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首页> 外文期刊>Environmental Science & Technology >High-Throughput Analysis of Enzymatic Hydrolysis of Biodegradable Polyesters by Monitoring Cohydrolysis of a Polyester-Embedded Fluorogenic Probe
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High-Throughput Analysis of Enzymatic Hydrolysis of Biodegradable Polyesters by Monitoring Cohydrolysis of a Polyester-Embedded Fluorogenic Probe

机译：高通量分析可生物降解聚酯的酶水解，通过监测嵌入聚酯的荧光探针的共水解

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摘要

Biodegradable polyesters have the potential to replace nondegradable, persistent polymers in numerous applications and thereby alleviate plastic accumulation in the environment. Herein, we present an analytical approach to study enzymatic hydrolysis of polyesters, the key step in their overall biodegradation process. The approach is based on embedding fluorescein dilaurate (FDL), a fluorogenic ester substrate, into the polyester matrix and on monitoring the enzymatic cohydrolysis of FDL to fluorescein during enzymatic hydrolysis of the polyester. We validated the approach against established techniques using FDL-containing poly(butylene adipate) films and Fusarium solani cutinase (FsC). Implemented on a microplate reader platform, the FDL-based approach enabled sensitive and high-throughput analysis of the enzymatic hydrolysis of eight aliphatic polyesters by two fungal esterases (FsC and Rhizopus oryzae lipase) at different temperatures. While hydrolysis rates for both enzymes increased with decreasing differences between the polyester melting temperatures and the experimental temperatures, this trend was more pronounced for the lipase than the cutinase. These trends in rates could be ascribed to a combination of temperature-dependent polyester chain flexibility and accessibility of the enzyme active site. The work highlights the capability of the FDL-based approach to be utilized in both screening and mechanistic studies of enzymatic polyester hydrolysis.
机译：可生物降解的聚酯有潜力在许多应用中替代不可降解的持久性聚合物，从而减轻环境中的塑料积累。在这里，我们提出一种分析方法来研究聚酯的酶促水解，这是其整体生物降解过程的关键步骤。该方法是基于将荧光素二月桂酸酯荧光素（FDL）嵌入聚酯基体中，并基于在聚酯酶水解过程中监测FDL酶促共水解为荧光素。我们使用含FDL的聚己二酸丁二酯薄膜和茄形镰刀菌角质酶（FsC）验证了该方法是否与既定技术相对立。在微孔板读取器平台上实施的基于FDL的方法能够在不同温度下通过两种真菌酯酶（FsC和米根霉脂肪酶）对八种脂肪族聚酯的酶解进行灵敏且高通量的分析。尽管两种酶的水解速率随着聚酯熔融温度和实验温度之间差异的减小而增加，但对于脂肪酶而言，这种趋势比角质酶更明显。速率的这些趋势可以归因于温度依赖性聚酯链的柔韧性和酶活性位点可及性的结合。这项工作强调了基于FDL的方法可用于酶促聚酯水解的筛选和机理研究的能力。

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  • 来源
    《Environmental Science & Technology》 |2017年第8期|4358-4367|共10页
  • 作者

    Michael Thomas Zumstein; Hans-Peter E. Kohler; Kristopher McNeill; Michael Sander;

  • 作者单位

    Institute of Biogeochemistry and Pollutant Dynamics, ETH Zurich, 8092 Zurich, Switzerland;

    Environmental Biochemistry Group, Environmental Microbiology, Swiss Federal Institute of Aquatic Science and Technology (Eawag), 8600 Duebendorf, Switzerland;

    Institute of Biogeochemistry and Pollutant Dynamics, ETH Zurich, 8092 Zurich, Switzerland;

    Institute of Biogeochemistry and Pollutant Dynamics, ETH Zurich, 8092 Zurich, Switzerland;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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