Leveraging liquid-liquid phase separation and volume modulation to regulate the enzymatic activity of formate dehydrogenase

IF 3.3 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biophysical chemistry Pub Date : 2023-10-29 DOI:10.1016/j.bpc.2023.107128

Lena Ostermeier , Moreno Ascani , Nicolás Gajardo-Parra , Gabriele Sadowski , Christoph Held , Roland Winter

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Abstract

Engineering of reaction media is an exciting alternative for modulating kinetic properties of biocatalytic reactions. We addressed the combined effect of an aqueous two-phase system (ATPS) and high hydrostatic pressure on the kinetics of the Candida boidinii formate dehydrogenase-catalyzed oxidation of formate to CO₂. Pressurization was found to lead to an increase of the binding affinity (decrease of K_M, respectively) and a decrease of the turnover number, k_cat. The experimental approach was supported using thermodynamic modeling with the electrolyte Perturbed-Chain Statistical Associating Fluid Theory (ePC-SAFT) equation of state to predict the liquid-liquid phase separation and the molecular crowding effect of the ATPS on the kinetic properties. The ePC-SAFT was able to quantitatively predict the K_M-values of the substrate in both phases at 1 bar as well as up to a pressure of 1000 bar. The framework presented enables significant advances in bioprocess engineering, including the design of processes with significantly fewer experiments and trial-and-error approaches.

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利用液-液相分离和体积调节来调节甲酸脱氢酶的酶活性。

反应介质工程是调节生物催化反应动力学性质的一种令人兴奋的替代方案。我们讨论了双水相系统（ATPS）和高静水压对布氏假丝酵母甲酸脱氢酶催化甲酸盐氧化为CO2的动力学的综合影响。加压可导致结合亲和力的增加（分别为KM的减少）和周转数kcat的减少。使用电解质微扰链统计缔合流体理论（ePC-SAFT）状态方程的热力学建模来预测液-液相分离和ATPS对动力学性质的分子拥挤效应，从而支持了实验方法。ePC SAFT能够定量预测在1巴以及高达1000巴的压力下两相中的基底的KM值。所提出的框架使生物过程工程取得了重大进展，包括用更少的实验和试错方法设计过程。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biophysical chemistry 生物-生化与分子生物学

CiteScore

6.10

自引率

10.50%

发文量

121

审稿时长

20 days

期刊介绍： Biophysical Chemistry publishes original work and reviews in the areas of chemistry and physics directly impacting biological phenomena. Quantitative analysis of the properties of biological macromolecules, biologically active molecules, macromolecular assemblies and cell components in terms of kinetics, thermodynamics, spatio-temporal organization, NMR and X-ray structural biology, as well as single-molecule detection represent a major focus of the journal. Theoretical and computational treatments of biomacromolecular systems, macromolecular interactions, regulatory control and systems biology are also of interest to the journal.