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食品与发酵工业  2018, Vol. 44 Issue (12): 30-35    DOI: 10.13995/j.cnki.11-1802/ts.017372
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
酶解褐藻寡糖的分离制备及其脂蛋白调节作用
陈岩君1,窦文芳1,李恒1,史劲松1,许正宏2,3*
1(江南大学 药学院,糖化学与生物技术教育部重点实验室,江苏 无锡,214122)
2(江南大学 粮食发酵工艺与技术国家工程实验室,江苏 无锡,214122)
3(江南大学 生物工程学院,江苏 无锡,214122)
Isolation and lipoprotein regulation activity of alginate oligosaccharides
CHEN Yan-jun1, DOU Wen-fang1, LI Heng1, SHI Jin-song1, XU Zheng-hong2,3*
1(Key Laboratory of Carbohydrate Chemistry and Biotechnology, School of Pharmaceutical Science, Jiangnan University,Wuxi 214122, China)
2(National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China)
3(School of biotechnology, Jiangnan University, Wuxi 214122, China)
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摘要 采用Isoptericola halotolerans CGMCC 5336褐藻胶裂解酶制备褐藻寡糖,通过液质联用技术(liquid chromatograph-mass spectrometer, LC-MS)对其组成进行初步分析,后经过凝胶色谱柱对其进行分离、纯化,得到4种组分,分别命名为Fr1、Fr2、Fr3、Fr4。通过LC-MS分析各组分纯度并确定分子量,使用G-25凝胶柱进行脱盐。通过RT-PCR和Western Blot技术,研究所获得的4个组分对低密度脂蛋白受体(low-density-lipoprotein receptor,LDLR)基因和蛋白水平的调节作用以及对前蛋白转化酶枯草溶菌素9(proprotein convertase subtilisin/kexin type 9,PCSK9)蛋白水平的调节作用。结果表明,Fr1中含有2种聚合度的寡糖,为五糖和六糖;Fr2、Fr3和Fr4均为聚合度均一的寡糖,分别为四糖、三糖和二糖。4个组分中,Fr1、Fr3、Fr4均能显著上调LDLR的蛋白水平,其中Fr3和Fr4的活性尤为显著。Fr3能下调PCSK9的蛋白水平,推测Fr3对LDLR蛋白表达的上调作用可能与PCSK9有关。而Fr4对LDLR的上调作用与PCSK9无关,可能受固醇反应原件结合蛋白-2(SREBP-2)的调节。结果表明,Fr3和Fr4可开发作为食品添加剂用于降低血浆胆固醇。
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陈岩君
窦文芳
李恒
史劲松
许正宏
关键词:  褐藻寡糖  凝胶分离  低密度脂蛋白受体(low-density-lipoprotein receptor,LDLR)  前蛋白转化酶枯草溶菌素9(proprotein convertase subtilisin/kexin type 9,PCSK9)    
Abstract: Alginate oligosaccharides were prepared by hydrolyzing alginate with algae lyase from Isoptericola halotolerans CGMCC 5336. The components were analyzed by liquid chromatography-mass spectrometry and purified by gel chromatography. Four main components, named Fr1, Fr2, Fr3, and Fr4, were purified and identified. Their effects on the transcription and translation of low-density-lipoprotein receptor (LDLR) gene and pro-protein convertase subtilisin/kexin type 9 (PCSK9) were investigated. The results showed that Fr1 contained two oligosaccharides, pento- and hexo-oligosaccharides; Fr2, Fr3 and Fr4 were tetra-, tri- and di-oligosaccharides. Among them, Fr1, Fr3, and Fr4 significantly up-regulated the level of LDLR while Fr3 down-regulated the level of PCSK9, indicating that the up-regulated LDLR by Fr3 might be related to PCSK9. The up-regulation of LDLR by Fr4 was not related to PCSK9, however, might be regulated by the sterol response binding protein-2 (SREBP-2). These indicated that Fr3 and Fr4 may be further developed as new food additives for lowering the plasma cholesterol level.
Key words:  alginate oligosaccharides    gel separation    low-density lipoprotein receptor (LDLR)    proprotein convertase subtilisin/kexin type 9 (PCSK9)
收稿日期:  2018-03-24                出版日期:  2018-12-25      发布日期:  2019-01-22      期的出版日期:  2018-12-25
作者简介:  硕士研究生(许正宏教授为通讯作者,E-mail:zhenghxu@jiangnan.edu.cn)。
引用本文:    
陈岩君,窦文芳,李恒,等. 酶解褐藻寡糖的分离制备及其脂蛋白调节作用[J]. 食品与发酵工业, 2018, 44(12): 30-35.
CHEN Yan-jun,DOU Wen-fang,LI Heng,et al. Isolation and lipoprotein regulation activity of alginate oligosaccharides[J]. Food and Fermentation Industries, 2018, 44(12): 30-35.
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http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.017372  或          http://sf1970.cnif.cn/CN/Y2018/V44/I12/30
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