A powerful analytical method for the identification of biologically meaningful metabolic subpathways. Subpathway-GM integrates ‘interesting genes’ and ‘interesting metabolites’ related to the study condition (e.g. disease) into the corresponding enzyme and metabolite nodes (referred to as signature nodes) within the metabolic pathway. We then analyzed lenient distance similarities of signature nodes within the pathway structure to locate key metabolic cascade subpathway regions. Finally, a hypergeometric test was used to evaluate the enrichment significance of these subpathway regions.