Recently, researchers from the Feed Enzyme Engineering Research Team of IAS and Hangzhou Levinthal Biotech Ltd worked together to developed an innovative tool to improve enzyme thermostability based on the short board theory and the Zero-Shot Hamiltonian model. The findings are published in the journal of Advanced Science.

Understanding the mechanism underlying thermostabilization in naturally stable enzymes and enhancing the thermostability of unstable enzymes are crucial aspects in enzyme engineering. Despite the development of various engineering methods, there remains substantial scope for improvement.

Fig.1 Diagram illustrates the three levels of the “short board”.

In this study, a novel concept termed as the “short board” theory is proposed, which conceptualizes proteins as barrels with each component representing a jagged board. Notably, optimizing modifications to the shortest board yields optimal enhancements in terms of thermostability performance. To validate this theory, α-amylase, an industrial bulk enzyme with multiple domains, is employed as a model enzyme. The existence of “short boards” and their impact on thermostability modification are demonstrated at the domain, residue, and atomic levels through experimental confirmation using domain substitution. Furthermore, a novel thermostable design and prediction model called Zero-Shot Hamiltonian (ZSH) is established and evaluated on α-amylase. This coevolutionary approach based on thermostability and deep learning exhibits remarkable success exclusively when applied to enzymes with fixed short boards. The integration of the “short board” theory with the ZSH model presents an innovative tool for enhancing enzymatic thermostability.

The online version available at：

https://onlinelibrary.wiley.com/doi/10.1002/advs.202402441