／ｌ、不明物■が１６ ｇ／
ｌであった。The analysis results of the obtained supernatant liquid (approximately 19.97 g) showed that erythritol was 213117 A' and glycerin was 17.5 g.
/ノ, unknown substance I is 18 g/l, unknown substance ■ is 16 g/l
It was l.
この上澄液をダイヤイオンＷＫ−２０（三菱化成工業株
式会社商品名）のＮａ型樹脂を充填した塔に通して軟化
処理をしたのち、溶存物質濃度が４５重量％になるまで
濃縮した。その濃縮量は約１０．２１であった。This supernatant liquid was softened by passing through a column filled with Na-type resin of Diaion WK-20 (trade name of Mitsubishi Chemical Industries, Ltd.), and then concentrated until the concentration of dissolved substances became 45% by weight. The concentration amount was about 10.21.
次いで、ダイヤイオンＵＢＫ−５３０（三菱化成工業株
式会社商品名）のＮａ型樹脂０．６２１を塔（直径１１
インチ（呼び径）×高さ１００（ｈａ＋）に充填した分
離塔を用いて、上記の濃縮液及び水を、０．４９６ノ／
ｈｒの一定の通液速度で、次の四段階に分けてそれぞれ
通液した。Next, a column (diameter 11
Using a separation column packed in inches (nominal diameter) x height 100 (ha+), the above concentrate and water were separated at 0.496 nm/h.
The liquid was passed through each of the following four stages at a constant liquid passing rate of hr.
第一段階：
前記の濃度４５重量％の濃縮液７０−を塔頂から供給し
、塔底から塔底流出液を７０ｍ流出させた。この塔底流
出液は、前回の第四段階に引続き塔内液を塔底方向に移
動させることによって樹脂から溶離した液であるので、
エリスリトールを主成分とする画分てあった。First stage: The concentrated liquid 70- with a concentration of 45% by weight was supplied from the top of the column, and the bottom effluent was allowed to flow out 70 m from the bottom of the column. This tower bottom effluent is the liquid eluted from the resin by moving the tower internal liquid toward the tower bottom following the previous fourth stage.
There was a fraction whose main component was erythritol.
第二段階：
塔内の液を塔底からポンプで塔頂に１５．７分間（１３
０ｍＡ’分）循環させた。Second stage: The liquid in the column is pumped from the bottom of the column to the top of the column for 15.7 minutes (13
0 mA'min).
第三段階：
塔頂から水１４３ゴを供給し、塔底からの流出液（１４
３ｍｌ）を・ぐ−ジした。この場合の塔底からの流出液
は各種塩類、着色成分及び不明物■を含有する液である
ので上記のようにパージした。Third stage: 143 g of water is supplied from the top of the column, and the effluent from the bottom of the column (14 g.
3 ml) was poured. In this case, the effluent from the bottom of the column contained various salts, coloring components, and unknown substance (1), and was therefore purged as described above.
第四段階：
塔頂から水１８０−を供給し、塔底から１８０ｍ１の流
出液を取出した。この流出液はエリスリトールを主成分
として含有する画分であるので、第１段階での塔底流出
液と混合した。Fourth stage: 180 mL of water was supplied from the top of the column, and 180 ml of effluent was taken from the bottom of the column. Since this effluent is a fraction containing erythritol as a main component, it was mixed with the bottom effluent from the first stage.
以上の四段階の処理を１サイクルとし、第四段階後には
再び第１段階に戻るサイクルを、前記の上澄液の濃縮液
（１０，２Ａりがなくなるまで繰返して、エリスリトー
ルを主成分とする画分３６．３１を得た。The above four-stage treatment is one cycle, and after the fourth stage, the cycle of returning to the first stage is repeated until the supernatant concentrate (10,2A) is removed, and the main component is erythritol. Fraction 36.31 was obtained.
その画分の分析結果は、エリスリトールが１１６ｊ；！
／ｌ、グリセリンが９．６ｇ／Ａ！、不明物■が２．６
ｇ／ｌであシ、その脱塩率が９５チ、脱色率が８７チ、
エリスリトール回収率が９９．９％、不明物■のパージ
率が７０チであった。また、その画分は着色度（Ａ４２
０１ ｃｍセル、以下、同様）が０．３２、電気伝導度
が１６７μＳ／ｅｍであった。The analysis results of that fraction showed that erythritol was 116j;!
/l, glycerin is 9.6g/A! , unknown item ■ is 2.6
g/l, its desalination rate is 95cm, its decolorization rate is 87cm,
The recovery rate of erythritol was 99.9%, and the purge rate of unknown substance (2) was 70%. In addition, the fraction has a coloring degree (A42
01 cm cell (hereinafter the same) was 0.32, and the electrical conductivity was 167 μS/em.
