The Study of In vitro Phagocytic Activity of Soybean Meal Fermented with Bacillus subtilis TISTR 2664

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Kamonsri Nuankham
Wanvipa Chuenchom
Pattarawadee Kengkwasingh
Neungnut Chaiyawan
Viroj Tachapuripunya
Watcharee Kunyalung


This study aimed to investigate the components and activity of bioactive peptides derived from Bacillus subtilis TISTR 2664 fermentation of soybean meal to stimulate phagocytosis. Soybean meal obtained from a soybean oil factory was used as a raw material for fermentation. It contained 44.65% protein, 1.28% oil, 5.54% dietary fiber, 12.08% moisture and 6.40% ash. When fermented with B. subtilis TISTR2664 and examined by SDS-PAGE, most of the protein size remained in the range of 5-29 kilo Dalton (kDa). The 8,981 types of peptides in the fermented soybean meal were investigated by Liquid Chromatography-Mass Spectrometry (LC-Q-TOF-MS/MS). Ten biologically active peptides having the following properties were predicted: anticancer, antifungal, antihypertensive antimicrobial antiparasitic anti-inflammatory antivirus cell communication, drug delivery, and tumor homing. Interestingly, the peptides with antihypertensive properties were found in the highest amounts. The results of this prediction will be useful for planning for further laboratory trials of the peptide derived from fermented soybean meal to lead to its utilization. A preliminary laboratory investigation into the toxicity of fermented soybean meal on RAW264.7 mouse macrophages was performed using the MTT assay. No cytotoxicity was observed at less than or equal to 25 milligrams per milliliters (mg/ml), with more than 70% viability of RAW264.7 cells. Moreover, sample concentrations ranging from 3.13 to 25 mg/ml were investigated. The results showed that at a 12.5% concentration, the highest phagocytic activity was induced at 93.27±0.50%, which was significantly higher than that of the positive control (p < 0.01). Nitric oxide production at 48.53±0.35 micromolar (µM) was non-significantly different from the positive control at all tested sample concentrations. The result of this study can be used as a guideline for fermented soybean meal research in immune stimulation for further development as a health supplement or as an added economic value.

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