Table 6.
The compositions, types of assay, and detecting wavelengths of the enzyme inhibitory assays.
| Assay | Enzyme | Substrate | Indicator | Inhibitor | Assay Type | Detecting Wavelength |
|---|---|---|---|---|---|---|
| Lipase | 100 µL of 0.01 mg/mL Candida rugosa lipase (type VII, ≥700 unit/mg) | 50 µL of 0.2 mM 5-5′-dithiobis(2-nitrobenzoic-N-phenacyl-4,5-dimethyyhiazolium bromide) | 10 µL of 16 mM DTNB | 40 µL of extract | Kinetics | 412 nm |
| α-Amylase | 100 µL of 30 mg/mL porcine pancreatic α-amylase (type VII, ≥ 10 unit/mg) | 50 µL of 30 mM p-nitrophenyl-α-d-maltopentaoside | 50 µL of extract | Kinetics | 405 nm | |
| α-Glucosidase | 100 µL of 0.1 U/mL Saccharomyces cerevisiae α-glucosidase (type 1, ≥10 U/mg protein) | 50 µL of 2 mM p-nitrophenyl-α-D-glucopyranoside | 50 µL of extract | Kinetics | 405 nm | |
| DPP-IV | 100 µL of 0.01 U/mL human dipeptidyl peptidase-IV (recombinant, ≥ 10 units/mg) | 50 µL of 6 mM Gly-Pro-p-nitroanilide hydrochloride | 50 µL of extract | Kinetics | 405 nm | |
| AChE | 100 μL of 20 ng Electrophorus electricus AChE (1,000 units/mg) | 40 μL of 0.8 mM acetylthiocholine | 10 µL of 16 mM DTNB | 40 µL of extract | Kinetics | 412 nm |
| BChE | 100 µL of 0.5 µg/mL equine serum BChE (≥ 10 units/mg) | 50 µL of 0.4 mM butyrylthiocholine | 10 µL of 16 mM DTNB | 40 µL of extract | Kinetics | 412 nm |
| BACE-1 | a BACE-1 FRET assay kit | 20 µL of extract | End-point | λex = 320 nm, λem = 405 nm | ||
| ACE | 3 µL of 0.5 U/mL rabbit lung ACE (≥ 2 unit/mg) | 30 µL of 3 mM hippuryl-histidyl-leucine | 15 µL of 20 mg/mL PDA | 50 µL of extract | End-point | λex = 360 nm, λem = 485 nm |
DPP-IV: dipeptidyl peptidase-IV; AChE: acetylcholinesterase; BChE: butyrylcholinesterase; BACE-1: β-secretase; ACE: angiotensin-converting enzyme; FRET: fluorescence resonance energy transfer; DTNB: 5,5′-dithiobis(2-nitrobenzoic acid); PDA: o-phthaldialdehyde.