Optimization of Soybean Meal Fermentation for Aqua-Feed with <i>Bacillus subtilis natto</i> Using the Response Surface Methodology

This study aimed to improve the nutritional value of soybean meal (SBM) by solid-state fermentation (SSF) using <i>Bacillus subtilis natto</i> (<i>B. s. natto</i>) to overcome the limitations of SBM usage in aquafeed. The response surface methodology (RSM) was employed to explore the relationships of fermentation conditions, such as temperature, time, water-substrate ratio, and layer thickness, on the degree of protein hydrolysis (DH) and the crude protein (CP) content. The optimum conditions for achieving the higher DH (15.96%) and CP (55.76%) were 43.82 °C, 62.32 h, 1.08 of water-substrate ratio, and a layer thickness of 2.02 cm. CP and DH in the fermented soybean meal (FSM) increased by 9.8% and 177.1%, respectively, and crude fiber decreased by 14.1% compared to SBM. The protein dispersibility index (PDI) decreased by 29.8%, while KOH protein solubility (KPS) was significantly increased by 17.4%. Flavonoids and total phenolic acid content in FSM were increased by 231.0% and 309.4%, respectively. Neutral protease activity (NPA) also reached a high level (1723.6 U g⁻¹). Total essential amino acids (EAA) in FSM increased by 12.2%, higher than the 10.8% increase of total non-essential amino acids (NEAA), while the total free amino acids content was 12.76 times higher than that of SBM. Major anti-nutritional factors in SBM were significantly reduced during the process, and almost all SBM protein macromolecules were decomposed. Together with the cost-effectiveness of SSF, <i>B. s. natto</i>-fermented SBM products have great potential to improve the plant composition and replace high-cost ingredients in aquafeed, contributing to food security and environmental sustainability.