State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, P.R. China.
J Microbiol Biotechnol. 2021 Jun 28;31(6):855-866. doi: 10.4014/jmb.2102.02006.
The effects of various carbon sources on mycelial growth and polysaccharide synthesis of the medicinal fungus in liquid fermentation were investigated. After 12-d fermentation, mycelial biomass, polysaccharide yield, and polysaccharide content were significantly higher in Glc+Lac group (glucose and lactose used as combined carbon source) than in other groups. Crude polysaccharides (CIOPs) and the derivative neutral polysaccharides (NIOPs) were obtained from mycelia fermented using Glc, fructose (Fru), Lac, or Glc+Lac as carbon source. Molecular weights of four NIOPs (termed as NIOPG, NIOPF, NIOPL, and NIOPGL) were respectively 780.90, 1105.00, 25.32, and 10.28 kDa. Monosaccharide composition analyses revealed that NIOPs were composed of Glc, Man, and Gal at different molar ratios. The NIOPs were classified as α-type heteropolysaccharides with 1→2, 1→3, 1→4, 1→6 linkages in differing proportions. In in vitro cell proliferation assays, viability of RAW264.7 macrophages was more strongly enhanced by NIOPL or NIOPGL than by NIOPG or NIOPF, and proliferation of HeLa or S180 tumor cells was more strongly inhibited by NIOPG or NIOPGL than by NIOPF or NIOPL, indicating that immune-enhancing and anti-tumor activities of NIOPs were substantially affected by carbon source. qRT-PCR analysis revealed that expression levels of phosphoglucose isomerase (PGI) and UDP-Glc 4-epimerase (UGE), two key genes involved in polysaccharide synthesis, varied depending on carbon source. Our findings, taken together, clearly demonstrate that carbon source plays an essential role in determining structure and activities of polysaccharides by regulating expression of key genes in polysaccharide biosynthetic pathway.
研究了不同碳源对药用真菌液体发酵中菌丝生长和多糖合成的影响。经过 12 天发酵,Glc+Lac 组(葡萄糖和乳糖作为组合碳源)的菌丝生物量、多糖得率和多糖含量明显高于其他组。从以 Glc、果糖(Fru)、Lac 或 Glc+Lac 作为碳源发酵的菌丝中获得了粗多糖(CIOPs)和衍生中性多糖(NIOPs)。四种 NIOPs(分别命名为 NIOPG、NIOPF、NIOPL 和 NIOPGL)的分子量分别为 780.90、1105.00、25.32 和 10.28 kDa。单糖组成分析表明,NIOPs 由不同摩尔比的 Glc、Man 和 Gal 组成。NIOPs 被分类为α-型杂多糖,具有不同比例的 1→2、1→3、1→4 和 1→6 键。在体外细胞增殖试验中,NIOPL 或 NIOPGL 比 NIOPG 或 NIOPF 更能增强 RAW264.7 巨噬细胞的活力,而 NIOPG 或 NIOPGL 比 NIOPF 或 NIOPL 更能抑制 HeLa 或 S180 肿瘤细胞的增殖,表明 NIOPs 的免疫增强和抗肿瘤活性受到碳源的显著影响。qRT-PCR 分析表明,参与多糖合成的两个关键基因磷酸葡萄糖异构酶(PGI)和 UDP-Glc 4-差向异构酶(UGE)的表达水平随碳源而变化。综合我们的研究结果,清楚地表明碳源通过调节多糖生物合成途径中的关键基因的表达,对多糖的结构和活性起着至关重要的作用。