Modification of Acaudina Molpadioides Hydrolysates by Plastein Reaction and Preparation of ACE Inhibitory Peptides†

doi:10.7503/cjcu20130951

Abstract

Abstract:

The effect of modification by plastein reaction on the angiotensin converting enzyme(ACE) inhibitory activity of Acaudina molpadioides hydrolysates and its peptides was researched to show that the ACE inhibitory activity of modified peptide decreased definitely, compared with the activity of original hydrolysates. The stability of the modified peptides to simulated gastrointestinal digestion was tested in vitro. The result showed that the ACE inhibitory activities of the peptides could be enhanced by digestive enzyme. Toxicity test showed no toxic to the MDCK cells while concentration was less than 10 mg/mL, and low concentration could promote the cells’ growth. A novel ACE inhibitory peptide was isolated from modified product using different methods including ultrafiltration, Sephadex G-15 gel filtration and RP-HPLC. The purified ACE inhibitory peptide was sequenced as NQNFVQYTTNT with an IC₅₀ value of 3.67 μmol/L. The accurate sites of ACE that the peptide bind to were determined through the computer assisted software SYBYL, and UV spectra analysis proved that some changes could be observed in the binding of ACE and peptides.

Key words: Acaudina molpadioides, Plastein reaction, ACE inhibitory peptides, Isolation and purification

CLC Number:

O629.72

TrendMD:

SHEN Qingqing, ZENG Mingyong, ZHAO Yuanhui. Modification of Acaudina Molpadioides Hydrolysates by Plastein Reaction and Preparation of ACE Inhibitory Peptides^†[J]. Chem. J. Chinese Universities, 2014, 35(5): 965.

Figures/Tables 6

Fig.1 Isolation and purification of modified Acaudina molpadioides peptides by plastein reaction and the IC50 values of different components^(A) a—h are 8 groups of modified peptides isolated by Sephadex G-15 chromatograph; (A') IC50 values of a—h components(P<0.01); (B) component d is further purified by RP-HPLC on Zorbax SB-C18 column(5 μm, 9.4 mm×250 mm); (B') IC50 values of components d1—d7(P<0.05).

Fig.2 Purity and identification of modified peptides and the sequence profile of purified fraction d1

Fig.3 ACE-binding peptide by docking algorithm of computer aided molecular design

Fig.4 UV-Vis absorption analysis of the ACE-binding peptide(a), peptide(b) and ACE(c)

Fig.5 Cytotoxicity experiments of modified peptide on MDCK cells

Table 1 Impact of gastrointestinal proteases of modified hydrolysates by plastein reaction on ACE inhibitory activity and free amino groups

Sample	Free amino group/(mmol·L^-1)	IC₅₀/(μg·mL^-1)
Control	3.06±0.03	122±7.2^a
Pepsin	3.32±0.06	56±8.4^b
Pepsin+Chymotrypsin	3.49±0.05	66±6.7^b^,^c
Pepsin+Chymotrypsin+Trypsin	3.54±0.03	74±5.7^c

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