灵芝作为一种典型的食药用真菌,其多糖是主要的活性物质之一,应用广泛。在多糖合成的途径中,糖基转移酶发挥着重要的作用。为探究葡萄糖基转移酶基因GL24971的过表达对灵芝多糖的影响,通过聚乙二醇转化法将过表达质粒转化至原生质体,用潮霉素抗性筛选,得到转化菌株。结果表明,转化菌株T1与T2的单位菌体胞外多糖最大产量分别为0.07、0.06 mg/mg,分别提高了132.4%、70.0%,而胞内多糖产量均为0.03 mg/mg,分别降低了10.7%、37.4%;其次,几丁质和β-1,3-葡聚糖含量降低;单糖组成中,半乳糖的比例减少,甘露糖的比例增加,但仍然以葡萄糖为主。研究结果为灵芝多糖产量的提高提供了新的思路,有利于促进多糖合成途径的进一步完善,也为后续研究灵芝中其余的糖基转移酶提供了参考。
In order to investigate the effect on polysaccharide synthesis by overexpression of glucosyltransferase GL24971, the overexpression plasmid was transformed into G. lucidum protoplast. The results showed that the maximum production of extracellular polysaccharide in T1 and T2 was 0.07 and 0.06 mg/mg, which increased by 132.4% and 70.0%, respectively. The yield of intracellular polysaccharide of T1 and T2 were both 0.03 mg/mg, which decreased by 10.7% and 37.4%, respectively. Secondly, the content of chitin and β-1, 3-glucan decreased. The results of the monosaccharide composition analysis showed that the proportion of galactose was decreased with the increased mannose proportion, in which glucose was still the main component. These results provide new ideas for the enhancement of G. lucidum polysaccharide production and further clarify the polysaccharide synthesis pathway.
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