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Isolation of angiotensin I-converting enzyme inhibitor from pepsin hydrolysate of porcine hemoglobin

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Abstract

Angiotensin I-converting enzyme (ACE) inhibitors have been widely used as antihypertensive agents. However, most synthetic ACE inhibitors ineluctably have severe side effects. Researchers have focused on various ACE-inhibitory peptides derived from dietary food. In the present study, we reported peptides produced from porcine blood, an important food in Asian countries. Through enzymatic hydrolysis, we found that peptides from this animal compound have ACE-inhibitory effects. Porcine hemoglobin was hydrolyzed using ordinary proteases, including alcalase, trypsin, neutral, papain, protamex, and pepsin. Results showed that pepsin was the most efficient protease in producing active peptides, and the pepsin hydrolysate of porcine hemoglobin showed the highest activity (IC50 = 1.53 ± 0.03 mg/mL). Combining DA 201-C macroporous resin chromatography, Sephadex LH-20 gel chromatography, and reversed-phase high-performance liquid chromatography, the fraction 2-IV was purified from pepsin hydrolysis of porcine hemoglobin; this compound exhibited the highest ACE-inhibitory activity (IC50 = 0.02 ± 0.01 mg/mL). Through Edman degradation, we also found that the exact amino acid sequence of fraction 2-IV was Gln–Glu–Leu–Pro–Gly. The results indicated that porcine hemoglobin peptides possessed significant ACE-inhibitory effect in vitro, which is an important complement of the previous work.

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Abbreviations

ACE:

Angiotensin I-converting enzyme

Ang I:

Angiotensin I

Ang II:

Angiotensin II

DDW:

Double-distilled water

DH:

Degree of hydrolysis

HPLC:

High-performance liquid chromatography

IC50 :

50 % ACE inhibition

KKS:

Kallikrein–kinin system

RAS:

Rennin–angiotensin–aldosterone system

RP-HPLC:

Reversed-phase high-performance liquid chromatography

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Acknowledgments

The authors wish to thank Dr. Zhaohui Zhang from the Agricultural Biological Detection Laboratory, Chinese Academy of Agricultural Sciences, for the analysis of peptide sequence. This research was supported by a Grant (XDJK2009C055) from the Southwest University’s basic scientific research business expenses special, and the Laboratory of Quality and Safety Risk Assessment for Agro-products on Storage and Preservation (Chongqing), Ministry of Agriculture, China.

Conflict of interest

The authors declare no conflict of interests.

Compliance with Ethics Requirements

All procedures on animals were in strict accordance with the guide for the care and using of laboratory animals published by the US national Institutes of Health and was approved by the Institutional Animal Care Committee at the Southwest University.

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Authors and Affiliations

  1. Laboratory of Quality & Safety Risk Assessment for Agro-products on Storage and Preservation (Chongqing), Ministry of Agriculture, College of Food Science, Southwest University, Chongqing, 400715, China

    HuiLing Deng, Jiong Zheng, Fusheng Zhang, Yang Wang & Jianquan Kan

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  1. HuiLing Deng
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Correspondence to Jianquan Kan.

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Deng, H., Zheng, J., Zhang, F. et al. Isolation of angiotensin I-converting enzyme inhibitor from pepsin hydrolysate of porcine hemoglobin. Eur Food Res Technol 239, 933–940 (2014). https://doi.org/10.1007/s00217-014-2290-0

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  • DOI: https://doi.org/10.1007/s00217-014-2290-0

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