Lueangsakulthai Jiraporn, Phosri Santi, Theansungnoen Tinnakorn, Jangpromma Nisachon, Temsiripong Theeranan, Mckendrick John E, Khunkitti Watcharee, Klaynongsruang Sompong
Protein and Proteomics Research Center for Commercial and Industrial Purposes (ProCCI), Faculty of Science, Khon Kaen University, Khon Kaen, Thailand.
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand.
Biotechnol Appl Biochem. 2018 May;65(3):455-466. doi: 10.1002/bab.1628. Epub 2017 Dec 15.
Novel antioxidant and anti-inflammatory peptides were isolated from hydrolysates of Siamese crocodile (Crocodylus siamensis) hemoglobin. C. siamensis hemoglobin hydrolysates (CHHs) were obtained by pepsin digestion at different incubation times (2, 4, 6, and 8 H) at 37 °C and subjected to antioxidant and anti-inflammatory activity assessment. CHH obtained by 2-H hydrolysis (2H-CHH) showed the highest anti-inflammatory activity with respect to decreasing nitric oxide (NO) production, whereas the strongest antioxidant activity was found for 6-H hydrolysis (6H-CHH) against nitric oxide radicals. To evaluate the anti-inflammatory and antioxidant activity of individual peptide components, 2H-CHH and 6H-CHH were purified by semipreparative HPLC. Peptide fraction P57 isolated from 6H-CHH was found to exhibit the highest nitric oxide radical inhibition activity (32.0%). Moreover, purification of 2H-CHH yielded peptide fraction P16, which displayed a high efficacy in decreasing NO production of macrophage RAW 264.7 cells (83.2%) and significantly reduced proinflammatory cytokines and inflammatory mediators interleukin-6 (IL-6), interleukin-1 beta (IL-1β), and prostaglandin-E2 (PGE ) production to about 2.0, 0.3, and 1.9 ng/mL, respectively. Using LTQ orbitrap XL mass spectrometry, active peptide sequences were identified as antioxidant KIYFPHF (KF7), anti-inflammatory SAFNPHEKQ (SQ9), and IIHNEKVQAHGKKVL (IL15). Additionally, CHHs simulated gastric and intestinal in vitro digestion positively contributed to antioxidant and anti-inflammatory activity. Taken collectively, the results of this work demonstrate that CHHs contain several peptides with anti-inflammatory and antioxidant properties, which may prove valuable as treatment or supplement against diseases associated with inflammation and oxidative stress.
从暹罗鳄（Crocodylus siamensis）血红蛋白水解物中分离出了新型抗氧化和抗炎肽。通过在37℃下不同孵育时间（2、4、6和8小时）用胃蛋白酶消化获得暹罗鳄血红蛋白水解物（CHHs），并对其进行抗氧化和抗炎活性评估。通过2小时水解获得的CHH（2H-CHH）在降低一氧化氮（NO）生成方面表现出最高的抗炎活性，而在对抗一氧化氮自由基方面，6小时水解获得的CHH（6H-CHH）具有最强的抗氧化活性。为了评估单个肽组分的抗炎和抗氧化活性，通过半制备型高效液相色谱法对2H-CHH和6H-CHH进行了纯化。从6H-CHH中分离出的肽组分P57表现出最高的一氧化氮自由基抑制活性（32.0%）。此外，纯化2H-CHH得到肽组分P16，其在降低巨噬细胞RAW 264.7细胞的NO生成方面具有高效性（83.2%），并显著降低促炎细胞因子和炎症介质白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）和前列腺素-E2（PGE）的生成，分别降至约2.0、0.3和1.9 ng/mL。使用LTQ轨道阱XL质谱法，活性肽序列被鉴定为抗氧化肽KIYFPHF（KF7）、抗炎肽SAFNPHEKQ（SQ9）和IIHNEKVQAHGKKVL（IL15）。此外，CHHs模拟胃和肠道的体外消化对其抗氧化和抗炎活性有积极贡献。总体而言，这项工作的结果表明，CHHs含有几种具有抗炎和抗氧化特性的肽，这可能被证明在治疗或补充与炎症和氧化应激相关的疾病方面具有价值。
