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嗜麦芽寡养单胞菌蔗糖酶酶解生成纳米颗粒莱菔糖及其超声降解产物在人骨肉瘤细胞中的产生和生物活性。

Production and bioactivities of nanoparticulated and ultrasonic-degraded levan generated by Erwinia tasmaniensis levansucrase in human osteosarcoma cells.

机构信息

Department of Chemistry, Faculty of Science, Silpakorn University, Nakhon Pathom 73000, Thailand.

Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.

出版信息

Int J Biol Macromol. 2022 Nov 30;221:1121-1129. doi: 10.1016/j.ijbiomac.2022.09.096. Epub 2022 Sep 15.

DOI:10.1016/j.ijbiomac.2022.09.096
PMID:36115448
Abstract

Levan is a bioactive polysaccharide that can be synthesized by various microorganisms. In this study, the physicochemical properties and bioactivity of levan synthesized by recombinant levansucrase from Erwinia tasmaniensis were investigated. The synthesis conditions, including the enzyme concentration, substrate concentration, and temperature, were optimized. The obtained levan generally appeared as a cloudy suspension. However, it could transform into a hydrogel at concentrations exceeding 10 % (w/v). Then, ultrasonication was utilized to reduce the molecular weight and increase the bioavailability of levan. Dynamic light scattering (DLS) and gel permeation chromatography (GPC) indicated that the size of levan was significantly decreased by ultrasonication, whereas Fourier transform infrared spectroscopy, H-nuclear magnetic resonance, and X-ray powder diffraction revealed that the chemical structure of levan was not changed. Finally, the bioactivities of both levan forms were examined using human osteosarcoma (Saos-2) cells. The result clearly illustrated that sonicated levan had higher antiproliferative activity in Saos-2 cells than original levan. Sonicated levan also activated Toll-like receptor expression at the mRNA level. These findings suggested the important beneficial applications of sonicated levan for the development of cancer therapies.

摘要

低聚糖是一种具有生物活性的多糖,可由多种微生物合成。本研究采用重组塔玛氏欧文氏菌来源的蔗糖酶合成低聚糖,并对其理化性质和生物活性进行了研究。优化了合成条件,包括酶浓度、底物浓度和温度。所得低聚糖通常呈混浊悬浮液,但浓度超过 10%(w/v)时可转化为水凝胶。然后,采用超声处理降低低聚糖的分子量,提高其生物利用度。动态光散射(DLS)和凝胶渗透色谱(GPC)表明,超声处理显著降低了低聚糖的分子量,而傅里叶变换红外光谱、氢核磁共振和 X 射线粉末衍射表明,低聚糖的化学结构没有改变。最后,采用人骨肉瘤(Saos-2)细胞检测了两种低聚糖形式的生物活性。结果表明,超声处理的低聚糖在 Saos-2 细胞中的增殖抑制活性明显高于原低聚糖。超声处理的低聚糖还能在 mRNA 水平上激活 Toll 样受体的表达。这些发现表明,超声处理的低聚糖在癌症治疗的开发中有重要的有益应用。

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