RESUMO
This work aimed to propose a rapid method to screen the bioactive peptides with anti-α-glucosidase activity instead of traditional multiple laborious purification and identification procedures. 242 peptides binding to α-glycosidase were quickly screened and identified by bio-affinity ultrafiltration combined with LC-MS/MS from the double enzymatic hydrolysate of black beans. Top three peptides with notable anti-α-glucosidase activity, NNNPFKF, RADLPGVK and FLKEAFGV were further rapidly screened and ranked by the three artificial intelligence tools (three-AI-tool) BIOPEP database, PeptideRanker and molecular docking from the 242 peptides. Their IC50 values were in order as 4.20 ± 0.11 mg/mL, 2.83 ± 0.03 mg/mL, 1.32 ± 0.09 mg/mL, which was opposite to AI ranking, for the hydrophobicity index of the peptides was not included in the screening criteria. According to the kinetics, FT-IR, CD and ITC analyses, the binding of the three peptides to α-glucosidase is a spontaneous and irreversible endothermic reaction that results from hydrogen bonds and hydrophobic interactions, which mainly changes the α-helix structure of α-glucosidase. The peptide-activity can be evaluated vividly by AFM in vitro. In vivo, the screened FLKEAFGV and RADLPGVK can lower blood sugar levels as effectively as acarbose, they are expected to be an alternative to synthetic drugs for the treatment of Type 2 diabetes.
Assuntos
Inibidores de Glicosídeo Hidrolases , Peptídeos , alfa-Glucosidases , alfa-Glucosidases/metabolismo , Fabaceae/química , Inibidores de Glicosídeo Hidrolases/química , Inibidores de Glicosídeo Hidrolases/farmacologia , Cinética , Espectrometria de Massa com Cromatografia Líquida , Simulação de Acoplamento Molecular , Peptídeos/química , Peptídeos/farmacologia , Espectrometria de Massas em Tandem , Ultrafiltração/métodosRESUMO
Highly sensitive and stable measurement of methane (CH4) and acetylene (C2H2) based on a novel dual-channel off-beam quartz-enhanced photoacoustic spectroscopy and time-division multiplexing technique was realized by a compact 3D-printed gas cell with a size of 3 × 2 × 1 cm3. Two near-infrared distributed feedback diode lasers were employed to target the CH4 absorption line at 6046.9 cm-1 and the C2H2 absorption line at 6521.2 cm-1, respectively. Second-harmonic wavelength modulation spectroscopy method was used for photoacoustic signal recovery. A minimum detection level of â¼ 7.63 parts-per-million in volume (ppmv) for CH4 and a level of â¼ 17.47 ppmv for C2H2 were achieved with a 1 s lock-in integration time, leading to a normalized noise equivalent absorption (NNEA) coefficient of 7.24 × 10-8 cm-1·W·Hz-1 and 3.73 × 10-8 cm-1·W·Hz-1 for CH4 and C2H2, respectively. Allan-Werle deviation analysis was employed to evaluate the stability and the minimum detection limit (MDL) of the developed photoacoustic CH4/C2H2 dual-gas photoacoustic sensor. Owing to the high stability of the developed sensor system, an MDL of â¼ 0.73 ppmv and an MDL of â¼ 1.60 ppmv with a 100 s averaging time were achieved for CH4 and C2H2, respectively.