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Fine-Tuning of Using CRISPRi for Increasing 5-Aminolevulinic Acid Production in .利用CRISPRi对[具体对象]中5-氨基乙酰丙酸产量增加进行微调。 （注：原文中“in.”后面缺少具体内容）
Front Microbiol. 2019 Jul 31;10:1731. doi: 10.3389/fmicb.2019.01731. eCollection 2019.
2
Engineering of multiple modular pathways for high-yield production of 5-aminolevulinic acid in Escherichia coli.工程化多个模块化途径以提高大肠杆菌中 5-氨基乙酰丙酸的产量。
Bioresour Technol. 2019 Feb;274:353-360. doi: 10.1016/j.biortech.2018.12.004. Epub 2018 Dec 3.
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Pathway engineering in S9114 for 5-aminolevulinic acid production.用于5-氨基乙酰丙酸生产的S9114中的途径工程。
3 Biotech. 2018 May;8(5):247. doi: 10.1007/s13205-018-1267-2. Epub 2018 May 8.
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Re-direction of carbon flux to key precursor malonyl-CoA via artificial small RNAs in photosynthetic sp. PCC 6803.通过人工小RNA将光合细菌嗜热栖热放线菌PCC 6803中的碳通量重定向至关键前体丙二酰辅酶A。
Biotechnol Biofuels. 2018 Feb 5;11:26. doi: 10.1186/s13068-018-1032-0. eCollection 2018.
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Recent advances in production of 5-aminolevulinic acid using biological strategies.利用生物策略生产5-氨基乙酰丙酸的最新进展。
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N-terminal engineering of glutamyl-tRNA reductase with positive charge arginine to increase 5-aminolevulinic acid biosynthesis.通过带正电荷的精氨酸对谷氨酰胺-tRNA还原酶进行N端工程改造以增加5-氨基乙酰丙酸的生物合成。
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通过合成反义RNA下调hemB以提高[具体生物]中5-氨基乙酰丙酸的产量

Downregulating of hemB via synthetic antisense RNAs for improving 5-aminolevulinic acid production in .

作者信息

Ge Fanglan, Wen Dongmei, Ren Yao, Chen Guiying, He Bing, Li Xiaokun, Li Wei

机构信息

College of Life Sciences, Sichuan Normal University, Chengdu, 610068 People's Republic of China.

Key Laboratory for Utilization and Conservation of Bio-Resources of Education, Department of Sichuan Province, Chengdu, People's Republic of China.

出版信息

3 Biotech. 2021 May;11(5):230. doi: 10.1007/s13205-021-02733-8. Epub 2021 Apr 21.

DOI:10.1007/s13205-021-02733-8
PMID:33968574
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8060372/
Abstract

UNLABELLED

Aminolevulinic acid (ALA), a type of natural non-protein amino acid, is a key precursor for the biosynthesis of heme, and it has been broadly applied in medicine, agriculture. Several strategies have been applied to enhance ALA synthesis in bacteria. In the present study, we employed synthetic antisense RNAs (asRNAs) of (encodes ALA dehydratase) to weaken metabolic flux of ALA to porphobilinogen (PBG), and investigated their effect on ALA accumulation. For this purpose, we designed and constructed vectors pET28a-hemA-asRNA and pRSFDuet-hemA-asRNA to simultaneously express 5-ALA synthase (ALAS, encoded by ) and PTasRNAs (2 inverted repeat DNA sequences sandwiched with the antisense sequence of ), selecting the region ranging from - 57 nt upstream to + 139 nt downstream of the start codon of as a target. The qRT-PCR analysis showed that the mRNA levels of were decreased above 50% of the control levels, suggesting that the anti-hemB asRNA was functioning appropriately. ALA accumulation in the hemB weakened strains were 17.6% higher than that obtained using the control strains while accumulating less PBG. These results indicated that asRNAs can be used as a tool for regulating ALA accumulation in .

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1007/s13205-021-02733-8.

摘要

未标记

氨基乙酰丙酸（ALA）是一种天然非蛋白质氨基酸，是血红素生物合成的关键前体，已广泛应用于医学、农业领域。已采用多种策略来增强细菌中ALA的合成。在本研究中，我们利用编码ALA脱水酶的hemB的合成反义RNA（asRNAs）来减弱ALA向胆色素原（PBG）的代谢通量，并研究它们对ALA积累的影响。为此，我们设计并构建了载体pET28a-hemA-asRNA和pRSFDuet-hemA-asRNA，以同时表达5-ALA合酶（由hemA编码的ALAS）和PTasRNAs（2个反向重复DNA序列夹着hemB的反义序列），选择hemB起始密码子上游-57 nt至下游+139 nt的区域作为靶标。qRT-PCR分析表明，hemB的mRNA水平降低至对照水平的50%以上，表明抗hemB asRNA发挥了适当的作用。hemB弱化菌株中的ALA积累比对照菌株高17.6%，同时积累的PBG较少。这些结果表明，asRNAs可作为调节大肠杆菌中ALA积累的工具。

补充信息

在线版本包含可在10.1007/s13205-021-02733-8获取的补充材料。