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一种从重组菌废弃细胞中分离纯化葡聚糖的环保方法及纯化葡聚糖的生物活性表征

An environment-friendly approach to isolate and purify glucan from spent cells of recombinant and the bioactivity characterization of the purified glucan.

作者信息

Xing Yan, Chen Chaonan, Sun Wenlong, Zhang Bowei, Sang Yuanbin, Xiu Zhilong, Dong Yuesheng

机构信息

School of Life Science and Biotechnology Dalian University of Technology Dalian Liaoning P. R. China.

出版信息

Eng Life Sci. 2018 Jan 19;18(4):227-235. doi: 10.1002/elsc.201700125. eCollection 2018 Apr.

Abstract

While the methylotrophic yeast enables the industrial-scale biosynthesis of many recombinant products, large amount of nutrient-rich biomass emerged along this process. Polysaccharides, especially glucans that are abundant in the cell wall of , are yet to be better utilized owing to their various biological activities. However, the isolation and purification of cell wall glucan from has not been reported. In this study, we established an environment-friendly approach, including induced autolysis, hot-water treatment, ultrasonication, isopropanol extraction, and protease treatment, to isolate and purify glucan from the cell wall of . We achieved a purity of 85.3% and a yield of 11.7% for the purified glucan. Proteins, nucleic acids, lipids, and ash were efficiently removed during the purification. The activities of the purified glucan were investigated in mice fed with a high-fat diet. The purified glucan decreased the level of total cholesterol and triglycerides by 30.3 and 29.7%, respectively. This result suggested that the cell wall glucan of could be developed to a therapeutic agent for dyslipidemia. Our study proposed an environment-friendly and effective method to isolate and purify the glucan from , providing solid foundation for the high-value utilization of this yeast.

摘要

虽然甲基营养型酵母能够实现多种重组产品的工业化规模生物合成,但在此过程中会产生大量营养丰富的生物质。多糖,尤其是在其细胞壁中大量存在的葡聚糖,由于其多种生物活性,尚未得到更好的利用。然而,尚未有从其细胞壁中分离和纯化葡聚糖的报道。在本研究中,我们建立了一种环境友好的方法,包括诱导自溶、热水处理、超声处理、异丙醇提取和蛋白酶处理,以从其细胞壁中分离和纯化葡聚糖。我们获得的纯化葡聚糖纯度为85.3%,产率为11.7%。在纯化过程中,蛋白质、核酸、脂质和灰分被有效去除。对喂食高脂饮食的小鼠研究了纯化葡聚糖的活性。纯化葡聚糖使总胆固醇和甘油三酯水平分别降低了30.3%和29.7%。这一结果表明,其细胞壁葡聚糖可开发成为治疗血脂异常的药物。我们的研究提出了一种环境友好且有效的方法来从其细胞壁中分离和纯化葡聚糖,为该酵母的高值利用提供了坚实基础。

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