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[Display omitted] • UDB and DB at different pyrolysis temperatures played contrasting roles. • DB800-amended group displayed superior nitrogen removal performance. • DB at lower pyrolysis temperature imposed suppression. • Properties of dissolved organic matter released by DB were tracked at molecular level. Biochar has attracted increasingly attention in the biological wastewater treatment owing to its unique environmental behavior. However, the differential effects of undissolved and dissolved fractions of biochar (hereafter named as UDB and DB, respectively) on the anaerobic ammonium oxidation (anammox) process remain unclear. The study evaluated the nitrogen removal performance of anammox system in response to UDB and DB prepared at different pyrolysis temperatures (300 °C, 550 °C and 800 °C). The DB800-amended group displayed superior nitrogen removal performance with the total nitrogen removal efficiency (TNRE) of 92.8 ± 1.4 %. Compared with CK, TNRE was changed by +5.0 % (UDB300), +0.4 % (UDB550) and −22.3 % (UDB800) for UDB-amended groups, and by −16.0 % (DB300), −9.4 % (DB550), and +9.2 % (DB800) for DB-amended groups, respectively. The differences were closely related to the physicochemical properties of UDB and DB. Particularly, dissolved organic matter released by DB were tracked at molecular level, which revealed that CHO occupied the highest proportion of 70.3 % and lignin-like and carboxylic rich alicyclic molecules were dominant in DB800. This study would facilitate the understanding of the differences between the nitrogen removal performance of UDB and DB-amended anammox, thus providing deeper insights for the comprehensive assessment of the impacts of biochar on the anammox process. [ABSTRACT FROM AUTHOR]