Separation of ACE Inhibitory Peptides from Casein Hydrolysate and Analysis of Their Amino Acid Sequences

doi:10.7506/spkx1002-6630-201524028

Abstract

Abstract:

The objectives of this study were to separate angiotensin converting enzyme (ACE) inhibitory peptides from
casein hydrolysate produced by sequential hydrolysis of casein with pepsin followed by trypsin and to analyze their
amino acid sequences. The hydrolysate was concentrated by ultrafiltration and separated by Sephadex G-15 column
chromatography into three components. Components Ⅱ and Ⅲ, which had higher inhibitory effect on ACE activity, were
collected. Ion chromatography was used to analyze amino acid composition of fragments from the two components. LCMS/
MS was used to analyze amino acid sequence of fragments, solid-phase synthesis was used to synthetize short-chain
peptides, and a spectrophotometric method was used to test the inhibitory effect of hydrolysate on ACE activity. Results
showed that component Ⅱ contained eight amino acids including valine, serine, proline, leucine, phenylalanine, glutamatic
acid, asparaginic acid, and tyrosine; while component Ⅲ contained trace amounts of serine and tyrosine. Eight fragments
were obtained from both components by LC-MC/MC analysis, including three ACE inhibitory fragments αs2-f (56-57):YS,
αs2-f (98-107):YQKFPQYLQY and κ-f (52-61):INNQFLPYPY with half maximal inhibitory concentration (IC50) of 11.89,
11.75 and 421 μg/mL, respectively.

Key words: bovine casein, pepsin, trypsin, angiotensin converting enzyme (ACE), inhibitory peptide

CLC Number:

TS252.42

HU Zhihe1,2, XIA Lei1, SUN Zhengang3, WU Wenqi3, FENG Yongqiang3, XUE Lu1,2, LIU Xujin1, JIA Ying1. Separation of ACE Inhibitory Peptides from Casein Hydrolysate and Analysis of Their Amino Acid Sequences[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201524028.

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Separation of ACE Inhibitory Peptides from Casein Hydrolysate and Analysis of Their Amino Acid Sequences

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