灵芝细胞中异源表达透明颤菌血红蛋白基因提高胞外多糖的产量
发布时间：2018-07-15 10:11
【摘要】：在灵芝(Ganoderma lingzhi)中异源表达透明颤菌血红蛋白(Vitreoscilla hemoglobin,VHb)基因,采用一氧化碳差异波谱分析的方法检测转基因菌株的生物活性,并对野生型菌株和工程菌株进行发酵,通过荧光定量PCR对灵芝多糖生物合成途径上的葡萄糖磷酸变位酶(α-phosphoglucomutase,PGM)、尿苷二磷酸葡萄糖焦磷酸化酶(uridinediphosphate glucose pyrophosphorylase,UGP)和β-1,3-葡聚糖合酶(β-1,3-glucan synthase,GLS)3个基因的转录水平进行分析。研究表明:透明颤菌血红蛋白基因(Vitreoscilla hemoglobin gene,vgb)在灵芝中能够成功表达,并且具有生物活性;工程菌株中胞外多糖的最高产量达0.83g/L,比野生型菌株提高了88.6%;与野生型菌株相比,工程菌株中多糖生物合成途径上PGM、UGP和GLS基因的相对表达量分别是1.52、1.55和3.85。异源表达透明颤菌血红蛋白基因是提高灵芝胞外多糖产量的一种有效方法。
[Abstract]:Vitreoscilla hemoglobin VHb gene was expressed in Ganoderma lingzhi. The biological activity of transgenic strain was detected by carbon monoxide differential spectrum analysis, and the wild type strain and engineering strain were fermented. The transcription levels of three genes, 伪 -phosphoglucomutase (PGM), uridine diphosphate glucose pyrophosphorylase (uridinediphosphate glucose) and 尾 -1n 3-glucan synthase (GLS), in the biosynthesis pathway of Ganoderma lucidum were analyzed by fluorescence quantitative polymerase chain reaction (FQ-PCR). The results showed that Vitreoscilla hemoglobin gene VGB could be successfully expressed in Ganoderma lucidum and had biological activity, the highest yield of exopolysaccharide in engineering strain was 0.83g / L, which was 88.6g / L higher than that of wild-type strain. The relative expression levels of PGMU UGP and GLS genes in the polysaccharide biosynthesis pathway were 1.521.55 and 3.85, respectively. Heterologous expression of fibrillation hemoglobin gene is an effective method to increase the production of Ganoderma lucidum extracellular polysaccharides.
【作者单位】： 昆明理工大学生命科学与技术学院;昆明理工大学理学院;
【基金】：国家自然科学基金项目(31360495和21566016)资助
【分类号】：S567.31
，
本文编号：2123728
[Abstract]:Vitreoscilla hemoglobin VHb gene was expressed in Ganoderma lingzhi. The biological activity of transgenic strain was detected by carbon monoxide differential spectrum analysis, and the wild type strain and engineering strain were fermented. The transcription levels of three genes, 伪 -phosphoglucomutase (PGM), uridine diphosphate glucose pyrophosphorylase (uridinediphosphate glucose) and 尾 -1n 3-glucan synthase (GLS), in the biosynthesis pathway of Ganoderma lucidum were analyzed by fluorescence quantitative polymerase chain reaction (FQ-PCR). The results showed that Vitreoscilla hemoglobin gene VGB could be successfully expressed in Ganoderma lucidum and had biological activity, the highest yield of exopolysaccharide in engineering strain was 0.83g / L, which was 88.6g / L higher than that of wild-type strain. The relative expression levels of PGMU UGP and GLS genes in the polysaccharide biosynthesis pathway were 1.521.55 and 3.85, respectively. Heterologous expression of fibrillation hemoglobin gene is an effective method to increase the production of Ganoderma lucidum extracellular polysaccharides.
【作者单位】： 昆明理工大学生命科学与技术学院;昆明理工大学理学院;
【基金】：国家自然科学基金项目(31360495和21566016)资助
【分类号】：S567.31
，
本文编号：2123728
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