碳源对manA基因突变大肠杆菌代谢的影响
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北京市教委长城学者培养计划项目(CIT&TCD20140315)


Effects of Carbon Sources on the Metabolism of manA Gene Mutant Strain in Escherichia coli
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    摘要:

    ManA基因编码的甘露糖-6-磷酸异构酶在大肠杆菌中催化D-甘露糖和D-果糖的异构化,促进大肠杆菌对碳源的代谢吸收。本文通过研究manA基因突变大肠杆菌对碳源的利用和编码糖代谢基因情况,探讨甘露糖-6-磷酸异构酶对大肠杆菌糖代谢的影响。采用Ⅱ型内含子逆转录突变方法构建manA基因突变大肠杆菌,分析manA基因突变大肠杆菌对不同碳源的利用情况和manA基因突变对大肠杆菌糖代谢相关基因表达的影响,结果显示,大肠杆菌BL21(DE3)~(ΔmanA)以甘露糖、果糖为碳源时,菌株生长受到显著抑制;以淀粉为碳源时,BL21(DE3)~(ΔmanA)菌株的生长显著优于野生型大肠杆菌;以葡萄糖为碳源时,manA基因突变对大肠杆菌的生长无显著影响。通过基因表达分析,发现大肠杆菌BL21(DE3)~(ΔmanA)中甘露糖代谢相关基因的表达显著性降低;果糖代谢途径中6-磷酸果糖激酶Ⅰ亚基的编码基因(pfkA)显著下调表达;水解淀粉的α-淀粉酶编码基因(malS)显著性上调表达。ManA基因突变影响大肠杆菌甘露糖、果糖和淀粉代谢途径中相关基因的表达,从而影响大肠杆菌对碳源的利用。

    Abstract:

    Mannose-6-phosphate isomerase encoded by manA gene catalyzes the isomerization of D-mannose and D-fructose in Escherichia coli and promotes the metabolic uptake of carbon sources in Escherichia coli. In this paper, the study of carbon source utilization and gene encoding carbon metabolism in manA mutant strain aims to investigate the effect of mannose-6-phosphate isomerase on the metabolism of Escherichia coli. ManA gene mutant strain was constructed by group II intron reverse transcriptase mutation method. The utilization of manA gene mutant strain on different carbon sources and the effect of manA gene mutant strain on the gene expression of carbon metabolism in Escherichia coli were analyzed. The results showed that the growth of Escherichia coli BL21(DE3)ΔmanA strain was significantly inhibited when the mannose and starch were used as the carbon source, but the growth of BL21(DE3)ΔmanA strain was significantly better than that of wild type strain when starch was used as carbon source. The mutation of manA gene has no significant effect on the growth of BL21(DE3)ΔmanA strain in glucose. Further study gene expression showed that the expression of mannose-related genes in BL21(DE3)ΔmanA strain was significantly decreased. The gene (pfkA) of the 6-phosphoglucose kinase I subunit was significantly down-regulated in the fructose metabolic pathway. The expression of the gene (malS) of the α--amylase of the hydrolyzed starch was up-regulated. In conclusion, The mutation of manA gene affected the gene expression in the mannose, fructose and starch metabolic pathways, thus affecting the utilization of carbon sources in Escherichia coli.

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李晓霞;王欢;孙京利;张红星;谢远红;.碳源对manA基因突变大肠杆菌代谢的影响[J].中国食品学报,2020,20(2):103-110

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  • 在线发布日期: 2020-03-07
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