English

KinForm: Kinetics Informed Feature Optimised Representation Models for Enzyme $k_{cat}$ and $K_{M}$ Prediction

Quantitative Methods 2025-07-22 v1 Machine Learning

Abstract

Kinetic parameters such as the turnover number (kcatk_{cat}) and Michaelis constant (KMK_{\mathrm{M}}) are essential for modelling enzymatic activity but experimental data remains limited in scale and diversity. Previous methods for predicting enzyme kinetics typically use mean-pooled residue embeddings from a single protein language model to represent the protein. We present KinForm, a machine learning framework designed to improve predictive accuracy and generalisation for kinetic parameters by optimising protein feature representations. KinForm combines several residue-level embeddings (Evolutionary Scale Modeling Cambrian, Evolutionary Scale Modeling 2, and ProtT5-XL-UniRef50), taken from empirically selected intermediate transformer layers and applies weighted pooling based on per-residue binding-site probability. To counter the resulting high dimensionality, we apply dimensionality reduction using principal--component analysis (PCA) on concatenated protein features, and rebalance the training data via a similarity-based oversampling strategy. KinForm outperforms baseline methods on two benchmark datasets. Improvements are most pronounced in low sequence similarity bins. We observe improvements from binding-site probability pooling, intermediate-layer selection, PCA, and oversampling of low-identity proteins. We also find that removing sequence overlap between folds provides a more realistic evaluation of generalisation and should be the standard over random splitting when benchmarking kinetic prediction models.

Keywords

Cite

@article{arxiv.2507.14639,
  title  = {KinForm: Kinetics Informed Feature Optimised Representation Models for Enzyme $k_{cat}$ and $K_{M}$ Prediction},
  author = {Saleh Alwer and Ronan Fleming},
  journal= {arXiv preprint arXiv:2507.14639},
  year   = {2025}
}
R2 v1 2026-07-01T04:09:20.868Z