JP5818305B2 - 油脂生産能の高い微生物株の選抜方法 - Google Patents
油脂生産能の高い微生物株の選抜方法 Download PDFInfo
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
糖変換率=(油脂生産量/消費糖量)
油脂含有率=(油脂生産量/乾燥菌体重量)
野外から分離したLipomyces starkeyi(受領番号NITE AP-1070)を液体YM培地(1% Glucose, 0.5% peptone, 0.3% Yeast extract, 0.3% malt extract)で25℃、3日間、L字管(5mL)で振とう培養後、集菌して蒸留水で洗浄した。これを蒸留水に懸濁し、撹拌下、生存率約10%のUV処理条件(すなわち、Sanyo社製クリーンベンチ(MCV-710ATS)UV殺菌等下(50cm)で90秒)でUV処理を行なった。処理後の菌体をYM寒天プレート(1% Glucose, 0.5% peptone, 0.3% Yeast extract, 0.3% malt extract, 1.5% agar)上に播種して25℃、約7日間培養し、生育したコロニーの中から多糖の少ない株を取得した。完全に分離されていないコロニーについては、数回寒天培地に継いだ後、明らかに菌体外多糖形成の見られないコロニーを候補株として単離した。約120枚のプレートから52株を取得した。これら菌株をL字管(5mL)にて培養してOD・乾燥菌体重量・油脂生産量・残糖量を測定し、油脂含有率・糖変換率を算出した。油脂生産能力が高い株をさらに選抜するため、候補菌株数を絞り2次スクリーニングを行なった。
実施例1で選抜した変異株及び親株について、キシロースを炭素源とする培地(YX培地;Xylose10%, Yeast extract1%)75 mlで培養し(150 rpm、25℃)、油脂生産能力を変異前の親株と比較した。その結果、変異株8-4で最も油脂生産効率の向上が認められた。同じ条件で培養した親株との油脂生産量の比較を図3に示す。
実施例1で選抜した変異株および親株の培養液の遠心分離後の上清の粘度を、小型振動式粘度計CJV5000 (株式会社 エー・アンド・デイ) を用いて測定したところ、変異株の培養液上清の粘度は著しく低下していた。そこで、それぞれの培養液を遠沈管に入れ、3000rpm(1200G)、20分の条件で遠心分離したところ、親株の培養液では沈殿がほとんど見られないのに対し、変異株はほぼ全ての菌体が沈殿した(図4)。変異株の沈降性が著しく向上していることが確認された。
Claims (4)
- 油脂生産能を有するリポミセス属酵母を培養し、細胞外多糖の生産量が少ない株を選抜することを含む、油脂生産能の高いリポミセス属酵母の候補株の選抜方法。
- リポミセス属酵母を変異処理し、次いで細胞外多糖の生産量が少ない株を選抜する請求項1記載の方法。
- 請求項1又は2記載の方法により、油脂生産能の高いリポミセス属酵母の候補株を選抜し、選抜された株を増殖させることを含む、油脂生産能の高いリポミセス属酵母株の製造方法。
- 請求項1又は2記載の方法を1次スクリーニングとして実施し、さらに2次スクリーニングを実施することを含む、請求項3記載の方法。
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JP5588578B2 (ja) * | 2012-05-17 | 2014-09-10 | 東洋紡株式会社 | 担子菌酵母変異体 |
JP2021058103A (ja) * | 2019-10-03 | 2021-04-15 | 学校法人帝京大学 | 油脂生産酵母の油脂蓄積誘導培地 |
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JP3553128B2 (ja) * | 1994-05-11 | 2004-08-11 | 焼津水産化学工業株式会社 | リポマイセス属に属する菌体外多糖高生産株の育種法 |
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