Enzyme-assisted extraction of anti-inflammatory compounds from habanero chili pepper (Capsicum chinense) seeds

Abstract

Capsaicinoids are the main bioactive compounds extracted from chili pepper seeds (CPSs) but other bioactive compounds such as phenolic compounds may be found. Enzyme-assisted extraction (EAE) improves the extraction of bioactive compounds from fruits and seeds. The aim of this study was to establish the cellulase-assisted extraction conditions of capsaicinoids and phenolic compounds from Habanero CPSs (Capsicum chinense) and to evaluate the anti-inflammatory activity of the obtained extracts on murine macrophages. EAE was performed using different temperatures (T1 = 30°C, T2 = 45°C and T3 = 60°C), enzyme concentrations (E1 = 2,500 UI/L and E2 = 250 UI/L), and extraction time periods (0-150 min). Total phenolic compounds were quantified using the Folin-Ciocalteu assay, capsaicin (CAP) and dihydrocapsaicin (DHC) contents were evaluated by HPLC, and anti-inflammatory activity was performed with Griess assay on murine macrophage RAW 264.7 cell culture. The highest phenolic compound content (337.96 mg GAE/L) was achieved at 30°C, 2,500 UI/L, and 150 min of extraction. The highest CAP content (310.23 μg/ml) was obtained at 45°C with 250 UI/L for 150 min, while for DHC (167.72 μg/ml), the conditions were 60°C, 2,500 UI/L, and 120 min. The highest anti-inflammatory response was obtained when 60°C, E2, and 150 min were used for the extraction, and nitric oxide (NO) production was reduced to 22.56%. Based on the results obtained in this research, EAE allowed the recovery of compounds with anti-inflammatory activity from CPS using water as a solvent. There was a correlation between the extraction of CAP and DHC. But although a moderate direct correlation between the concentration of capsaicinoids and total phenolic compounds (TPCs) and an inverse correlation of the presence of the bioactive compounds (TPC, CAP, and DHC) with the NO synthesis, these were not statistically significant. We demonstrated that Habanero seeds are an important raw material to recover anti-inflammatory compounds beyond capsaicinoids using water in EAE.

Interaction Between Salicylic or Ferulic Acid and Cellulase Used to Produce Phenolic Compounds Aggregates from “Jalapeño” (Capsicum AnnumL.) Chili Pepper Seeds

“…At 180 min, TPC was like the one obtained at 90 min (34.20 and 36.01 mg/L). Similar nonlinear results were obtained by Cortés-Ferré et al for Habanero chili pepper seeds. For ferulic and salicylic acids, the maximum TPC was also reached at 90 min and no significant difference was observed in comparison to the control group.…”

“…Cytotoxicity assay was performed using the methodology reported by Cortés-Ferré et al using CellTiter 96® Aqueous One Solution Cell Proliferation Assay Kit (Promega, Madison, WI, USA) following manufacturer’s instructions. Subsequently, 100 μL of cell culture was inoculated at a density of 1 x 10 4 cells per well into 96-well plates and incubated at 37 °C for 24 h. After incubation time, cells were pretreated with 50 μL of JPS extracts at 0.01% (v/v) for 24 h before adding 50 μL of growth media (for control wells) or 50 μL of 10 μg/mL lipopolysaccharides (LPS) from Salmonella enterica serotype typhimurium (Sigma-Aldrich, St. Lous, MO, USA).…”

“…Nitrite in the macrophage culture medium was measured to estimate the immune response. NO synthesis in cell-free culture supernatants was measured using Griess reagent 14 according to Corteś-Ferréet al 13 Griess System Reagents (Promega, Madison, WI, USA) were used according to the manufacturer's instructions (20 μL of reagent A and 20 μL of reagent B), incubated at room temperature for 10 min for each reagent, and absorbance was measured at 540 nm using a microplate reader (BioTek Cytation 5, BioTek Instruments, Winooski, VT, USA).…”

Genetic Regulation, Environmental Cues, and Extraction Methods for Higher Yield of Secondary Metabolites in Capsicum

“…Cellulase-assisted extraction in water at 40−45 °C for a period of 150 min was best suited at a concentration of 2500 and 250 U L −1 for maximum phenolic compounds and capsaicinoids yield, respectively, from "Habanero" seeds. 206 Five times more capsaicinoids and carotenoids were extracted in the cellulase-treated "escabeche chili" (C. baccatum) powder than the control while using an extraction solvent mixture of hexane, acetone, and ethanol. 207 Treatment of "Tabasco Pepper" with 1 mL kg −1 pectinase and cellulase each increased the extract yield by 17.5% and enhanced the spice and flavor of the pepper sauce.…”

In vitro Evaluation of Anti-Inflammatory Activity of “Habanero” Chili Pepper (Capsicum chinense) Seeds Extracts Pretreated with Cellulase

“…Cortés-Ferré et al (2022) demonstrated that Enzyme Assisted Extraction (EAE) allowed the recovery of compounds with anti-in ammatory activity from CPS using water as solvent. Although signi cant differences were observed in the CPS extracts obtained at different times, temperature and cellulase concentration, no statistically signi cant correlation was found between capsaicinoids or total phenolic content (TPC) with the NO synthesis [11].…”