JP6841412B2 - オロット酸カリウム塩含有組成物及びその製造方法 - Google Patents
オロット酸カリウム塩含有組成物及びその製造方法 Download PDFInfo
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- JP6841412B2 JP6841412B2 JP2016238125A JP2016238125A JP6841412B2 JP 6841412 B2 JP6841412 B2 JP 6841412B2 JP 2016238125 A JP2016238125 A JP 2016238125A JP 2016238125 A JP2016238125 A JP 2016238125A JP 6841412 B2 JP6841412 B2 JP 6841412B2
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- Prior art keywords
- potassium
- acid
- ollotate
- edible
- salt
- 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.)
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Links
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- 229910052700 potassium Inorganic materials 0.000 title claims description 50
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- 239000000203 mixture Substances 0.000 title claims description 40
- 238000004519 manufacturing process Methods 0.000 title claims description 23
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- PXQPEWDEAKTCGB-UHFFFAOYSA-N orotic acid Chemical compound OC(=O)C1=CC(=O)NC(=O)N1 PXQPEWDEAKTCGB-UHFFFAOYSA-N 0.000 claims description 25
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 15
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- 241000894006 Bacteria Species 0.000 claims description 12
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- DHBUISJCVRMTAZ-UHFFFAOYSA-M potassium;2,4-dioxo-1h-pyrimidine-6-carboxylate Chemical compound [K+].[O-]C(=O)C1=CC(=O)NC(=O)N1 DHBUISJCVRMTAZ-UHFFFAOYSA-M 0.000 claims description 7
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- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 8
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Description
[1]オロット酸産生能を有する納豆菌を、カリウムイオンの存在下で液体培養し、培養後の液体培養物を固液分離して固体部分を採取することを特徴とする食用オロット酸カリウム塩組成物の製造方法。
[2]カリウムイオンが硫酸カリウム又は水酸化カリウムに由来することを特徴とする[1]記載の製造方法。
[3]固液分離が遠心分離又は膜分離による固液分離であることを特徴とする[1]又は[2]記載の製造方法。
[4]固液分離後の固体部分を乾燥することを特徴とする[1]〜[3]のいずれかに記載の製造方法。
[5]食用オロット酸カリウム塩組成物が106CFU/g以上の納豆菌の芽胞又は死菌体を含むことを特徴とする[5]記載の製造方法。
[6]オロット酸カリウム塩とオロット酸産生能を有する納豆菌の芽胞又は死菌体とを含む食用オロット酸カリウム塩組成物。
[7]10質量%以上のオロット酸がカリウム塩を含むことを特徴とする[6]記載の食用オロット酸カリウム塩組成物。
[8]106CFU/g以上の納豆菌の(a)芽胞、(b)死菌体及び/又は不活性化胞子のいずれかを含むことを特徴とする[6]又[7]記載の食用オロット酸カリウム塩組成物。
[9]更に、納豆菌培養成分を含むことを特徴とする[8]記載の食用オロット酸カリウム塩組成物。
[10][6]〜[9]のいずれかに記載の食用オロット酸カリウム塩組成物を含むことを特徴とするサプリメント。
必要に応じて、残存培地成分を、菌体とオロット酸の分離後の洗浄により除ける。また、上記納豆菌を含むオロット酸カリウム塩組成物をオートクレーブ殺菌し、上記納豆菌を死菌体とし、上記納豆菌が形成した芽胞を不活性化胞子にできる。
Bacillus subtilis ver Natto NBRC-3335株をBacillus用培地(Tryptone1%、Yeast Extract0.3%、NaCl0.5%、pH7.0)で30℃にて一夜培養後、同培地に1%(v/v)接種し30℃にて振盪培養した。対数期(OD660≒0.5)に達した菌体を遠心分離(3、000rpm、10分)で集菌し、50mMトリスマレイン酸緩衝液(pH6.5)で洗浄後、100μg/Lのニトロソグアニジン(NTG)を含む同緩衝液中で30分処理を行った。洗浄後に、6アザウラシル1mg/mLを添加した下記表1に示す最小寒天培地に塗布し、37℃にて4日間培養し、6アザウラシルの耐性株を50株取得した。
得られた6アザウラシルの耐性株50株を下記表2に示す種培養培地で培養後、生産培地で4日間、30℃にて培養を行った結果、No.39株が、ウリジン3g/L、ウラシル7g/Lを蓄積した。
No.39株について実施例1と同様のNTG処理を行い、5フルオロウラシル1mg/mL、ウラシル50μg/mLを添加した最小寒天培地に塗布し、37℃にて4日間培養し、得られたコロニーを前記最小寒天培地とウラシル50μg/mLを添加した最小培地にレプリカし、ウラシル要求性を示す株10株を取得した。これらウラシル要求性株10株を実施例1の生産培地で培養を行った結果、すべての菌株でオロット酸の生産性が確認され、6株がオロット酸単独、4株がオロット酸とオロチジンの併産株であった。TG−7株がオロット酸16g/Lと最も高い生産性を示した。この際、保存培養液中にオロット酸の結晶の析出も認められた。
オロット酸産生能を有するTG−7株を、マルトース2%、ペプトン1%、酵母エキス0.3%、肉エキス1%を含む種培養液中で30℃で1晩培養後、1%(v/v)の種培養液を、グルコース8%、コーンスティープリカー3%、尿素2%、ウラシル5mg/L、炭酸カルシウム2%及び硫酸カリウム2%を含む生産培地30mlに摂取し、30℃で4日間培養した。培地のpH調整は水酸化カリウムで行った。培養終了後、液体培養物中に、オロット酸のフリー体1水和物換算で18g/Lのオロット酸カリウム塩の蓄積を確認した。液体培養物を3000rpmで10分間遠心分離した。遠心分離後の上清中に溶解しているオロット酸は0.5g/Lであった。
本発明の製造方法は、食用オロット酸カリウム塩組成物を、エタノールアミン等の溶媒を使用せず、通常の食品用乳酸菌の製造設備を利用して安価、簡便に製造できる。
Claims (9)
- オロット酸産生能を有する納豆菌を、カリウムイオンの存在下で液体培養し、培養後の液体培養物を固液分離して固体部分を採取することを特徴とする食用オロット酸カリウム塩組成物の製造方法。
- カリウムイオンが硫酸カリウム又は水酸化カリウムに由来することを特徴とする請求項1記載の製造方法。
- 固液分離が遠心分離又は膜分離による固液分離であることを特徴とする請求項1又は2記載の製造方法。
- 固液分離後の固体部分を乾燥することを特徴とする請求項1〜3のいずれかに記載の製造方法。
- 食用オロット酸カリウム塩組成物が106CFU/g以上の納豆菌の芽胞又は死菌体を含むことを特徴とする請求項5記載の製造方法。
- オロット酸カリウム塩とオロット酸産生能を有する納豆菌の芽胞又は死菌体とを含み、組成物全体に対して10質量%以上がオロット酸カリウム塩であることを特徴とする食用オロット酸カリウム塩組成物。
- 106CFU/g以上の納豆菌の(a)芽胞、(b)死菌体及び/又は不活性化胞子のいずれかを含むことを特徴とする請求項6記載の食用オロット酸カリウム塩組成物。
- 更に、納豆菌培養成分を含むことを特徴とする請求項7記載の食用オロット酸カリウム塩組成物。
- 請求項6〜8のいずれかに記載の食用オロット酸カリウム塩組成物を含むことを特徴とするサプリメント。
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