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To what extent are different levels of expertise reflected in the functional connectivity of the brain? We addressed this question by using resting-state functional magnetic resonance imaging (fMRI) in mathematicians versus non-mathematicians. To this end, we investigated how the two groups of participants differ in the correlation of their spontaneous blood oxygen level-dependent fluctuations across the whole brain regions during resting state. Moreover, by using the classification algorithm in machine learning, we investigated whether the resting-state fMRI networks between mathematicians and non-mathematicians were distinguished depending on features of functional connectivity. We showed diverging involvement of the frontal–thalamic–temporal connections for mathematicians and the medial–frontal areas to precuneus and the lateral orbital gyrus to thalamus connections for non-mathematicians. Moreover, mathematicians who had higher scores in mathematical knowledge showed a weaker connection strength between the left and right caudate nucleus, demonstrating the connections’ characteristics related to mathematical expertise. Separate functional networks between the two groups were validated with a maximum classification accuracy of 91.19% using the distinct resting-state fMRI-based functional connectivity features. We suggest the advantageous role of preconfigured resting-state functional connectivity, as well as the neural efficiency for experts’ successful performance.