Overall analyses of the reactions catalyzed by acetohydroxyacid synthase/acetolactate synthase using a precolumn derivatization-HPLC method

Xian, Hui; Tian, Jin-Meng; Wang, Xin; Zhang, Zhenqian; Zhao, Yi; Li, Heng

doi:10.1016/j.ab.2022.114980

Cited by 5 publications

(1 citation statement)

References 32 publications

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“…HPLC is suitable for the separation and purification of physiologically active giant molecule substances, such as proteins, enzymes, hormones, monoamino acid, nucleic acids, etc. [41].…”

Section: Chromatography Analysis Technologymentioning

confidence: 99%

Methods and Advances in the Design, Testing and Development of In Vitro Diagnostic Instruments

Wang

et al. 2023

Processes

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With the continuous improvement of medical testing and instrumentation engineering technologies, the design, testing and development methods of in vitro diagnostic instruments are developing rapidly. In vitro diagnostic instruments are also gradually developing into a class of typical high-end medical equipment. The design of in vitro diagnostic instruments involves a variety of medical diagnostic methods and biochemical, physical and other related technologies, and its development process involves complex system engineering. This paper systematically organizes and summarizes the design, testing and development methods of in vitro diagnostic instruments and their development in recent years, focusing on summarizing the related technologies and core aspects of the R&D process, and analyzes the development trend of the in vitro diagnostic instrument market.

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“…HPLC is suitable for the separation and purification of physiologically active giant molecule substances, such as proteins, enzymes, hormones, monoamino acid, nucleic acids, etc. [41].…”

Section: Chromatography Analysis Technologymentioning

confidence: 99%

Methods and Advances in the Design, Testing and Development of In Vitro Diagnostic Instruments

Wang

et al. 2023

Processes

View full text Add to dashboard Cite

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An easy and sensitive assay for acetohydroxyacid synthases based on the simultaneous detection of substrates and products in a single step

Engelhardt,

Ebeling,

Kaltenegger

et al. 2024

Anal Bioanal Chem

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Acetohydroxyacid synthase (AHAS, EC 2.2.1.6) catalyzes the first step in the synthesis of the branched-chain amino acids valine, leucine, and isoleucine, pathways being present in plants and microorganisms, but not in animals. Thus, AHAS is an important target for numerous herbicides and, more recently, for the development of antimicrobial agents. The need to develop new and optimized herbicides and pharmaceuticals requires a detailed understanding of the biochemistry of AHAS. AHAS transfers an activated two-carbon moiety derived from pyruvate to either pyruvate or 2-oxobutyrate as acceptor substrates, forming 2-acetolactate or 2-acetohydroxy-2-butyrate, respectively. Various methods have been described in the literature to biochemically characterize AHAS with respect to substrate preferences, substrate specificity, or kinetic parameters. However, the simultaneous detection and quantification of substrates and unstable products of the AHAS-catalyzed reaction have always been a challenge. Using AHAS isoform II from Escherichia coli, we have developed a sensitive assay for AHAS-catalyzed reactions that uses derivatization with ethyl chloroformate to stabilize and volatilize all reactants in the aqueous solution and detect them by gas chromatography coupled to flame ionization detection or mass spectrometry. This assay allows us to characterize the product formation in reactions in single and dual substrate reactions and the substrate specificity of AHAS, and to reinterpret previous biochemical observations. This assay is not limited to the AHAS-catalyzed reactions, but should be applicable to studies of many metabolic pathways. Graphical Abstract

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Re-investigation of in vitro activity of acetohydroxyacid synthase I holoenzyme from Escherichia coli

Wang,

Sun,

Zheng

et al. 2024

Archives of Biochemistry and Biophysics

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Overall analyses of the reactions catalyzed by acetohydroxyacid synthase/acetolactate synthase using a precolumn derivatization-HPLC method

Cited by 5 publications

References 32 publications

Methods and Advances in the Design, Testing and Development of In Vitro Diagnostic Instruments

Methods and Advances in the Design, Testing and Development of In Vitro Diagnostic Instruments

An easy and sensitive assay for acetohydroxyacid synthases based on the simultaneous detection of substrates and products in a single step

Re-investigation of in vitro activity of acetohydroxyacid synthase I holoenzyme from Escherichia coli

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