Optimizing enzyme inhibition analysis: precise estimation with a single inhibitor concentration

Abstract Enzyme inhibition analysis is essential in drug development and food processing, necessitating precise estimation of inhibition constants. Traditionally, these constants are estimated through experiments using multiple substrate and inhibitor concentrations, but inconsistencies across studi...

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Main Authors: Hyeong Jun Jang, Yun Min Song, Jang Su Jeon, Hwi-yeol Yun, Sang Kyum Kim, Jae Kyoung Kim
Format: Article
Language:English
Published: Nature Portfolio 2025-06-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60468-z
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author Hyeong Jun Jang
Yun Min Song
Jang Su Jeon
Hwi-yeol Yun
Sang Kyum Kim
Jae Kyoung Kim
author_facet Hyeong Jun Jang
Yun Min Song
Jang Su Jeon
Hwi-yeol Yun
Sang Kyum Kim
Jae Kyoung Kim
author_sort Hyeong Jun Jang
collection DOAJ
description Abstract Enzyme inhibition analysis is essential in drug development and food processing, necessitating precise estimation of inhibition constants. Traditionally, these constants are estimated through experiments using multiple substrate and inhibitor concentrations, but inconsistencies across studies highlight a need for a more systematic approach to set experimental designs across all types of enzyme inhibition. Here, we address this by analyzing the error landscape of estimations in various experimental designs. We find that nearly half of the conventional data is dispensable and even introduces bias. Instead, by incorporating the relationship between IC50 and inhibition constants into the fitting process, we find that using a single inhibitor concentration greater than IC50 suffices for precise estimation. This IC50-based optimal approach, which we name 50-BOA, substantially reduces (>75%) the number of experiments required while ensuring precision and accuracy. Additionally, we provide a user-friendly package that implements the 50-BOA.
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issn 2041-1723
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publishDate 2025-06-01
publisher Nature Portfolio
record_format Article
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spelling doaj-art-b1d29c258243433dbb70f13c4388c03f2025-08-20T02:05:38ZengNature PortfolioNature Communications2041-17232025-06-0116111410.1038/s41467-025-60468-zOptimizing enzyme inhibition analysis: precise estimation with a single inhibitor concentrationHyeong Jun Jang0Yun Min Song1Jang Su Jeon2Hwi-yeol Yun3Sang Kyum Kim4Jae Kyoung Kim5School of Transdisciplinary Studies, KAISTBiomedical Mathematics Group, Pioneer Research Center for Mathematical and Computational Sciences, Institute for Basic ScienceCollege of Pharmacy, Chungnam National UniversityCollege of Pharmacy, Chungnam National UniversityCollege of Pharmacy, Chungnam National UniversityBiomedical Mathematics Group, Pioneer Research Center for Mathematical and Computational Sciences, Institute for Basic ScienceAbstract Enzyme inhibition analysis is essential in drug development and food processing, necessitating precise estimation of inhibition constants. Traditionally, these constants are estimated through experiments using multiple substrate and inhibitor concentrations, but inconsistencies across studies highlight a need for a more systematic approach to set experimental designs across all types of enzyme inhibition. Here, we address this by analyzing the error landscape of estimations in various experimental designs. We find that nearly half of the conventional data is dispensable and even introduces bias. Instead, by incorporating the relationship between IC50 and inhibition constants into the fitting process, we find that using a single inhibitor concentration greater than IC50 suffices for precise estimation. This IC50-based optimal approach, which we name 50-BOA, substantially reduces (>75%) the number of experiments required while ensuring precision and accuracy. Additionally, we provide a user-friendly package that implements the 50-BOA.https://doi.org/10.1038/s41467-025-60468-z
spellingShingle Hyeong Jun Jang
Yun Min Song
Jang Su Jeon
Hwi-yeol Yun
Sang Kyum Kim
Jae Kyoung Kim
Optimizing enzyme inhibition analysis: precise estimation with a single inhibitor concentration
Nature Communications
title Optimizing enzyme inhibition analysis: precise estimation with a single inhibitor concentration
title_full Optimizing enzyme inhibition analysis: precise estimation with a single inhibitor concentration
title_fullStr Optimizing enzyme inhibition analysis: precise estimation with a single inhibitor concentration
title_full_unstemmed Optimizing enzyme inhibition analysis: precise estimation with a single inhibitor concentration
title_short Optimizing enzyme inhibition analysis: precise estimation with a single inhibitor concentration
title_sort optimizing enzyme inhibition analysis precise estimation with a single inhibitor concentration
url https://doi.org/10.1038/s41467-025-60468-z
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