Rational protein design is a powerful strategy,not only for revealing the structure and function relationship of natural metalloproteins,but also for creating artificial metalloproteins with improved properties and functions.Myoglobin(Mb),a small heme protein created by nature with diverse functions,has been shown to be an ideal scaffold for rational protein design.The progress reviewed herein includes fine-tuning its native functions of O2binding and transport,peroxidase activity and nitrite reductase(NIR)activity,and rational expanding its functionalities to peroxygenase,heme-copper oxidase(HCO),nitric oxide reductase(NOR),as well as hydroxylamine reductase.These studies have enhanced our understanding of how metalloproteins work in nature,and provided insights for rational design of functional metalloproteins for practical applications in the future.