• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

通过固定化提高芳基丙二酸脱羧酶的过程稳定性用于生物催化合成洛芬

Improvement of the Process Stability of Arylmalonate Decarboxylase by Immobilization for Biocatalytic Profen Synthesis.

作者信息

Aßmann Miriam, Mügge Carolin, Gaßmeyer Sarah Katharina, Enoki Junichi, Hilterhaus Lutz, Kourist Robert, Liese Andreas, Kara Selin

机构信息

Institute of Technical Biocatalysis, Hamburg University of Technology, Hamburg, Germany.

Junior Research Group for Microbial Biotechnology, Ruhr-University Bochum, Bochum, Germany.

出版信息

Front Microbiol. 2017 Mar 16;8:448. doi: 10.3389/fmicb.2017.00448. eCollection 2017.

DOI:10.3389/fmicb.2017.00448
PMID:28360905
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5352704/
Abstract

The enzyme arylmalonate decarboxylase (AMDase) enables the selective synthesis of enantiopure ()-arylpropinates in a simple single-step decarboxylation of dicarboxylic acid precursors. However, the poor enzyme stability with a half-life time of about 1.2 h under process conditions is a serious limitation of the productivity, which results in a need for high catalyst loads. By immobilization on an amino C2 acrylate carrier the operational stability of the ()-selective AMDase variant G74C/M159L/C188G/V43I/A125P/V156L was increased to a half-life of about 8.6 days, which represents a 158-fold improvement. Further optimization was achieved by simple immobilization of the cell lysate to eliminate the cost- and time intensive enzyme purification step.

摘要

芳基丙二酸脱羧酶（AMDase）能够通过二羧酸前体的简单单步脱羧反应选择性合成对映体纯的（）-芳基丙酸酯。然而，在工艺条件下该酶稳定性较差，半衰期约为1.2小时，这严重限制了生产率，导致需要高催化剂负载量。通过固定在氨基C2丙烯酸酯载体上，（）-选择性AMDase变体G74C/M159L/C188G/V43I/A125P/V156L的操作稳定性提高到约8.6天的半衰期，这代表了158倍的改进。通过简单固定细胞裂解物以消除成本高且耗时的酶纯化步骤实现了进一步优化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/d26fb30e9a1e/fmicb-08-00448-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/3be9becf3f2f/fmicb-08-00448-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/30f3f273a2c8/fmicb-08-00448-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/37afef543d77/fmicb-08-00448-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/dc3f4332c815/fmicb-08-00448-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/7f6a6273b67f/fmicb-08-00448-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/1b29d288a7cc/fmicb-08-00448-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/d26fb30e9a1e/fmicb-08-00448-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/3be9becf3f2f/fmicb-08-00448-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/30f3f273a2c8/fmicb-08-00448-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/37afef543d77/fmicb-08-00448-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/dc3f4332c815/fmicb-08-00448-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/7f6a6273b67f/fmicb-08-00448-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/1b29d288a7cc/fmicb-08-00448-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/5352704/d26fb30e9a1e/fmicb-08-00448-g007.jpg

相似文献

1
Improvement of the Process Stability of Arylmalonate Decarboxylase by Immobilization for Biocatalytic Profen Synthesis.通过固定化提高芳基丙二酸脱羧酶的过程稳定性用于生物催化合成洛芬
Front Microbiol. 2017 Mar 16;8:448. doi: 10.3389/fmicb.2017.00448. eCollection 2017.
2
Engineered hydrophobic pocket of (S)-selective arylmalonate decarboxylase variant by simultaneous saturation mutagenesis to improve catalytic performance.通过同时饱和诱变构建（S）-选择性芳基丙二酸脱羧酶变体的疏水性口袋以提高催化性能。
Biosci Biotechnol Biochem. 2015;79(12):1965-71. doi: 10.1080/09168451.2015.1060844. Epub 2015 Jun 26.
3
Chemoenzymatic Cascade Synthesis of Optically Pure Alkanoic Acids by Using Engineered Arylmalonate Decarboxylase Variants.通过使用工程化的芳基丙二酸酶变体进行手性醇的酶促级联合成来合成光学纯的烷酸。
Chemistry. 2019 Apr 1;25(19):5071-5076. doi: 10.1002/chem.201806339. Epub 2019 Mar 12.
4
Arylmalonate decarboxylase-a highly selective bacterial biocatalyst with unknown function.芳基丙二酸盐脱羧酶——一种具有未知功能的高选择性细菌生物催化剂。
Appl Microbiol Biotechnol. 2016 Oct;100(20):8621-31. doi: 10.1007/s00253-016-7778-z. Epub 2016 Aug 26.
5
Dramatically improved catalytic activity of an artificial (S)-selective arylmalonate decarboxylase by structure-guided directed evolution.通过结构指导的定向进化，极大地提高了人工（S）选择性芳基丙二酸脱羧酶的催化活性。
Chem Commun (Camb). 2011 Jul 14;47(26):7503-5. doi: 10.1039/c1cc11953b. Epub 2011 May 11.
6
Purification and characterization of arylmalonate decarboxylase from Achromobacter sp. KU1311.无色杆菌属菌株KU1311中芳基丙二酸脱羧酶的纯化与特性分析
J Biosci Bioeng. 2007 Oct;104(4):263-7. doi: 10.1263/jbb.104.263.
7
Inversion of enantioselectivity of asymmetric biocatalytic decarboxylation by site-directed mutagenesis based on the reaction mechanism.基于反应机理通过定点诱变实现不对称生物催化脱羧对映选择性的反转。
Chem Commun (Camb). 2005 Feb 21(7):877-9. doi: 10.1039/b416398b. Epub 2005 Jan 10.
8
Active-site mobility revealed by the crystal structure of arylmalonate decarboxylase from Bordetella bronchiseptica.支气管败血波氏杆菌芳基丙二酸脱羧酶晶体结构揭示的活性位点流动性
J Mol Biol. 2008 Mar 21;377(2):386-94. doi: 10.1016/j.jmb.2007.12.069. Epub 2008 Jan 5.
9
Engineering the promiscuous racemase activity of an arylmalonate decarboxylase.工程化芳基丙二酸盐脱羧酶的混杂外消旋酶活性。
Chemistry. 2011 Jan 10;17(2):557-63. doi: 10.1002/chem.201001924. Epub 2010 Nov 9.
10
Erratum: Preparation of Poly(pentafluorophenyl acrylate) Functionalized SiO2 Beads for Protein Purification.勘误：用于蛋白质纯化的聚（丙烯酸五氟苯酯）功能化二氧化硅微珠的制备
J Vis Exp. 2019 Apr 30(146). doi: 10.3791/6328.

