Halogenating Enzymes Produced by Marine Algae and Microbes---Structures and Functions

海藻和微生物产生的卤化酶——结构和功能

• 批准号: 04044118
• 负责人: IZUMI Yoshikazu
• 金额: $ 8.13万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04044118/
• 关键词: Halogenating enzyme; Vanadium enzyme; Primary structure of an enzyme; Marinebiotechnology; Halogenated organic compound; Macro-algae; Coralline algae; 比較生化学的研究

Many physiologically active, halogenated compounds have been found in marine algae. Haloperoxidases are known to be involved in the biosynthesis of these halometabolites. Our two groups found that some algae and microorganisms produced a large amount of haloperoxidase at the same time and independently. However, concerning algae producing the enzyme, Corallina pilulifera could be available only around Japan, and Ascophyllum nodosum could be only in the North Sea. This circumstance have led to this joint research.Bromoperoxidase was purified from the crude extract of C. pilulifera to complete homogeneity. The content of the enzyme in the alga was as much as 3% the total protein of the crude extract. The enzyme had a molecular mass of ca. 790 kDa and was composed of twelve subunits of identical molecular mass of 64 kDa. The enzyme as isolated contained 4 mol of vanadium per mol of bromoperoxidase (790 kDa). Also the enzyme was found to contain Fe^<3+> and Mg^<2+> abundantly. Ferric ion m … More arkedly shortened the time required for the full activation of the apoenzyme by vanadate. We also studied the application of the enzyme to the production of various halogenated compounds.By the researchers at Amsterdam University the bromoperoxidase from A.nodosum was purified to homogeneity. The enzyme had a molecular mass of 67 kDa, and contained 1 mol of vanadium per mol of bromoperoxidase. It has been revealed that the enzyme was stable even in 40% acetone, methanol, and n-propanol for a month at a room temperature.The chloroperoxidase from the fungi, Curvularia inaequalis, was also purified to homogeneity by the researchers in Amsterdam University. It has shown that histidine residue was involved in the binding site of vanadium for the enzyme.When these enzymes were digested by proteases and compared one another by the electrophoresis, it seemed that they had some similarity. Concerning the enzyme of C. pilulifera, 253 amino acid sequences have been identified. Furthermore, the basic conditions were examined to crystallize the enzyme of C. pilulifera in collaboration with the research group at Imperial College in U.K. Less

在海藻中发现了许多具有生理活性的卤代化合物。已知卤代过氧化物酶参与这些卤代代谢物的生物合成。我们的两个小组发现，一些藻类和微生物同时并独立地产生大量的卤代过氧化物酶。然而，关于产生这种酶的藻类，Cortophyllum pilulifera只能在日本附近找到，而Ascophyllum nodosum只能在北海找到。本研究从C. pillulifera完全同质。酶的含量高达粗提物总蛋白的3%。该酶的分子量约为100。790 kDa，由12个相同的分子量为64 kDa的亚基组成。所分离的酶含有4摩尔钒/摩尔溴过氧化物酶（790 kDa）。同时发现该酶含有丰富的Fe^<3+>和Mg^<2+>.铁离子M ...更多信息 大大缩短了钒酸盐完全激活脱辅基酶所需的时间。我们还研究了该酶在生产各种卤代化合物中的应用。阿姆斯特丹大学的研究人员将来自A.nodosum的溴过氧化物酶纯化至均一。该酶的分子量为67 kDa，每摩尔溴过氧化物酶含有1摩尔钒。研究表明，该酶在室温下，在40%丙酮、甲醇和正丙醇中稳定1个月。阿姆斯特丹大学的研究人员还将来自真菌Curvularia inaequalis的氯过氧化物酶纯化至均一。结果表明，组氨酸残基参与了该酶与钒的结合位点，经蛋白酶酶解和电泳比较，它们似乎具有一定的相似性。对C. pillulifera中，已经鉴定了253个氨基酸序列。此外，还对C. pilulifera与英国帝国理工学院的研究小组合作。少

项目成果

Toyokazu Yoshida: "Enzymatic assay for L-serine and glyoxylate involving the enzymes in the serine pathway of a methylotroph" Analytical Biochemistry. 208. 296-299 (1993)

Toyokazu Yoshida：“L-丝氨酸和乙醛酸的酶法测定涉及甲基营养菌丝氨酸途径中的酶”分析生物化学。

和泉好計: "バナジウムを補欠分子族にもつ酵素ブロモペルオキシダーゼ" Biomedical Research on Trace Elements. 3. 49-50 (1992)

Yoshikei Izumi：“以钒为辅基的溴过氧化物酶”，微量元素生物医学研究，3. 49-50 (1992)。

Yoshikazu Izumi: "L-Serine production by a methylotroph and its related enzymes" Applied Microbiology Biotechnology. 39. 427-432 (1993)

Yoshikazu Izumi：“甲基营养菌及其相关酶生产 L-丝氨酸”应用微生物学生物技术。

和泉 好計: "バナジウむを補欠分子族にもつ酵素ブロモペルオキシダーゼ" Biomedical Research on Trace Elements. 3. 49-50 (1992)

Yoshikazu Izumi：“以钒为辅基的溴过氧化物酶”微量元素生物医学研究3. 49-50 (1992)。

Johannes W.P.M.van Schijndel: "The vanadium chloroperoxidase from the fungus,Curvularia inaequalis-Evidence for the involvement of a histidine residue in the binding of vanadate" FEBS Letter. 336. 239-242 (1993)

Johannes W.P.M.van Schijndel：“来自真菌 Curvularia inaequalis 的钒氯过氧化物酶 - 组氨酸残基参与钒酸盐结合的证据”FEBS Letter。