Revealing of free radical scavenging and angiotensin I-converting enzyme inhibitor potency of pigmented rice seed protein

SUSILOWATI, Erlin; SANJAYA, Bella Rhea Lavifa; NUGRAHA, Ari Satia; UBAIDILLAH, Mohammad; SISWOYO, Tri Agus

doi:10.1590/fst.66520

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Original Article • Food Sci. Technol (Campinas) 42 • 2022 • https://doi.org/10.1590/fst.66520 copy

Revealing of free radical scavenging and angiotensin I-converting enzyme inhibitor potency of pigmented rice seed protein

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

The potential source of bioactive protein from pigmented rice seed was investigated by buffered extracting from nine varieties (Merah-SP, Gogo Niti-2, Merah Wangi, Super Manggis, Lamongan-1, Mota, Ketan Hitam-2, Beureum Taleus, and Aek Sibundong) and non-pigmented rice (IR-64) as a control. The potent contributor to free radical scavenging of extracted proteins was evaluated by analyzing their free amino acid composition. The free radical scavenging and angiotensin-I converting enzyme (ACE-I) inhibitory activity of these proteins were analyzed in-vitro. The free radical scavenging activity was analyzed using various standard methods, including radical cation 2,2-azinobis-3-ethylbenzothiazoline-6-sulphonate acid (ABTS^•+), hydroxyl radical (OH^•) and defense against radical-mediated DNA damages by hydroxyl. Total free amino acid content ranged between 77.7-181.2 g/100 g protein, and among them, Ketan Hitam-2 displayed the highest abundant antioxidant amino acid grouping (23.23%) and shown significantly higher ABTS^•+ activity (IC₅₀=8.64 µg/mL) and OH^• activity (IC₅₀=20.33 µg/mL). Moreover, Ketam Hitam-2 also exhibited notable ACE-I inhibitory activity (IC₅₀=6.20 µg/mL) and protected hydroxyl-induced oxidative damage to DNA. The in-vitro systems for free radical scavenging and ACE-I inhibitory were used to acquire the data. The potency of Ketan Hitam-2 seed protein could be utilized as a natural nutraceuticals compound.

Keywords:
ACE-I inhibitory; free radical scavenging; pigmented rice; protein

Amino acid ^* * g/100 g protein;	Variety										Mean	SE	CV (%)
Amino acid ^* * g/100 g protein;	A	B	C	D	E	F	G	H	I	J	Mean	SE	CV (%)
L-Arginine^ Antioxidant amino acid grouping.	9.34	14.22	9.72	12.61	11.76	16.60	10.36	9.96	7.48	10.56	11.26	0.83	23.44
L-Aspartic acid	7.56	10.96	8.61	10.24	9.35	15.72	7.73	7.89	5.90	8.56	9.25	0.85	29.02
L-Cysteine**	0.40	0.67	0.49	0.63	0.53	0.80	0.65	0.55	0.40	0.57	0.57	0.04	21.85
L-Glutamic acid	16.18	24.43	19.02	23.02	20.71	33.32	16.38	16.89	12.40	18.58	20.09	1.84	28.96
L-Histidine**	2.89	4.57	3.38	4.10	3.52	5.20	3.23	3.47	2.58	3.60	3.65	0.25	21.34
L-Lysine**	3.75	4.55	5.30	5.32	4.33	6.71	3.77	3.95	2.86	4.64	4.52	0.34	23.71
L-Serine	6.37	9.61	7.55	8.82	7.58	10.45	6.45	7.26	5.41	7.72	7.72	0.49	19.91
L-Threonine	4.19	6.73	5.07	5.96	5.81	7.40	4.64	4.71	3.64	5.30	5.35	0.36	21.56
L-Alanine	5.53	8.23	6.61	7.70	6.72	10.33	5.76	5.82	4.29	6.42	6.74	0.53	24.93
L-Phenylalanine	5.76	9.64	6.13	8.90	7.90	12.40	6.62	6.84	5.13	7.10	7.64	0.69	28.40
L-Tyrosine**	4.23	6.73	4.06	6.62	5.77	8.42	4.98	5.70	3.51	4.75	5.48	0.47	27.14
Glycine	5.45	8.22	6.29	7.34	6.47	8.90	5.68	5.79	4.42	6.42	6.50	0.42	20.57
L-Isoleucine	4.59	6.89	5.16	6.46	5.73	8.07	4.67	4.64	3.52	5.19	5.49	0.42	24.20
L-Leucine	9.19	14.19	10.36	13.20	11.91	16.55	9.44	9.19	7.08	10.47	11.16	0.89	25.25
L-Methionine**	0.01	0.86	0.78	0.51	0.68	0.70	0.96	0.62	0.37	0.02	0.55	0.10	59.52
L-Valine	6.68	10.07	7.63	9.22	8.41	11.52	6.97	6.60	5.09	7.57	7.98	0.60	23.60
L-Proline	4.69	6.96	5.42	6.44	5.72	8.13	4.80	4.80	3.62	5.39	5.60	0.41	23.12
TAA (g/100 g Protein)	96.81	147.53	111.58	137.09	122.9	181.22	103.09	104.68	77.7	112.86	119.55	9.29	24.58
TAAAnt (g/100 g Protein)	20.62	31.60	23.73	29.79	26.59	38.43	23.95	24.25	17.20	24.14	26.03	1.89	23.01
TAAAnt /TAA (%)	21.30	21.42	21.27	21.73	21.64	21.21	23.23	23.17	22.14	21.39	21.85	0.24	3.49

