Purification of Antioxidant Peptides from Natto Supernatant and Study on Its Activity

doi:10.13560/j.cnki.biotech.bull.1985.2021-0379

Abstract

Abstract:

High antioxidant polypeptides were extracted from fermented natto,the supernatant of the natto were isolated and purified by SephadexG-50 and RP HPLC,and their structures were identified by electrospray ionization tandem mass spectrometry. The results showed that 3 components（F1,F2 and F3）from the polypeptide mixture of fermented natto were obtained after the separation and purification of the antioxidant peptide via Sephadex G-50 gel. The antioxidant activity of the component F3 was the strongest,the total reduction power reached（8.4+0.6）mmol/g,higher than that of other components. F3 was separated and purified by semi preparative RP-HPLC,and 4 components（G1,G2,G3,and G4）were obtained. The G1 and G3 in the highest contents were selected for determining DPPH scavenging rate,and G1’s DPPH free radical scavenging rate reached 32.67% The antioxidant peptides in G1 were sequenced and identified by ESI-MS/MS. and the 10 antioxidant peptides with the highest content and the strongest activity were obtained. The activity of peptide SFEWVLEH was the strongest verified by chemical synthesis method. The DPPH scavenging rate was 46.66% ± 0.96%.

Key words: natto, antioxidant peptide, isolation and purification, identification

ZHANG Feng-wen, ZHOU Li-ya, DONG Chao, SHI Yan-mao. Purification of Antioxidant Peptides from Natto Supernatant and Study on Its Activity[J]. Biotechnology Bulletin, 2022, 38(2): 158-165.

Figures/Tables 10

References 21

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时间Time/min	流动相B Mobile phase B%
0-5	5-10
5-35	10-80
35-45	80-100
45-50	100-100
50-55	100-80
55-75	80-10
75-80	10-5

时间Time/min	流动相B Mobile phase B%
0-5	5-10
5-35	10-80
35-45	80-100
45-50	100-100
50-55	100-80
55-75	80-10
75-80	10-5

组分 Fraction	蛋白或多肽浓度 Protein or peptide concentration/（μg·mL^-1）	FRAP值 FRAP value/ （μmol·L^-1）	每克蛋白的还原力 Reducing power per gram protein/ （mmol·g^-1）
F1	237±7^A	419±10.97^A	3.9±0.5^C
F2	84.4±7.6^B	408±49.5^A	5±0.2^B
F3	50±2^B	422±12.6^A	8.4±0.6^A

组分 Fraction	蛋白或多肽浓度 Protein or peptide concentration/（μg·mL^-1）	FRAP值 FRAP value/ （μmol·L^-1）	每克蛋白的还原力 Reducing power per gram protein/ （mmol·g^-1）
F1	237±7^A	419±10.97^A	3.9±0.5^C
F2	84.4±7.6^B	408±49.5^A	5±0.2^B
F3	50±2^B	422±12.6^A	8.4±0.6^A

编号 Code	Peptide < ProteinMetrics Confidential >	Observed （M+H）	Calc. mass（M+H）
1	RAELSEDDVFVIPA	1560.785	1560.790
2	LSEDDVFVIPA	1204.607	1204.610
3	DVFRAIPSEVL	1245.681	1245.684
4	FEEINRVLL	1132.633	1132.636
5	SLLNALPEEVIQH	1462.787	1462.790
6	LAVLI	528.375	528.376
7	SFEWVLEH	1046.493	1046.494
8	GIFGM	540.248	540.249
9	QVFL	506.296	506.297
10	FPF	410.207	410.207