本文關(guān)鍵詞： 雙齒圍沙蠶 抗凝血肽 堿性蛋白酶 分離純化 抗血栓　出處：《食品科學(xué)》2017年23期 　論文類型：期刊論文

【摘要】：目的:探討雙齒圍沙蠶抗凝血肽(anticoagulant peptides from Perinereis aibuhitensis,PAAP)的制備及其抗血栓作用。方法:以抗凝活力為檢測(cè)指標(biāo),篩選最佳酶對(duì)沙蠶進(jìn)行酶解。酶解產(chǎn)物經(jīng)超濾、陰離子交換層析、凝膠過濾層析和反相高效液相色譜法分離純化,得到具有最強(qiáng)抗凝活力的沙蠶寡肽,將此肽命名為PAAP并測(cè)定其急性毒性作用。采用角叉菜膠所致的小鼠尾部血栓形成實(shí)驗(yàn)、三氯化鐵所致的大鼠動(dòng)脈血栓形成實(shí)驗(yàn)及二磷酸腺苷(adenonisine disphosphate,ADP)誘導(dǎo)的血小板聚集實(shí)驗(yàn),研究PAAP的抗血栓作用。結(jié)果:選取堿性蛋白酶作為最佳酶對(duì)沙蠶進(jìn)行酶解,經(jīng)過分離純化后得到一條氨基酸序列:脯氨酸-纈氨酸-谷氨酸-精氨酸-賴氨酸(Pro-Val-Glu-Arg-Lys)。經(jīng)測(cè)定,該寡肽的體外抗凝活力可達(dá)(1 320.0±20.3)U/g。PAAP的最大耐受量為11.0 g/(kg·d)且未出現(xiàn)小鼠異常及死亡現(xiàn)象,體內(nèi)實(shí)驗(yàn)表明PAAP對(duì)小鼠尾部血栓的形成、大鼠動(dòng)脈血栓的形成及ADP誘導(dǎo)的血小板聚集均有明顯抑制作用。結(jié)論:從雙齒圍沙蠶中提取出的抗凝肽PAAP具有抗血栓作用。
[Abstract]:Objective: to study anticoagulant peptides from Perinereis aibuhitensis. The preparation and antithrombotic effect of PAAP.Methods: the best enzyme was selected to hydrolyze silkworm by ultrafiltration and anion exchange chromatography with anticoagulant activity as detection index. Gel filtration chromatography and reversed phase high performance liquid chromatography (RP-HPLC) were used to isolate and purify oligopeptides from silkworm, which had the strongest anticoagulant activity. The peptide was named PAAP and its acute toxicity was determined. The tail thrombosis of mice induced by carrageenin was studied. Thrombosis induced by ferric trichloride in rats and platelet aggregation induced by adenonisine disphosphate ADP (adenosine diphosphate) in rats. The antithrombotic effect of PAAP was studied. Results: alkaline protease was selected as the best enzyme to hydrolyze silkworm silkworm. A amino acid sequence, Pro-Val-Glu-Arg-Lysine, was isolated and purified. The amino acid sequence, Pro-Val-Glu-Arg-Lysn, was determined. The anticoagulant activity of the oligopeptide in vitro was 1320.0 鹵20.3U / g 路PAAP, the maximum tolerance was 11.0 g / kg 路d) and no abnormal or death occurred in mice. In vivo experiments showed that PAAP induced thrombosis in the tail of mice. Conclusion: the anticoagulant peptide PAAP extracted from the silkworm has antithrombotic effect.
【作者單位】： 浙江海洋大學(xué)食品與醫(yī)藥學(xué)院浙江省海洋生物醫(yī)用制品重點(diǎn)工程技術(shù)研究中心;浙江省海洋水產(chǎn)研究所;
【基金】：國(guó)家自然科學(xué)基金青年科學(xué)基金項(xiàng)目(81001393) 2015年度國(guó)家星火計(jì)劃項(xiàng)目(2015GA700044) 國(guó)家海洋局重大計(jì)劃項(xiàng)目(2015862) 浙江省科技廳重大科技專項(xiàng)計(jì)劃項(xiàng)目(2013C03036) 浙江省自然科學(xué)基金項(xiàng)目(LS15H30001);浙江省自然科學(xué)基金青年資金項(xiàng)目(LQ16H300001) 舟山市公益類科技項(xiàng)目(2015C31012)
【分類號(hào)】：R943;R965
【正文快照】： 血栓形成是缺血性心臟病、缺血性中風(fēng)及靜脈栓塞量,高速組織搗碎機(jī)勻漿,加3倍體積預(yù)冷的磷酸鹽常見的病理學(xué)基礎(chǔ),嚴(yán)重危害人類健康且近年來還有漸緩沖液(0.02 mol/L,p H 7.0),4℃靜置2 h以上。增之勢(shì)[1-3]。抑制血小板的聚集與釋放、抑制凝血酶系8 000 r/min離心15 min,收集上

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