Soil aminopeptidase activities under 145-year crop rotation and fertility practices in the North Central US

Jasdeep Singh, Jessie Arabely Navas Soto, Rosa Elena Ibarra Lόpez, Andrew J. Margenot

Research output: Contribution to journalArticlepeer-review


Aminopeptidases catalyze the hydrolysis of peptides into constituent amino acids and are therefore a key step in nitrogen (N) depolymerization and downstream mineralization. Differences in carbon (C):N as well as sulfur (S) content of amino acids present the possibility of amino acid-specific aminopeptidase activities revealing distinct information on N and non-N processes in soils. However, chromogenic assays of soil aminopeptidases using para-nitroaniline (pNA) substrates are underutilized and have largely focused on just two aminopeptidases to-date. We evaluated activities of eight amino acid-specific aminopeptidases using pNA substrates for arginine (ARG), glycine (GLY), alanine (ALA), lysine (LYS), glutamic acid (GLU), methionine (MET), proline (PRO) and leucine (LEU) aminopeptidases in a high-resolution sampling (30 samples per 0.01 ha−1 treatment plot) of nine 145-year treatment plots resulting from a factorial combination of three crop rotations and three fertility treatments on a Argiudoll in North Central United States. Across treatments, we identified non-detectable activities of ARG and PRO, and detectable activities of GLY, ALA, LYS, GLU, MET, and LEU ranging 0.2 to 400 nmol pNA h−1 g−1. Except for LYS and GLU activities, aminopeptidase activities were homogeneously distributed within treatments (n=30 locations per plot), and exhibited a mean coefficient of variation (CV) of 9 to 30 % and spatial range of 4.1 to 9.0 m. Activities of ALA and MET had the lowest CV and greatest contribution in explaining overall variability of aminopeptidase activities and were more sensitive to treatments compared to GLY and LEU activity. Multivariate analysis revealed a positive correlation among aminopeptidase activities, with major variation explained by β-glucosidase activity followed by pH and total N (TN). Absolute aminopeptidase activities generally reflected soil organic C (SOC) and TN concentrations, with greater activities under diverse rotation with organic fertilization (manure) and lower activities under continuous maize without fertilization. However, aminopeptidase activities normalized to SOC concentration were greater in soils without fertilization relative to organic fertilization. This study provides benchmark values of soil aminopeptidase activities assayed by pNA substrates using the world's second oldest continuous agricultural experiment, including six aminopeptidases that have not been previously assayed with these substrates. Given their low magnitude of variability, high spatial correlation, and sensitivity to management, assays of aminopeptidase activities – particularly MET and ALA activities – could be fruitful tools for better understanding soil N cycling. Future work should evaluate assay optimization and expand to additional amino acid-specific substrates using pNA-based substrates.

Original languageEnglish (US)
Article number116703
StatePublished - Dec 2023


  • Aminopeptidases
  • Depolymerization
  • Exopeptidases
  • Nitrogen cycling
  • Soil enzymes

ASJC Scopus subject areas

  • Soil Science


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