黑曲霉SH-2基因组中注释的7个α-葡萄糖苷酶基因的鉴定与表达分析

发布时间：2018-03-09 15:41

本文选题：黑曲霉　切入点：α-葡萄糖苷酶　出处：《现代食品科技》2017年06期 　论文类型：期刊论文

【摘要】：已公布的黑曲霉CBS513.88基因组中注释了7个α-葡萄糖苷酶基因信息,但其中主要高活性基因一直未报道。本文通过同源重组构建了黑曲霉SH-2菌株7个α-葡萄糖苷酶基因的系列多重敲除株。α-葡萄糖苷酶活性测定结果显示仅agd B敲除株的酶活相较于野生株SH-2降低36%,而其余多重敲除株菌株酶活影响较小。以敲除糖化酶、淀粉酶背景的黑曲霉SH-2-3菌株为宿主,分别敲除和过量表达基因agd B及与已报道的黑曲霉α-葡萄糖苷酶基因序列相似度最高的基因agd A,结果显示agd B敲除株生长速度在不含葡萄糖的培养条件下最慢,并随着葡萄糖添加量增加;agd B过量表达株酶活提高到野生型的10倍。本研究结果说明目前黑曲霉SH-2基因组中所注释的7个α-葡萄糖苷酶基因中仅agd B为主要高活性α-葡萄糖苷酶基因(占总体酶活36%),黑曲霉SH-2中α-葡萄糖苷酶基因仍需进一步探索。
[Abstract]:Seven 伪 -glucosidase genes were annotated in the CBS513.88 genome of Aspergillus Niger. But the major high activity genes have not been reported. In this paper, we constructed 7 multiple knockout strains of 伪 -glucosidase gene from Aspergillus Niger SH-2 strain by homologous recombination. The results of 伪 -glucosidase activity test showed that only agd B knockout was obtained. The enzyme activity of the strain was 36% lower than that of the wild strain, while the enzyme activity of the other multiple knockout strains was less than that of the wild strain. Amylase background of Aspergillus Niger SH-2-3 strain as host, The knockout and overexpression gene agd B and the gene agd A, which has the highest similarity to the reported 伪 -glucosidase gene of Aspergillus Niger, respectively. The results showed that the growth rate of agd B knockout plant was the slowest in the culture condition without glucose. With the increase of glucose supplementation, the enzyme activity of the overexpression strain of agd B was increased by 10 times to that of the wild type. The results show that only agd B is the main high activity of 7 伪 -glucosidase genes annotated in the genome of Aspergillus Niger at present. The 伪-glucosidase gene (which accounts for 36% of the total enzyme activity) needs to be further explored in Aspergillus Niger (SH-2).
【作者单位】：华南理工大学生物科学与工程学院;
【基金】：广东省科技计划项目(2016A050503016、S2012030006235、2016A010105004) 华南理工大学中央高校基本科研业务费资助项目(2015ZP032、2015ZZ040)
【分类号】：Q936

【相似文献】