JP5420153B2 - 乳酸菌によるアルドン酸の製造方法 - Google Patents
乳酸菌によるアルドン酸の製造方法 Download PDFInfo
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- JP5420153B2 JP5420153B2 JP2007088046A JP2007088046A JP5420153B2 JP 5420153 B2 JP5420153 B2 JP 5420153B2 JP 2007088046 A JP2007088046 A JP 2007088046A JP 2007088046 A JP2007088046 A JP 2007088046A JP 5420153 B2 JP5420153 B2 JP 5420153B2
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- Prior art keywords
- acid
- lactic acid
- culture
- lactobionic acid
- aldonic
- Prior art date
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- 239000000701 coagulant Substances 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
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- 229910052802 copper Inorganic materials 0.000 description 1
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- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical compound CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 description 1
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- 229940029339 inulin Drugs 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 229960004903 invert sugar Drugs 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
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- FSICMNGKCHFHGP-AMTLMPIISA-N lactobiono-1,5-lactone Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(=O)[C@H](O)[C@H]1O FSICMNGKCHFHGP-AMTLMPIISA-N 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 235000020191 long-life milk Nutrition 0.000 description 1
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- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
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- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 229910052750 molybdenum Inorganic materials 0.000 description 1
- 239000011733 molybdenum Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
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- 238000007254 oxidation reaction Methods 0.000 description 1
- BJRNKVDFDLYUGJ-UHFFFAOYSA-N p-hydroxyphenyl beta-D-alloside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-UHFFFAOYSA-N 0.000 description 1
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- 235000020232 peanut Nutrition 0.000 description 1
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- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
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- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 235000011962 puddings Nutrition 0.000 description 1
- 150000003856 quaternary ammonium compounds Chemical class 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000001105 regulatory effect Effects 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
- 150000003839 salts Chemical group 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
- 239000010822 slaughterhouse waste Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
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- 238000000859 sublimation Methods 0.000 description 1
