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We develop a method for bifunctional regulation on the surface micro-morphology and nitrogen-doping configuration of graphene-based nanosheets by annealing chitin and graphene oxide. Systematic studies reveal that annealing temperature is a key to determine micro-morphology and type of nitrogen-doping of the graphene matrix, and 2D multi-wrinkled graphene (CS-rGO-600) with enriched active nitrogen species and moderate defects can be obtained successfully under an optimized annealing temperature. When used as a "green" electrocatalyst for oxygen reduction reaction (ORR), CS-rGO-600 exhibits superior performance, in terms of more outstanding electrode activity and durability, higher output power density in fuel cell system compared to that of Pt/C. These could be attributed to the fact that appropriate annealing treatment is not only conducive to exogenous nitrogen enrichment and oriented conversion but also beneficial to micro-structure construction. The synergistic effects between multi-wrinkled 2D structure and active sites endow CS-rGO-600 with superior ORR performance in the fuel cell. • CS-rGO-X was prepared via pyrolysis of biomass waste and GO. • Appropriate pyrolysis is conducive to N enrichment and structure construction. • Synergism between structure and active sites endow CS-rGO-600 with superior activity. • CS-rGO-600 with highest E onset (0.059 V) and P max (0.665 mWcm−2). [ABSTRACT FROM AUTHOR]