Findinf of novel decarboxylases which catalyzes carbon dioxide fixation and establishment of the precise conversion techniques of carbon dioxide

催化二氧化碳固定的新型脱羧酶的发现及二氧化碳精准转化技术的建立

• 批准号: 12559003
• 负责人: NAGASAWA Toru
• 金额: $ 7.3万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12559003/
• 关键词: Pytrole-2-carboxylate decarboxylase; Indole-3-carboxylate decarboxylase; Carbon dioxide fixation; CO_2固定; 脱炭酸酵素; 可逆的脱炭酸酵素; 炭酸固定酵素

Due to the abundance of CO2 as the major greenhouse gas and C-C-bond-formlng properties, CO2-flxing enzymes are of high interest. Recently, it is revealed that a certain group of decarboxylases can catalyze carboxylation by their reverse reaction. We have found that pyrrole-2-carboxylase efficiently catalyzes the carboxylation of pyrrole to pyrrole-2-carboxylic acid. In addition, recently we also found that indole-3carboxylate decarboxylase catalyzes the carboxylation of indole to indole-3-carboxylate. Pyrrole-2-carboxylate decarboxylase from BacMus megaterium PRY2910 attains a balanced reaction quilibrium with an equilibrium constant of 0.3-0.4 M. Therefore, saturating amounts of KHCO3 ( 3 M ) were used leading to a shift of the reaction equilibrium towards pyrrole-2-carboxylic acid. The increased pressure supported the reverse reaction productivity 2.5-fold compared to atmospheric pressure. In a batch reactin, 300 mM pyrrole'were converted to 230 mM (25.5 g/1) pyrrole-2-carboxylate. The productivity was increased to 325 mM ( 36. 1 g/1 ) by feeding 150 mM pyrrole after 3 h to a reaction with initially 250 mM pyrrole. The bioconversion yield was limited to 77-81 % due to the reaction equilibrium. Pyrrole-2-carboxylate decarboxylase and indole-3-carboxylate decarboxlase were purified and characterized. We could find some similarity. Their gene analyzes were also carrried out.

由于CO2作为主要温室气体的丰度和C-C键形成特性，CO2-flxing酶受到高度关注。最近，研究发现某些脱羧酶可以通过其逆反应催化羧化反应。我们已经发现吡咯-2-羧化酶有效地催化吡咯羧化为吡咯-2-羧酸。另外，最近我们还发现吲哚-3-羧酸脱羧酶催化吲哚羧化为吲哚-3-羧酸。来自BacMus megaterium PRY 2910的吡咯-2-羧酸脱羧酶达到平衡反应平衡，平衡常数为0.3-0.4 M。因此，使用饱和量的KHCO 3（3 M），导致反应平衡向吡咯-2-羧酸移动。与大气压相比，增加的压力支持2.5倍的逆反应产率。在分批反应中，将300 mM吡咯转化为230 mM（25.5 g/l）吡咯-2-羧酸酯。生产率提高到325 mM（36. 1g/l），在3小时后将150 mM吡咯加入到最初为250 mM吡咯的反应中。由于反应平衡，生物转化产率被限制在77- 81%。吡咯-2-羧酸脱羧酶和吲哚-3-羧酸脱羧酶的纯化和表征。我们可以找到一些相似之处。他们的基因分析也进行了。

项目成果

M.Wieser et al.: "Carbon Dioxide Fixation by Reversible Pyrrole-2-carboxylate Decarboxylase and its Application"J.Mol.Catal.B : Enzyatic.. 11(4-6). 179-184 (2001)

M.Wieser 等人：“可逆吡咯-2-羧酸脱羧酶的二氧化碳固定及其应用”J.Mol.Catal.B：酶学.. 11(4-6)。

Wieser,M. et al.: "Carbon Dioxide Fixation by Reversible Pyrrole-2-carboxylate Decarboxylase and its Application"Journal of Molecular Catalysis B:Enzymatic. 11 ( 4-6 ). 179-184 (2001)

维塞尔，M.

Matsuda,T. et al.: "Conversion of pyrrole-2-carboxylate by cells of Bacillus megaterium in supercritical C02"Chemical Communications. 2194-2195 (2001)

松田，T.

T.Yoshida & T.Nagasawa: "Enzymatic Functionalization of Aromatic N-Heterocycles : Hydroxylation and Carboxylation"J.Biosci.Bioeng.. 89(2). 111-118 (2000)

吉田T

Yoshida,T. et al.: "Enzymatic Functional ization of Aromatic N-Heterocycles: Hydroxylation and Carboxylation"Jouranl of Bioscience and Bioengineering. 89 ( 2 ). 111-118 (2000)

吉田，T.