School of Biotechnology, International University, Ho Chi Minh City, Vietnam.
Vietnam National University, Ho Chi Minh City, Vietnam.
BMC Biotechnol. 2021 Mar 27;21(1):28. doi: 10.1186/s12896-021-00688-5.
Microbial polysaccharides have been reported to possess remarkable bioactivities. Physarum polycephalum is a species of slime mold for which the microplasmodia are capable of rapid growth and can produce a significant amount of cell wall-less biomass. There has been a limited understanding of the polysaccharides produced by microplasmodia of slime molds, including P. polycephalum. Thus, the primary objectives of this research were first to chemically characterize the exopolysaccharides (EPS) and intracellular polysaccharides (IPS) of P. polycephalum microplasmodia and then to evaluate their cytotoxicity against several cancer cell lines.
The yields of the crude EPS (4.43 ± 0.44 g/l) and partially purified (deproteinated) EPS (2.95 ± 0.85 g/l) were comparable (p > 0.05) with the respective crude IPS (3.46 ± 0.36 g/l) and partially purified IPS (2.45 ± 0.36 g/l). The average molecular weight of the EPS and IPS were 14,762 kDa and 1788 kDa. The major monomer of the EPS was galactose (80.22%), while that of the IPS was glucose (84.46%). Both crude and purified IPS samples showed significantly higher cytotoxicity toward Hela cells, especially the purified sample and none of the IPSs inhibited normal cells. Only 38.42 ± 2.84% Hela cells remained viable when treated with the partially purified IPS (1 mg/ml). However, although only 34.76 ± 6.58% MCF-7 cells were viable when exposed to the crude IPS, but the partially purified IPS displayed non-toxicity to MCF-7 cells. This suggested that the cytotoxicity toward MCF-7 would come from some component associated with the crude IPS sample (e.g. proteins, peptides or ion metals) and the purification process would have either completely removed or reduced amount of that component. Cell cycle analysis by flow cytometry suggested that the mechanism of the toxicity of the crude IPS toward MCF-7 and the partially purified IPS toward Hela cells was due to apoptosis.
The EPS and IPS of P. polycephalum microplasmodia had different chemical properties including carbohydrate, protein and total sulfate group contents, monosaccharide composition and molecular weights, which led to different cytotoxicity activities. The crude and partially purified IPSs would be potential materials for further study relating to cancer treatment.
微生物多糖具有显著的生物活性。多头绒泡菌是一种粘菌,其微变形虫能够快速生长,并能产生大量无细胞壁的生物质。人们对粘菌微变形虫产生的多糖,包括多头绒泡菌多糖的了解有限。因此,本研究的主要目的首先是对多头绒泡菌微变形虫的胞外多糖(EPS)和细胞内多糖(IPS)进行化学表征,然后评估其对几种癌细胞系的细胞毒性。
粗 EPS(4.43±0.44g/L)和部分纯化(去蛋白)EPS(2.95±0.85g/L)的产率与粗 IPS(3.46±0.36g/L)和部分纯化 IPS(2.45±0.36g/L)相当(p>0.05)。EPS 和 IPS 的平均分子量分别为 14762kDa 和 1788kDa。EPS 的主要单糖是半乳糖(80.22%),而 IPS 的主要单糖是葡萄糖(84.46%)。粗提和纯化 IPS 样品对 Hela 细胞均显示出显著的细胞毒性,尤其是纯化样品,而 IPS 对正常细胞没有抑制作用。当用部分纯化 IPS(1mg/ml)处理时,只有 38.42±2.84%的 Hela 细胞存活。然而,尽管用粗 IPS 处理 MCF-7 细胞时,只有 34.76±6.58%的 MCF-7 细胞存活,但部分纯化的 IPS 对 MCF-7 细胞无毒性。这表明,对 MCF-7 的细胞毒性可能来自于粗 IPS 样品中某些与蛋白质、肽或离子金属相关的成分,而纯化过程可能完全去除或减少了该成分的含量。流式细胞术的细胞周期分析表明,粗 IPS 对 MCF-7 和部分纯化 IPS 对 Hela 细胞的毒性机制是凋亡。
多头绒泡菌微变形虫的 EPS 和 IPS 具有不同的化学性质,包括碳水化合物、蛋白质和总硫酸根含量、单糖组成和分子量,这导致了不同的细胞毒性活性。粗提和部分纯化的 IPS 可能是进一步研究癌症治疗的潜在材料。