In vitro and in silico interaction of porcine α-amylase with Vicia faba crude seed extract and evaluation of antidiabetic activity

Figures & data

Figure 1. Showing minimized energy structure. (A) Porcine α- amylase; (B) Acarbose; (C) Gallic acid; (D) Catechin; (E) Epicatechin; (F) Proanthocyanidin.

Table 1. Chemoprofiling of Vicia faba seed extracts in different solvents.

S.No	Phytochemicals	Hexane	Chloroform	methanol	Acetone	Ethanol	Ethyl acetate	Water
1	Alkaloids	−	+	+	+	+	+	+
2	Tannins	−	+	++	+	+++	+	+
3	Saponins	−	−	-	-	-	-	+
4	Phenolics compounds	−	+	+++	+	+	+	+
5	Flavonoids	+	+	+	+	+	+	+
6	Terpenoids	−	++	−	+	+	−	−
7	Glycosides	-	-	+	+	+	+	+
8	Carbohydrates	+	+	+	+	+	+	+
9	Proteins	−	+	+	+	+	+	+
10	Reducing sugar	+	+	+	+	+	+	+

Figure 2. Inhibitory potency of Vicia faba seed extracts against α-amylase activity.

Table 2. IC50 values for α-amylase inhibitory potential of Vicia faba seed extracts.

S.no	Extracts	Alpha-amylase IC50 (mg/mL)
1	Acetone	2.94 ± 0 .07
2	Methanol	5.27 ± 0.25
3	Ethanol	6.58 ±  0.37
4	Chloroform	8.23 ± 0.72
5	Water	9.25 ±  0.85

Figure 3. Mode of inhibition of α-amylase by acetone and methanol seed extract (Michaelis- Menten plot).

Figure 4. Mode of inhibition of α-amylase by acetone and methanol seed extract (Lineweaver-Burk plot).

Table 3. Kinetic analysis of Vicia faba seed extract with respect to reference molecule.

S.no	Extract	Km (mM)	Vmax(mM/min)
1	Acetone	0.56	0.14
2	Methanol	0.50	0.16
3	Control	0.40	0.208

Figure 5. FTIR analysis of (A) fraction 4 (B) fraction 5 showing different functional groups.

Table 4. Total sugar.total phenol content of acetonic seed extract fraction.

Different fractions	Total sugar( µg/ml)	Total phenol(µM)	% inhibition
F1	6	32	4.5
F2	30	29	9
F3	45	58	11
F4	51	75	15
F5	62	85	28

Figure 6. HPLC analysis of (A) tannic acid and (B) acetonic seed fraction 5.

Figure 7. Lig plot showing hydrogen bonding and hdrophobic interaction in different ligand, (A) Acarbose; (B) Epicatechein; (C) Gallic acid; (D) Proanthocyanidin; (E) Catechin with porcine α- amylase.

Figure 8. Showing autodock tool analysis of different types of interaction in different ligands. (A) Catechin; (B) Epicatechin; (C) Acarbose; (D) Proanthocyanidin; (E) Gallic acid with porcine α- amylase.

Table 5. Docking analysis of different ligands with porcine α amylase.

S.no	Compound	Binding energy	Number of hydrogen binding	Amino acid participated in hydrogen bonding
1	Acarbose	−2.29	4	Asp197,Asp300,Glu233,Asp300
2	Gallic acid	−2.43	3	His201,Asp197,Asp300
3	Proanthocyanidin	−6.39	5	Asp300,Glu233,His201,Glu240,Tyr195
4	Catechein	−6.56	5	Glu233,Arg195,His201,Ile235,Asp197
5	Epicatechein	−6.45	5	Glu233,Arg195,His299,Ile235,Asp197