Comparative Proteomic Analysis of Bacillus subtilis and Aspergillus niger in Black Soldier Fly Co-Fermentation

Black soldier fly larvae have gained popularity as an organic waste bio-conversional tool and fodder protein replacement in recent decades. It can consume all kinds of animal feces, kitchen waste and agricultural waste with great efficiency and transform them into high-value insect protein, fatty acids, and amino acids, which makes the larva a good substitute for costly fish meal and bean pulp in animal diets. However, excess chitin in the larva skin limits its application as an animal feed additive, consequently, employing fermentation with zymocytes to remove the chitin is necessary. In this study, we raised black soldier fly larvae (BSFL) with different carbon sources, such as chicken feces, straws and glucose, and examined the growth condition; we applied Bacillus subtilis and Aspergillus niger to co-ferment BSFL paste to analyze its nutrition changes. Data revealed that among the four kinds of cultures, the body weight of the corn powder group increased most rapidly; the wood chip group was the most underweight; however, it increased faster than others before day 4, and contained the least fat. Label-free quantitative proteomic analysis revealed that the expression of multiple enzymes from B. subtilis and A. niger involved in polysaccharide hydrolysis, amino acid biosynthesis and fatty acid metabolism, such as peptidase of S8 family, maltogenic alpha-amylase, oligo-1,6-glucosidase and lysophospholipase like protein changed significantly compared to the control group. Production detection showed that free amino acids, acid-soluble proteins, and short-chain fatty acids increased after fermentation; 13 out of 17 amino acids were increased and total free amino acids were increased from 0.08 g/100 g to 0.3 g/100 g; organic acids increased by 4.81 to 17 fold through fermentation, respectively; the actual protein content declined from 3.03 g/100 g to 1.81 g/100 g, the peptide content increased from 1.3 g/100 g to 2.46 g/100 g, the chitin degradation rate was 40.3%, and fat decreased 30% (p < 0.05). These findings might provide important information for future applications of black soldier fly larvae in different carbon waste recycling measures and material for animal feed/organic fertilizer after fermentation.