Microbiology Department, College of Life Sciences, Nanjing Agricultural University, Key Laboratory of Agricultural Environmental MicrobiologM, yinistry of Agriculture, Nanjing, Jiangsu, China.
J Basic Microbiol. 2021 Mar;61(3):253-264. doi: 10.1002/jobm.202000622. Epub 2021 Feb 5.
The heme oxygenase gene has antioxidant and cytoprotective effects in organisms, but no related research has been conducted in Ganoderma lucidum. For the first time, we cloned the HMX1 gene in G. lucidum. The CDS is 1092 bp in length and encodes 363 amino acids. The HMX1 protein was prokaryotically expressed and purified, and the enzyme activity of the purified protein was measured. The value of K was 0.699 μM, and V was 81.9 nmol BV h nmol protein. By constructing the silencing vector pAN7-dual-HMX1i, the transformants HMX1i1 and HMX1i2 were obtained. Compared with the wild-type (WT), the average growth rate of HMX1i1 and HMX1i2 decreased by 31% and 23%, respectively, and the mycelium biomass decreased by 53% and 48%, respectively. Compared with the WT, the extracellular polysaccharide content of HMX1i1 and HMX1i2 increased by 59% and 51%, and the intracellular polysaccharide content increased by 24% and 22%, respectively. These results indicate that the HMX1 gene affects mycelial growth and polysaccharide synthesis in G. lucidum.
血红素加氧酶基因在生物体内具有抗氧化和细胞保护作用,但灵芝中尚未开展相关研究。我们首次在灵芝中克隆了 HMX1 基因。CDS 长 1092bp,编码 363 个氨基酸。HMX1 蛋白在原核细胞中表达和纯化,并测定了纯化蛋白的酶活性。K 值为 0.699μM,V 值为 81.9nmolBV h nmol 蛋白。通过构建沉默载体 pAN7-dual-HMX1i,获得了转化子 HMX1i1 和 HMX1i2。与野生型(WT)相比,HMX1i1 和 HMX1i2 的平均生长速率分别降低了 31%和 23%,菌丝体生物量分别降低了 53%和 48%。与 WT 相比,HMX1i1 和 HMX1i2 的胞外多糖含量分别增加了 59%和 51%,胞内多糖含量分别增加了 24%和 22%。这些结果表明 HMX1 基因影响灵芝菌丝体生长和多糖合成。