引用本文的文献

1
Ground-State Destabilization by Active-Site Hydrophobicity Controls the Selectivity of a Cofactor-Free Decarboxylase.活性位点疏水性对基态失稳的控制作用决定了一种无辅因子脱羧酶的选择性。
J Am Chem Soc. 2020 Nov 25;142(47):20216-20231. doi: 10.1021/jacs.0c10701. Epub 2020 Nov 12.
2
ign of Sustainable Chemoenzymatic Organic Transformations in for the Synthesis of 1,2-Disubstituted Aromatic Olefins.用于合成1,2 - 二取代芳族烯烃的可持续化学酶促有机转化的迹象。
Front Chem. 2020 Mar 6;8:139. doi: 10.3389/fchem.2020.00139. eCollection 2020.
3
Highly efficient production of chiral amines in batch and continuous flow by immobilized ω-transaminases on controlled porosity glass metal-ion affinity carrier.

本文引用的文献

1
Enzyme immobilization: an overview on techniques and support materials.酶固定化：技术与载体材料综述
3 Biotech. 2013 Feb;3(1):1-9. doi: 10.1007/s13205-012-0071-7. Epub 2012 Jun 6.
2
An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes.酶固定化技术及固定化酶表面分析技术综述。
Biotechnol Biotechnol Equip. 2015 Mar 4;29(2):205-220. doi: 10.1080/13102818.2015.1008192. Epub 2015 Feb 17.
3
A general protein purification and immobilization method on controlled porosity glass: biocatalytic applications.
在控制孔玻璃金属离子亲和载体上固定化ω-转氨酶，在分批和连续流中高效生产手性胺。
J Biotechnol. 2019 Feb 10;291:52-60. doi: 10.1016/j.jbiotec.2018.12.001. Epub 2018 Dec 11.
一种基于可控孔隙率玻璃的通用蛋白质纯化与固定化方法：生物催化应用
Chem Commun (Camb). 2014 Aug 21;50(65):9134-7. doi: 10.1039/c4cc02605e.
4
Site directed immobilization of glucose-6-phosphate dehydrogenase via thiol-disulfide interchange: influence on catalytic activity of cysteines introduced at different positions.通过巯基-二硫键交换实现葡萄糖-6-磷酸脱氢酶的定向固定化：对不同位置引入的半胱氨酸对催化活性的影响。
J Biotechnol. 2013 Aug 10;167(1):1-7. doi: 10.1016/j.jbiotec.2013.06.002. Epub 2013 Jun 14.
5
Chiral drugs: an overview.手性药物：概述
Int J Biomed Sci. 2006 Jun;2(2):85-100.
6
Evaluation of immobilized enzymes for industrial applications.固定化酶在工业中的应用评价。
Chem Soc Rev. 2013 Aug 7;42(15):6236-49. doi: 10.1039/c3cs35511j.
7
Engineering the third wave of biocatalysis.工程化第三波生物催化。
Nature. 2012 May 9;485(7397):185-94. doi: 10.1038/nature11117.
8
Engineering the promiscuous racemase activity of an arylmalonate decarboxylase.工程化芳基丙二酸盐脱羧酶的混杂外消旋酶活性。
Chemistry. 2011 Jan 10;17(2):557-63. doi: 10.1002/chem.201001924. Epub 2010 Nov 9.
9
Site-selective immobilisation of functional enzymes on to polystyrene nanoparticles.将功能酶选择性固定在聚苯乙烯纳米颗粒上。
Org Biomol Chem. 2010 Feb 21;8(4):782-7. doi: 10.1039/b916773k. Epub 2009 Dec 23.
10
Structure-guided directed evolution of alkenyl and arylmalonate decarboxylases.基于结构导向的烯基和芳基丙二酸脱羧酶定向进化
Angew Chem Int Ed Engl. 2009;48(41):7691-4. doi: 10.1002/anie.200904112.