Selective hydroxylation of L-proline and derivatives with proline hydroxylases - An approach to reactivity und mechanism

L-脯氨酸及其衍生物与脯氨酸羟化酶的选择性羟基化 - 反应性和机制的方法

• 批准号: 246035025
• 负责人: Dr. Wolfgang Hüttel
• 金额: --
• 依托单位: Arbeitsgruppe Physikalische Chemie/ Biophysikalische Chemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/246035025?language=en
• 关键词: Selective; hydroxylation; proline; derivatives; hydroxylases

Prolinehydroxylases, which belong to the enzyme class of the alpha-ketoglutarate dependent dioxygenases, are an ideal model for investigations in enzyme-catalyzed CH-activation reactions. First, enzymes with different regio- and stereoselectivities are available, so that factors directing regio- and stereoselectivities can be determined more easily. Second, hydroxyprolines and their derivatives are valuable chiral building blocks for the synthesis of pharmaceuticals and other active agents, which are only difficult to access by means of classical organic chemistry. The prolinehydroxylases known so far shall be characterized thoroughly by applying different methodological approaches, which, in combination, allow a deeper understanding of the catalytic activity and the causes of different selectivities. To investigate substrate specificity and catalytic selectivity, the active center of the prolinhydroxylases will be investigated by electron paramagnetic resonance and other spectroscopic methods. In parallel, the catalytic properties of the enzymes will be characterized using an assay with natural and non-natural substrates. By means of mutagenesis variants will be generated and included into the investigation. The aim is to understand the factors determining selectivity and substrate specificity of the prolinehydroxylases to make it possible to modify the catalytic properties of the enzymes in rational approaches. In this way new and biotechnologically usable CH-activating biocatalysts shall be generated.

脯氨酸羟化酶属于α-酮戊二酸依赖性双加氧酶，是研究酶催化CH-活化反应的理想模型。首先，具有不同区域和立体选择性的酶是可用的，使得可以更容易地确定指导区域和立体选择性的因素。其次，羟基脯氨酸及其衍生物是用于合成药物和其他活性剂的有价值的手性结构单元，其仅难以通过经典有机化学获得。迄今为止已知的脯氨酸羟化酶应通过应用不同的方法学途径来彻底表征，这些方法学途径相结合，可以更深入地了解催化活性和不同选择性的原因。为了研究底物特异性和催化选择性，将通过电子顺磁共振和其他光谱方法研究脯氨酸羟化酶的活性中心。同时，将使用天然和非天然底物的测定来表征酶的催化性质。将通过诱变产生变体并纳入研究中。目的是了解的因素，决定选择性和底物特异性的脯氨酸羟化酶，使其能够修改的催化性能的酶在合理的方法。以这种方式，将产生新的和生物技术上可用的CH活化生物催化剂。

项目成果

Cryptic Production of trans-3-Hydroxyproline in Echinocandin B Biosynthesis

• DOI: 10.1128/aem.02370-17
• 发表时间: 2018-01
• 期刊: Applied and Environmental Microbiology
• 影响因子: 4.4
• 作者: Johanna Mattay;S. Houwaart;Wolfgang Hüttel

The Clinically Used Iron Chelator Deferasirox is an Inhibitor of Epigenetic JumonjiC Domain-Containing Histone Demethylases.

• DOI: 10.1021/acschembio.9b00289
• 发表时间: 2019-02
• 期刊: ACS chemical biology
• 影响因子: 4
• 作者: Martin Roatsch;I. Hoffmann;Martine I. Abboud;R. L. Hancock;Hanna Tarhonskaya;Kuo-Feng Hsu;Sarah E Wilkins;Tzu-Lan Yeh;K. Lippl;Kerstin Serrer;I. Moneke;Theresa D. Ahrens;D. Robaa;Sandra Wenzler;Nicolas P. F. Barthes;Henriette Franz;W. Sippl;S. Lassmann;S. Diederichs;E. Schleicher;C. Schofield;A. Kawamura;R. Schüle;M. Jung

The toolbox of Auricularia auricula-judae dye-decolorizing peroxidase - Identification of three new potential substrate-interaction sites.

黑木耳染料脱色过氧化物酶的工具箱 - 三个新的潜在底物相互作用位点的鉴定

• DOI: 10.1016/j.abb.2014.12.016
• 发表时间: 2015
• 期刊: Archives of biochemistry and biophysics
• 影响因子: 3.9
• 作者: Strittmatter;Serrer;Ullrich;Hofrichter;Piontek;Schleicher;Plattner
• 通讯作者: Plattner