酶解影响的腹足纲动物源生物活性肽的抗炎特性

Anti-inflammatory Properties of Bioactive Peptide Derived from Gastropod Influenced by Enzymatic Hydrolysis.

作者信息

Joshi Ila, Sudhakar Sekar, Nazeer Rasool Abdul

机构信息

Department of Biotechnology, School of Bioengineering, SRM University, Kattankulathur, Chennai, Tamilnadu, 603 203, India.

出版信息

Appl Biochem Biotechnol. 2016 Nov;180(6):1128-1140. doi: 10.1007/s12010-016-2156-y. Epub 2016 Jun 11.

DOI:10.1007/s12010-016-2156-y

PMID:27287997

Abstract

The visceral mass of the gastropod, Harpa ventricosa was hydrolysed using trypsin, alcalase and pepsin for 12 h to produce protein hydrolysates. Subsequently, the active hydrolysate was observed in the 3rd hour of tryptic hydrolysate (29.17 ± 0.62 and 34.85 ± 0.55 %) using human red blood cell (HRBC) membrane stabilization and albumin denaturation (AD) assays. The active hydrolysate was fractionated by membrane filtration unit, where <10-kDa fraction revealed better anti-inflammatory activity with IC value 6.27 ± 0.05 and 5.38 ± 0.02 mg/ml for HRBC and AD assays, respectively. Additionally, the active fraction contains essential and non-essential (aspartic acid, arginine, glutamic acid and leucine) amino acids and, sequentially, the active fraction was further purified using consecutive chromatography, in which fraction C-II exhibited strong anti-inflammatory activity (HRBC 56.02 ± 0.52 and AD 50.71 ± 1.10 % assays). The non-toxic, low molecular weight (690.2 Da) hexapeptide (Ala-Lys-Gly-Thr-Trp-Lys) suppressed the nitric oxide (NO) and pro-cytokine production in a dose-dependent manner on THP-1 cell lines.

摘要

用胰蛋白酶、碱性蛋白酶和胃蛋白酶对腹足纲动物竖琴螺的内脏团进行12小时水解，以产生蛋白水解物。随后，使用人红细胞（HRBC）膜稳定化和白蛋白变性（AD）试验，在胰蛋白酶水解物的第3小时观察到活性水解物（分别为29.17±0.62%和34.85±0.55%）。活性水解物通过膜过滤装置进行分级分离，其中分子量小于10 kDa的级分显示出更好的抗炎活性，HRBC试验和AD试验的IC值分别为6.27±0.05和5.38±0.02 mg/ml。此外，活性级分含有必需氨基酸和非必需氨基酸（天冬氨酸、精氨酸、谷氨酸和亮氨酸），随后，活性级分通过连续色谱法进一步纯化，其中级分C-II表现出较强的抗炎活性（HRBC试验为56.02±0.52%，AD试验为50.71±1.10%）。无毒、低分子量（690.2 Da）的六肽（Ala-Lys-Gly-Thr-Trp-Lys）对THP-1细胞系中一氧化氮（NO）和前细胞因子的产生具有剂量依赖性抑制作用。