PRODUCTION OF TETRAPYRROLE COENZYMES BY A METHANOGEN

通过产甲烷菌生产四吡咯辅酶

• 批准号: 62560109
• 负责人: NISHIO Naomichi
• 金额: $ 1.22万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560109/
• 关键词: Methanogen; Methanosarcina barkeri; Corrinoid; F_<430>; cytochrome; 5-Aminolevulinic acid; Fixed-bed reactor; 固定床連続培養; 5-アミノレブリン酸; ALA; 固定床バイオリアクター; 2-オキソグルタン酸; 4、6-ジオキソヘプタン酸; ビタミンB-12; F430; δ-アミノレブリン酸; コバルト欠除コリノイド; テトラピロール化合物; ヘム

A methanogen, Methanosarcina barkeri contains three tetrapyrroles, i.e., Cocontaining corrinoids, Ni-containing factor F_<430> and Fe-containing cytochroms (hemes) which are biosynthesized via 5-aminolevulinic acid(ALA). In this work, the selective formation of these tetrapyrroles and ALA, a potent herbicide were carried out. The results obtained were as follows.1. Effects of Co^<2+>, Ni^<2+>, Fe^<2+> on the selective synthesis of respective tetrapyrroels were examined in batch culture. The increase of Co^<2+> in the medium fixing the low concentrations of Ni^<2+> and F^<2+> in the medium significantly stimulated the extracellular corrinoids syntheses, while F_<430> and heme were kept almost unchanged. Similar stimulative synthesis of extracellular F_<430> or heme was observed as a function of increasing Ni^<2+> or Fe^<2+> in the medium suggesting that the syntheses of these three tetrapyrroles might be regulated with the respective matal. Then, continuous production of tetrapyrroles i … More n a fixed-bed reactor(supporter: porous diatomaceous clay) was carried out a a dilution rate, 10.1 day^<-1>. The effluent discharged from the top of reactor contained the excreted tetrapyrroles whose concentration was depended upon Co^<2+>, Ni^<2+>, Fe^<2+> in the feed medium. The maximum productivies from the roactor(1-1 bases) were 52 m [corrinoids]/day, 24 m[F_<430>]/day and 8 m [heme]/day.2. To accumulate ALA into the culture medium, some ALA analogues were tested as inhibitor of ALA dehydratase. It was found that ALA was accumulated when levulinic acid or 4,6-dioxoheptanoic acid was added, the latted was more effective even at a much lower concentration. It was found that 2-oxoglutarate reacted as a precursor on the formation of ALA indicating that the production ALA increased 83 m from 37 m with the addition of 30 mM of 2-oxoglutarate compared with the control experiment. The continuous productions of ALA from methanol in fixed-bed reactors was tested indicating that the maximum production rate was 400 m/day. Less

Methanosarcina barkeri 是一种产甲烷菌，含有三种四吡咯，即含类咕啉、含镍因子 F_<430> 和含铁细胞色素（血红素），它们是通过 5-氨基乙酰丙酸（ALA）生物合成的。在这项工作中，选择性地形成了这些四吡咯和 ALA（一种有效的除草剂）。得到的结果如下： 1．在分批培养中检查了Co 2+ 、Ni 2+ 、Fe 2+ 对各自四吡咯选择性合成的影响。培养基中Co^<2+>的增加固定了培养基中低浓度的Ni^<2+>和F^<2+>，显着刺激了细胞外咕啉的合成，而F_<430>和血红素几乎保持不变。观察到细胞外F_ 430 或血红素的类似刺激合成作为培养基中Ni 2+ 或Fe 2+ 增加的函数，这表明这三种四吡咯的合成可能受到各自金属的调节。然后，在固定床反应器(载体：多孔硅藻土)中以10.1天^<-1>的稀释率连续生产四吡咯。从反应器顶部排出的流出物含有排出的四吡咯，其浓度取决于进料介质中的Co 2+ 、Ni 2+ 、Fe 2+ 。反应器（1-1个碱基）的最大生产率为52m[类咕啉]/天，24m[F_<430>]/天和8m[血红素]/天。2．为了将 ALA 积累到培养基中，一些 ALA 类似物被测试为 ALA 脱水酶抑制剂。结果发现，当添加乙酰丙酸或4,6-二氧庚酸时，ALA会积累，即使在较低的浓度下，ALA也更有效。结果发现，2-酮戊二酸作为 ALA 形成的前体进行反应，表明与对照实验相比，添加 30 mM 2-酮戊二酸时，ALA 产量从 37 m 增加了 83 m。对固定床反应器中甲醇连续生产ALA进行了测试，最大产率为400 m/天。较少的

项目成果

DONG LIN: "PRODUCTION OF 5-AMINOLEVULINIC ACID BY METHANOGENS" J. FERMENT. BIOENG.

董林：“产甲烷菌生产 5-氨基酮戊酸”J. FERMENT。

林東: Apple.Microbiol.Biotechnol.30. (1989)

东林：Apple.Microbiol.Biotechnol.30（1989）。

Tapan Kumar Mazumder: Appl. Microbiol. Biotechnol.26. 511-516 (1987)

Tapan Kumar Mazumder：应用。

Dong Lin: Appl. Environ. Microbiol.

林东：申请。

NAOMICHI NISHIO: "EXTRACELLULAR PRODUCTION OF CORRINOIDS FROM METHANOL BY METHANOSARCINA BARKERI FUSARO (DSM 804)" VITAMINS(JAPAN). 62. 657-667 (1988)

Naomichi Nishio：“通过 METHANOSARCINA BARKERI FUSARO (DSM 804) 从甲醇中生产类咕啉”维生素（日本）。