JP2021126061A - 新規乳酸菌用培地 - Google Patents
新規乳酸菌用培地 Download PDFInfo
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- JP2021126061A JP2021126061A JP2020021764A JP2020021764A JP2021126061A JP 2021126061 A JP2021126061 A JP 2021126061A JP 2020021764 A JP2020021764 A JP 2020021764A JP 2020021764 A JP2020021764 A JP 2020021764A JP 2021126061 A JP2021126061 A JP 2021126061A
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- lactic acid
- medium
- acid bacteria
- lactobacillus
- bacteria according
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- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
酵母エキス 0.1〜10%、好ましくは0.5〜5%
発酵大麦エキス 0.1〜15%、好ましくは1〜10%
乳化剤 0.01〜1%、好ましくは0.05〜0.5%
糖類 0.01〜10%、好ましくは0.05〜5%
水 残量
酵母エキス 2%
発酵大麦エキス 5%
モノオレイン酸デカグリセリン 0.1%
グルコース 2%
水 残量
培地の調製および乳酸菌の培養:
下記表1に記載の成分を水に混合、溶解させ、次いで、pHを水酸化ナトリウムで6.7に調整した後、殺菌して、培地を調製した(以下、「MBG培地」という)。この培地に凍結保存されたラクトバチルス・カゼイYIT9029(以下、「LcS」という)を1%(生菌数としては1.0×105〜1.0×107(cfu/ml)程度)接種し、37℃で24時間静置培養した。培養後の生菌数(cfu/ml)を測定した結果を表1に示す。増殖性の良否の基準として、生菌数5.0×108(cfu/ml)以上とした。
グリセロールストックの作製及び評価:
(1)菌体または培養液のグリセロールストックの調製
MBG培地で前培養したLcSの培養液を、MBG培地に0.1%接種後、37℃で24時間静置培養した(本培養)。次いで、培養液を遠心し、遠心上清を除去して菌体を得て、滅菌処理したグリセロールに懸濁し、終濃度12.5%とした(菌体サンプル)。
また、氷冷した本培養液を採取し、滅菌処理をしたグリセロールと混合し、終濃度12.5%とした(培養液サンプル)。
これらをクライオチューブに1mlずつ分注し、ドライアイスエタノールバス内で急速冷凍した。出来上がったグリセロールストックは、−70℃で保存した。
(1)で得た2種類のグリセロールストックについて、生菌数測定を行った。生菌数はグリセロールストックを生理食塩水(0.85%NaCl溶液)で希釈後、MRS寒天培地(BD社、DifcoTM Lactobacilli MRS agar)にスパイラルプレーターEDDY JET2(IUL Instruments)を用いて塗布し、37℃、2日間以上好気培養した。得られたコロニー数をコロニーカウンター(Qcount)で測定した。なお、本培養後及び凍結前の生菌数についても同様にして測定した。また以下の式により生残率(%)を求めた。
(生残率)
生残率(%)=(凍結後の生菌数/凍結前の生菌数)×100
Lcs以外の菌種でのグリセロールストックの作製及び評価:
(1)菌体のグリセロールストックの調製
実施例2と同様の方法で、ラクトバチルス・ガセリ(Lactobacillus gasseri)、ラクトバチルス・ブレビス(Lactobacillus brevis)、ラクトバチルス・プランタラム(Lactobacillus plantarum)の菌体のグリセロールストックを調整した。
実施例2と同様の方法で、凍結前、凍結後における菌体のグリセロールストックの生菌数を調べ、生残率を求めた。なお、ラクトバチルス・ガセリのみ嫌気条件下で培養した。
グリセロールストックのスターターとしての培養性状評価:
脱脂粉乳(雪印株式会社)を用いて10%脱脂粉乳培地を調製した。また調整豆乳(株式会社ヤクルト本社)を3倍希釈し、ブドウ糖(扶桑薬品工業株式会社)を1%添加して豆乳培地を調製した。実施例2で調製した菌体及び培養液のグリセロールストックを室温で溶解し、脱脂粉乳培地または豆乳培地に0.1%接種後、攪拌し、37℃で培養した。培養は2連で行った(培養(1)及び(2))。培養時間は、脱脂粉乳培地では48時間、豆乳培地では24時間とした。培養後及び4℃で所定日数保存後、生菌数、酸度及びpHを以下のようにして測定した。脱脂粉乳培地での結果を表3(培養後)及び4(4℃14日保存後)、豆乳培地での結果を表5(培養後)及び6(10℃10日保存後)に示す。
(生菌数測定)