- 230000008022 sublimation Effects 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Dairy Products (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
(形態的特性)
培養温度:30℃
形状:球菌
大きさ:0.9―1.0μm
運動性の有無:なし
胞子の形成:なし
(培養的性質)
色調:クリーム色
コロニー形態:円形
***状態:レンズ状
周縁:全縁
表面:スムーズ
透明度:不透明
粘稠度:バター様
(生理学的性質)
グラム染色:+
カタラーゼ:−
オキシダーゼ:−
O/F試験:+
40℃での生育:−
4%NaClでの生育:−
(発酵試験)
グリセロール:−
エリスリトール:−
D-アラビノース:−
L-アラビノース:−
リボース:−
D-キシロース:−
L-キシロース:−
アドニトール:−
β-メチル-D-キシロシド:−
ガラクトース:+
グルコース:+
フラクトース:+
マンノース:+
ソルボース:−
ラムノース:−
ズルシトール:−
イノシトール:−
マンニトール:−
α-メチル-D-マンノシド:−
α-メチル-D-グルコシド:−
N-アセチルグルコサミン:+
アミグダリン:−
アルブチン:−
エスクリン:+
サリシン:+
セロビオース:+
マルトース:+
ラクトース:+
メリビオース:−
サッカロース:−
トレハロース:+
イヌリン:−
メレチトース:−
ラフィノース:−
でんぷん:−
グリコーゲン:−
キシリトール:−
ゲンチオビオース:+
D-ツラノース:−
D-リキソース:−
D-タガトース:−
D-フコース:−
L-フコース:−
D-アラビトール:−
L-アラビトール:−
グルコネート:−
2-ケトグルコネート:−
5-ケトグルコネート:−
上記のような本菌株の菌学的性質と、16S rDNA塩基配列をGenBank/DDBJ/EMBLにより相同性検索した結果から、本菌株はL. lactisに帰属し、亜種レベルではL. lactis subsp. cremorisに帰属する可能性が最も高いと考えられた。しかしながら、後述するようにラクトビオン酸の産生能の点で公知のL. lactis subsp. Cremorisと相異が認められたため、新菌株として寄託を行なった。
HPLC分析条件
カラムAsahipak NH2P−50(昭和電工社製)
溶離液 アセトニトリル:40mMクエン酸−NaH2PO4緩衝液(pH5.0)=60:40(体積比)
条件 温度:40℃
流速:0.8mL/mL
検出器:示差屈折計
〔菌体の調製〕
試験管(18 mm×200 mm)に、ラクトース0.5%、グルコース0.5%、酵母エキス0.5%、ポリペプトン0.5%、硫酸マグネシウム0.1%(pH7.0)を含む培地3mLを分注し、121℃で20分間殺菌した。その試験管にラクトコッカス・ラクティス・サブスピーシーズ・クレモリスKYG−33(FERM P−21207)(実施例1)及び独立行政法人製品評価技術基盤機構や東京大学分子細胞生物学研究所から入手した乳酸菌(実施例2、3、4)の一白金耳を植菌し、27℃で3日間振とう培養(220rpm)した。その後、培養液を遠心し菌体を回収した。
〔ラクトビオン酸の生産〕
上記の方法で得られた菌体の全量を2%ラクトース1mLに再懸濁し、27度で3日間振とうし、反応液中のラクトビオン酸を測定した。結果を表1に示す。
実施例1と同様にして、ラクトコッカス・ラクティス・サブスピーシーズ・クレモリスKYG−33(FERM P−21207)の菌体を調製し、ヘミアセタール水酸基を有する糖12種(D−マンノース、D−ガラクトース、D−リボース、D−キシロース、L−アラビノース、セロビオース、マルトース、マルトトリオース、マルトテトラオース、マルトペンタオース、ラクトース)の2%水溶液1mLに再懸濁し、27度で3日間振とうした。反応液中に含まれる各種アルドン酸の定量結果を表2に示した。
〔前培養〕
試験管(18 mm×200 mm)に、グルコース0.5%、酵母エキス0.5%、ポリペプトン0.5%、硫酸マグネシウム0.1%(pH7.0)を含む培地3mLを分注し、121℃で20分間殺菌した。その試験管に一白金耳のラクトコッカス・ラクティス・サブスピーシーズ・クレモリスKYG−33(FERM P−21207)を植菌し、27℃で1晩振とう培養(220rpm)した。次に上記組成培地を1L分注し121℃で20分間殺菌した3L三角フラスコに上記試験管培養液を植菌し30℃で3日間振とう培養(220rpm)した。
ジャーファーメンターによる培養を行った。グルコース1.5%、ラクトース15%、酵母エキス0.5%、ポリペプトン0.5%、硫酸マグネシウム0.1%、炭酸カルシウム7.5%を含む培地(pH7.0)を20L調製し121℃で20分間殺菌した。これに上記前培養液1Lを植菌し、30℃で深部撹拌培養(300回転、1vvm)をした。培養液中に蓄積したラクトビオン酸濃度を図1に示す。
〔乳酸菌の培養〕
500mLの三角フラスコに、Lactobacilli MRS Broth(デフィコ社)100mLを入れ121℃で20分間殺菌した。そこに一白金耳のラクトコッカス・ラクティス・サブスピーシーズ・クレモリスKYG−33(FERM P−21207)(実施例7)、ラクトコッカス・ラクティス・サブスピーシーズ・クレモリス NBRC100676(比較例1)ラクトコッカス・ラクティス・サブスピーシーズ・ラクティス IAM10070(比較例2)を植菌し27℃で2日間静置培養した。
培養した乳酸菌を脱脂粉乳培地に添加し凍結乾燥により粉末状にすることにより、粉末状の種菌を得た。
〔ラクトビオン酸高含有発酵乳の作製〕
殺菌済みの容器の中に、殺菌済みの新鮮な牛乳100mlを入れた。そこに、調製したヨーグルト種菌を10g添加した。蓋をして閉め、27℃で24時間放置した。このようにして作製された発酵乳をサンプリングしラクトビオン酸含量を測定した。発酵乳中のラクトビオン酸含量を表3に示した。
Claims (3)
- ラクトースに、ラクトバチルス属に属する乳酸菌の菌体を接触させ、ラクトースのヘミアセタール水酸基を酸化しラクトビオン酸を生成させた後に、ラクトビオン酸を分離することを特徴とするラクトビオン酸の製造方法。
- ラクトバチルス属に属する乳酸菌を、ラクトースを含有した培地で培養することにより、培養液中にラクトビオン酸を蓄積させ、該培養液からラクトビオン酸を分離することを特徴とするラクトビオン酸の製造方法。
- ラクトバチルス属に属する乳酸菌が、ラクトバチルス・デルブルッキー・サブスピーシーズ・デルブルッキーIAM12067である、請求項1又は2記載のラクトビオン酸の製造方法。
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