TY - JOUR
T1 - Protease inhibitors purified from the canola meal extracts of two genetically diverse genotypes exhibit antidiabetic and antihypertension properties
AU - Hussain, Saira
AU - Rehman, Ata
AU - Luckett, David
AU - Naqvi, S. M Saqlan
AU - Blanchard, Christopher
PY - 2021/4/4
Y1 - 2021/4/4
N2 - Valorization of vegetable oil waste residues is gaining importance due to their high protein and polyphenol contents. Protease inhibitors (PIs), proteins from these abundantly available waste residues, have recently gained importance in treating chronic diseases. This research aimed to use canola meal of genetically diverse Brassica napus genotypes, BLN-3347 and Rivette, to identify PIs with diverse functionalities in therapeutic and pharmacological applications. The canola meal PI purification steps involved: native PAGE and trypsin inhibition activity, followed by ammonium sulfate fractionation, anion exchange, gel filtration, and reverse-phase chromatography. The purified PI preparations were characterized using SDS-PAGE, isoelectric focusing (IEF), and N terminal sequencing. SDS-PAGE analysis of PI preparations under native reducing and non reducing conditions revealed three polymorphic PIs in each genotype. The corresponding IEF of the genotype BLN-3347, exhibited three acidic isoforms with isoelectric points (pI) of 4.6, 4.0, and 3.9, while Rivette possessed three isoforms, exhibiting two basic forms of pI 8.65 and 9.9, and one acidic of pI 6.55. Purified PI preparations from both the genotypes displayed dipeptidyl peptidase-IV (DPP-IV)and angiotens in-converting enzyme (ACE) inhibition activities; the BLN-3347 PI preparation exhibited a strong inhibitory effect with lower IC50 values (DPP-IV 37.42 μg/mL; ACE 129 μg/mL) than that from Rivette (DPP-IV 67.97 μg/mL; ACE 376.2 μg/mL). In addition to potential human therapy,these highly polymorphic PIs, which can inhibit damaging serine proteases secreted by canola plant pathogens, have the potential to be used by canola plant breeders to seek qualitative trait locus(QTLs) linked to genes conferring resistance to canola diseases.
AB - Valorization of vegetable oil waste residues is gaining importance due to their high protein and polyphenol contents. Protease inhibitors (PIs), proteins from these abundantly available waste residues, have recently gained importance in treating chronic diseases. This research aimed to use canola meal of genetically diverse Brassica napus genotypes, BLN-3347 and Rivette, to identify PIs with diverse functionalities in therapeutic and pharmacological applications. The canola meal PI purification steps involved: native PAGE and trypsin inhibition activity, followed by ammonium sulfate fractionation, anion exchange, gel filtration, and reverse-phase chromatography. The purified PI preparations were characterized using SDS-PAGE, isoelectric focusing (IEF), and N terminal sequencing. SDS-PAGE analysis of PI preparations under native reducing and non reducing conditions revealed three polymorphic PIs in each genotype. The corresponding IEF of the genotype BLN-3347, exhibited three acidic isoforms with isoelectric points (pI) of 4.6, 4.0, and 3.9, while Rivette possessed three isoforms, exhibiting two basic forms of pI 8.65 and 9.9, and one acidic of pI 6.55. Purified PI preparations from both the genotypes displayed dipeptidyl peptidase-IV (DPP-IV)and angiotens in-converting enzyme (ACE) inhibition activities; the BLN-3347 PI preparation exhibited a strong inhibitory effect with lower IC50 values (DPP-IV 37.42 μg/mL; ACE 129 μg/mL) than that from Rivette (DPP-IV 67.97 μg/mL; ACE 376.2 μg/mL). In addition to potential human therapy,these highly polymorphic PIs, which can inhibit damaging serine proteases secreted by canola plant pathogens, have the potential to be used by canola plant breeders to seek qualitative trait locus(QTLs) linked to genes conferring resistance to canola diseases.
KW - canola meal
KW - protease inhibitor
KW - Dipeptidyl peptidase IV
KW - angiotensin converting enzyme
U2 - 10.3390/molecules26072078
DO - 10.3390/molecules26072078
M3 - Article
C2 - 33916639
SN - 1420-3049
VL - 26
SP - 1
EP - 16
JO - Molecules
JF - Molecules
IS - 7
M1 - 2078
ER -