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从牦牛乳中分离的干酪乳杆菌 SB27 胞外多糖的理化性质及抗肿瘤活性

Physicochemical characterization and antitumour activity of exopolysaccharides produced by Lactobacillus casei SB27 from yak milk.

机构信息

School of Chemistry and Chemical Engineering, Harbin Institute of Technology, Harbin, 150090, China.

School of Chemistry and Chemical Engineering, Harbin Institute of Technology, Harbin, 150090, China; College of Food Science and Engineering, Ocean University of China, Qingdao, 266003, China.

出版信息

Carbohydr Polym. 2017 Sep 1;171:307-315. doi: 10.1016/j.carbpol.2017.03.018. Epub 2017 Mar 8.

Abstract

Two high molecular weight fractions (LW1 and LW2) of exopolysaccharides (EPSs) produced by Lactobacillus casei SB27 were isolated from yak milk obtained from the Gansu Tibetan area of China. GC-MS, FTIR spectroscopy, methylation analysis and FE-SEM analysis were performed to elucidate the physicochemical characterization of these two fractions, and their in vitro antitumour activities were also evaluated. The molecular weights (Mws) of LW1 and LW2 as determined by HPGPC were 25.10 and 12.34kDa, respectively. Monosaccharide composition analysis revealed that LW1 and LW2 were mainly composed of galactose (52.4% and 57.4%, mol%) and glucose (29.1% and 22.2%, mol%), respectively. Methylation results showed that the main chain of LW1 likely involves (1→4)-linked Galp and (1→4)-linked Glcp with its side chains being (1→4,6)-linked Galp through the O-6 position connected to the backbone, whereas the main chain of LW2 likely involves (1→4)-linked Galp and (1→4)-linked Glcp with its side chains being (1→3)-Galp through the O-6 position of (1→3,6)-Galp linked to the main chain. Evaluation of the microcosmic morphology, as revealed by FE-SEM analysis of the two EPS fractions, showed a sheet-like appearance with a folded surface and a compact structure. The results from in vitro antitumour tests indicated that both LW1 and LW2 could significantly inhibit the proliferation of HT-29 colorectal cancer cells and up-regulated the expressions of Bad, Bax, Caspase-3 and -8 genes. Finally, TEM images revealed the apoptotic morphological changes of HT-29 cells induced by LW1 and LW2. Our results suggested that LW1 and LW2 possess potential not only for use in functional food products but also as a source of natural antitumour drugs.

摘要

从中国甘肃藏区牦牛乳中分离到两株干酪乳杆菌 SB27 产生的高分子量胞外多糖(EPS)的两个部分(LW1 和 LW2)。通过 GC-MS、FTIR 光谱、甲基化分析和 FE-SEM 分析,对这两个部分的理化特性进行了阐明,并对其体外抗肿瘤活性进行了评价。HPGPC 测定的 LW1 和 LW2 的分子量(Mw)分别为 25.10 和 12.34kDa。单糖组成分析表明,LW1 和 LW2 主要由半乳糖(52.4%和 57.4%,摩尔%)和葡萄糖(29.1%和 22.2%,摩尔%)组成。甲基化结果表明,LW1 的主链可能涉及(1→4)-连接的半乳糖(Galp)和(1→4)-连接的葡萄糖(Glcp),其侧链通过 O-6 位与主链相连,为(1→4,6)-连接的半乳糖(Galp),而 LW2 的主链可能涉及(1→4)-连接的半乳糖(Galp)和(1→4)-连接的葡萄糖(Glcp),其侧链通过 O-6 位与主链相连,为(1→3)-半乳糖(Galp)。通过 FE-SEM 分析两种 EPS 部分的微观形态,结果表明两部分均呈片状,表面折叠,结构紧凑。体外抗肿瘤试验结果表明,LW1 和 LW2 均可显著抑制 HT-29 结肠癌细胞的增殖,并上调 Bad、Bax、Caspase-3 和 -8 基因的表达。最后,TEM 图像显示了 LW1 和 LW2 诱导 HT-29 细胞的凋亡形态变化。研究结果表明,LW1 和 LW2 不仅具有作为功能性食品的潜力,而且具有作为天然抗肿瘤药物的来源的潜力。

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