次いで、上記のようにして得られたエリスリトールを主
成分として含有する画分（３６，３１）に、粉末油戻を
０．１重量％加えて脱色処理をしたところ、着色度が０
，１６にまで低下した。さらに、この脱色処理液を、常
法にしたがってＨ型の強酸性カチオン交換樹脂（三菱化
成工業株式会社商品名ダイヤイオン５ＫＩＢ）を充填し
た塔、ＯＨ型の弱塩基性アニオン交換樹脂（三菱化成工
業株式会社商品名 ダイヤイオンＷＡ３０）を充填した
塔、及び前記のＨ型樹脂及びＯＲ型強塩基性アニオン交
換樹脂（三菱化成工業株式会社商品名 ダイヤイオンＰ
Ａ４０８）を充填した混床塔に順次に通して、さらに脱
塩・脱色したのち、減圧下、８０℃で固形分が６１重量
％になるまで濃縮した。Next, when the fractions (36, 31) containing erythritol as a main component obtained as described above were decolorized by adding 0.1% by weight of powdered oil reconstituted powder, the degree of coloration was 0.
, it dropped to 16. Furthermore, this decolorizing solution was added to a column filled with an H-type strongly acidic cation exchange resin (trade name Diaion 5KIB, manufactured by Mitsubishi Chemical Industries, Ltd.) and an OH-type weakly basic anion exchange resin (Mitsubishi Chemical Industries, Ltd. Co., Ltd. (trade name: Diaion WA30), and the above-mentioned H-type resin and OR-type strongly basic anion exchange resin (Mitsubishi Chemical Industries, Ltd. (trade name: Diaion P)).
The mixture was sequentially passed through a mixed bed tower filled with A408) for further desalination and decolorization, and then concentrated under reduced pressure at 80°C until the solid content was 61% by weight.
次いで、この８０℃の濃縮液を２時間かけて１５℃まで
徐冷し、さらに１５℃に２時間保持して結晶を成長させ
た。生成結晶を炉別し、得られた湿潤結晶をその０．６
倍量の１５℃の水で洗浄し、乾燥させたところ、純度１
００チのエリスリトール結晶２．４０ｋｇが得られた。Next, this 80°C concentrated solution was gradually cooled down to 15°C over 2 hours, and further maintained at 15°C for 2 hours to grow crystals. The produced crystals are separated by furnace, and the obtained wet crystals are 0.6
When washed with twice the amount of water at 15°C and dried, the purity was 1.
2.40 kg of erythritol crystals were obtained.
比較例１
実施例２で得られた培養液の上澄液（着色６．２、電気
伝導度７．０ｍｓ／ｃｍ）を脱色するために粉末油戻を
５重量％添加したが、着色度が０．３５にまでしか低下
できなかった。さらに、その脱色後の上澄液を、常法に
したがって実施例２におけると同様のイオン交換樹脂に
よる脱塩処理をしたところ、イオン交換樹脂は、実施例
２の場合の２０倍量を必要とした。Comparative Example 1 In order to decolorize the supernatant liquid of the culture solution obtained in Example 2 (coloration 6.2, electrical conductivity 7.0ms/cm), 5% by weight of powdered oil reconstituted was added, but the degree of coloration was It could only be reduced to 0.35. Furthermore, when the supernatant liquid after decolorization was desalted using an ion exchange resin similar to that in Example 2 according to a conventional method, the amount of ion exchange resin required was 20 times that in Example 2. did.
さらに、その脱色・脱塩処理した上澄液を８０℃の減圧
下で固形分濃度５０重量％まで濃縮してから、実施例２
におけると同様の条件を用いて冷却・晶析させ、結晶分
離、水洗、乾燥をして純度１００チの結晶ｉ、５ｏｋ１
７を得だ。この場合の５０重量％濃縮液中のエリスリト
ールに対する晶析収率は４３．８％であった。Furthermore, the decolorized and desalted supernatant liquid was concentrated under reduced pressure at 80°C to a solid content concentration of 50% by weight, and then
Cool and crystallize using the same conditions as in , crystal separation, water washing, and drying to obtain crystals with a purity of 100%, 5ok1.
I got a 7. In this case, the crystallization yield for erythritol in the 50% by weight concentrate was 43.8%.
また、製品エリスリトール結晶収率を高めるために、８
０℃の減圧下での濃縮時の固形分濃度を６０重量％に高
めたところ、製品エリスリトール結晶の純度が９９．８
％に低下した。また、濃縮液濃度５０重量％及び６０重
−ｉ％のいずれの条件で得られた結晶も、１０重量％の
水溶液にすると白濁が認められた。In addition, in order to increase the yield of product erythritol crystals, 8
When the solid content concentration during concentration under reduced pressure at 0°C was increased to 60% by weight, the purity of the product erythritol crystals was 99.8.
