Computational redesign of the heme acquisition protein HasA for enhanced thermostability while retaining its ability to bind synthetic metalloporphyrins

Abstract

Abstract Heme acquisition protein HasA was computationally redesigned using GAOptimizer and LigandMPNN to obtain thermostable variants that retained their metalloporphyrin-binding ability. The most thermostable variant, 2nd300gen, exhibited an approximately 17.5 ℃ higher Tm value than the wild type while maintaining its structure and function. This design strategy effectively enhanced the robustness of HasA and demonstrates its potential for broadening the applicability of HasA-based artificial protein assemblies.