Aspergillus oryzae 3.042 and Aspergillus sojae 3.495 are crucial starters for fermented soybean foods since their abundant secreted enzymes. This study aimed to compare the differences in protein secretion between A. oryzae 3.042 and A. sojae 3.495 during the soy sauce koji fermentation and the effect on volatile metabolites to un-derstand the fermentation characteristics of the strains better. Label-free proteomics detected 210 differentially expressed proteins (DEPs) enriched in amino acid metabolism and protein folding, sorting and degradation pathways. Subsequently, extracellular enzyme analysis showed that three peptidases, including peptide hydro-lase, dipeptidyl aminopeptidase, and peptidase S41, were up-regulated in A. sojae 3.495. Seven carbohydrases, including alpha-galactosidase, endo-arabinase, beta-glucosidase, alpha-galactosidase, alpha-glucuronidase, arabinan-endo 1,5-alpha-L-arabinase, and endo-1,4-beta-xylanase were up-regulated in A. oryzae 3.042, contributing to the difference in enzyme activity. Significantly different extracellular enzymes influenced the content of volatile alcohols, alde-hydes and esters such as (R, R)-2,3-butanediol, 1-hexanol, hexanal, decanal, ethyl L-lactate and methyl myristate in both strains, which affected the type of aroma of koji. Overall, this study revealed the differences in molecular mechanisms between A. oryzae 3.042 and A. sojae 3.495 under solid-state fermentation, providing a reference for targeted enhancement strains.