%. In addition, when the crystals obtained under both the conditions of concentrated solution concentration of 50% by weight and 60% by weight were made into a 10% by weight aqueous solution, white turbidity was observed.
（ｃ）発明の効果
本発明の分離・回収方法は、エリスリトール含有培養液
から高純度のエリスリトール結晶を高い晶析収率で容易
に分離・回収することができる。(c) Effects of the Invention The separation/recovery method of the present invention allows highly pure erythritol crystals to be easily separated and recovered from an erythritol-containing culture solution at a high crystallization yield.
第１図は実施例１における分離塔から溶離液の水に溶解
して流出する流出液物質と流出液量の関係を示したもの
であシ、Ａは流出液量と液の着色度との関係を、Ｂは流
出液量と各物質の濃度との関係を図示したものである。
特許出願人 三菱化成工業株式会社
日研化学株式会社Figure 1 shows the relationship between the amount of effluent and the amount of effluent that dissolves in the eluent water and flows out from the separation tower in Example 1, and A is the relationship between the amount of effluent and the degree of coloration of the liquid. B shows the relationship between the amount of effluent and the concentration of each substance. Patent applicant Mitsubishi Chemical Industries, Ltd. Nikken Chemical Co., Ltd.
Claims (1)
で培養して得られた培養液から菌体を除去した上澄液を
、アルカリ金属型若しくはアンモニウム型の強酸性カチ
オン交換樹脂を充填した分離塔に通し、次いで水で溶離
流出させ、その流出液からエリスリトールを主成分とし
て含有する画分を分取し、該画分よりエリスリトールを
回収することを特徴とするエリスリトール含有培養液か
らエリスリトールを分離・回収する方法。 ２）流出液からエリスリトールとグリセリンとを主とし
て含有する画分を分取し、その画分から晶析によってエ
リスリトール結晶を回収する特許請求の範囲第１項記載
の方法。 ３）上澄液を分離塔に通すに先立って、上澄液中の硬度
成分をアルカリ金属型カチオン交換樹脂を用いて除いて
おく特許請求の範囲第１項又は第２項記載の方法。 ４）強酸性カチオン交換樹脂がＮａ型の樹脂である特許
請求の範囲第１項、第２項又は第３項記載の方法。[Scope of Claims] 1) The supernatant liquid obtained by removing the bacterial cells from the culture solution obtained by culturing erythritol-producing bacteria in an aqueous medium under aerobic conditions is Erythritol is passed through a separation tower filled with a cation exchange resin, then eluted with water, a fraction containing erythritol as a main component is separated from the effluent, and erythritol is recovered from the fraction. A method for separating and recovering erythritol from a culture solution containing it. 2) The method according to claim 1, wherein a fraction mainly containing erythritol and glycerin is separated from the effluent, and erythritol crystals are recovered from the fraction by crystallization. 3) The method according to claim 1 or 2, wherein hard components in the supernatant are removed using an alkali metal cation exchange resin before the supernatant is passed through the separation column. 4) The method according to claim 1, 2 or 3, wherein the strongly acidic cation exchange resin is a Na type resin.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63021347A JPH0734748B2 (en) | 1988-02-02 | 1988-02-02 | Method for separating and recovering erythritol from culture medium containing erythritol |
| EP19890101623 EP0327016B1 (en) | 1988-02-02 | 1989-01-31 | Process for isolating and recovering erythritol from culture medium containing the same |
| DE1989611158 DE68911158T2 (en) | 1988-02-02 | 1989-01-31 | Process for the separation and extraction of erythritol from a culture medium containing it. |
| US07/304,794 US4906569A (en) | 1988-02-02 | 1989-01-31 | Process for isolating and recovering erthritol from culture medium containing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63021347A JPH0734748B2 (en) | 1988-02-02 | 1988-02-02 | Method for separating and recovering erythritol from culture medium containing erythritol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH01199583A true JPH01199583A (en) | 1989-08-10 |
| JPH0734748B2 JPH0734748B2 (en) | 1995-04-19 |
Family
ID=12052557
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63021347A Expired - Lifetime JPH0734748B2 (en) | 1988-02-02 | 1988-02-02 | Method for separating and recovering erythritol from culture medium containing erythritol |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0734748B2 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009057286A (en) * | 2007-08-29 | 2009-03-19 | Japan Organo Co Ltd | Method for purifying alcohol containing cationic impurity |
-
1988
- 1988-02-02 JP JP63021347A patent/JPH0734748B2/en not_active Expired - Lifetime
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009057286A (en) * | 2007-08-29 | 2009-03-19 | Japan Organo Co Ltd | Method for purifying alcohol containing cationic impurity |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0734748B2 (en) | 1995-04-19 |
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