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中链长度聚羟基脂肪酸酯降解酶的降解动力学:石英晶体微天平研究

Degradation kinetics of medium chain length Polyhydroxyalkanoate degrading enzyme: a quartz crystal microbalance study.

作者信息

Millan Fabien, Hanik Nils

机构信息

Institute of Life Technologies, School of Engineering, University of Applied Science and Arts Western Switzerland, Sion, Switzerland.

出版信息

Front Bioeng Biotechnol. 2023 Dec 14;11:1303267. doi: 10.3389/fbioe.2023.1303267. eCollection 2023.

Abstract

This study investigates the enzymatic degradation processes of different classes of polyhydroxyalkanoates (PHAs), a group of biopolymers naturally synthesized by various microorganisms. Medium chain length PHAs (mcl-PHAs) are distinguished biopolymers due to their biodegradability and diverse material properties. Using quartz crystal microbalance measurements as a valuable tool for accurate real-time monitoring of the enzymatic degradation process, the research provides detailed kinetic data, describing the interaction between enzymes and substrates during the enzymatic degradation process. Thin films of poly-3-hydroxybutyrate (PHB) and polyhydroxyoctanoate copolymer (PHO), containing molar fractions of about 84% 3-hydroxyoctanoate and 16% 3-hydroxyhexanoate, were exposed to scl-depolymerases from LMG 2207 and recombinant mcl-depolymerase produced in DH5α harboring the plasmid pMAD8, respectively. Analyses based on a heterogeneous kinetic model for the polymer degradation indicated a six-fold stronger adsorption equilibrium constant of mcl-depolymerase to PHO. Conversely, the degradation rate constant was approximately twice as high for scl-depolymerases acting on PHB. Finally, the study highlights the differences in enzyme-substrate interactions and degradation mechanisms between the investigated scl- and mcl-PHAs.

摘要

本研究调查了不同类别的聚羟基脂肪酸酯(PHA)的酶促降解过程,PHA是一类由各种微生物天然合成的生物聚合物。中链长度PHA(mcl-PHA)因其生物可降解性和多样的材料特性而成为独特的生物聚合物。利用石英晶体微天平测量作为精确实时监测酶促降解过程的宝贵工具,该研究提供了详细的动力学数据,描述了酶促降解过程中酶与底物之间的相互作用。分别将含有约84% 3-羟基辛酸酯和16% 3-羟基己酸酯摩尔分数的聚-3-羟基丁酸酯(PHB)和聚羟基辛酸酯共聚物(PHO)薄膜暴露于来自LMG 2207的短链解聚酶和在携带质粒pMAD8的DH5α中产生的重组中链解聚酶。基于聚合物降解的非均相动力学模型的分析表明,中链解聚酶对PHO的吸附平衡常数强六倍。相反,作用于PHB的短链解聚酶的降解速率常数约为其两倍。最后,该研究突出了所研究的短链和中链PHA之间酶-底物相互作用和降解机制的差异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7149/10756687/b8c6cedfe692/fbioe-11-1303267-g001.jpg

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