{"title":"了解通过高通量筛选发现的赤霉菌突变体在合成衣康酸过程中的代谢节点","authors":"","doi":"10.1016/j.procbio.2024.09.003","DOIUrl":null,"url":null,"abstract":"<div><p><em>Aspergillus terreus</em> is the most important strain for the industrial production of itaconic acid. Few works have reported the critical metabolic nodes in the synthesis of itaconic acid in <em>A. terreus</em>. Herein a high-throughput screening method was established to screen an efficient mutant <em>A. terreus</em> 6–12#, which could produce 28.6 g/L itaconic acid with a productivity of 0.36 g/g glucose, about 66 % and 57 % higher than that of the parent strain. On this basis, the expression of the key enzymes was analyzed centering on the glycolytic pathway, the precursor synthesis pathway, the itaconic acid synthesis and degradation pathways, and the membrane transport system. It was proven that the expressions of enzymes favoring itaconic acid synthesis in each of these modules were significantly elevated in the mutant. But the enzymes CAD, MTT, MFS in the itaconic acid synthesis pathway and the membrane transport system displayed low elevation levels, while the degradation pathway was activated with the improvement of itaconic acid, which was considered the possible limited step in itaconic acid synthesis in <em>A. terreus</em>. These findings provided potential metabolic modification targets for further acquisition of excellent <em>A. terreus</em> cell factories and helped to facilitate the industrial production of itaconic acid.</p></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.7000,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Understanding the metabolic nodes in itaconic acid synthesis of an Aspergillus terreus mutant identified by high-throughput screening\",\"authors\":\"\",\"doi\":\"10.1016/j.procbio.2024.09.003\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p><em>Aspergillus terreus</em> is the most important strain for the industrial production of itaconic acid. Few works have reported the critical metabolic nodes in the synthesis of itaconic acid in <em>A. terreus</em>. Herein a high-throughput screening method was established to screen an efficient mutant <em>A. terreus</em> 6–12#, which could produce 28.6 g/L itaconic acid with a productivity of 0.36 g/g glucose, about 66 % and 57 % higher than that of the parent strain. On this basis, the expression of the key enzymes was analyzed centering on the glycolytic pathway, the precursor synthesis pathway, the itaconic acid synthesis and degradation pathways, and the membrane transport system. It was proven that the expressions of enzymes favoring itaconic acid synthesis in each of these modules were significantly elevated in the mutant. But the enzymes CAD, MTT, MFS in the itaconic acid synthesis pathway and the membrane transport system displayed low elevation levels, while the degradation pathway was activated with the improvement of itaconic acid, which was considered the possible limited step in itaconic acid synthesis in <em>A. terreus</em>. These findings provided potential metabolic modification targets for further acquisition of excellent <em>A. terreus</em> cell factories and helped to facilitate the industrial production of itaconic acid.</p></div>\",\"PeriodicalId\":20811,\"journal\":{\"name\":\"Process Biochemistry\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2024-09-07\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Process Biochemistry\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1359511324002927\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Process Biochemistry","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1359511324002927","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
土曲霉是工业生产衣康酸最重要的菌株。很少有研究报道了土曲霉合成衣康酸的关键代谢节点。本文建立了一种高通量筛选方法,筛选出了一种高效突变体 A. terreus 6-12#,它能生产 28.6 g/L 的衣康酸,生产率为 0.36 g/g 葡萄糖,比亲本菌株分别高出约 66 % 和 57 %。在此基础上,以糖酵解途径、前体合成途径、衣康酸合成和降解途径以及膜运输系统为中心,分析了关键酶的表达情况。结果表明,突变体中有利于衣康酸合成的酶的表达量均显著增加。但衣康酸合成途径和膜转运系统中的酶 CAD、MTT、MFS 的表达水平较低,而降解途径则随着衣康酸的改善而被激活,这被认为是 A. terreus 中衣康酸合成的可能限制步骤。这些发现为进一步获得优良的赤霉菌细胞工厂提供了潜在的代谢改造目标,有助于促进衣康酸的工业化生产。
Understanding the metabolic nodes in itaconic acid synthesis of an Aspergillus terreus mutant identified by high-throughput screening
Aspergillus terreus is the most important strain for the industrial production of itaconic acid. Few works have reported the critical metabolic nodes in the synthesis of itaconic acid in A. terreus. Herein a high-throughput screening method was established to screen an efficient mutant A. terreus 6–12#, which could produce 28.6 g/L itaconic acid with a productivity of 0.36 g/g glucose, about 66 % and 57 % higher than that of the parent strain. On this basis, the expression of the key enzymes was analyzed centering on the glycolytic pathway, the precursor synthesis pathway, the itaconic acid synthesis and degradation pathways, and the membrane transport system. It was proven that the expressions of enzymes favoring itaconic acid synthesis in each of these modules were significantly elevated in the mutant. But the enzymes CAD, MTT, MFS in the itaconic acid synthesis pathway and the membrane transport system displayed low elevation levels, while the degradation pathway was activated with the improvement of itaconic acid, which was considered the possible limited step in itaconic acid synthesis in A. terreus. These findings provided potential metabolic modification targets for further acquisition of excellent A. terreus cell factories and helped to facilitate the industrial production of itaconic acid.
期刊介绍:
Process Biochemistry is an application-orientated research journal devoted to reporting advances with originality and novelty, in the science and technology of the processes involving bioactive molecules and living organisms. These processes concern the production of useful metabolites or materials, or the removal of toxic compounds using tools and methods of current biology and engineering. Its main areas of interest include novel bioprocesses and enabling technologies (such as nanobiotechnology, tissue engineering, directed evolution, metabolic engineering, systems biology, and synthetic biology) applicable in food (nutraceutical), healthcare (medical, pharmaceutical, cosmetic), energy (biofuels), environmental, and biorefinery industries and their underlying biological and engineering principles.