培養物を良く攪拌し、生理食塩水で希釈後、MRS寒天培地(BD社製)にスパイラルプレーターEDDY JET2(IUL Instruments)で塗布した。これを好気下で37℃、2日間以上培養した。得られたコロニーをコロニーカウンター(Qcount)で測定し、1ml当りのコロニー数(cfu)を求めた。増殖性の良否の基準は、脱脂粉乳培地では1.0×109(cfu/ml)以上、豆乳培地では1.0×109(cfu/ml)以上とした。
(酸度測定)
各サンプルを良く攪拌後、6g採取しRO水(27.3ml)を添加して酸度測定用サンプル(固形分濃度18.02%)とした。酸度滴定装置(COM−1700平沼産業)にて、pH8.5に達するまでに用いた0.1N NaOHの量(ml)を測定した。得られた値をサンプル9g当たりに換算し酸度(ml/9g)とした。増殖性の良否の基準は脱脂粉乳培地では8.5(ml/9g)以上、豆乳培地では8.5(ml/9g)以上とした。
(pH測定)
培養物をよく攪拌後、ポータブルpHメーター LaquaAct(HORIBA社製)を用いて測定した。
Claims (15)
- 酵母エキス及び発酵大麦エキスを含有する乳酸菌用培地。
- 酵母エキスがビール酵母エキスである請求項1記載の乳酸菌用培地。
- 発酵大麦エキスが、大麦焼酎蒸留残液である請求項1または2記載の乳酸菌用培地。
- 更に、乳化剤を含有する請求項1〜3のいずれかの項記載の乳酸菌用培地。
- 乳化剤が、モノオレイン酸デカグリセリンである請求項4記載の乳酸菌用培地。
- 更に、糖類を含有する請求項1〜5のいずれかの項記載の乳酸菌用培地。
- 糖類が、グルコースである請求項6項記載の乳酸菌用培地。
- 乳成分を含まないものである請求項1〜7のいずれかの項記載の乳酸菌用培地。
- 大豆成分を含まないものである請求項1〜8のいずれかの項記載の乳酸菌用培地。
- 乳酸菌の凍結保存用である請求項1〜9のいずれかの項記載の乳酸菌用培地。
- ラクトバチルス属乳酸菌用である請求項1〜10のいずれかの項記載の乳酸菌用培地。
- ラクトバチルス属乳酸菌が、ラクトバチルス・カゼイ、ラクトバチルス・ガセリ、ラクトバチルス・ブレビスまたはラクトバチルス・プランタラムである請求項11記載の乳酸菌用培地。
- 乳酸菌を、請求項1〜12のいずれかの項記載の乳酸菌用培地で培養することを特徴とする乳酸菌の培養方法。
- 乳酸菌を、請求項1〜12のいずれかの項記載の乳酸菌用培地で培養して得られる乳酸菌培養物。
- 請求項14記載の乳酸菌培養物を含有する飲食品。
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JPH10327852A (ja) * | 1997-05-28 | 1998-12-15 | Sanwa Shiyurui Kk | 焼酎蒸留残液から微生物増殖促進成分を分離濃縮する方法 |
JP2009209132A (ja) * | 2008-02-05 | 2009-09-17 | Kitii Corp | 新菌株ラクトバチルス・クリスパタスkt−11、kt−23、およびkt−25を用いた抗アレルギー用組成物 |
JP2010004787A (ja) * | 2008-06-26 | 2010-01-14 | Kaneka Corp | ラクトバチルス・ブレビス菌の培養方法 |
WO2010113816A1 (ja) * | 2009-03-30 | 2010-10-07 | 明治乳業株式会社 | 乳酸菌の培養法および発酵乳の製造法 |
JP2015167474A (ja) * | 2014-03-04 | 2015-09-28 | 学校法人日本大学 | ラクトバチルス属乳酸菌培養用食品グレード培地、ラクトバチルス属乳酸菌の培養方法及びラクトバチルス属乳酸菌培養物の製造方法 |
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2020
- 2020-02-12 JP JP2020021764A patent/JP2021126061A/ja active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
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JPH10327852A (ja) * | 1997-05-28 | 1998-12-15 | Sanwa Shiyurui Kk | 焼酎蒸留残液から微生物増殖促進成分を分離濃縮する方法 |
JP2009209132A (ja) * | 2008-02-05 | 2009-09-17 | Kitii Corp | 新菌株ラクトバチルス・クリスパタスkt−11、kt−23、およびkt−25を用いた抗アレルギー用組成物 |
JP2010004787A (ja) * | 2008-06-26 | 2010-01-14 | Kaneka Corp | ラクトバチルス・ブレビス菌の培養方法 |
WO2010113816A1 (ja) * | 2009-03-30 | 2010-10-07 | 明治乳業株式会社 | 乳酸菌の培養法および発酵乳の製造法 |
JP2015167474A (ja) * | 2014-03-04 | 2015-09-28 | 学校法人日本大学 | ラクトバチルス属乳酸菌培養用食品グレード培地、ラクトバチルス属乳酸菌の培養方法及びラクトバチルス属乳酸菌培養物の製造方法 |
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