Variety	Free Radical ( ^* * IC50 value (µg/mL) is the effective concentration of antioxidants to inhibit 50% radical ABTS and hydroxyl. Glutathione (G-SH) was used as a positive control. All values are expressed as the mean of three replicates ± standard deviations (SD), the Duncan’s multiple range tests with a significance level of p<0.05 were followed. The different letters in the column are represented significantly different. IC₅₀, µg/mL)
Variety	ABTS^•+ (µg/mL)	Hydroxyl (µg/mL)
Merah-SP	45.66 ± 0.93^c	25.30 ± 0.53^d
Gogo Niti-2	47.34 ± 1.15^c	33.08 ± 0.26^a
Merah Wangi	44.63 ± 1.09^c	23.43 ± 0.15^f
Super Manggis	43.99 ± 0.24^c	22.11 ± 0.17^g
Lamongan-1	61.62 ± 1.02^b	24.44 ± 0.15^e
Mota	43.61 ± 0.34^c	30.30 ± 0.19^b
Ketan Hitam-2	8.64 ± 0.68^e	20.33 ± 0.23ⁱ
Beureum Taleus	36.87 ± 0.35^d	20.90 ± 0.18^h
Aek Sibundong	63.66 ± 1.94^b	20.35 ± 0.17ⁱ
IR-64	72.53 ± 1.67^a	29.47 ± 0.23^c
G-SH	0.22 ± 0.08^f	8.24 ± 0.12^j

Variety	ACE-I inhibitor ( ^* * IC50 value (µg/mL) is the effective concentration of antioxidants to inhibit 50% ACE-I activity. Captopril was used as a positive control of ACE-I. All values are expressed as the mean of three replicates ± standard deviations (SD), the Duncan’s multiple range tests with a significance level of p<0.05 were followed. The different letters in the column are represented significantly different. IC₅₀, µg/mL)
Merah-SP	15.75 ± 0.04^f
Gogo Niti-2	7.66 ± 0.63^e
Merah Wangi	6.39 ± 0.06^c
Super Manggis	6.76 ± 0.05^d
Lamongan-1	6.31 ± 0.02^c
Mota	7.05 ± 0.02^e
Ketan Hitam-2	6.20 ± 0.05^b
Beureum Taleus	17.30 ± 0.11^g
Aek Sibundong	26.60 ± 0.14ⁱ
IR-64	21.63 ± 0.02^h
Captopril	2.08 ± 0.20^a

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[1] *Corresponding author: triagus.faperta@unej.ac.id, siswoyo.triagus@gmail.com