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No.51 (2004/12) >

 
Title :液体培養による紅麹菌のカルボキシペプチダーゼの生産
Title alternative :Production of Carboxypeptidase by Monascus purpureus in Submerged Culture
Authors :刘, 芳
橘, 信二郎
安田, 正昭
Authors alternative :Liu, Fang
Tachibana, Shinjiro
Yasuda, Masaaki
Issue Date :1-Dec-2004
Abstract :カルボキシペプヂダーゼは,タンパク質やペプチドのカルボキシル末端からアミノ酸を順次遊離する酵素である.紅麹菌(Monascus属カビ)は,中国では紅腐乳や紅酒,沖縄ではとうふようの製造に古くから使用されてきた食品微生物である.我々は,これまでにとうふようの食品科学的特性を明らかにしており,本発酵食品のアミノ酸系呈昧成分の生成にカルボキシペプテダーゼが重要な役割を演じていると考えられる.本研究では,とうふよう製造および呈味形成に重要であると考えられる紅麹菌の生産するカルボキシペプチダーゼに着目し,本酵素活性の高い優良菌株のスクリーニング,本酵素生産の培地の組成と培養条件の確立を目的とした.まず,紅麹菌28菌株を用いて,カルボキシペプテダーゼ高生産菌株の1次スクリーニングを行い,本酵素活性が高い6菌株を選択した.次に,2次スクリーニングを行い,本酵素活性及び菌体生育の最も良好なMonascus purpひreus IFO4478をカルボキシペプチダーゼ高産生優良菌株として選択した.続いて,本菌株を用いてカルボキシペプテダーゼの最適生産条件を調べた.その結果,本酵素生産の最適培養組成と培養条件は次のように設定した.(1)培地組成:グルコース1%(w/v),分離大豆タンパク質2%(w/v),MgSO_4・7H_2O 0.5%(w/v),KH_2PO_4 0.5%(w/v),ミネラル溶液A,B and C各0.1%(v/v)及び培地の初pHが6.0, (2)培養条件:28℃,150rum,6日間振とう培養.
In this study, the screening test on twenty eight strains of Monascus for the production of carboxypeptidasse (CPase) in submerged culture was performed, Monascus purpureus IFO 4478 was indicated to be the best producer of CPase. The culture medium compositions and cultivation conditions for the production of CPase by this strain during submerged culture in shake flasks were optimized. Subsequently the process was scaled up to 5 L jar fermentor. The results of carbon source test showed that glucose was the best inducer of CPase among the eleven kinds of carbon sources tested with the addition of 1% (w/v). For the nitrogen source test, soybean protein isolate (SPI) was the best inducer of CPase among the seventeen kinds of nitrogen sources tested with the addition of 2% (w/v). Moreover, the mineral salts solutions with the addition of each 0.1% (v/v) could induce the production of CPase, and when the initial pH of the culture medium was 6.0, the maximal CPase activity was obtained. In addition, the effects of shaking speed, incubation temperature and incubation time on CPase production were also investigated. The results indicated that when M. purpureus IFO 4478 was cultivated in a reciprocal shake incubator with a shaking speed 150 rpm and at 28°C, the maximal CPase activity was obtained after 6 days. Therefore, the optimal culture medium for the CPase production was: glucose 1.0% (w/v), SPI 2.0% (w/v), MgSO4 ・ 7H2O 0. 5% (w/v), KH2PO4 0.5 % (w/v), mineral salt solutions 0.1% (v/v) (pH 6.0), and the optimal cultivation condition was: at 150 rpm and 28°C for 6 days.
Type Local :紀要論文
ISSN :0370-4246
Publisher :琉球大学農学部
URI :http://hdl.handle.net/20.500.12000/3577
Citation :琉球大学農学部学術報告 = The Science Bulletin of the Faculty of Agriculture. University of the Ryukyus no.51 p.131 -138
Appears in Collections:No.51 (2004/12)

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